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Onset of expression of the alpha subunit of Ca2+/calmodulin-dependent protein kinase II and a novel related protein in the developing retina

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Abstract

Calcium-calmodulin-dependent protein kinase II is an abundant protein in the nervous system and has been associated with many aspects of neuronal function, including events related to synaptic transmission. The purpose of this study is to correlate the onset of expression of this kinase with a specific developmental event in retinal morphogenesis using a monoclonal antibody to the 50-kDa alpha-subunit.

Microscopy showed the antigen to be associated with the plexiform layers of the retina. Western blots demonstrated that the onset of expression of the alpha-subunit coincided in time with the initial formation of the plexiform layers. However, the onset of expression of the 50-kDa alpha-subunit was preceded by the earlier embryonic appearance of a related 82.5-kDa antigen that was recognized by the antibody. The amount of this latter protein declined as the amount of the alpha-subunit increased in retinal homogenates. Although this related 82.5 kDa protein disappeared from blots of retinal homogenates after embryonic d 14, it could be detected in concentrated supernatant fractions isolated from the retinae of hatched chicks. Microscopy showed that a subset of retinal cells and their processes contained this antigen in early embryonic chicks. Finally, the 50 kDa alpha-subunit of kinase II and the 82.5 kDa novel antigen were shown to be separable by differential centrifugation.

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References

  • Benson D. L., Isackson P. J., Hendry S. H., and Jones E. G. (1991) Differential gene expression for glutamic acid decarboxylase and type II calcium-calmodulin-dependent protein kinase in basal ganglia, thalamus, and hypothalamus of the monkey.J. Neurosci. 11, 1540–1564.

    PubMed  CAS  Google Scholar 

  • Bronstein J. M., Wasterlain C. G., and Farber D. B. (1988a) A retinal calmodulin-dependent kinase: calcium/calmodulin-stimulated and -inhibited states.J. Neurochem. 50, 1438–1446.

    Article  PubMed  CAS  Google Scholar 

  • Bronstein J. M., Wasterlain C. G., Bok D., Lasher R., and Farber D. B. (1988b) Localization of retinal calmodulin kinase.Exp. Eye Res. 47, 391–402.

    Article  PubMed  CAS  Google Scholar 

  • Bronstein J., Wasterlain C. G., Lasher R., and Farber D. B. (1989) Dark-induced changes in activity and compartmentalization of retinal calmodulin kinase in the rat.Brain Res. 495, 83–88.

    Article  PubMed  CAS  Google Scholar 

  • Erondu N. E. and Kennedy M. B. (1985) Regional distribution of type II Ca2+/calmodulin-dependent protein kinase in rat brain.J. Neurosci. 5, 3270–3277.

    PubMed  CAS  Google Scholar 

  • Goldenring J. R., Gonzalez B., McGuire J. S., and Lorenzo R. J. (1983) Purification and characterization of a calmodulin-dependent kinase from rat brain cytosol able to phosphorylate tubulin and microtubule-associated proteins.J. Biol. Chem. 258, 12,632–12,640.

    CAS  Google Scholar 

  • Goldenring J. R., McGuire J. S. Jr., and DeLorenzo R. J. (1984a) Identification of the major post-synaptic density protein as homologous with the major calmodulin binding subunit of a calmodulin dependent protein kinase.J. Neurochem. 42, 1077–1084.

    Article  PubMed  CAS  Google Scholar 

  • Goldenring J. R., Casanova J. E., and DeLorenzo R. J. (1984b) Tubulin-associated calmodulin-dependent kinase: evidence for an endogenous complex of tubulin with a calcium-calmodulin-dependent kinase.J. Neurochem. 43, 1669–1679.

    Article  PubMed  CAS  Google Scholar 

  • Gupta R. P., Lapadula D. M., and Abou-Donia M. B. (1992) Ca2+/calmodulin-dependent protein kinase II from hen brain.Biochem. Pharmacol. 43, 1975–1988.

    Article  PubMed  CAS  Google Scholar 

  • Hamburger V. and Hamilton H. L. (1951) A series of normal stages in the development of the chick embryo.J. Morph. 88, 49–92.

    Article  Google Scholar 

  • Hanks S. K. and Quinn A. M. (1991) Protein kinase catalytic domain sequence data base: identification of conserved features of primary structure and classification of family members.Methods Enzymol. 200, 38–62.

    Article  PubMed  CAS  Google Scholar 

  • Hendry S. H. and Kennedy M. B. (1986) Immunoreactivity for a calmodulin-dependent protein kinase is selectively increased in macaque striate cortex after monocular deprivation.Proc. Natl. Acad. Sci. USA 83, 1536–1541.

    Article  PubMed  CAS  Google Scholar 

  • Hughes W. F. and LaVelle A. (1974) On the synaptogenic sequence in the chick retina.Anat. Rec. 179, 297–302.

    Article  PubMed  CAS  Google Scholar 

  • Jones E. G., Huntley G. W., and Benson D. L. (1994) Alpha calcium/calmodulin-dependent protein kinase II selectively expressed in a subpopulation of excitatory neurons in monkey sensory-motor cortex: comparison with GAD-67 expression.J. Neurosci. 14, 611–629.

    PubMed  CAS  Google Scholar 

  • Kanaseki T., Ikeuchi Y., Suiura H., and Yamauchi T. (1991) Structural features of Ca2+/calmodulin-dependent protein kinase revealed by electron microscopy.J. Cell Biol. 115, 1049–1060.

    Article  PubMed  CAS  Google Scholar 

  • Kato M., Hagiwara M., and Hodaka H. (1992) Identification of a 80 KDa calmodulin-dependent kinase by renaturation blotting assay.Biochem. J. 281, 339–342.

    PubMed  CAS  Google Scholar 

  • Kelly P. T., McGuiness T. L., and Greengard P. (1984) Evidence that the major post-synaptic density protein is a component of a calcium-calmodulin dependent protein kinase.Proc. Natl. Acad. Sci. USA 81, 945–949.

    Article  PubMed  CAS  Google Scholar 

  • Kelly P. T. and Vernon P. (1985) Changes in the subcellular distribution of calmodulin-kinase II during brain development.Brain Res. 350, 211–224.

    PubMed  CAS  Google Scholar 

  • Kennedy M. B. and Greengard P. (1981) Two calcium/calmodulin-dependent protein kinases which are highly concentrated in brain phosphorylate Protein I at distinct sites.Proc. Natl. Acad. Sci. USA 78, 1293–1297.

    Article  PubMed  CAS  Google Scholar 

  • Kennedy M. B., Bennett M. K., and Erondu N. E. (1983) Biochemical and immunochemical evidence that the major post synaptic density protein is α-subunit of a calmodulin-dependent protein kinase.Proc. Natl. Acad. Sci. USA 80, 7357–7361.

    Article  PubMed  CAS  Google Scholar 

  • Kuret J. and Schulman H. (1984) Purification and characterization of a Ca2+/calmodulin-dependent protein kinase from rat brain.Biochemistry 23, 5495–5504.

    Article  PubMed  CAS  Google Scholar 

  • Laemmli U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–685.

    Article  PubMed  CAS  Google Scholar 

  • Liu N., Wei X., and Cooper N. G. F. (1995) Ca2+/calmodulin-dependent kinase II in developing chick retina.Invest. Ophthalmol. Vis. Sci. Suppl. 36, S59.

    Google Scholar 

  • Liu N. and Cooper N. G. F. (1994) Purification and characterization of the Ca2+/calmodulin-dependent protein kinase II from chicken forebrain.J. Mol. Neurosci. 5, 193–206.

    Article  PubMed  CAS  Google Scholar 

  • Llinas R., McGuiness T. L., Leonard C. S., Sugimori M., and Greengard P. (1985) Intraterminal injection of synapsin I or calcium/calmodulin-dependent protein kinase II alters transmitter release at the squid giant synapse.Proc. Natl. Acad. Sci. USA 82, 3035–3039.

    Article  PubMed  CAS  Google Scholar 

  • McGuiness T. L., Lai Y., Greengard P., Woodgett J. R., and Cohen P. (1983) A multifunctional calmodulin-dependent protein kinase. Similarities between skeletal muscle glycogen synthase kinase and a brain synapsin I kinase.FEBS Lett. 163, 329–334.

    Article  Google Scholar 

  • McGuiness T. L., Lai Y., and Greengard P. (1985) Ca2+/calmodulin-dependent protein kinase II. Isoenzymic forms from rat forebrain and cerebellum.J. Biol. Chem. 260, 1696–1704.

    Google Scholar 

  • Miller S. G. and Kennedy M. B. (1985) Distinct forebrain and cerebellar isozymes of type II Ca2+/calmodulin-dependent protein kinase associate differently with the postsynaptic density fraction.J. Biol. Chem. 260, 9039–9046.

    PubMed  CAS  Google Scholar 

  • Ochiishi T., Terashima T., Sugiura H., and Yamauchi T. (1994) Immunohistochemical localization of Ca2+/calmodulin-dependent protein kinase II in the rat retina.Brain Res. 634, 257–265.

    Article  PubMed  CAS  Google Scholar 

  • Ouimet C. C., Miller P. E., Hemmings H. C. Jr., Walaas S. I., and Greengard P. (1984) DARPP-32, a dopamine- and adenosine 3′:5′-monophosphate-regulated phosphoprotein enriched in dopamine-innervated brain regions. III. Immunocytochemical localization.J. Neurosci. 4, 111–124.

    PubMed  CAS  Google Scholar 

  • Rostas J. A. P., Brent V. A., Heath J. W., Neame R. L. B., Powis D. A., Weinberger R. P., and Dunkley P. R. (1986) The subcellular distribution of a membrane bound calmodulin-stimulated protein kinase.Neurochem. Res. 11, 253–268.

    Article  PubMed  CAS  Google Scholar 

  • Rostas J. A. P., Brent V. A., Seccombe M., Weinberger R. P., and Dunkley P. R. (1989) Purification and characterization of calmodulin-stimulated protein kinase II from two-day and adult chicken forebrain.J. Mol. Neurosci. 1, 93–104.

    PubMed  CAS  Google Scholar 

  • Sahyoun N., LeVine H. III, McDonald O. B., and Cuatrecasas P. (1986) Specific postsynaptic density proteins bind tubulin and calmodulin-dependent protein kinase II.J. Biol. Chem. 261, 12,339–12,344.

    CAS  Google Scholar 

  • Sakagami H. and Kondo H. (1993) Differential expression of mRNAs encoding γ and δ subunits of Ca2+/calmodulin-dependent protein kinase type II (Cam kinase II) in the mature and postnatally developing brain.Mol. Brain Res. 20, 51–63.

    Article  PubMed  CAS  Google Scholar 

  • Scholtz W. K., Baitinger C., Schulman H., and Kelly P. T. (1988) Developmental changes in Ca2+/calmodulin-dependent protein kinase II in cultures of hippocampal pyramidal neurons and astrocytes.J. Neurosci. 8, 1039–1051.

    Google Scholar 

  • Schulman H. (1988) The multifunctional Ca2+/calmodulin-dependent protein kinase, inAdvances in Second Messenger and Phosphoprotein Research, vol. 22 (Greengard P. and Robinson R., eds.), Raven, New York, pp. 39–112.

    Google Scholar 

  • Tobimatsu T. and Fujisawa H. (1989) Tissue-specific expression of four types of rat calmodulin-dependent protein kinase II mRNAs.J. Biol. Chem. 264, 17,907–17,912.

    CAS  Google Scholar 

  • Towbin H., Staehelin T., and Gordon J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proc. Natl. Acad. Sci. USA 76, 4350–4354.

    Article  PubMed  CAS  Google Scholar 

  • Yamauchi T. and Fujisawa H. (1983) Disassembly of microtubules by the action of calmodulin-dependent protein kinase (kinase II) which occurs only in brain tissues.Biochem. Biophys. Res. Commun. 110, 287–291.

    Article  PubMed  CAS  Google Scholar 

  • Yamauchi T., Ohsako S., and Deguchi T. (1989) Expression and characterization of calmodulin-dependent protein kinase II from cloned cDNAs in Chinese hamster ovary cells.J. Biol. Chem. 264, 19,108–19,116.

    CAS  Google Scholar 

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Cooper, N.G.F., Wei, X. & Liu, N. Onset of expression of the alpha subunit of Ca2+/calmodulin-dependent protein kinase II and a novel related protein in the developing retina. J Mol Neurosci 6, 75–89 (1995). https://doi.org/10.1007/BF02736768

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  • DOI: https://doi.org/10.1007/BF02736768

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