Downregulation of Centaurin gamma1A increases synaptic transmission at Drosophila larval neuromuscular junctions.

Adequate regulation of synaptic transmission is critical for appropriate neural circuit functioning. Although a number of molecules involved in synaptic neurotransmission have been identified, the molecular mechanisms regulating neurotransmission are not fully understood. Here, we focused on Centaurin gamma1A (CenG1A) and examined its role in synaptic transmission regulation using Drosophila larval neuromuscular junctions. CenG1A is a member of the Centaurin family, which contains Pleckstrin homology, ADP ribosylation factor GTPase-activating protein, and ankyrin repeat domains. Due to the existence of these functional domains, CenG1A is proposed to be involved in the process of synaptic release; however, no evidence for this has been found to date. In this study, we investigated the potential role for CenG1A in the process of synaptic release by performing intracellular recordings in larval muscle cells. We found that neurotransmitter release from presynaptic cells was enhanced in cenG1A mutants. This effect was also observed in larvae with reduced CenG1A function in either presynaptic or postsynaptic cells. In addition, we revealed that suppressing CenG1A function in postsynaptic muscle cells led to an increase in the probability of neurotransmitter release, whereas its suppression in presynaptic neurons led to an increase in neurotransmitter release probability and an increase in the number of synaptic vesicles. These results suggested that CenG1A functions at both presynaptic and postsynaptic sites as a negative regulator of neurotransmitter release. Our study provided evidence for a key role of CenG1A in proper synaptic transmission at neuromuscular junctions.

Lola regulates glutamate receptor expression at the Drosophila neuromuscular junction.

Communication between pre- and post-synaptic cells is a key process in the development and modulation of synapses. Reciprocal induction between pre- and postsynaptic cells involves regulation of gene transcription, yet the underlying genetic program remains largely unknown. To investigate how innervation-dependent gene expression in postsynaptic cells supports synaptic differentiation, we performed comparative microarray analysis of Drosophila muscles before and after innervation, and of prospero mutants, which show a delay in motor axon outgrowth. We identified 84 candidate genes that are potentially up- or downregulated in response to innervation. By systematic functional analysis, we found that one of the downregulated genes, longitudinals lacking (lola), which encodes a BTB-Zn-finger transcription factor, is required for proper expression of glutamate receptors. When the function of lola was knocked down in muscles by RNAi, the abundance of glutamate receptors (GluRs), GluRIIA, GluRIIB and GluRIII, as well as that of p-21 activated kinase (PAK), was greatly reduced at the neuromuscular junctions (NMJs). Recordings of the synaptic response revealed a decrease in postsynaptic quantal size, consistent with the reduction in GluR levels. Lola appears to regulate the expression of GluRs and PAK at the level of transcription, because the amount of mRNAs encoding these molecules was also reduced in the mutants. The transcriptional level of lola, in turn, is downregulated by increased neural activity. We propose that Lola coordinates expression of multiple postsynaptic components by transcriptional regulation.