Danio rerio Transcriptome or Gene expression
Optic flow processing is critical for the visual control of body and eye movements in many animals. Rotational and translational binocular optic flow patterns need to be clearly distinguished to induce different behavior outputs. However, the specific neuron types and their connectivity involved in this computation remain unclear. Here, we developed a method to link the functional labeling using a photoconvertible calcium indicator called CaMPARI2 and single-cell RNA-sequencing (CaMPARI-seq) to investigate the transcriptional profile of the pretectum, a center for processing optic flow in larval zebrafish. Using this technique, we identified a pretectal cluster expressing tcf7l2, which can be further classified into molecularly distinct subclusters. In vivo calcium imaging and cell ablation revealed that nkx1.2lb-positive pretectal neurons are commissural inhibitory neurons required for the optomotor response but not for the optokinetic response. Our genetic and functional dissection using CaMPARI-seq uncovered the neuronal organization essential for dissociating different optic flow-dependent behaviors. Overall design: To identify genetic markers for optic flow responsive cells, we conducted single-cell RNA sequencing analysis. We used zebrafish larvae pan-neuronally expressing NLS-CaMPARI2 to label neurons during optic flow processing. CaMPARI2 labeling was performing by deliverying light as zebrafish larvave respond to either moving or stationary optic flow stimuli. After dissection and dissociation of the brain, only active neurons (CaMPARI, red fluorescent) were isolated for sequencing.