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J Neurophysiol (August 17, 2005). doi:10.1152/jn.01193.2004
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01193.2004v1
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Submitted on November 19, 2004
Accepted on August 7, 2005

Imaging of Ca2+ dynamics within the presynaptic terminals of salamander rod photoreceptors

Ernest C. Steele, Jr.*, Xiaoming Chen, P. Michael Iuvone, and Peter R. MacLeish

* To whom correspondence should be addressed. E-mail: esteele{at}msm.edu.

While the overall importance of Ca2+ as a mediator of cell signaling and neurotransmitter release has long been appreciated, the details of Ca2+ dynamics within vertebrate rod photoreceptors are just beginning to be elucidated. Even less is known regarding Ca2+ dynamics within the rod presynaptic terminal compartment. Using fura-2 to report changes in intracellular Ca2+, we imaged the responses of enzymatically dissociated rod photoreceptors stimulated with a brief depolarizing puff of KCl (30 mM pipette concentration). In the vast majority of cells, the response was a large increase in Ca2+ levels in the terminal compartment, but not in the soma. In contrast, rods exhibited a large elevation in somatic Ca2+ levels when depolarized with a brief puff of 100 mM KCl (pipette concentration). These data are consistent with previously reported differences in Ca2+ buffering mechanisms within the somatic and terminal compartments. Additionally, they may reflect the presence of Ca2+ channels having distinct properties within the membranes of the two compartments.Consistent with this hypothesis, fluorescent immunocytochemistry using an antibody against the L-type Ca2+ channel Cav1.2 ({alpha}1C) subunit and semi-quantitative confocal microscopy revealed a high concentration of immunoreactivity in the membranes of terminals of intact rods compared to the somata. Further investigations using enzymatically dissociated preparations of intact rod photoreceptors retaining their presynaptic terminals will allow further testing of these and other hypotheses regarding the compartmentalized regulation of Ca2+dynamics within rod photoreceptors.







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