human cb1 receptor  (Revvity)

Journal: International Journal of Molecular Sciences

Article Title: Cannabinoid Receptor Type 1 Regulates Drug Reward Behavior via Glutamate Decarboxylase 67 Transcription

doi: 10.3390/ijms221910486

Figure Lengend Snippet: Effects of CB1 knockdown on GAD67 expression in primary cultured neurons and METH-induced CPP. ( A ) Vehicle group was treated with vehicle (5% DMSO, 5% tween-20, 90% PBS) as the same amount of JWH-210 for 24 h. JWH-210 group was treated with 0.1 μM JWH-210 for 24 h. Mock + JWH-210 and CB1 overexpression (OE) + JWH-210 groups were treated with mock or CB1 ORF lentiviral particles (5 MOI) for 24 h before treatment with vehicle or 0.1 μM JWH-210 for 24 h. Immunostaining of CB1, Myc, and GAD67 was performed with specific antibodies (Scale bar: 200 μm). ( B ) Cultured neurons were treated with control or CB1 shRNA lentiviral particles. mRNA levels of ( C ) CB1 and ( D ) GAD67 were confirmed by using qPCR with each specific primer and normalized to the relative amplification of GAPDH. Data are expressed as the mean ± S.E. ( n = 6; Student’s t -test; * p < 0.05 and ** p < 0.01 vs. vehicle group). ( E ) After CB1 knockdown, 0.3 mg/kg METH-induced CPP scores (s) was measured by CPP test. Data are expressed as the mean ± S.E. ( n = 8) and were analyzed using two-way ANOVA followed by Bonferroni post-hoc t -test (* p < 0.05 and ** p < 0.01 vs. each saline group; # ## p < 0.001 vs. Control shRNA/METH group). OE: overexpression. TUJ1: beta-tubulin III. METH: methamphetamine.

Article Snippet: CB1 human open reading frame (ORF) clone lentiviral particles (c-Myc tagged) were obtained from Origene (Rockville, MD, USA) for CB1 overexpression.

Techniques: Expressing, Cell Culture, Over Expression, Immunostaining, shRNA, Amplification