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J Physiol Volume 586, Number 16, 3949-3962, August 15, 2008 DOI: 10.1113/jphysiol.2008.151191
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NEUROSCIENCE

Regulation of quantal shape by Rab3A: evidence for a fusion pore-dependent mechanism

Xueyong Wang1,2, Ramachandran Thiagarajan1,2, Qingbo Wang1,2, Teclemichael Tewolde1, Mark M. Rich1,2 and Kathrin L. Engisch1,2

1 Department of Physiology, Emory University School of Medicine, Atlanta, GA 30322, USA
2 Department of Neuroscience, Cell Biology and Physiology, Boonshoft School of Medicine, Wright State University, Dayton, OH 45435, USA

The function of Rab3A, a small GTPase located on synaptic vesicles, is not well understood. Studies in the Rab3A–/– mouse support a role in activity-dependent plasticity, but have not reported any effects on spontaneously occurring miniature synaptic currents, except that there is a decrease in resting frequency at the neuromuscular junction. Therefore we were surprised to find an increase in the occurrence of mEPCs with abnormally long half-widths at the neuromuscular junctions of Rab3A–/– mice. The abnormal miniature endplate currents (mEPCs), which have significantly greater charge than the average mEPCs for the same fibres, could arise from larger vesicles. However, the type of mEPC most increased in Rab3A–/– mice has a slow rise, which suggests it is not the result of full collapse fusion. To test if the slow mEPCs increased after loss of Rab3A could be due to malfunctioning fusion pores, we used carbon fibre amperometry to record pre-spike feet, which have been shown to correspond to the initial opening of a narrow fusion pore, in adrenal chromaffin cells of wild-type and Rab3A–/– mice. We found that small amplitude pre-spike feet with abnormally long durations were increased in Rab3A–/– cells. The correspondence between mEPC and amperometric data supports our interpretation that slow rising, long half-width mEPCs are caused by reduced diameter fusion pores that remain open longer. These data could be explained by a direct action of Rab3A on the fusion pore, or by Rab3A-dependent control of vesicles with unusual fusion pore characteristics.

(Received 15 January 2008; accepted after revision 20 June 2008; first published online 26 June 2008)
Corresponding author K. Engisch: Department of Neuroscience, Cell Biology and Physiology, 014 M&M Building, Boonshoft School of Medicine, Wright State University, 3640 Colonel Glenn Hwy., Dayton, OH 45435, USA. Email: kathrin.engisch{at}wright.edu


X. Wand and R. Thiagarajan contributed equally to this work.




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