A Syntaxin 1, Gαo, and N-Type Calcium Channel Complex at a Presynaptic Nerve Terminal: Analysis by Quantitative Immunocolocalization

By Qi Li, A. Lau, Terence J. Morris, Lin Guo, C. Fordyce, E. F. Stanley. Journal of Neuroscience, 2004

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Presynaptic CaV2.2 (N-type) calcium channels are subject to modulation by interaction with syntaxin 1 and by a syntaxin 1-sensitive GαO G-protein pathway. We used biochemical analysis of neuronal tissue lysates and a new quantitative test of colocalization by intensity correlation analysis at the giant calyx-type presynaptic terminal of the chick ciliary ganglion to explore the association of CaV2.2 with syntaxin 1 and GαO. CaV2.2 could be localized by immunocytochemistry (antibody Ab571) in puncta on the release site aspect of the presynaptic terminal and close to synaptic vesicle clouds. Syntaxin 1 coimmunoprecipitated with CaV2.2 from chick brain and chick ciliary ganglia and was widely distributed on the presynaptic terminal membrane. A fraction of the total syntaxin 1 colocalized with the CaV2.2 puncta, whereas the bulk colocalized with MUNC18-1. GαO, whether in its trimeric or monomeric state, did not coimmunoprecipitate with CaV2.2, MUNC18-1, or syntaxin 1. However, the G-protein exhibited a punctate staining on the calyx membrane with an intensity that varied in synchrony with that for both Ca channels and syntaxin 1 but only weakly with MUNC18-1. Thus, syntaxin 1 appears to be a component of two separate complexes at the presynaptic terminal, a minor one at the transmitter release site with CaV2.2 and GαO, as well as in large clusters remote from the release site with MUNC18-1. These syntaxin 1 protein complexes may play distinct roles in presynaptic biology.

Subfields: Cell Biology, Cellular and Molecular Neuroscience, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/e606aaada0621493a6fecbf0b94f16470b6a1117

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Citations in the corpus

A guided tour into subcellular colocalization analysis in light microscopy (2006, methodology): “In principle this is done using a modification of the intensity correlation analysis method (Li et al., 2004).”
A single pair of neurons links sleep to memory consolidation in Drosophila melanogaster (2015, methodology): “For the images presented in Figure 3, Intensity Correlation Analysis (ICA) was also performed to assess spatial colocalization of staining between channels (Li et al., 2004).”
Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. (2009, methodology): “Quantification was performed using the intensity correlation analysis method published in Li et al. (2004), and as implemented in the IMAGEJ plugin of the MBF IMAGEJ bundle (http:// www.macbiophotonics.ca/imagej/).”
Tau oligomers impair memory and induce synaptic and mitochondrial dysfunction in wild-type mice (2011, background): “This was confirmed by ICA of the signal (Figure 6H), as described in [ 51 ], suggesting a direct interaction between tau oligomers and mitochondria.”
Quantifying colocalization by correlation: The Pearson correlation coefficient is superior to the Mander's overlap coefficient (2010, methodology): “quantify colocalization, the intensity correlation quotient (ICQ) (6,7) and the Spearman rank correlation (SRC) (8,9), both derived from the PCC.”