Extended Data Fig. 9: Opto-stimulation of WT and grm5a-/-grm5b-/- chx10 interneurons.
From: Activity-driven myelin sheath growth is mediated by mGluR5
A: Opto-simulation of a WT Tg(chx10:Gal4;UAS:ChRimsonR-tdTomato) animal with a 595 nm fiber optic coupled laser induces a characteristic motor response. B: Opto-simulation of a grm5a-/-grm5b-/- Tg(chx10:Gal4;UAS:ChRimsonR-tdTomato) animal with a 595 nm fiber optic coupled laser also induces a characteristic motor response. C: Images of chx10:Gal4;UAS:axonGCaMP7s in an animal treated with the neuromuscular blocker Mivacurium Chloride and subsequently with the Na+ channel blocker MS222. C’: dF/F trace of axonal GCaMP7s with Mivacurium, with opto-stimulations indicated by red lines. C”: dF/F trace of axonal GCaMP7s with MS222, with opto-stimulations indicated by red lines. D: Opto-stimulation of a WT Tg(chx10:Gal4;UAS:ChRimsonR-tdTomato;UAS:axonGCaMP7s) animal and corresponding time-lapse recording of axonal GCaMP7s signal following two separate stimulation steps (Optostim 1 (yellow) and Optostim2 (orange)) of two distinct ROIs. E: Mean dF/F traces of axonal GCaMP7s following two to three opto-stimulations per fish (N = 6). F: Individual dF/F traces of all axonal GCaMP7s following opto-stimulation of animals noted in E (n = 31). G: Representative image of the double transgene Tg(chx10:Gal4;UAS:ChRimsonR-tdTomato;UAS:axonGCaMP7s). Scale bar, 10 µm. H: Latency between optical stimulation and max. amplitude dF/F of the axonal GCaMP7s signal after the first, second and third stimulation. I. Relative frequency of myelin sheaths exhibiting a myelin Ca2+ transient with and without opto-stimulation.
