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Localization of Na/HCO3 cotransporter NBCn1 (SLC4A7) in rat hippocampus
Author(s) -
Park Hae Jeong,
Rajbhandari Ira,
Choi Inyeong
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.790.10
Subject(s) - colocalization , subiculum , dentate gyrus , hippocampus , hippocampal formation , neocortex , cerebellum , neuroscience , biology , entorhinal cortex , olfactory bulb , granular layer , chemistry , microbiology and biotechnology , central nervous system
In an effort to facilitate the colocalization study, we developed a new NBCn1 antibody by immunizing a guinea pig with the BSA‐linked C‐terminal 14 amino acids of rat NBCn1. In membrane preparation from adult rat brain, the antibody recognized a protein of ~150 kDa, similar to NBCn1 protein detected with a rabbit polyclonal antibody against the same antigenic domain. The immunoreactive band disappeared when the antibody was preabsorbed with the antigenic peptides. The antibody labeled plasma membranes of HEK293 cells transfected with NBCn1. Immunochemistry of rat brain slices showed that NBCn1 immunofluorescence was localized in neurons of the regions including hippocampus, prefrontal cortex, neocortex, cerebellum and olfactory bulb. In the hippocampus, NBCn1 was found in the pyramidal layer of the hippocampal proper and the granular layer of the dentate gyrus. The fluorescence was also dispersed in the molecular layer (particularly stratum lacunosum‐moleculare). Double‐label immunofluorescence of NBCn1 and NMDA receptor subunit NR2A was performed to determine the colocalization of these proteins in the hippocampus. Both NBCn1 and NR2A were detected in the pyramidal layer and the granular layer, while NR2A additionally labeled subiculum. Controls without primary antibodies showed no discernible labeling. These data suggest that NBCn1 and NR2A are colocalized in some neurons of rat hippocampus.