Immunocytocheniical Localization of the a Subspecies of Protein Kinase
Proc. Nati. Acad. Sci. USA Vol. 87, pp. 3195-3199, April 1990 Neurobiology Immunocytocheniical localization of the a subspecies of protein kinase C in rat brain (in situ hybridization histochemistry) ATSUKO ITO*, NAOAKI SAITO*, MIDORI HIRATA*, AKIKO KOSE*, TAKESHI TsUJINO*, CHIKA YOSHIHARA*, Kouji OGITAt, AKIRA KISHIMOTOt, YASUTOMI NISHIZUKAt, AND CHIKAKO TANAKA** Departments of *Pharmacology and tBiochemistry, Kobe University School of Medicine, Kobe 650, Japan Contributed by Yasutomi Nishizuka, January 2, 1990 ABSTRACT The distribution ofthe a subspecies ofprotein types, with limited intracellular localization. The present kinase C (PKC) in rat brain was demonstrated immunocy- studies were undertaken to identify a-PKC in the rat brain by tochemically by using polyclonal antibodies raised against a using immunocytochemistry, and the results show that this synthetic oligopeptide corresponding to the carboxyl-terminal PKC subspecies is enriched in particular cell types. sequence of a-PKC. The a-PKC-specific immunoreactivity was widely but discretely distributed in both gray and white MATERIAL AND matter. The immunoreactivity was associated predominantly METHODS with neurons, particularly with perikaryon, dendrite, or axon, Preparation of Antibodies Against a-PKC. The carboxyl- but little was seen in the nucleus. Glial cells expressed this PKC terminal portion of a-PKC (residues 662-672; Gln-Phe-Val- subspecies poorly, if at all. The highest density of immuno- His-Pro-Ile-Leu-Gln-Ser-Ala-Val) was selected as a se- reactivity was seen in the olfactory bulb, septohippocampal quence specific to a-PKC. The oligopeptide was coupled to nucleus, indusium griseum, islands of Calleja, intermediate keyhole limpet with m-maleimidobenzoic acid N-hydrox- part of the lateral septal nucleus, and Ammon's horn.
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