Journal: ACS Omega

Article Title: Electroacupuncture Attenuates Cyclophosphamide-Induced Bladder Pain by Restoring Autophagy in Spinal Inhibitory Interneurons

doi: 10.1021/acsomega.6c01060

Figure Lengend Snippet: Time-dependent increase in oxidative DNA damage in the spinal dorsal horn and its cellular distribution. (A) Representative images of 8-OHdG immunofluorescence in the spinal dorsal horn of control rats and CYP-treated rats at 4, 7, and 15 days. (B) Quantification of 8-OHdG integrated density. ( n = 5 rats/group; per-rat averages of 4–5 sections; Kruskal–Wallis test followed by Dunn’s multiple comparisons test.) (C) Representative double immunofluorescence images of 8-OHdG (red) with GFAP, Iba1, or NeuN (green) in CYP-treated rats at 4, 7, and 15 days (left). Quantification of the proportion of double-labeled cells among GFAP + , Iba1 + , or NeuN + cells (middle). Colocalization was also quantified by Pearson’s correlation coefficient ( r ) (right). ( n = 5 rats/time point; per-rat averages of 4–5 sections; Kruskal–Wallis test followed by Dunn’s multiple comparisons test.). (D) Representative images of 8-OHdG (red) and GAD67-GFP (green) in the spinal dorsal horn of control mice and CYP-treated GAD67-GFP mice at 15 days after CYP injection. Quantification of the proportion of double-labeled cells among GAD67-GFP-positive neurons (middle) and Pearson’s correlation coefficient ( r ) for 8-OHdG and GAD67-GFP colocalization (right). ( n = 5 mice/group; per-mouse averages of 4–5 sections; Mann–Whitney U test.) Scale bar: 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: GAD67-GFP mice were acquired from The Jackson Laboratory and were used for immunofluorescence identification of inhibitory neurons.

Techniques: Immunofluorescence, Control, Labeling, Injection, MANN-WHITNEY

Journal: ACS Omega

Article Title: Electroacupuncture Attenuates Cyclophosphamide-Induced Bladder Pain by Restoring Autophagy in Spinal Inhibitory Interneurons

doi: 10.1021/acsomega.6c01060

Figure Lengend Snippet: Autophagy impairment is mainly observed in neurons and involves inhibitory neurons. (A) Representative double immunofluorescence images of p62 (red) with Iba1, GFAP, or NeuN (green) in the spinal dorsal horn of rats at 4, 7, and 15 days after CYP injection (left). Quantification of the proportion of double-labeled cells among GFAP-positive, Iba1-positive, or NeuN-positive cells (middle) and Pearson’s correlation coefficient ( r ) for colocalization between p62 and each marker (right). ( n = 5 rats/time point; per-rat averages of 4–5 sections; Kruskal–Wallis test followed by Dunn’s multiple comparisons test.) (B) Representative transmission electron microscopy (TEM) images of neurons in the spinal dorsal horn from control and CYP-treated rats (left), with quantification of the number of autophagosomes (right). Low-magnification images show overview fields, and high-magnification images show the corresponding zoom-ins of the boxed regions. Green arrows indicate ribosomes, and white arrows indicate autophagosomes/phagophores. Scale bars: 2 μm (low magnification) and 1 μm (high magnification). ( n = 6 rats/group; per-rat averages of 4–5 sections; Mann–Whitney U test.) (C) Representative immunofluorescence images showing p62 (red) and GAD67-GFP (green) in the spinal dorsal horn of control and CYP-treated mice at 15 days after CYP injection (left). Quantification of the proportion of double-labeled cells among GAD67-GFP-positive neurons (middle) and Pearson’s correlation coefficient ( r ) for colocalization between p62 and GAD67-GFP (right). ( n = 5 mice/group; per-mouse averages of 4–5 sections; Mann–Whitney U test.) Scale bar: 100 μm for immunofluorescence images. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: GAD67-GFP mice were acquired from The Jackson Laboratory and were used for immunofluorescence identification of inhibitory neurons.

Techniques: Immunofluorescence, Injection, Labeling, Marker, Transmission Assay, Electron Microscopy, Control, MANN-WHITNEY

Journal: Science Advances

Article Title: A causal coding variant regulating alternative splicing of DOC2A at 16p.11.2 GWAS locus influences susceptibility to schizophrenia

doi: 10.1126/sciadv.adw7667

Figure Lengend Snippet: ( A ) Representative traces of mEPSC in primary hippocampal neuron infected with control (empty vector), DOC2A Full-Length , or DOC2A ∆Val217-Pro218 AAV. ( B and C ) Quantification of mEPSC amplitude and frequency. N ≥ 29 neurons. ( D ) Representative traces of mIPSC in in primary hippocampal neuron infected with control, DOC2A Full-Length , or DOC2A ∆Val217-Pro218 AAV. ( E and F ) Quantification of mIPSC amplitude and frequency. N ≥ 20 neurons. ( G ) Representative gels of Western blot. ( H to K ) Quantification of synaptophysin, VAMP-2, GAD65, and GAD67 expression in hippocampus tissue overexpressing control, DOC2A Full-Length , or DOC2A ∆Val217-Pro218 . N = 3 in each group. Full Western blotting images are shown in fig. S16. Data are presented as the means ± SEM; * P < 0.05, ** P < 0.01, and *** P < 0.001. Error bars indicate the SEM.

Article Snippet: The primary antibodies were as follows: FLAG (Cell Signaling Technology, no. 8146S; 1:1000) and DOC2A (Invitrogen, no. PA5-31345; 1:1000), GAD67 (Proteintech, no. 10408-1-AP; 1:5000), GAD65 (Cell Signaling Technology, no. 5843T; 1:1000), synaptophysin (Sigma-Aldrich, no. S5768; 1:500), VAMP-2 (Synaptic Systems, no. 104211; 1:2000), α-tubulin (Proteintech, no. 66031-1-Ig; 1:20,000), and GAPDH (glyceraldehyde-3-phosphate dehydrogenase; Proteintech, no. 10494-1-AP; 1:20,000).

Techniques: Infection, Control, Plasmid Preparation, Western Blot, Expressing