Monoclonal antibodies against rat brain protein kinase C and their application to immunocytochemistry in nervous tissues

Kitano, Tatsuro; Hashimoto, Takashi; Kikkawa, Ushio; Ase, Katsuhiko; Saito, Naoaki; Tanaka, Chikako; Ichimori, Yuzo; Tsukamoto, Kikuo; Nishizuka, Yasutomi

doi:10.1523/jneurosci.07-05-01520.1987

Cited by 70 publications

(45 citation statements)

References 30 publications

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“…More precisely, the materials reactive with the antibody, CKpVSPII-a, surrounded the Purkinje cell bodies as well as their dendrites, most likely representing the numerous nerve terminals synapsing with Purkinje cells. This distribution pattern can now be seen to contrast markedly with that of y-subspecies, which is present in the cell bodies, dendrites, and axons of Purkinje cells (Hashimoto et al, 1988;Saito et al, 1987).…”

Section: Resultsmentioning

confidence: 88%

“…At this level of resolution, the pattern for @II-subspecies was apparently similar to that of y-subspecies. The monoclonal antibody CKmIy-97, previously prepared (Kitano et al, 1987), specifically recognizes Type I protein kinase C (y-subspecies) as described (Hashimoto et al, 1988). The y-subspecies was present in the molecular layer and Purkinje cell layer, though no immunoreactivity is observed in the granular layer or white matter of the cerebellar cortex.…”

Section: Resultsmentioning

confidence: 99%

“…One week after the final immunization, the rabbits were bled, and each antibody was purified from the serum by affinity column chromatography on Sepharose CL-4B coupled with each oligopeptide. The monoclonal antibody CKmIy-97 was raised originally against a mixture of Type I, II, and III protein kinase C (Kitano et al, 1987). Subsequent analysis with these protein kinase C fractions has revealed that the antibody specifically reacts with Type I protein kinase C (y-subspecies) (Hashimoto et al, 1987).…”

Section: Methodsmentioning

confidence: 99%

“…Each type of protein kinase C, which was obtained by hydroxyapatite column chromatography from rat brain or transfected COS 7 cells, was applied to SDS-PAGE and transferred to a nitrocellulose filter as described (Kitano et al, 1987). After reaction with each antibody, the nitrocellulose filter was incubated with biotinylated second antibody and subsequently with avidin-biotinylated HRP complex (Vectastain, Vector Laboratories).…”

Section: Methodsmentioning

confidence: 99%

“…Immnohistochemical studies were carried out as described (Kitano et al, 1987;Saito et al, 1988). Anesthetized rats were perfused through the left ventricle with 200 ml of periodate-lysine-pamformaldehyde fixative (McLean and Nakane, 1974), and the brain was removed.…”

Section: Methodsmentioning

confidence: 99%

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Distinct cellular expression of beta I- and beta II-subspecies of protein kinase C in rat cerebellum

Ase

Saito²,

Shearman³

et al. 1988

J. Neurosci.

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Immunohistochemical and biochemical studies with subspecies-specific antibodies have revealed that beta I- and beta II-subspecies of protein kinase C, which result from alternative splicing of a single RNA transcript, show different regional expression in rat CNS. In the cerebellar cortex, beta I-subspecies is localized mainly in the granular layer, whereas beta II-subspecies is found predominantly in the molecular layer, most apparently in the presynaptic nerve endings that terminate at Purkinje cells. These distribution patterns are in sharp contrast to that of gamma-subspecies, which is most abundant within the Purkinje cells. The different patterns of expression imply that the multiple subspecies of protein kinase C may each have a specific function in modulating the neuronal activity of particular cell types.

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Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Distinct cellular expression of beta I- and beta II-subspecies of protein kinase C in rat cerebellum

Ase

Saito²,

Shearman³

et al. 1988

J. Neurosci.

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Regional and cellular distribution of protein kinase C in rat cerebellar purkinje cells

2000

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Protein kinase C (PCK) is a family of isoforms that are implicated in subcellular signal transduction. The authors investigated the distribution of several PKC isoforms (PKC-alpha, PKC-beta, PKC-gamma, PKC-delta, and PKC-epsilon) within major cerebellar cell types as well as cerebellar projection target neurons, including Purkinje neurons, cerebellar nuclear neurons, and secondary vestibular neurons. PKC-alpha, PKC-beta, PKC-gamma, PKC-delta, and PKC-epsilon are found within the cerebellum. Of these isoforms, PKC-gamma and PKC-delta are highly expressed in Purkinje cells. PKC-gamma is expressed in all Purkinje cells, whereas the expression of PKC-delta is restricted to sagittal bands of Purkinje cells in the posterior cerebellar cortex. In the lower folia of the uvula and nodulus, Purkinje cell expression of PKC-delta is uniformly high, and the sagittal banding for PKC-delta expression is absent. Within the cerebellar nuclei, PKC-delta-immunolabeled axons terminate within the medial aspect of the caudal half of the ipsilateral interpositus nucleus. PKC delta-immunolabeled axons also terminated within the caudal medial and descending vestibular nuclei (MVN and DVN, respectively), the parasolitary nucleus (Psol), and the nucleus prepositus hypoglossi (NPH). PKC-gamma-immunolabeled axons terminated in all of the cerebellar nuclei as well as in the lateral and superior vestibular nuclei and the MVN, DVN, Psol, and NPH. The projection patterns of PKC-immunolabeled Purkinje cells were confirmed by lesion-depletion studies in which unilateral uvula-nodular lesions caused depletion of PKC-immunolabeled terminals ipsilateral to the lesion in the vestibular complex. These data identify circuitry that is unique to cerebellar-vestibular interactions.

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Immunocytochemical localization of protein kinase C subspecies in the rat spinal cord: Light and electron microscopic study

Mori

Kose

Tsujino

et al. 1990

J of Comparative Neurology

125

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Distinct expression of protein kinase C (PKC) subspecies in the central nervous system suggested that each subspecies has a distinct neural function in the processing and modulation of a variety of physiological responses to external signals. In this study, the cellular and subcellular distributions of beta I-, beta II- and gamma-subspecies of PKC were demonstrated by using subspecies-specific antibodies in the rat spinal cord. By light microscopy both gamma- and beta II-subspecies immunoreactivities were found only in neurons of the substantia gelatinosa and axons of the dorsal corticospinal tract in the spinal cord. Use of a double staining method, however, revealed that beta II-subspecies immunoreactivity was localized in the outer part of the lamina II, whereas gamma-subspecies immunoreactivity was found in the inner part of lamina II. Immunoreactive neurons containing beta I-subspecies were scattered in the substantia gelatinosa. Beta I-subspecies immunoreactivity varied in neuronal types. Furthermore, electron microscopic analysis clearly showed the subcellular distribution of these subspecies to be different from one another. Dense gamma-subspecies immunoreactivity was found in the cytoplasm except within cell organelles of the perikarya and dendrites. Some nuclei were stained as strongly as the cytoplasm and others were stained less heavily. The nucleoli had faint or no immunoreactivity. Reaction products of beta II-subspecies were located against the inner plasma membrane but not seen in the nuclei or nucleoli. Beta I-subspecies immunoreactivity appeared to be associated with the Golgi complex. No immunoreactive products of any PKC subspecies were detected in the presynaptic terminals. The different patterns of expression described above imply that individual PKC subspecies may have a specific function in modulating the neuronal activity in the different neurons of the spinal cord.

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Monoclonal antibodies against rat brain protein kinase C and their application to immunocytochemistry in nervous tissues

Cited by 70 publications

References 30 publications

Distinct cellular expression of beta I- and beta II-subspecies of protein kinase C in rat cerebellum

Distinct cellular expression of beta I- and beta II-subspecies of protein kinase C in rat cerebellum

Regional and cellular distribution of protein kinase C in rat cerebellar purkinje cells

Immunocytochemical localization of protein kinase C subspecies in the rat spinal cord: Light and electron microscopic study

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