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Differential Expression Analysis Identifies Candidate Synaptogenic Molecules for Wiring Direction-Selective Circuits

Joshua M Tworig1, Ryan D Morrie1, Karina Bistrong2

  • 1Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California 94720.

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
|March 21, 2024
PubMed
Summary

Investigating retinal direction-selective circuits, this study explored molecular cues for specific synaptic connections. Cerebellin-4 (Cbln4) showed a minor role in developing direction selectivity in starburst amacrine cell circuits.

Keywords:
RNA-seqdirection selectivityretinaretinal ganglion celltwo-photon calcium imaging

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Molecular Biology

Background:

  • Neural circuits exhibit specific synaptic connections, exemplified by retinal direction-selective (DS) circuits.
  • Starburst amacrine cells (SACs) form selective connections with DS retinal ganglion cell (DSGC) subtypes, crucial for motion detection.

Purpose of the Study:

  • To investigate the molecular basis of specific synaptic wiring in DS retinal circuits.
  • To identify unique gene expression profiles in DSGC subtypes that may guide SAC connectivity.
  • To test the role of candidate molecules, such as cerebellin-4 (Cbln4), in the development of DS tuning.

Main Methods:

  • Paired recordings in isolated mouse retinas to determine synapse formation timing (postnatal day 10).
  • RNA sequencing and differential expression analysis of isolated ON-OFF DSGCs.
  • Conditional knockout of Cbln4 and subsequent two-photon calcium imaging and whole-cell voltage-clamp recordings.

Main Results:

  • Postnatal day 10 is identified as a critical period for asymmetric synapse formation in DS circuits.
  • Transcriptomic analysis revealed candidate molecules potentially involved in direction-specific wiring.
  • Mice lacking Cbln4 showed a slight deficit in directional tuning in ventral-preferring DSGCs, but no significant change in inhibitory inputs was detected.

Conclusions:

  • Cbln4 does not appear to primarily act through a cell-autonomous mechanism to instruct DS circuit wiring.
  • The study identified novel candidate factors that warrant further investigation into the molecular mechanisms of synaptic specificity in the DS circuit.