Rabbit anti cb1

Two offspring of VIP-IRES-cre mice crossed with Ai14 reporter mice were transcardially perfused with 4% PFA in 0.1 m PB. The brain was removed from the skull and resectioned into 50-μm-thick horizontal slices. To estimate the GABAergic inputs received by tdTomato-expressing VIP+ INs, sections containing the BLA were processed for immunostaining with the following antibodies: guinea pig anti-VGAT (Synaptic Systems; 1:1000), rat anti-RFP (Chromotek; 1:1000), and rabbit anti-CB1 (Cayman Chemicals; 1:1000). VGAT was visualized with Alexa Fluor 488-conjugated donkey anti-guinea pig (Jackson ImmunoResearch; 1:500), tdTomato signal was enhanced by Cy3-conjugated donkey anti-rat (Jackson ImmunoResearch; 1:500), and CB1 was visualized with Cy5-conjugated donkey anti-rabbit antibody (Jackson ImmunoResearch; 1:500). Confocal images were obtained using a Nikon C2 confocal microscope (Plan Apo VC 20× objective; numerical aperture, 0.75; z-step size, 1 μm; xy, 0.62 μm/pixel). The image analysis was performed using Neurolucida Explorer.

Chemicals were of the purest analytical grade. MEM, glutamine, PES, DMSO, collagenase, DNase1, trypsin, BPA (Bisphenol A), ABTS (2,2′-azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt), and hyaluronidase were purchased from Sigma (St. Louis, MO, USA). Rabbit anti-CB₁, anti-CB₂, anti-GPR55, anti-FAAH, anti-DAGL-α, anti-DAGL-β, anti-MAGL, anti-NAPE-PLD polyclonal antibodies, and KT109 were purchased from Cayman Chemicals (Ann Arbor, MI, USA). Mouse anti-actin antibody was obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and rabbit anti-TRPV1 polyclonal antibody was from OriGene Technologies (Rockville, MD, USA). Rabbit anti-inhibin B polyclonal antibody was obtained from Elabscience Biotechnology (Houston, TX, USA), and rabbit anti-transferrin polyclonal antibody was from Proteintech Group (Chicago, IL, USA). Iodoresinferatoxin was obtained from Tocris Cookson (Bristol, UK), whereas SR144528 was a kind gift from Sanofi-Synthelabo (Montpellier, France).

The chemicals used in this study were obtained from the following companies: secondary goat anti-mouse IgG conjugated to HRP (sc-2005) from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and goat anti-rabbit IgG conjugated to horseradish peroxidase (HRP), (cat. 111-035-003) from Thermo Fisher Scientific (Waltham, MA, USA). The mouse-to-mouse HRP ready-to-use kit was obtained from ScyTek Laboratories, Inc. (Logan, UT, USA). All of the other reagents and ABTS (2,20-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)-diammonium salt) were purchased from Sigma (St. Louis, MO, USA) and were of the purest analytical grade. Rabbit anti-CB₁ (cat. 101500), anti-CB₂ (cat. 101550), anti-GPR55 (cat. 10224), anti-FAAH (cat. 101600), anti-MAGL (cat. 100035), and anti-NAPE-PLD (cat. 10305) polyclonal antibodies were obtained from Cayman Chemicals (Ann Arbor, MI, USA). Rabbit anti-DAGL-α (cat. PA5-23765) and anti-DAGL-β (PA5-26331) polyclonal antibodies were obtained from Invitrogen (Thermo Fisher Scientific; Waltham, MA, USA). The rabbit anti-TRPV1 (cat. TA336871) polyclonal antibody was obtained from OriGene Technologies (Rockville, MD, USA).