High-content image-based pooled screens reveal regulators of synaptogenesis

Area of Science:

• Neuroscience
• Molecular Biology
• Cell Biology

Background:

• Synapse formation is crucial for nervous system function but its regulatory mechanisms are not fully understood.
• The specific roles of these mechanisms in different synapse types require further definition.

Purpose of the Study:

• To uncover novel regulators of synapse induction using a large-scale screening approach.
• To investigate the modulators of neuroligin-1, a key molecule in organizing synapses.

Main Methods:

• Utilized a scalable optical pooled screening platform for high-content analysis.
• Analyzed over two million single-cell phenotypic profiles to identify candidate regulators.
• Investigated the roles of PTEN and DAG1 in neuroligin-1-mediated synaptogenesis.

Main Results:

• Identified 102 candidate regulators of neuroligin-1 involved in cell adhesion, cytoskeletal dynamics, and signaling.
• Demonstrated that PTEN and DAG1 promote neuroligin-1's role in presynaptic assembly.
• Found that DAG1 selectively regulates inhibitory synapses.

Conclusions:

• Established a scalable screening method for studying cell-cell interactions and synaptogenesis.
• Uncovered specific molecular players, PTEN and DAG1, that modulate synapse formation.
• Highlighted the selective role of DAG1 in inhibitory synapse regulation.