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aavdj ef1a mcherry ires cre  (Addgene inc)


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    Addgene inc aavdj ef1a mcherry ires cre
    Fig. 3. vlPAG activity is required for systemic morphine antinociception and drives spinal NA–dependent antinociception. (A) Bilateral vlPAG AAV-DIO-hM4Di- <t>mCherry</t> injections in Vglut2-cre mice with representative image of viral expression. Scale bar, 500 μm. (B) Left: Hot plate withdrawal latencies of vlPAGVglut2-cre::hM4Di mice administered CNO (3 mg/kg i.p.) versus saline without (light green bars) and with morphine (5 mg/kg s.c.) (dark green bars) (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 13 mice; saline versus CNO effect, P < 0.0001, F1,12 = 56.93). Right: Withdrawal latencies of non–virus-injected controls (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 9 mice; saline versus CNO effect, P = 0.32, F1,8 = 1.143). (C) Bilateral vlPAG AAV-DIO-hM3Dq- mCherry injections in Vglut2-cre mice, combinations of systemic CNO with intrathecal antagonists, and representative image of viral expression. Scale bar, 500 μm. (D) Left: Withdrawal latencies after systemic saline or CNO and intrathecal saline, phentolamine (5 μg), or naltrexone (5 μg) (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P < 0.0001, Friedman statistic = 26.40). Right: Von Frey mechanical thresholds (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P = 0.0001, Friedman statistic = 21.00). (E) Representative images of c-Fos immunohistochemistry in TH-positive LC neurons of vlPAGVglut2-cre::hM3Dq mice after systemic injection of saline (top) or CNO (bottom). Scale bars, 150 μm. Right: % of TH-positive LC neurons that colocalize with green c-Fos signal (five to eight images analyzed per mouse; n = 6 saline, n = 5 CNO, two-sided Mann-Whitney test). Data in each graph reported as means ± SEM.
    Aavdj Ef1a Mcherry Ires Cre, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/aavdj+ef1a+mcherry+ires+cre/pAAV-Ef1a-mCherry-IRES-Cre+(Plasmid+%2355632)/pm38669335-345-31-36
    Average 96 stars, based on 114 article reviews
    aavdj ef1a mcherry ires cre - by Bioz Stars, 2026-10
    96/100 stars

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    1) Product Images from "Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation."

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.

    Journal: Science advances

    doi: 10.1126/sciadv.adj9581

    Fig. 3. vlPAG activity is required for systemic morphine antinociception and drives spinal NA–dependent antinociception. (A) Bilateral vlPAG AAV-DIO-hM4Di- mCherry injections in Vglut2-cre mice with representative image of viral expression. Scale bar, 500 μm. (B) Left: Hot plate withdrawal latencies of vlPAGVglut2-cre::hM4Di mice administered CNO (3 mg/kg i.p.) versus saline without (light green bars) and with morphine (5 mg/kg s.c.) (dark green bars) (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 13 mice; saline versus CNO effect, P < 0.0001, F1,12 = 56.93). Right: Withdrawal latencies of non–virus-injected controls (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 9 mice; saline versus CNO effect, P = 0.32, F1,8 = 1.143). (C) Bilateral vlPAG AAV-DIO-hM3Dq- mCherry injections in Vglut2-cre mice, combinations of systemic CNO with intrathecal antagonists, and representative image of viral expression. Scale bar, 500 μm. (D) Left: Withdrawal latencies after systemic saline or CNO and intrathecal saline, phentolamine (5 μg), or naltrexone (5 μg) (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P < 0.0001, Friedman statistic = 26.40). Right: Von Frey mechanical thresholds (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P = 0.0001, Friedman statistic = 21.00). (E) Representative images of c-Fos immunohistochemistry in TH-positive LC neurons of vlPAGVglut2-cre::hM3Dq mice after systemic injection of saline (top) or CNO (bottom). Scale bars, 150 μm. Right: % of TH-positive LC neurons that colocalize with green c-Fos signal (five to eight images analyzed per mouse; n = 6 saline, n = 5 CNO, two-sided Mann-Whitney test). Data in each graph reported as means ± SEM.
    Figure Legend Snippet: Fig. 3. vlPAG activity is required for systemic morphine antinociception and drives spinal NA–dependent antinociception. (A) Bilateral vlPAG AAV-DIO-hM4Di- mCherry injections in Vglut2-cre mice with representative image of viral expression. Scale bar, 500 μm. (B) Left: Hot plate withdrawal latencies of vlPAGVglut2-cre::hM4Di mice administered CNO (3 mg/kg i.p.) versus saline without (light green bars) and with morphine (5 mg/kg s.c.) (dark green bars) (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 13 mice; saline versus CNO effect, P < 0.0001, F1,12 = 56.93). Right: Withdrawal latencies of non–virus-injected controls (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 9 mice; saline versus CNO effect, P = 0.32, F1,8 = 1.143). (C) Bilateral vlPAG AAV-DIO-hM3Dq- mCherry injections in Vglut2-cre mice, combinations of systemic CNO with intrathecal antagonists, and representative image of viral expression. Scale bar, 500 μm. (D) Left: Withdrawal latencies after systemic saline or CNO and intrathecal saline, phentolamine (5 μg), or naltrexone (5 μg) (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P < 0.0001, Friedman statistic = 26.40). Right: Von Frey mechanical thresholds (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P = 0.0001, Friedman statistic = 21.00). (E) Representative images of c-Fos immunohistochemistry in TH-positive LC neurons of vlPAGVglut2-cre::hM3Dq mice after systemic injection of saline (top) or CNO (bottom). Scale bars, 150 μm. Right: % of TH-positive LC neurons that colocalize with green c-Fos signal (five to eight images analyzed per mouse; n = 6 saline, n = 5 CNO, two-sided Mann-Whitney test). Data in each graph reported as means ± SEM.

    Techniques Used: Activity Assay, Expressing, Saline, Virus, Injection, Immunohistochemistry, MANN-WHITNEY

    Fig. 4. Anatomical characterization of inputs to the LC from the vlPAG. (A) vlPAG injection of AAV-DIO-tdTom in Vglut2- or Vgat-cre mice. (B) Vglut2-cre injection site. (C) vlPAGVglut2-cre terminals (magenta) and TH (green) in the peri-LC. Bar, Barrington’s nucleus. (D) Vgat-cre injection site. (E) vlPAGVgat-cre terminals. Scale bars, 300 μm [(B) to (E)]. (F) Quantification of magenta and green pixel intensity in the peri-LC of vlPAGVglut2-cre::tdTom mice normalized by z score (n = 6 LC slices from three mice). (G) Same as (F) for vlPAGVgat-cre::tdTom. (H) Left: Injections of AAVretro-cre in the LC and AAV-DIO-mCherry in the vlPAG of wild-type mice. Middle: Image of mCherry+ vlPAG neurons. Right: Resulting terminals in the LC and RVM. Scale bars, 500 μm. (I) Same as (H) for vlPAG→RVM neurons. Scale bars, 500 μm. (J) Left: Orthogonal recombinase strategy to label vlPAG neurons that project to the RVM and LC. Middle: Representative image of mCherry (magenta) and YFP (green). Arrows indicate double-labeled (white) neu- rons. Scale bar, 300 μm. Right: Quantification of mCherry and YFP in the vlPAG.
    Figure Legend Snippet: Fig. 4. Anatomical characterization of inputs to the LC from the vlPAG. (A) vlPAG injection of AAV-DIO-tdTom in Vglut2- or Vgat-cre mice. (B) Vglut2-cre injection site. (C) vlPAGVglut2-cre terminals (magenta) and TH (green) in the peri-LC. Bar, Barrington’s nucleus. (D) Vgat-cre injection site. (E) vlPAGVgat-cre terminals. Scale bars, 300 μm [(B) to (E)]. (F) Quantification of magenta and green pixel intensity in the peri-LC of vlPAGVglut2-cre::tdTom mice normalized by z score (n = 6 LC slices from three mice). (G) Same as (F) for vlPAGVgat-cre::tdTom. (H) Left: Injections of AAVretro-cre in the LC and AAV-DIO-mCherry in the vlPAG of wild-type mice. Middle: Image of mCherry+ vlPAG neurons. Right: Resulting terminals in the LC and RVM. Scale bars, 500 μm. (I) Same as (H) for vlPAG→RVM neurons. Scale bars, 500 μm. (J) Left: Orthogonal recombinase strategy to label vlPAG neurons that project to the RVM and LC. Middle: Representative image of mCherry (magenta) and YFP (green). Arrows indicate double-labeled (white) neu- rons. Scale bar, 300 μm. Right: Quantification of mCherry and YFP in the vlPAG.

    Techniques Used: Injection, Labeling

    Fig. 8. Pathway-specific modulation of vlPAG and RVM terminals in the LC modulates nociceptive behavior. (A) Left: Bilateral cannula placement over the LC in uninjected control mice. Right: Hot plate withdrawal latencies of control mice microinfused in the LC with saline (150 nl) versus CNO (3 μM, 150 nl) without (light gray) and with morphine (5 mg/kg s.c.) (dark gray; two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 8 mice; saline versus CNO effect, P = 0.4178, F1,7 = 0.7412). (B) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the vlPAG of Vglut2-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion with saline versus CNO without (light green) and with morphine (5 mg/kg s.c.) (dark green; two-way repeated-measures ANOVA with Sidak’s multiple comparisons test; n = 12 mice; saline versus CNO effect, P = 0.0022, F1,11 = 15.72). (C) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the RVM of Vgat-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion of saline versus CNO (blue bars, n = 12 mice), microinfusion of CNO with intrathecal injections of saline versus phentolamine (5 μg, light blue bars, n = 9 mice), and microinfusion of saline versus CNO with morphine (5 mg/kg s.c.) (dark blue, n = 12 mice; mixed-effects analysis with matching across row and Tukey’s multiple comparisons test, P < 0.0001, F2.560,25.09 = 27.36). (D) Circuit diagram of DPMS inputs to the LC and their opioid sensitivity. Data in each graph reported as means ± SEM.
    Figure Legend Snippet: Fig. 8. Pathway-specific modulation of vlPAG and RVM terminals in the LC modulates nociceptive behavior. (A) Left: Bilateral cannula placement over the LC in uninjected control mice. Right: Hot plate withdrawal latencies of control mice microinfused in the LC with saline (150 nl) versus CNO (3 μM, 150 nl) without (light gray) and with morphine (5 mg/kg s.c.) (dark gray; two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 8 mice; saline versus CNO effect, P = 0.4178, F1,7 = 0.7412). (B) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the vlPAG of Vglut2-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion with saline versus CNO without (light green) and with morphine (5 mg/kg s.c.) (dark green; two-way repeated-measures ANOVA with Sidak’s multiple comparisons test; n = 12 mice; saline versus CNO effect, P = 0.0022, F1,11 = 15.72). (C) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the RVM of Vgat-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion of saline versus CNO (blue bars, n = 12 mice), microinfusion of CNO with intrathecal injections of saline versus phentolamine (5 μg, light blue bars, n = 9 mice), and microinfusion of saline versus CNO with morphine (5 mg/kg s.c.) (dark blue, n = 12 mice; mixed-effects analysis with matching across row and Tukey’s multiple comparisons test, P < 0.0001, F2.560,25.09 = 27.36). (D) Circuit diagram of DPMS inputs to the LC and their opioid sensitivity. Data in each graph reported as means ± SEM.

    Techniques Used: Control, Saline, Injection

    Related Articles

    Activity Assay:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Expressing:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Saline:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Virus:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Injection:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Immunohistochemistry:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    MANN-WHITNEY:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Labeling:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.

    Control:

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.
    Article Snippet: The following viruses were purchased from Addgene: AAV8- hsynDIO- hM4Di- mCherry (Addgene 44362, titer 2.3 × 1013), AAV8hsyn- DIO- hM3Dq- mCherry (Addgene 44361, titer 2.1 × 1013), AAVretro- hsyn- cre (Addgene 105553, titer 2.1 × 1013), AAVretro- Ef1aFlpO (Addgene 55637, titer 1.3 × 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013).× 1013), AAV1- syn- FLEX- jGCaMP8s (Addgene 162377, titer 2.3 × 1013), AAV8- Ef1a- Con/Fon- GCaMP6m (Addgene 137119, titer 2.4 × 1013), AAVretro- Ef1a- DIO- FlpO (Addgene 87306, titer 1.6 × 1013), and AAVretro- hSyn- DIO- EGFP (Addgene 50457, titer 1.4 × 1013). ... The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).. AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.AAV1- FLEX- ef1ataCasp3- TEVP (titer 2.1 × 1012) and AAV1- CAG- FLEX- tdTomato (titer 7.6 × 1012) were both purchased from the UNC Vector Core. scAAV1- hSyn- cre (titer 2.8 × 1013) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-hM4Di-mCherry (Addgene 44362, titer 2.3 × 10 13 ), AAV8-hsyn-DIO-hM3Dq-mCherry (Addgene 44361, titer 2.1 × 10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1 × 10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ).55637, titer 1.3 × 10 13 ), AAV1-syn-FLEX-jGCaMP8s (Addgene 162377, titer 2.3 × 10 13 ), AAV8-Ef1a-Con/Fon-GCaMP6m (Addgene 137119, titer 2.4 × 10 13 ), AAVretro-Ef1a-DIO-FlpO (Addgene 87306, titer 1.6 × 10 13 ), and AAVretro-hSyn-DIO-EGFP (Addgene 50457, titer 1.4 × 10 13 ). ... The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5 × 10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2 × 10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3 × 10 12 ), and AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55 × 10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1 × 10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6 × 10 12 ) were both purchased from the UNC Vector Core. scAAV1-hSyn-cre (titer 2.8 × 10 13 ) was purchased from WZBiosciences.

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation
    Article Snippet: The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).The following viruses were purchased from Addgene: AAV8-hsyn-DIO-HM4Di-mCherry, (Addgene 44362, titer 2.3×10 13 ), AAV8-hsyn-DIO-HM3Dq-mCherry (Addgene 44361, titer 2.1×10 13 ), AAVretro-hsyn-cre (Addgene 105553, titer 2.1×10 13 ), AAVretro-Ef1a-FlpO (Addgene 55637, titer 1.3×10 13 ).. The following were made and titered in-house using Addgene plasmids: AAVDJ-hsyn-DIO-mCherry (Addgene 50459, titer 3.5×10 12 ), AAVDJ-ef1a-fDIO-EYFP (Addgene 55641, titer 2×10 11 ), AAVDJ-Ef1a-mCherry-IRES-cre (Addgene 55632, titer 4.3×10 12 ), AAVDJ-ef1a-DIO-ChR2(H134R)-mCherry (Addgene 20297, titer 2.55×10 13 ).. AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.AAV1-FLEX-ef1a-taCasp3-TEVP (titer 2.1×10 12 ) and AAV1-CAG-FLEX-tdTomato (titer 7.6×10 12 ) were both purchased from the UNC Vector Core.



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    aavdj ef1a mcherry ires cre  (Addgene inc)
    96
    Addgene inc aavdj ef1a mcherry ires cre
    Fig. 3. vlPAG activity is required for systemic morphine antinociception and drives spinal NA–dependent antinociception. (A) Bilateral vlPAG AAV-DIO-hM4Di- <t>mCherry</t> injections in Vglut2-cre mice with representative image of viral expression. Scale bar, 500 μm. (B) Left: Hot plate withdrawal latencies of vlPAGVglut2-cre::hM4Di mice administered CNO (3 mg/kg i.p.) versus saline without (light green bars) and with morphine (5 mg/kg s.c.) (dark green bars) (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 13 mice; saline versus CNO effect, P < 0.0001, F1,12 = 56.93). Right: Withdrawal latencies of non–virus-injected controls (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 9 mice; saline versus CNO effect, P = 0.32, F1,8 = 1.143). (C) Bilateral vlPAG AAV-DIO-hM3Dq- mCherry injections in Vglut2-cre mice, combinations of systemic CNO with intrathecal antagonists, and representative image of viral expression. Scale bar, 500 μm. (D) Left: Withdrawal latencies after systemic saline or CNO and intrathecal saline, phentolamine (5 μg), or naltrexone (5 μg) (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P < 0.0001, Friedman statistic = 26.40). Right: Von Frey mechanical thresholds (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P = 0.0001, Friedman statistic = 21.00). (E) Representative images of c-Fos immunohistochemistry in TH-positive LC neurons of vlPAGVglut2-cre::hM3Dq mice after systemic injection of saline (top) or CNO (bottom). Scale bars, 150 μm. Right: % of TH-positive LC neurons that colocalize with green c-Fos signal (five to eight images analyzed per mouse; n = 6 saline, n = 5 CNO, two-sided Mann-Whitney test). Data in each graph reported as means ± SEM.
    Aavdj Ef1a Mcherry Ires Cre, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/aavdj+ef1a+mcherry+ires+cre/pAAV-Ef1a-mCherry-IRES-Cre+(Plasmid+%2355632)/pm38669335-345-31-36
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    Fig. 3. vlPAG activity is required for systemic morphine antinociception and drives spinal NA–dependent antinociception. (A) Bilateral vlPAG AAV-DIO-hM4Di- mCherry injections in Vglut2-cre mice with representative image of viral expression. Scale bar, 500 μm. (B) Left: Hot plate withdrawal latencies of vlPAGVglut2-cre::hM4Di mice administered CNO (3 mg/kg i.p.) versus saline without (light green bars) and with morphine (5 mg/kg s.c.) (dark green bars) (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 13 mice; saline versus CNO effect, P < 0.0001, F1,12 = 56.93). Right: Withdrawal latencies of non–virus-injected controls (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 9 mice; saline versus CNO effect, P = 0.32, F1,8 = 1.143). (C) Bilateral vlPAG AAV-DIO-hM3Dq- mCherry injections in Vglut2-cre mice, combinations of systemic CNO with intrathecal antagonists, and representative image of viral expression. Scale bar, 500 μm. (D) Left: Withdrawal latencies after systemic saline or CNO and intrathecal saline, phentolamine (5 μg), or naltrexone (5 μg) (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P < 0.0001, Friedman statistic = 26.40). Right: Von Frey mechanical thresholds (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P = 0.0001, Friedman statistic = 21.00). (E) Representative images of c-Fos immunohistochemistry in TH-positive LC neurons of vlPAGVglut2-cre::hM3Dq mice after systemic injection of saline (top) or CNO (bottom). Scale bars, 150 μm. Right: % of TH-positive LC neurons that colocalize with green c-Fos signal (five to eight images analyzed per mouse; n = 6 saline, n = 5 CNO, two-sided Mann-Whitney test). Data in each graph reported as means ± SEM.

    Journal: Science advances

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.

    doi: 10.1126/sciadv.adj9581

    Figure Lengend Snippet: Fig. 3. vlPAG activity is required for systemic morphine antinociception and drives spinal NA–dependent antinociception. (A) Bilateral vlPAG AAV-DIO-hM4Di- mCherry injections in Vglut2-cre mice with representative image of viral expression. Scale bar, 500 μm. (B) Left: Hot plate withdrawal latencies of vlPAGVglut2-cre::hM4Di mice administered CNO (3 mg/kg i.p.) versus saline without (light green bars) and with morphine (5 mg/kg s.c.) (dark green bars) (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 13 mice; saline versus CNO effect, P < 0.0001, F1,12 = 56.93). Right: Withdrawal latencies of non–virus-injected controls (two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 9 mice; saline versus CNO effect, P = 0.32, F1,8 = 1.143). (C) Bilateral vlPAG AAV-DIO-hM3Dq- mCherry injections in Vglut2-cre mice, combinations of systemic CNO with intrathecal antagonists, and representative image of viral expression. Scale bar, 500 μm. (D) Left: Withdrawal latencies after systemic saline or CNO and intrathecal saline, phentolamine (5 μg), or naltrexone (5 μg) (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P < 0.0001, Friedman statistic = 26.40). Right: Von Frey mechanical thresholds (Friedman test with Dunn’s multiple comparisons test, n = 12 subjects, P = 0.0001, Friedman statistic = 21.00). (E) Representative images of c-Fos immunohistochemistry in TH-positive LC neurons of vlPAGVglut2-cre::hM3Dq mice after systemic injection of saline (top) or CNO (bottom). Scale bars, 150 μm. Right: % of TH-positive LC neurons that colocalize with green c-Fos signal (five to eight images analyzed per mouse; n = 6 saline, n = 5 CNO, two-sided Mann-Whitney test). Data in each graph reported as means ± SEM.

    Article Snippet: The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).

    Techniques: Activity Assay, Expressing, Saline, Virus, Injection, Immunohistochemistry, MANN-WHITNEY

    Fig. 4. Anatomical characterization of inputs to the LC from the vlPAG. (A) vlPAG injection of AAV-DIO-tdTom in Vglut2- or Vgat-cre mice. (B) Vglut2-cre injection site. (C) vlPAGVglut2-cre terminals (magenta) and TH (green) in the peri-LC. Bar, Barrington’s nucleus. (D) Vgat-cre injection site. (E) vlPAGVgat-cre terminals. Scale bars, 300 μm [(B) to (E)]. (F) Quantification of magenta and green pixel intensity in the peri-LC of vlPAGVglut2-cre::tdTom mice normalized by z score (n = 6 LC slices from three mice). (G) Same as (F) for vlPAGVgat-cre::tdTom. (H) Left: Injections of AAVretro-cre in the LC and AAV-DIO-mCherry in the vlPAG of wild-type mice. Middle: Image of mCherry+ vlPAG neurons. Right: Resulting terminals in the LC and RVM. Scale bars, 500 μm. (I) Same as (H) for vlPAG→RVM neurons. Scale bars, 500 μm. (J) Left: Orthogonal recombinase strategy to label vlPAG neurons that project to the RVM and LC. Middle: Representative image of mCherry (magenta) and YFP (green). Arrows indicate double-labeled (white) neu- rons. Scale bar, 300 μm. Right: Quantification of mCherry and YFP in the vlPAG.

    Journal: Science advances

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.

    doi: 10.1126/sciadv.adj9581

    Figure Lengend Snippet: Fig. 4. Anatomical characterization of inputs to the LC from the vlPAG. (A) vlPAG injection of AAV-DIO-tdTom in Vglut2- or Vgat-cre mice. (B) Vglut2-cre injection site. (C) vlPAGVglut2-cre terminals (magenta) and TH (green) in the peri-LC. Bar, Barrington’s nucleus. (D) Vgat-cre injection site. (E) vlPAGVgat-cre terminals. Scale bars, 300 μm [(B) to (E)]. (F) Quantification of magenta and green pixel intensity in the peri-LC of vlPAGVglut2-cre::tdTom mice normalized by z score (n = 6 LC slices from three mice). (G) Same as (F) for vlPAGVgat-cre::tdTom. (H) Left: Injections of AAVretro-cre in the LC and AAV-DIO-mCherry in the vlPAG of wild-type mice. Middle: Image of mCherry+ vlPAG neurons. Right: Resulting terminals in the LC and RVM. Scale bars, 500 μm. (I) Same as (H) for vlPAG→RVM neurons. Scale bars, 500 μm. (J) Left: Orthogonal recombinase strategy to label vlPAG neurons that project to the RVM and LC. Middle: Representative image of mCherry (magenta) and YFP (green). Arrows indicate double-labeled (white) neu- rons. Scale bar, 300 μm. Right: Quantification of mCherry and YFP in the vlPAG.

    Article Snippet: The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).

    Techniques: Injection, Labeling

    Fig. 8. Pathway-specific modulation of vlPAG and RVM terminals in the LC modulates nociceptive behavior. (A) Left: Bilateral cannula placement over the LC in uninjected control mice. Right: Hot plate withdrawal latencies of control mice microinfused in the LC with saline (150 nl) versus CNO (3 μM, 150 nl) without (light gray) and with morphine (5 mg/kg s.c.) (dark gray; two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 8 mice; saline versus CNO effect, P = 0.4178, F1,7 = 0.7412). (B) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the vlPAG of Vglut2-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion with saline versus CNO without (light green) and with morphine (5 mg/kg s.c.) (dark green; two-way repeated-measures ANOVA with Sidak’s multiple comparisons test; n = 12 mice; saline versus CNO effect, P = 0.0022, F1,11 = 15.72). (C) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the RVM of Vgat-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion of saline versus CNO (blue bars, n = 12 mice), microinfusion of CNO with intrathecal injections of saline versus phentolamine (5 μg, light blue bars, n = 9 mice), and microinfusion of saline versus CNO with morphine (5 mg/kg s.c.) (dark blue, n = 12 mice; mixed-effects analysis with matching across row and Tukey’s multiple comparisons test, P < 0.0001, F2.560,25.09 = 27.36). (D) Circuit diagram of DPMS inputs to the LC and their opioid sensitivity. Data in each graph reported as means ± SEM.

    Journal: Science advances

    Article Title: Inputs to the locus coeruleus from the periaqueductal gray and rostroventral medulla shape opioid-mediated descending pain modulation.

    doi: 10.1126/sciadv.adj9581

    Figure Lengend Snippet: Fig. 8. Pathway-specific modulation of vlPAG and RVM terminals in the LC modulates nociceptive behavior. (A) Left: Bilateral cannula placement over the LC in uninjected control mice. Right: Hot plate withdrawal latencies of control mice microinfused in the LC with saline (150 nl) versus CNO (3 μM, 150 nl) without (light gray) and with morphine (5 mg/kg s.c.) (dark gray; two-way repeated-measures ANOVA with Tukey’s multiple comparisons test; n = 8 mice; saline versus CNO effect, P = 0.4178, F1,7 = 0.7412). (B) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the vlPAG of Vglut2-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion with saline versus CNO without (light green) and with morphine (5 mg/kg s.c.) (dark green; two-way repeated-measures ANOVA with Sidak’s multiple comparisons test; n = 12 mice; saline versus CNO effect, P = 0.0022, F1,11 = 15.72). (C) Left: Bilateral viral injection of AAV-DIO-hM4Di-mCherry in the RVM of Vgat-cre mice with bilateral cannula placement over the LC. Right: Hot plate withdrawal latencies after microinfusion of saline versus CNO (blue bars, n = 12 mice), microinfusion of CNO with intrathecal injections of saline versus phentolamine (5 μg, light blue bars, n = 9 mice), and microinfusion of saline versus CNO with morphine (5 mg/kg s.c.) (dark blue, n = 12 mice; mixed-effects analysis with matching across row and Tukey’s multiple comparisons test, P < 0.0001, F2.560,25.09 = 27.36). (D) Circuit diagram of DPMS inputs to the LC and their opioid sensitivity. Data in each graph reported as means ± SEM.

    Article Snippet: The following were made and titered in- house using Addgene plasmids: AAVDJ- hsyn- DIO- mCherry (Addgene 50459, titer 3.5 × 1012), AAVDJ- ef1a- fDIO- EYFP (Addgene 55641, titer 2 × 1011), AAVDJ- Ef1a- mCherry- IRES- cre (Addgene 55632, titer 4.3 × 1012), and AAVDJ- ef1a- DIO- ChR2(H134R)mCherry (Addgene 20297, titer 2.55 × 1013).

    Techniques: Control, Saline, Injection