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The Journal of Neuroscience, April 15, 1999, 19(8):2919-2928
A 29 kDa Intracellular Chloride Channel p64H1 Is Associated with
Large Dense-Core Vesicles in Rat Hippocampal Neurons
Jen-Zen
Chuang1,
Teresa
A.
Milner2,
Meicai
Zhu1, and
Ching-Hwa
Sung1, 3
1 Department of Ophthalmology, The Margaret M. Dyson
Vision Research Institute, 2 Department of Neurology and
Neuroscience, Division of Neurobiology, and 3 Department of
Cell Biology and Anatomy, Weill Medical College of Cornell University,
New York, New York 10021
A novel class of intracellular chloride channels, the p64 family,
has been found on several types of vesicles. These channels, acting in
concert with the electrogenic proton pump, regulate the pH of the
vesicle interior, which is critical for vesicular function. Here we
describe the molecular cloning of p64H1, a p64 homolog, from both human
and cow. Northern blot analysis showed that p64H1 is expressed
abundantly in brain and retina, whereas the other members of
this family (e.g., p64 and NCC27) are expressed only at low levels in
these tissues. Immunohistochemical analysis of p64H1 in rat brain,
using an affinity-purified antibody, revealed a high level of
expression in the limbic system the hippocampal formation, the
amygdala, the hypothalamus, and the septum. Immunoelectron microscopic
analysis of p64H1 in hippocampal neurons demonstrated a striking
association between p64H1 and large dense-core vesicles (LDCVs) and
microtubules. In contrast, very low p64H1 labeling was found in
perikarya or associated with small synaptic vesicles (SSVs) in axonal
profiles. Immunoblot analysis confirmed that p64H1 is colocalized with
heavy membrane fractions containing LDCVs rather than the fractions
containing SSVs. These results suggest that p64H1-mediated
Cl permeability may be involved in the maintenance
of low internal pH in LDCVs and in the maturation of LDCVs and the
biogenesis of functional neuropeptides.
Key words:
p64; p64H1; NCC27; intracellular chloride channel; large
dense-core vesicle; neuropeptide; hippocampus; microtubules
Copyright © 1999 Society for Neuroscience 0270-6474/99/1982919-10$05.00/0
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