The role of MAP1A light chain 2 in synaptic surface retention of Cav2.2 channels in hippocampal neurons.
Ca(v)2.2 channels are localized at nerve terminals where they play a critical role in neurotransmission. However, the determinant that controls surface retention of these channels has not been identified. Here, we report that presynaptic surface localization of Ca(v)2.2 is mediated through its interaction with light chain 2 (LC2) of microtubule-associated protein MAP1A. Deletion of a 23-residue binding domain within the Ca(v)2.2 C terminus resulted in reduced synaptic distribution of the mutant channels. Using an antibody generated against an extracellular epitope of Ca(v)2.2, we demonstrate that interfering the interaction with LC2 reduced surface expression of endogenous Ca(v)2.2 at presynaptic boutons. In addition, the disruption of LC2-Ca(v)2.2 coupling reduced Ca(2+)-influx into nerve terminals through Ca(v)2.2 and impaired activity-dependent FM4-64 uptake. The treatments of neurons with Latrunculin A to disrupt actin filaments resulted in reduced density of surface Ca(v)2.2-positive boutons. Furthermore, LC2NT, a LC2 truncated mutant lacking the actin-binding domain, could not rescue Ca(v)2.2 surface expression after suppressing LC2 expression with RNAi. Because actin filaments are major cytomatric components at the presynaptic boutons, these observations suggest a mechanism by which LC2 provides anchoring of surface Ca(v)2.2 to the actin cytoskeleton, thus contributing to presynaptic function.
Leenders
A G Miriam
AG
Synaptic Function Section, The Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892-3701, USA.
Lin
Lin
L
Huang
Li-Dong
LD
Gerwin
Claudia
C
Lu
Pei-Hua
PH
Sheng
Zu-Hang
ZH
D000818:Animals; D020864:Calcium Channels, N-Type / Q000378:metabolism / Q000502:physiology*; D002478:Cells, Cultured; D006624:Hippocampus / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism / Q000502:physiology*; D009474:Neurons / Q000737:chemistry / Q000502:physiology*; D017729:Presynaptic Terminals / Q000737:chemistry / Q000502:physiology; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013499:Surface Properties; D013569:Synapses / Q000737:chemistry / Q000502:physiology*
C503279:Cacna1b protein, rat; D020864:Calcium Channels, N-Type; C490546:Map1a protein, rat; D008869:Microtubule-Associated Proteins