Journal: eNeuro

Article Title: A Focal Inactivation and Computational Study of Ventrolateral Periaqueductal Gray and Deep Mesencephalic Reticular Nucleus Involvement in Sleep State Switching and Bistability

doi: 10.1523/ENEURO.0451-19.2020

Figure Lengend Snippet: Location of microperfusion sites and prediction of the anatomic extent of muscimol-mediated inhibition. A , Example site of microperfusion located at the boundary of the vlPAG and the DpMe. The black arrow indicates the most ventral point of the lesion left by the microdialysis probe. B , Probe locations sites for all 25 rats (blue rectangles = muscimol group; red rectangles = control group) located between the anterioposterior level defined by the caudal pole of the paratrochlear nucleus to the caudal extent of the superior cerebellar peduncle decussation. All probes were implanted on the right side; probes positions are shown on both sides for clarity purposes. C , Results of our simulation-based estimate of average PSI across all experiments in stereotaxic coordinate space.

Article Snippet: Between ZT5 and ZT7.5, ACSF microperfusion was maintained in the time-control group, while the drug-group received microperfusion of the GABA A receptor agonist muscimol (Tocris Bioscience; 85 μ m in ACSF; microdialyzed solution).

Techniques: Inhibition, Control

Journal: International Journal of Molecular Sciences

Article Title: A Novel Sulfonamide, 4-FS, Reduces Ethanol Drinking and Physical Withdrawal Associated With Ethanol Dependence

doi: 10.3390/ijms21124411

Figure Lengend Snippet: ( a , b , e , f ) Schematic of tissue punches from the dorsal hippocampus ( a , b ) and ventral hippocampus ( e , f ). ( c , g ) sample western blots from dorsal ( c ) and ventral ( g ) hippocampus tissue punches. ( d , h ) Densitometric analysis of tissue for levels of GluA1, GluN2A, GluN2B, ratio of GluN2A/2B and GABA A from the dorsal ( d ) and ventral ( h ) hippocampus tissue homogenate. n = 8 4-FS ED rats, n = 7 4-FS CIE-ED rats. Values are mean ± S.E.M. expressed as % control, where control is represented by the dotted line in each graph. * p < 0.05, compared to controls.

Article Snippet: The membranes were incubated with the following primary antibodies: total AMPA Receptor 1 antibody (GluA1; rabbit monoclonal; 1:500, cell signaling technology, Danvers, MA, USA, cat#13185, molecular weight 100 kDa), total glutamate (NMDA) receptor subunit 2A antibody (GluN2A; rabbit polyclonal, 1:200, Santa Cruz Biotechnology Dallas, TX, USA, cat# sc-9056, molecular weight 170 kDa), total glutamate (NMDA) receptor subunit 2B antibody (GluN2B; 1:200, Santa Cruz Biotechnology, Dallas, TX, USA, cat. no. sc-9057, molecular weight 180 kDa) and total GABA A receptor antibody, PhosphoSolutions, Aurora, CO, USA, cat. no. 850-GA6, molecular weight 60 kDa).

Techniques: Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: GABA A receptor subunit composition regulates circadian rhythms in rest-wake and synchrony among cells in the suprachiasmatic nucleus.

doi: 10.1073/pnas.2400339121

Figure Lengend Snippet: Fig. 1. Knock-down or mutation of γ2 or δ GABAA receptor subunits reduced the amplitude of daily rhythms and decreased nocturnal activity in wheel-running behavior. Representative actograms for mice treated with shRNA nontargeted controls (A, NT, n = 17) or targeting either γ2 (B, KD, n = 19) or δ (C, KD, n = 12) GABAA receptor subunits and mutation experiments from WT (F, n = 14) and δ KI (G and H, n = 17) recorded in LD for ~10 d and then released in DD for ~11 to 20 d. Activity profiles for group mean ± SEM in LD for KD (D) and mutants (I) and in DD for KD (E) and mutants (J). Locomotor analysis for group mean ± SEM in LD (K, for KD, and M, for KI) and DD (L, for KD, and N, for KI) showed a decrease in activity during the dark phase independent of light conditions. Panel K, F(2,40) = 4.2, P = 0.02 in LD, panel L, F(2,40) = 5.1, P = 0.01 in DD for KD, one-way ANOVA and panel M, t(28) = 4.9, P < 0.0001 in LD and panel N, t(30) = 2.6, P = 0.01 in DD for mutations, t test.

Article Snippet: The initial screening consisted of overexpressing (OVX) Gabrg2, the GABAA γ2 (Addgene, #49170) gene, or Gabrd, the gene encoding δ (Applied Biological Materials, #AAV0872429) receptor subunits in HEK- 293 cells (ATCC).

Techniques: Knockdown, Mutagenesis, Activity Assay, shRNA

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: GABA A receptor subunit composition regulates circadian rhythms in rest-wake and synchrony among cells in the suprachiasmatic nucleus.

doi: 10.1073/pnas.2400339121

Figure Lengend Snippet: Fig. 3. Knock-down or mutation of γ2 or δ GABAA receptor subunits reduced synchrony among circadian cells in the SCN. (A, B, and C) Raster plots depicting PER2 expression over 6 d from cells in a representative SCN explant treated either with NT, γ2 KD, and δ KD (n = 6) or cultured from WT, γ2 KI, or δ KI mice, respectively (G, H, and I). Ensemble (mean ± SEM) PER2 expression from SCN explants treated with NT (n = 4), γ2 KD (n = 4), and δ KD (n = 6, panel D) or from WT (n = 4), γ2 KI (n = 5), or δ KI (n = 7, panel J) mice. Loss or mutation of γ2 or δ GABA receptor subunits reduced daily synchrony indexes (E and K; mean ± SEM) and increased the period distributions among circadian cells (F and L) in the isolated SCN. For panel E, Group F(2,60) = 47, P < 0.0001; Days (5,60) = 1.7, P = 0.1; Group*Days F(10,60) = 0.4, P = 0.9. For panel K, Group F(2,48) = 20.5, P < 0.0001; Days F(5,48) = 1.8, P = 0.1; Group*Days F(10,48) = 0.5, P = 0.8, two-way mixed ANOVA. For panel F, F(2, 65) = 45.3 one-way ANOVA; NT period SD 1.3, period CV 5.4; γ2 KD period SD 1.5, period CV 5.7; δKD period SD 1.9, period CV 7.2, Bartlett’s test P < 0.0001. For panel L, F(2, 68) = 41, one-way ANOVA, WT period SD 0.3, period CV 1.3; γ2 KI period SD 1.9, period CV 7.9; δKI, period SD 1.1, period CV 4.4, Barlett’s test P < 0.0001. SD = standard deviation, CV = coefficient of variance.

Article Snippet: The initial screening consisted of overexpressing (OVX) Gabrg2, the GABAA γ2 (Addgene, #49170) gene, or Gabrd, the gene encoding δ (Applied Biological Materials, #AAV0872429) receptor subunits in HEK- 293 cells (ATCC).

Techniques: Knockdown, Mutagenesis, Expressing, Cell Culture, Isolation, Standard Deviation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: GABA A receptor subunit composition regulates circadian rhythms in rest-wake and synchrony among cells in the suprachiasmatic nucleus.

doi: 10.1073/pnas.2400339121

Figure Lengend Snippet: Fig. 5. Both γ2 and δ GABAA subunits are regulating synchrony among SCN cells. Cell raster plots from a representative single SCN explant for δ OV + γ2 KD and γ2 OV + δ KD (A and B) or δ OV + γ2 KI and γ2 OV + δ KI (F and G). Ensemble mean ± SEM PER2 expression traces from the same color-coded single SCN explants for KD (C; δ OV + γ2 KD, n = 2 and γ2 OV + δ KD, n = 2) and mutations (H; δOV + γ2 KI, n = 3 and γ2 OV + δ KI, n = 3). (D and I) Group mean ± SEM sync indices showing that overexpression of one GABA receptor rescues desynchrony induced by KD or mutations of the complementary GABA receptor. (E and J) Overexpression of γ2 or δ receptors restore period distributions in the KD (E) and mutations (J) groups. Panel D, Group F(2,36) = 1.6, P = 0.2; Days F(5,36) = 0.1, P = 0.9; Group*Days F(10,36) = 0.1, P = 0.9. Panel I, Group F(2,25) = 2.4, P = 0.3; Days F(4,25) = 0.2, P = 0.9; Group*Days F(8,25) = 0.04, P = 0.9, two-way mixed ANOVA.

Article Snippet: The initial screening consisted of overexpressing (OVX) Gabrg2, the GABAA γ2 (Addgene, #49170) gene, or Gabrd, the gene encoding δ (Applied Biological Materials, #AAV0872429) receptor subunits in HEK- 293 cells (ATCC).

Techniques: Expressing, Over Expression