Drosophila-Cdh1 (Rap/Fzr) a regulatory subunit of APC/C is required for synaptic morphology, synaptic transmission and locomotion.
The assembly of functional synapses requires the orchestration of the synthesis and degradation of a multitude of proteins. Protein degradation and modification by the conserved ubiquitination pathway has emerged as a key cellular regulatory mechanism during nervous system development and function (Kwabe and Brose, 2011). The anaphase promoting complex/cyclosome (APC/C) is a multi-subunit ubiquitin ligase complex primarily characterized for its role in the regulation of mitosis (Peters, 2002). In recent years, a role for APC/C in nervous system development and function has been rapidly emerging (Stegmuller and Bonni, 2005; Li et al., 2008). In the mammalian central nervous system the activator subunit, APC/C-Cdh1, has been shown to be a regulator of axon growth and dendrite morphogenesis (Konishi et al., 2004). In the Drosophila peripheral nervous system (PNS), APC2, a ligase subunit of the APC/C complex has been shown to regulate synaptic bouton size and activity (van Roessel et al., 2004). To investigate the role of APC/C-Cdh1 at the synapse we examined loss-of-function mutants of Rap/Fzr (Retina aberrant in pattern/Fizzy related), a Drosophila homolog of the mammalian Cdh1 during the development of the larval neuromuscular junction in Drosophila. Our cell biological, ultrastructural, electrophysiological, and behavioral data showed that rap/fzr loss-of-function mutations lead to changes in synaptic structure and function as well as locomotion defects. Data presented here show changes in size and morphology of synaptic boutons, and, muscle tissue organization. Electrophysiological experiments show that loss-of-function mutants exhibit increased frequency of spontaneous miniature synaptic potentials, indicating a higher rate of spontaneous synaptic vesicle fusion events. In addition, larval locomotion and peristaltic movement were also impaired. These findings suggest a role for Drosophila APC/C-Cdh1 mediated ubiquitination in regulating synaptic morphology, function and integrity of muscle structure in the peripheral nervous system.
Department of Biology, City College of New York, and The Graduate Center of CUNY, New York, NY 10031, United States; Department of Pathology and Cell Biology, Columbia University, 630 West 168th Street, New York, NY 10032, United States.
Alexandria
A
Wise
Emma
E
Schatoff
Julian
J
Flores
Shao-Ying
SY
Hua
Atsushi
A
Ueda
Chun-Fang
CF
Wu
Tadmiri
T
Venkatesh
D064173:Anaphase-Promoting Complex-Cyclosome / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D064200:Cdh1 Proteins / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology; D007814:Larva; D008124:Locomotion / Q000235:genetics / Q000502:physiology*; D046529:Microscopy, Electron, Transmission; D009119:Muscle Contraction / Q000235:genetics; D009135:Muscular Diseases / Q000235:genetics / Q000473:pathology; D009154:Mutation / Q000235:genetics; D009469:Neuromuscular Junction / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques; D017729:Presynaptic Terminals / Q000502:physiology* / Q000648:ultrastructure; D018091:Receptors, AMPA / Q000378:metabolism; D055366:Synaptic Potentials / Q000235:genetics*
D064200:Cdh1 Proteins; D029721:Drosophila Proteins; D018091:Receptors, AMPA; C577674:fzr protein, Drosophila; D064173:Anaphase-Promoting Complex-Cyclosome; C104722:glutamate receptor ionotropic, AMPA 2
Anaphase promoting complex/cyclosome; Cdh1; Developmental genetics; Drosophila; Neuromuscular junction; Synaptogenesis; Ubiquitination