Real time CPP

Published: 16 June 2020| Version 1 | DOI: 10.17632/cf8kzg53m2.1
Contributor:
Anna Terem

Description

Figure 4G- D1-cre Figure 5H- retroAAV-cre Figure S5D- retroAAV-cre control Surgery: At 8-12 weeks of age mice were stereotactically injected for real-time CPP (rtCPP) experiments. The first experimental cohort consisted of D1-Cre male mice (n=8) stereotactically injected with 80nl of AAV9-CAGGS-FLEX-ChR2-TdTomato (purchased from the UPENN virus core) to the claustrum region (LM: 2.82, RC: 1, DV: -3.7). The second experimental cohort consisted of WT c57/bl6 mice (n=6) stereotactically injected with 200nl retroAAV-CKII-iCre to the ACC (LM: ±0.25, RC: 1.1, DV: -1.9) and OFC (LM: ±1, RC: 2.55, DV: -2.4) and 80nl of AAV9-CAGGS-FLEX-ChR2-TdTomato stereotactically injected to the claustrum region (LM: ±2.82, RC: 1, DV: -3.7). The control group consisted of 4 WT c57/bl6 mice stereotactically injected with 200nl retroAAV-CKII-iCre to the ACC (LM: ±0.25, RC: 1.1, DV: -1.9) and OFC (LM: ±1, RC: 2.55, DV: -2.4) and 80nl of AAV9-CAGGS-FLEX-ChR2-TdTomato stereotactically injected to the claustrum region (LM:±2.82 , RC: 1, DV: -3.7). Silica multimode fiber optic cannula (200-μm core; 240-μm outer diameter) mounted in a 1.25-mm zirconia ferrule (Doric lenses) were slowly lowered into the brain bilaterally following virus injection (LM:±2.82 , RC: 1, DV: -3.65), and cemented in place such that the fiber tip was located above the claustrum. Optogenetics equipment: A blue laser (MBL-FN-437-200mW, CNI laser), was connected to 1x2 Intensity Division Fiberoptic Rotary Joint (FRJ_1x2i_SMA-2FC_0.22, Doric). Two 50cm Mono Fiberoptic Patchcord (MFP_200/230/900-0.37_0.5m_FC-ZF1.25(F), Doric) were connected to the implanted cannulae with a zirconia connection sleeve (1.25mm, Doric). Real-time CPP: A 3-chamber compartment was used. The north chamber had a smooth floor and walls textured with black and white vertical stripes, while the south chamber had a grid floor and white walls with black dots. The central chamber lacked discerning features and was meant to increase the division between the lateral chambers. Either the north or south chambers were paired with laser stimulation of the claustrum, counterbalanced within each experimental group. The mouse’s position was calculated in real time with Ethovision XT 11.5 software. The laser (5mW) was illuminated in a cycle of 1second on / 3second off for the duration that the mouse remained in the laser-paired chamber (as in (Kravitz and Kreitzer, 2012)). All experimental sessions were 20min long. For each real-time CPP experiment repeated measurements were performed on the same group of mice. No experimental mice were excluded from analysis.

Files