trpv1 mrna  (Human Protein Atlas)

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Manual Acupuncture at ST37 Modulates TRPV1 in Rats with Acute Visceral Hyperalgesia via Phosphatidylinositol 3-Kinase/Akt Pathway

doi: 10.1155/2021/5561999

Figure Lengend Snippet: Effects of MA stimulation on NGF, TrkA, PI3K, Akt, and TRPV1 mRNA expressions in the colon of rats with acute visceral hyperalgesia. Data are expressed as the mean ± SD ( n = 3 for each group. ∗ P < 0.05 vs. model group; & P < 0.05 vs. ST37 group; # P < 0.05 vs. ST37 + IGF-1 group).

Article Snippet: The primers for NGF, TrkA, PI3K, and TRPV1 mRNA detection were predesigned (GenScript, China) as follows: NGF F: 5ʹ- ACT GGA CTA AAC TTC AGC ATT CC-3ʹ, R: 5ʹ-GGG CAG CTA TTG GTT CAG CA-3ʹ; TrkA F: 5ʹ-GCC TAA CCA TCG TGA AGA GTG-3ʹ, R: 5ʹ-CCA AAG CAT TGG AGG AGA GAT-3ʹ; PI3K F: 5ʹ-CCC ACT TCT ATA GCC AAC AAC-3ʹ, R: 5ʹ-GAT ATC TCC CCA GTA CCA TT-3ʹ; Akt F: 5ʹ-CAC AGG TCG CTA CTA TGC CAT GAA G-3ʹ, R: 5ʹ- GCA GGA CAC GGT TCT CAG TAA GC-3ʹ; TRPV1, F: 5ʹ-CCC GGA AGA CAG ACA GCC TGA-3ʹ, R 5ʹ-TTC AAT GGC AAT GTG CAG TGC TG-3ʹ; and GAPDH F: 5ʹ-AGA TGG TGA AGG TCG GTG TGA-3ʹ, R: 5ʹ-CTG GAA GAT GGT GAT GGG TTT CC-3ʹ.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Manual Acupuncture at ST37 Modulates TRPV1 in Rats with Acute Visceral Hyperalgesia via Phosphatidylinositol 3-Kinase/Akt Pathway

doi: 10.1155/2021/5561999

Figure Lengend Snippet: Effects of MA stimulation on NGF, TrkA, PI3K, TRPV1, and p-Akt protein expressions in the colons of rats with acute visceral hyperalgesia. Representative blots for NGF, TrkA, PI3K, TRPV1, p-Akt, and densitometry analyses were performed (a–e). To confirm equal sample loading, β -actin was used for normalization (the level of the p-Akt was counted by the ratio of p-Akt/Akt). Data are expressed as the mean ± SD ( n = 6 for each group. ∗ P < 0.05 vs. model group; & P < 0.05 vs. ST37 group; # P < 0.05 vs. ST37 + IGF-1 group).

Article Snippet: The primers for NGF, TrkA, PI3K, and TRPV1 mRNA detection were predesigned (GenScript, China) as follows: NGF F: 5ʹ- ACT GGA CTA AAC TTC AGC ATT CC-3ʹ, R: 5ʹ-GGG CAG CTA TTG GTT CAG CA-3ʹ; TrkA F: 5ʹ-GCC TAA CCA TCG TGA AGA GTG-3ʹ, R: 5ʹ-CCA AAG CAT TGG AGG AGA GAT-3ʹ; PI3K F: 5ʹ-CCC ACT TCT ATA GCC AAC AAC-3ʹ, R: 5ʹ-GAT ATC TCC CCA GTA CCA TT-3ʹ; Akt F: 5ʹ-CAC AGG TCG CTA CTA TGC CAT GAA G-3ʹ, R: 5ʹ- GCA GGA CAC GGT TCT CAG TAA GC-3ʹ; TRPV1, F: 5ʹ-CCC GGA AGA CAG ACA GCC TGA-3ʹ, R 5ʹ-TTC AAT GGC AAT GTG CAG TGC TG-3ʹ; and GAPDH F: 5ʹ-AGA TGG TGA AGG TCG GTG TGA-3ʹ, R: 5ʹ-CTG GAA GAT GGT GAT GGG TTT CC-3ʹ.

Techniques:

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: TRPV1 Expression in Retinal Ganglion Cell Neurons. (A) Immuno-labeling for TRPV1 in mouse retina shows strong localization in the outer and inner plexiform layers (OPL and IPL, respectively) and in peri-nuclear compartments of RGC bodies co-labeled against phosphorylated neurofilaments (pNF). In macaque monkey retina (B), RGCs (large cells) demonstrate discrete pockets of TRPV1 localization (arrows), while in aged human retina (C) TRPV1 distributes throughout the RGC cytoplasm. Scale = 20 μm (A, B) or 10 μm (C).

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: Expressing, Immunolabeling, Labeling

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: Trpv1 mRNA in Retinal Ganglion Cell Neurons. (A) In situ hybridization using anti-sense (AS) sequence for RGC-specific Thy1.1 in whole-mounted C57 retina labels nearly every RGC compared to the control sense sequence (right). Anti-sense label for Trpv1 (B) in C57 labels fewer cells in ganglion cell layer (sense probe shown for comparison, right). (C) Fluorescent in situ hybridization probe for Trpv1 mRNA combined with immune-labeling for RGCs (pNF) and astrocytes (GFAP) shows strong anti-sense signal in roughly 70% of RGCs compared to the control sense sequence (right). (D) Quantitative RT-PCR detection of Trpv1 mRNA compared to 18s in individual naïve C57 retinas shows large inter-retina variability (mean ± sem is shown for comparison: 1.04 ± 0.17, n = 12). (E) Conventional RT-PCR product for Trpv1 mRNA in isolated RGCs maintained in culture (inset) shows significant increase following 24 hr exposure to elevated hydrostatic pressure (+70 mmHG; *, P < 0.001). By 48 hrs, expression had returned to ambient levels. Scale = 40 μm (B) or 20 μm (C).

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: In Situ Hybridization, Sequencing, Labeling, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Isolation, Expressing

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: TRPV1 Activation Increases RGC Intracellular Ca2+. (A) Fluo-4 conjugated Ca2+ accumulates in RGC cell bodies and neurites. Quantification with intensity map (right panel) shows a consistent network of neurites outlined by small puncta of low Ca2+ accumulation. (B) RGCs treated for 1 hour with 1 μM capsaicin (CAP) demonstrate increased Ca2+ in the cell body and primary neurites, with lower accumulation in background neurites (right panel). (C) RGCs treated for 1 hour with both 1 μM CAP and 10 nM of the TRPV1-specific antagonist IRTX show greatly diminished Ca2+ accumulation, especially in neurites, where Fluo-4 signal is almost completely absent. (D) Treatment with 10 nM IRTX alone eliminates most Ca2+ signal. E. Quantification of Fluo-4 signal indicates a significant increase with CAP compared to vehicle (*). IRTX significantly reduces signal relative to vehicle (+) and CAP-treated RGCs (‡). Symbols indicate P ≤ 0.05; error bars represent standard error. Pixel intensity calculated as average of total intensity from 15–20 independent fields. N = 3 preparations per condition. Scale = 100 μm (A-C) or 150 μm (D).

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: Activation Assay

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: Trpv1-/- Mice Express Truncated Form of Gene. Agarose gel showing PCR products from genotyping 4 Trpv1-/- and 2 C57 mice used in our experiments. Reaction products were 984 base pairs (bp) for C57 and 600 bp Trpv1-/- mice, as expected based on the truncated protein.

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: Agarose Gel Electrophoresis

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: TRPV1 Suppression Improves RGC Survival in Retinal Explants. (A) Confocal micrographs through ganglion cell layer of explants from C57 retina maintained ex vivo at either ambient or elevated (+70 mmHG) hydrostatic pressure for 48 hrs. Compared to vehicle (DMSO, top row), explants under pressure treated with the TRPV1-specific antagonist iodoresiniferatoxin (IRTX, 10 nM, bottom row) exhibit far fewer TUNEL+ nuclei (green) and no loss of Brn3a immune-labeled RGCs (red). (B) Explants from Trpv1-/- retina show similar density of Brn3a-labeled RGCs at 48 hrs of either ambient or elevated pressure and no increase in TUNEL with pressure.(C) Quantification of explants indicates a 25% reduction in Brn3a-labeled RGCs with pressure compared to ambient (*, p = 0.01). IRTX prevents the pressure-induced reduction in Brn3a-labeled RGCs compared to C57 (**, p = 0.02), but also reduces the number cells compared to vehicle at ambient pressure (‡, P < 0.001). EGTA (950 μM) reduces RGC loss in under pressure (**, p = 0.03), as does Trpv1-/- compared to C57 (**, p = 0.02). There was no difference between C57 and Trpv1-/- at ambient pressure (p = 0.4).(D) Fraction of TUNEL+ nuclei in the RGC layer of C57 explants is increased nearly 7-fold compared to ambient pressure (*, P < 0.001). IRTX treatment reduces TUNEL label compared to vehicle both at ambient (‡, p = 0.003) and elevated (**, P < 0.001) pressure. For explants under pressure, both EGTA and Trpv1-/- decrease TUNEL significantly compared to C57 with pressure (**P < 0.001). Scale = 20 μm (A,B).

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: Ex Vivo, TUNEL Assay, Labeling

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: TRPV1 Co-localizes with TRPV4. (A) Vertical sections of rat retina show localization of TRPV1 (left) and TRPV4 (right) in RGCs whose axons are strongly labeled for phosphorylated neurofilaments (pNF). The pNF fluorescent signal has been decreased purposefully due to its high levels in axons. Arrows indicate likely localization of TRPV4 in Müller glia end-feet. (B) In whole-mount of C57 retina, TRPV4 localizes to most cell bodies of the ganglion cell layer and to Müller glia end-feet in between them (arrows). GFAP-labeled astrocytes are shown for comparison. (C) Rat retina labeled for TRPV1 and TRPV4 shows strong co-localization (arrows) in pNF-labeled RGCs.

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: Labeling

Journal: Channels

Article Title: Activation of transient receptor potential vanilloid-1 (TRPV1) influences how retinal ganglion cell neurons respond to pressure-related stress

doi: 10.1080/19336950.2015.1009272

Figure Lengend Snippet: TRPV1 Forms a Protein Complex with TRPV4. (A) Western blots from brain and retina for TRPV4 and TRPV1 with full-length product in expected range near 100 kDa (dark arrowhead). TRPV1 also shows strong product near 75 kDa (open arrowhead), consistent with earlier work.20 Product for β-actin (*, run simultaneously) is shown for comparison. Smaller products (∼60 kDa) likely represent protein degradation. (B) Western blots (WB) for TRPV1 (left) and TRPV4 (right) following immuno-precipitation (IP) of TRPV4 or TRPV1, respectively, from brain and retina extracts. Co-immuno-precipitation product for the TRPV4/TRPV1 complex (bracket) is the same whether the immuno-precipitate is TRPV4 (left) or TRPV1 (right) and is slightly less heavy than the WB product for TRPV1 for brain in the absence of IP (arrowhead). (C) Western blots for TRPV1 (brain) following immuno-precipitation (IP) of TRPV4 shows same reaction product near 100 kDa as before. When supernatant (S) from the co-immuno-precipitation reaction is probed again for TRPV1, this product is absent (bracket), supporting specific binding of the 100 kDa form of TRPV1 to TRPV4. (D) Quantitative PCR measurements of Trpv1 mRNA in young (3–5 mo) and aged (8–10 mo) DBA2J retina relative to levels in age-matched D2 control retina; both calculated relative to 18s rRNA. Trpv1 mRNA correlates significantly with intraocular pressure for the 3–5 mo group (r2 = 0.656, P < 0.001) but not for the 8–10 mo group (r2 = 0.17, p = 0.17). Some data modified from recent work.35 In contrast, while expression of Trpv4 mRNA is higher than Trpv1 (E), levels are inversely correlated with increasing pressure for the 3–5 mo group (r2 = 0.79, P < 0.001), but not for the 8–10 mo group (r2 = 0.26, p = 0.091)

Article Snippet: Fluorescent in situ hybridization with immuno-labeling We generated Trpv1 mRNA probes for mouse tissue as described in our published protocol, using RNA extracted from C57BL/6 mouse brain (Qiagen Inc.. USA, Valencia, CA) and first-strand cDNA synthesis using Superscript III reverse transcriptase (Invitrogen) 35,82 An antisense probe recognizingTrpv1 mRNA was made against a nucleotide sequence present in mouse Trpv1 (nucleotides 226 to 500 of [GenBank: {"type":"entrez-nucleotide","attrs":{"text":"NM_001001445","term_id":"676260266","term_text":"NM_001001445"}} NM_001001445 ]).

Techniques: Western Blot, Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction, Modification, Expressing