Optogenetic Targeting of Interneurons in Mouse Cortex

Plasmid pAAV-EF1α-DIO-hChR2(H134R)-P2A-EYFP was generated by replacing the hChR2(C128A H134R) in pAAV-EF1α-DIO-hChR2(C128A H134R)-P2A-EYFP (Prakash et al., 2012 (link)) with the hChR2(H134R) from pAAV-EF1α-DIO-hChR2(H134R)-EYFP (Addgene #20298) and was deposited at Addgene (#139283). The recombinant AAV vectors were produced by the Gene Vector Core at Baylor College of Medicine. To express ChR2 in Pv or Sst interneurons, 200 nl of AAV9-EF1α-DIO-hChR2(H134R)-P2A-EYFP vectors (3 × 10¹³ genome copies/ml) were injected into the somatosensory cortex of Stxbp1^tm1d/+;Pv^Cre/+ and Stxbp1^+/+;Pv^Cre/+ or Stxbp1^tm1d/+;Sst^Cre/+ and Stxbp1^+/+;Sst^Cre/+ mice, respectively, at postnatal day 1–5 as previously described (Xue et al., 2014 (link); Messier et al., 2018 (link)) with an UltraMicroPump III and a Micro4 controller (World Precision Instruments).

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Chen W., Cai Z.L., Chao E.S., Chen H., Longley C.M., Hao S., Chao H.T., Kim J.H., Messier J.E., Zoghbi H.Y., Tang J., Swann J.W, & Xue M. (2020). Stxbp1/Munc18-1 haploinsufficiency impairs inhibition and mediates key neurological features of STXBP1 encephalopathy. eLife, 9, e48705.

Publication 2020

UltraMicroPump III Micro4 controller

Gene vector Genome Interneurons Medicine Mice Plasmid Somatosensory cortex Vectors

Corresponding Organization :

Other organizations : Baylor College of Medicine, Neurological Research Institute, Texas Children's Hospital, Robert and Janice McNair Foundation

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Variable analysis

independent variables

Genotype of the mice (Stxbp1tm1d/+ or Stxbp1+/+)
Cre recombinase expression (Pv-Cre/+ or Sst-Cre/+)

dependent variables

Expression of ChR2(H134R)-P2A-EYFP in Pv or Sst interneurons

control variables

AAV9-EF1α-DIO-hChR2(H134R)-P2A-EYFP vector concentration (3 × 1013 genome copies/ml)
Injection volume (200 nl)
Injection location (somatosensory cortex)
Injection timing (postnatal day 1–5)

controls

Positive control: Stxbp1+/+;Pv-Cre/+ or Stxbp1+/+;Sst-Cre/+ mice
Negative control: Not explicitly mentioned

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