brain tumor regulates neuromuscular synapse growth and endocytosis in Drosophila by suppressing Mad expression Short title: Brat regulates synaptic growth by suppressing Mad

Abstract

The precise regulation of synaptic growth is critical for the proper formation and plasticity of functional neural circuits. Identification and characterization of factors that regulate synaptic growth and function have been under intensive investigation. Here we report that brain tumor (brat), which was identified as a translational repressor in multiple biological processes, plays a crucial role at Drosophila neuromuscular junction (NMJ) synapses. Immunohistochemical analysis demonstrated that brat mutants exhibited synaptic overgrowth characterized by excess satellite boutons at NMJ terminals, while electron microscopy revealed increased synaptic vesicle size but reduced density at active zones compared to wild types. Spontaneous miniature excitatory junctional potential amplitudes were larger and evoked quantal content was lower at brat mutant NMJs. In agreement with the morphological and physiological phenotypes, loss of Brat resulted in reduced FM1-43 uptake and decreased immunostaining of the endocytic proteins AP180 and dynamin at the NMJ terminals, indicating that brat regulates synaptic endocytosis. Genetic analysis revealed that the actions of Brat at synapses are mediated through Mothers against decapentaplegic (Mad), the signal transduction effector of the bone morphogenetic protein (BMP) signaling pathway. Furthermore, biochemical analyses showed upregulated levels of Mad protein but normal mRNA levels in the larval brains of brat mutants, suggesting that Brat suppresses Mad translation. Consistently, knockdown of brat by RNA interference in Drosophila S2 cells also increased Mad protein level. These results together reveal an important and previously unidentified role for Brat in synaptic development and endocytosis mediated by suppression of BMP signaling.

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@inproceedings{Shi2014brainTR, title={brain tumor regulates neuromuscular synapse growth and endocytosis in Drosophila by suppressing Mad expression Short title: Brat regulates synaptic growth by suppressing Mad}, author={Wenwen Shi and Yan Chen and Guangming Gan and Dan Wang and Jinqi Ren and Qifu Wang and Zhiheng Xu and Wei Xie and Yong Q. Zhang and Chao Yang}, year={2014} }