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Immunohistochemical detection of calmodulin and calmodulin-dependent protein kinase II in the mouse testis

Published online by Cambridge University Press:  06 December 2000

M. Moriya ,
C. Katagiri ,
M. Ikebe  and
K. Yagi
M. Moriya
Affiliation:
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
C. Katagiri
Affiliation:
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
M. Ikebe
Affiliation:
Department of Physiology, University of Massachusetts Medical Center, Worcester, MA 01655, USA.
K. Yagi
Affiliation:
Agro-Science Research Laboratory, Hokkaisankyo, Kitahiroshima, Hokkaido 061-1111, Japan.
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Abstract

We reported previously that in mouse testis calmodulin-dependent protein phosphatase (calcineurin) is localised in the nuclei of round and elongating spermatids (Cell Tissue Res. 1995; 281: 273-81). In this study, we studied the immunohistochemical localisation of calcium/calmodulin-dependent protein kinase (CaM kinase II) using antibodies against CaM kinase IIγ from chicken gizzard and specific antibodies raised against the amino acid sequence Ileu480–Ala493 of this enzyme, and compared it with the distribution of calmodulin. Indirect immunofluorescence was most concentrated in early spermatocytes and localised in the outermost layer of seminiferous tubules where the calmodulin level was relatively low. Measurements of immuno-gold particle densities on electron micrographs revealed that CaM kinase II is transiently increased in the nucleus of zygotene spermatocytes. These observations suggest the involvement of CaM kinase II in the meiotic chromosomal pairing process. An extremely high concentration of calmodulin in spermatogenic cells undergoing meiosis may not be directly related to activation of calmodulin-dependent kinases and phosphatases.

Keywords

CalcineurinImmuno-electron microscopyImmunofluorescenceSpermatogenesisZygotene spermatocyte
Type
Research Article
Information
Zygote , Volume 8 , Issue 4 , November 2000 , pp. 303 - 314
Copyright
2000 Cambridge University Press

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