Immunohistochemical Localization of Ca2+/Calmodulin‐Dependent Protein Kinase II in Rat Brain and Various Tissues

Abstract: Polyclonal antibodies against Ca2+/calmodulin‐de‐pendent protein kinase II (CaM kinase II) of rat brain were prepared by immunizing rabbits and then purified by antigen‐affinity column. The antibodies which recognized both sub‐units of the enzyme with MrS 49K and 60K were used for the study on the distribution of CaM kinase II in formalin‐fixed, paraffin‐embedded tissues. In the brain, a light‐microscopic study demonstrated strong immunoreactivity in neu‐ronal somata and dendrites and weak immunoreactivity in nuclei. The densely stained regions included cerebral cortex, hippocampal formation, striatum, substantia nigra, and cer‐ebellar cortex. In substantia nigra, neurites were stained, but not neuronal somata. Electron microscopy revealed that the immunoreactive product was highly concentrated at the postsynaptic densities. In addition to neurons, weak immunoreactivity was also demonstrated in glial cells, such as as‐trocytes and ependymal cells of ventricles and epithelial cells of choroid plexus. In other tissues, strong immunoreactivity was observed in the islet of pancreas and moderate immunoreactivity in skeletal muscle and kidney tubules. Immunoreactivity was demonstrated in all of the tissues tested. The results suggest that CaM kinase II is widely distributed in the tissues.

[1]  K. Fukunaga,et al.  Phosphorylation and Inactivation of Brain Glycogen Synthase by a Multifunctional Calmodulin‐Dependent Protein Kinase , 1987, Journal of neurochemistry.

[2]  K. Fukunaga,et al.  MONOCLONAL ANTIBODY AGAINST A MULTIFUNCTIONAL CALMODULIN-DEPENDENT PROTEIN KINASE FROM RAT BRAIN AND THE TISSUE DISTRIBUTION OF THE ENZYME , 1986 .

[3]  E. Miyamoto,et al.  The distribution of calcineurin in rat brain by light and electron microscopic immunohistochemistry and enzyme-immunoassay , 1986, Brain Research.

[4]  M. Tanaka,et al.  Developmental Changes in Levels of Translatable mRNAs for Enolase Isozymes in Chicken Brain , 1986, Journal of neurochemistry.

[5]  K. Fukunaga,et al.  Purification and characterization of a multifunctional calmodulin-dependent protein kinase from canine myocardial cytosol. , 1986, Archives of biochemistry and biophysics.

[6]  K. Fukunaga,et al.  Regulation of the Actin‐Activated Mg‐ATPase of Brain Myosin via Phosphorylation by the Brain Ca2+, Calmodulin‐Dependent Protein Kinases , 1986, Journal of neurochemistry.

[7]  L. Rebhun,et al.  Purification and characterization of a sea urchin egg Ca2+-calmodulin-dependent kinase with myosin light chain phosphorylating activity. , 1986, The Journal of biological chemistry.

[8]  M. Kennedy,et al.  Regional distribution of type II Ca2+/calmodulin-dependent protein kinase in rat brain , 1985, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[9]  H. Schulman,et al.  Ca2+/calmodulin-dependent microtubule-associated protein 2 kinase: broad substrate specificity and multifunctional potential in diverse tissues. , 1985, Biochemistry.

[10]  P. Greengard,et al.  Protein Phosphorylation and Neuronal Function , 1985, Journal of neurochemistry.

[11]  K. Fukunaga,et al.  Dephosphorylation of Microtubule‐Associated Protein 2, τ Factor, and Tubulin by Calcineurin , 1985, Journal of neurochemistry.

[12]  P. Cuatrecasas,et al.  Cytoskeletal calmodulin-dependent protein kinase. Characterization, solubilization, and purification from rat brain. , 1985, The Journal of biological chemistry.

[13]  H. Schulman,et al.  Purification and characterization of a Ca2+/calmodulin-dependent protein kinase from rat brain. , 1984, Biochemistry.

[14]  P. Greengard,et al.  Immunocytochemical localization of calcium/calmodulin-dependent protein kinase II in rat brain. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[15]  M. Landt,et al.  Protein substrate specificity of a calmodulin-dependent protein kinase isolated from bovine heart. , 1984, Cell calcium.

[16]  K. Fukunaga,et al.  Phosphorylation and activation of calmodulin-sensitive cyclic nucleotide phosphodiesterase by a brain Ca2+, calmodulin-dependent protein kinase. , 1984, Life sciences.

[17]  P. Greengard,et al.  Mammalian brain phosphoproteins as substrates for calcineurin. , 1984, The Journal of biological chemistry.

[18]  P. Greengard,et al.  Calcium/calmodulin-dependent protein phosphorylation in the nervous system of Aplysia , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[19]  P. Cohen,et al.  Comparison of calmodulin‐dependent glycogen synthase kinase from skeletal muscle and calmodulin‐dependent protein kinase‐II from brain , 1984, FEBS letters.

[20]  J. Goldenring,et al.  Identification of the Major Postsynaptic Density Protein as Homologous with the Major Calmodulin‐Binding Subunit of a Calmodulin‐Dependent Protein Kinase , 1984, Journal of neurochemistry.

[21]  P. Greengard,et al.  Evidence that the major postsynaptic density protein is a component of a Ca2+/calmodulin-dependent protein kinase. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M K Bennett,et al.  Biochemical and immunochemical evidence that the "major postsynaptic density protein" is a subunit of a calmodulin-dependent protein kinase. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[23]  P. Greengard,et al.  A multifunctional calmodulin‐dependent protein kinase , 1983, FEBS letters.

[24]  M. T. Davison,et al.  The calmodulin-dependent glycogen synthase kinase from rabbit skeletal muscle. Purification, subunit structure and substrate specificity. , 1983, European journal of biochemistry.

[25]  M K Bennett,et al.  Purification and characterization of a calmodulin-dependent protein kinase that is highly concentrated in brain. , 1983, The Journal of biological chemistry.

[26]  B. González,et al.  Purification and characterization of a calmodulin-dependent kinase from rat brain cytosol able to phosphorylate tubulin and microtubule-associated proteins. , 1983, The Journal of biological chemistry.

[27]  J. Gershoni,et al.  Calmodulin-stimulated protein kinase activity from rat pancreas , 1983, The Journal of cell biology.

[28]  P. Greengard,et al.  Calmodulin-dependent protein kinase and associated substrates in Torpedo electric organ. , 1983, The Journal of biological chemistry.

[29]  P. Greengard,et al.  A calcium/calmodulin-dependent protein kinase from mammalian brain that phosphorylates Synapsin I: partial purification and characterization , 1983, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[30]  K. Fukunaga,et al.  Ca2+– and Calmodulin‐Dependent Phosphorylation of Microtubule‐Associated Protein 2 and t Factor, and Inhibition of Microtubule Assembly , 1983, Journal of neurochemistry.

[31]  T. Yamauchi,et al.  Purification and Cahracyterization of te Brain Calmodulin-Dependent Protein Kinase (Kinase II), Which Is involved in the Activtion of Tryptophan 5-Monooxygnase , 1983 .

[32]  K. Fukunaga,et al.  Calmodulin and Ca2+‐ and Calmodulin‐Dependent Protein Kinase in Rat Anterior Pituitary Gland , 1983, Journal of neurochemistry.

[33]  T. Soderling,et al.  Purification and characterization of rabbit liver calmodulin-dependent glycogen synthase kinase. , 1983, The Journal of biological chemistry.

[34]  T. Yamauchi,et al.  Phosphorylation of microtubule-associated protein 2 by calmodulin-dependent protein kinase (Kinase II) which occurs only in the brain tissues. , 1982, Biochemical and biophysical research communications.

[35]  K. Fukunaga,et al.  Purification and Characterization of a Ca2+‐ and Calmodulin‐Dependent Protein Kinase from Rat Brain , 1982, Journal of neurochemistry.

[36]  A. Depaoli-Roach,et al.  Purification and characterization of a rabbit liver calmodulin-dependent protein kinase able to phosphorylate glycogen synthase. , 1982, The Journal of biological chemistry.

[37]  P. Siekevitz,et al.  Function of calmodulin in postsynaptic densities. II. Presence of a calmodulin- activatable protein kinase activity , 1981, The Journal of cell biology.

[38]  T. Yamauchi,et al.  Tyrosine 3-monoxygenase is phosphorylated by Ca2+-, calmodulin-dependent protein kinase, followed by activation by activator protein. , 1981, Biochemical and biophysical research communications.

[39]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[40]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[41]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[42]  L. Sternberger,et al.  THE UNLABELED ANTIBODY ENZYME METHOD OF IMMUNOHISTOCHEMISTRY PREPARATION AND PROPERTIES OF SOLUBLE ANTIGEN-ANTIBODY COMPLEX (HORSERADISH PEROXIDASE-ANTIHORSERADISH PEROXIDASE) AND ITS USE IN IDENTIFICATION OF SPIROCHETES , 1970, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[43]  K. Fukunaga,et al.  Purification of the multifunctional calmodulin-dependent protein kinases from lung and liver, and the comparison to the brain enzyme. , 1988, Japanese journal of pharmacology.

[44]  K. Fukunaga,et al.  Identification of the multifunctional calmodulin-dependent protein kinase in the cytosol, sarcoplasmic reticulum, and sarcolemma of rabbit skeletal muscle. , 1988, Archives of biochemistry and biophysics.

[45]  E. Miyamoto Characterization of a Multifunctional Ca 2+ -Calmodulin-Dependent Protein Kinase in the Brain , 1986 .

[46]  畑田 泰行 Calmodulin and Ca[2+]- and calmodulin-dependent protein kinase in rat anterior pituitary gland , 1985 .

[47]  P. Greengard,et al.  Protein kinases in the brain. , 1985, Annual review of biochemistry.

[48]  P. Greengard,et al.  Calcium/Calmodulin-Dependent Protein Phosphorylation in the Nervous System , 1985 .

[49]  K. Fukunaga,et al.  A Ca 2+ -CALMODULIN-DEPENDENT PROTEIN KINASE IN THE PARTICULATE FRACTION OF RAT BRAIN AND ENDOGENOUS PHOSPHORYLATION OF PARTICULATE-BOUND SUBSTRATES , 1984 .

[50]  T. Yamauchi,et al.  Purification and characterization of the brain calmodulin-dependent protein kinase (kinase II), which is involved in the activation of tryptophan 5-monooxygenase. , 1983, European journal of biochemistry.