Clonally related, Notch-differentiated spinal neurons integrate into distinct circuits EPhys raw data

Shared lineage has diverse effects on patterns of neuronal connectivity. In mammalian cortex, excitatory sister neurons assemble into shared microcircuits, whereas throughout the Drosophila nervous system, Notch-differentiated sister neurons diverge into distinct circuits. Notch-differentiated sister neurons have been observed in vertebrate spinal cord and cerebellum, but whether they integrate into shared or distinct circuits remains unknown. Here we evaluate the connectivity between sister V2a/b neurons in the zebrafish spinal cord. Using an in vivo labeling approach, we identified pairs of sister V2a/b neurons born from individual Vsx1+ progenitors and observed that they have similar axonal trajectories and proximal somata. However, paired whole-cell electrophysiology and optogenetics revealed that sister V2a/b neurons receive input from distinct presynaptic sources, do not communicate with each other, and connect to largely distinct targets. These results resemble the divergent conn..., This is the raw data for the manuscript.
Cell-attached recordings were targeted to stochastically labeled WT fish with vsx1:Gal4 BAC and UAS:CoChR2-tdTomato plasmid to calibrate firing of vsx1:Gal4;UAS:CoChR2-tdTomato vsx1+ pairs. Whole-cell patch-clamp recordings were performed in Tg(chx10:loxP-dsRed-loxP:GFP) injected with vsx1:GFP and Tg(chx10:GFP;gata3:loxP-dsRed-loxP:GFP) larvae at 4-6 dpf for paired clonal V2a/b and non-clonal V2a/b recordings, respectively. Additionally, whole-cell patch-clamp recordings were performed in stochastically labeled WT fish with vsx1:Gal4 BAC and UAS:CoChR2-tdTomato in downstream targets. Larvae were immobilized with 0.1% α-bungarotoxin and fixed to a Sylgard lined Petri dish with custom-sharpened tungsten pins. Each larva was then transferred to a microscope (Scientifica SliceScope Pro) equipped with infrared differential interface contrast optics, epifluorescence, and immersion objectives (Olympus: 40X, 0.8 NA). One muscle segment overlaying the spi..., Data collected are .abf files that can be opened using Matlab and ClampFit.