Proceedings of Annual Meeting of the Physiological Society of Japan
Proceedings of Annual Meeting of the Physiological Society of Japan
Session ID : 1PHP-014
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Implication of chloride transporter, NKCC1, in neurite outgrowth
*Ken-ichi NakajimaHiroaki MiyazakiNaomi NiisatoYoshinori Marunaka
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Abstract
Dynamic organization of cytoskeletons, such as microtubules, actin filaments and intermediate filaments, is crucial for the neurite outgrowth in neuronal cells. On the other hand, it is suggested that changes of intracellular chloride ion concentration is involved in regulation of various cellular functions such as cell cycle, proliferation, adhesion, migration, and dynamics of cytoskeleton. Neuronal cells are known to cooperate the K+/Cl cotransporter (KCC) and/or the Cl-ATPase excreting Cl from the cells, and to incorporate Cl into the cells via the Na+/K+/2Cl cotransporter (NKCC). Further, it is also reported that the NKCC-like activity is up-regulated by NGF treatment in PC12 cells. In this study, we investigated the role of NKCC1 in NGF-induced neurite outgrowth in PC12D cells (a subclone of PC12 cells). Bumetanide, a specific inhibitor of NKCC, significantly inhibited the NGF-induced neurite outgrowth in PC12D cells. A similar phenomenon was observed in PC12D cells exposed to a low Cl concentration medium. Expression of NKCC1 protein was up-regulated by NGF treatment. Furthermore, knock down of NKCC1 by RNAi drastically diminished the NGF-induced neurite outgrowth. A study of GFP-tagged rNKCC1 transfection into PC12D for clarification of the cellular localization of NKCC1 indicated that the GFP-rNKCC1 was localized in growth cone during neurite outgrowth. Based upon these findings, we conclude that Cl uptake by NKCC1 localized in the growth cone promotes neurite outgrowth. [J Physiol Sci. 2007;57 Suppl:S121]
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© 2007 The Physiological Society of Japan
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