Journal: Translational Neurodegeneration

Article Title: Astrocytes and retrograde degeneration of nigrostriatal dopaminergic neurons in Parkinson’s disease: removing axonal debris

doi: 10.1186/s40035-021-00262-1

Figure Lengend Snippet: Antibodies used in the study

Article Snippet: VMaT2 , 1:1500 , Guinea Pig , Origene, EUD2801.

Techniques:

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: Immunohistochemical reagents

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques: Immunohistochemical staining

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: Immunohistochemical representation of P2X3, isolectin B4 (IB4), and substance P (SP) in nodose ganglion (NG) neurons. A: following staining of the immunohistochemical markers P2X3, SP, and IB4, the bar graph demonstrates that all three were well represented in the NG, with the majority being IB4+ (IB4 vs. P2X3, #P < 0.05; IB4 vs. SP, **P < 0.001; P2X3 vs. SP, *P < 0.01). Values are means ± SD; n = 3 rats and 6 ganglia (one-way ANOVA with Tukey post hoc t-tests). B: pie graph depicting all possible combinations of the molecular targets in the NG. Note that neurons that were IB4+ only were the most prevalent, and most NG neurons were labeled with at least one of the three cellular targets examined.

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques: Immunohistochemical staining, Staining, Labeling

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: Quadruple immunohistochemical staining in the NG. A: NeuN. B: IB4. C: P2X3. D: SP. E: merged. A confocal image displays the typical staining within the NG. NeuN was used to label all neurons. Different histochemical combinations include neurons that were IB4+, P2X3+, but SP− (white arrows), and neurons that were IB4−, P2X3+, and SP− (yellow arrows). Scale bar indicates 25 μm (×20 objective).

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques: Immunohistochemical staining, Staining

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: The effect of spinal cord injury (SCI) on number of NG neurons expressing the individual markers. Following SCI, there was an increase in P2X3-immunoreactivity (ir) in the transected group relative to intact/normal (SCI, 27.3 ± 4.8% vs. noninjured, 15.0 ± 3.3%, *P < 0.001) and a decrease in IB4 binding in the transected group relative to intact/normal (SCI, 23.7 ± 6.5% vs. noninjured, 33.3 ± 7.1%, #P < 0.05). No changes were apparent for SP. The “Other” category represents neurons that were NeuN+, but did not express or bind any of the markers. Values are means ± SD; n = 6 rats and 12 ganglia.

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques: Expressing, Binding Assay

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: The effect of SCI on P2X3 and IB4 in the NG. An example displaying P2X3-ir (A) and IB4 binding (C) in the NG and following chronic spinal cord transection injury at T8 (B and D, respectively) is shown. Note the presence of increased P2X3-ir and decreased IB4 binding post-SCI. Images of sections from both SCI and noninjured animals were stained and captured with the same protocols and at the same time (×10 objective).

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques: Binding Assay, Staining

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: Effect of SCI on bladder-traced NG neurons. The graph demonstrates that, out of the total percentage of either P2X3 or IB4 subsets after injury, more than one-half of the neurons were traced from bladder. Bladder innervating neurons in the P2X3+ subset represent 32.8 ± 1.1%, while, in the IB4+ subset, they represent 42.6 ± 5.1%. n = 3 rats and 6 ganglia.

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques:

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: The effect of spinal cord injury on the neurochemical properties of vagal sensory neurons

doi: 10.1152/ajpregu.00445.2014

Figure Lengend Snippet: P2X3-ir and IB4 binding in bladder-traced NG neurons after transection. A confocal image illustrates a DiI+ neuron in A that is also immunoreactive for P2X3 in B (white arrows). C: demonstration of the overlay. An image from the inverted Nikon microscope illustrates a DiI+ neuron in D that also binds IB4 in E (white arrowhead). F: demonstration of the overlay. One example of each is displayed. In both images, the scale bar indicates 25 μm (×20 objective).

Article Snippet: Secondary Detection Dilution/Vendor/Catalog No. P2X3 GP-anti-P2X3 1:1,000/Neuromics/GP10108 Dky anti-GP-AF488 1:100/Jackson/706-545-148 Biotinylated lectin from Bandeiraea simplicifolia isolectin B4 HRP-SA 1:500/Sigma-Aldrich/L2140 Tyramide-AF350 1:100/Molecular Probes/T20937TSA kit no. 27 SP RBT-anti-SP 1:1,000/Abcam/ab67006 Dky anti-RBT-Cy3 1:100/Jackson/711-165-152 NeuN MS-anti-NeuN 1:1,000/Chemicon/MAB377 Dky anti-MS-Cy5 1:100/Jackson/715-175-151 Open in a separate window SP, Substance P; GP, guinea pig; HRP, horseradish peroxidase; SA, strepavidin; RBT, rabbit; MS, mouse; Dky, donkey; AF, Alexa Fluor; TSA, tyramide signal amplification.

Techniques: Binding Assay, Microscopy

Journal: Scientific Reports

Article Title: Unraveling the role of natriuretic peptide clearance receptor (NPR3) in glomerular diseases

doi: 10.1038/s41598-024-61603-4

Figure Lengend Snippet: Characterization of NPR3 Pod-KO at baseline and after NTS-induced injury. ( A ) Generation of conditional knockout mice in which NPR3 is specifically ablated in podocytes using Cre–LoxP recombination system. Exon 3 is deleted upon NPHS2-Cre-mediated recombination (L/R = left/right genotyping primer). ( B ) Genotyping by ear preparation and PCR at 4 weeks of age. ( C ) Expression of Tomato-reporter gene (red) and Podocyte marker Nephrin (green), scale bar; 100 μm. ( D ) Urinary cGMP levels in WT and NPR3 Pod-KO mice, before and after NTS challenge (urine cGMP WT; n = 6, urine cGMP NPR3 Pod-KO ; n = 8, urine cGMP NTS WT; n = 5, urine cGMP NTS NPR3 Pod-KO ; n = 10). ( E ) Serum cGMP levels in WT and NPR3 Pod-KO mice, before and after NTS challenge (serum cGMP WT; n = 4, serum cGMP NPR3 Pod-KO ; n = 5, serum cGMP NTS WT; n = 9, serum cGMP NTS NPR3 Pod-KO ; n = 13). ( F ) Urinary albumin/ creatinine ratios (U-ACR) in WT and NPR3 Pod-KO after NTS-induced glomerulonephritis (WT; n = 5, NPR3 Pod-KO ; n = 6). ( G ) Histology and quantification of affected glomeruli in NTS-challenged mice. 30 randomly selected glomeruli were evaluated using PAS staining (WT; n = 4, NPR3 Pod-KO ; n = 5). ( H ) Electron microscopic findings and quantification of mean glomerular basement membrane (GBM) thickness and the number of slits per μm GBM in control and NTS challenged mice. Quantification was performed from transmission electron microscopy images (control; n = 5, WT; n = 4, NPR3 Pod-KO ; n = 4). ( I ) WT1 staining and quantification of positive cells in glomeruli, glomerular area and WT1 positive podocytes/glomerular area control and NTS challenged mice. Nephrin staining (green) was used for the quantification of glomerular area and WT1 (red) for number of podocytes. Quantification was performed from immunofluorescent images (control; n = 7, WT; n = 9, NPR3 Pod-KO ; n = 13). *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001, ****p ≤ 0.0001.

Article Snippet: Frozen and/or paraffin sections were incubated with NPR3 (ab97389, abcam, 1:1000), Synaptopodin (61094; Progen, 1:700), CD31 (303105; Biolegend, 1:500), PDGFRβ (MAB1263; R&D System, 1:1000), Nephrin (BP5030; Origene, 1:200), Wilms-Tumor1 (WT1) (ab89901; abcam, 1:100), ⍺-smooth muscle actin (A5228; Merck, 1:1000), tdTomato (TA150128; Origene, 1:50) at 4 °C overnight.

Techniques: Knock-Out, Expressing, Marker, Staining, Membrane, Control, Transmission Assay, Electron Microscopy

Journal: Scientific Reports

Article Title: Unraveling the role of natriuretic peptide clearance receptor (NPR3) in glomerular diseases

doi: 10.1038/s41598-024-61603-4

Figure Lengend Snippet: Treatment with NPR3 inhibitor via subcutaneous osmotic-pumps in mice with NTS-induced glomerular injury. ( A ) Schematic representation of the experimental treatment setup. ( B ) Urinary albumin/ creatinine ratios (U-ACR) in vehicle and NPR3 i-treated mice after induction of glomerulonephritis (n = 5 for both). ( C ) Histology and quantification of affected glomeruli in treated mice. 30 randomly selected glomeruli were evaluated using PAS and Masson Trichrome staining (vehicle; n = 8, NPR3i; n = 9). ( D ) WT1 staining and quantification of positive cells in glomeruli, glomerular area and WT-1 positive podocytes/glomerular area in control and treated mice. Nephrin staining (green) was used for the quantification of glomerular area and WT1 (red) for number of podocytes. Quantification was performed from immunofluorescent images; 20 randomly selected glomeruli were evaluated (WT; n = 6, vehicle; n = 9, NPR3i; n = 8). ( E ) Electron microscopic findings and quantification of mean glomerular basement membrane (GBM) thickness and the number of slits per μm GBM in control and NTS challenged mice. Quantification was performed from transmission electron microscopy images (control; n = 5, WT; n = 4, NPR3 Pod-KO ; n = 4). ( F ) Staining and quantification of fibrosis marker ⍺-sma positive area in kidney cortex of control and treated mice. Quantification was performed from immunofluorescent images; an average of 10–12 randomly selected cortical cross sections with visible glomeruli were evaluated (WT; n = 2, vehicle; n = 8, NPR3i; n = 9). Scale bars: 100 μm. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001, ****p ≤ 0.0001.

Article Snippet: Frozen and/or paraffin sections were incubated with NPR3 (ab97389, abcam, 1:1000), Synaptopodin (61094; Progen, 1:700), CD31 (303105; Biolegend, 1:500), PDGFRβ (MAB1263; R&D System, 1:1000), Nephrin (BP5030; Origene, 1:200), Wilms-Tumor1 (WT1) (ab89901; abcam, 1:100), ⍺-smooth muscle actin (A5228; Merck, 1:1000), tdTomato (TA150128; Origene, 1:50) at 4 °C overnight.

Techniques: Staining, Control, Membrane, Transmission Assay, Electron Microscopy, Marker

Journal: STAR Protocols

Article Title: Volumetric super-resolution imaging by serial ultrasectioning and stochastic optical reconstruction microscopy in mouse neural tissue

doi: 10.1016/j.xpro.2021.100971

Figure Lengend Snippet:

Article Snippet: Guinea pig polyclonal anti-Calbindin (1:100) , Synaptic Systems , Cat#214 005.

Techniques: Recombinant, Saline, Electron Microscopy, Plasmid Preparation, Software, Imaging, Control, Mass Measurement, Capsules, Microscopy, Staining, Adhesive, Spectrophotometry

Journal: eLife

Article Title: A proline-rich motif on VGLUT1 reduces synaptic vesicle super-pool and spontaneous release frequency

doi: 10.7554/eLife.50401

Figure Lengend Snippet:

Article Snippet: Antibody , VIAAT, Guinea pig, polyclonal , SYSY , Cat. 131004 RRID: AB_887873 , 1:1000.

Techniques: Transfection, Construct, Recombinant, Software

Journal: eLife

Article Title: Synaptotagmin-1 is the Ca 2+ sensor for fast striatal dopamine release

doi: 10.7554/eLife.58359

Figure Lengend Snippet:

Article Snippet: Antibody , Guinea pig polyclonal anti-Tyrosine hydroxylase (A111) , Synaptic Systems , Cat# 213 104, RRID: AB_2619897 , IHC (1:1000).

Techniques: Recombinant, Plasmid Preparation, Software