Creatine Kinase Activity in Postnatal Rat Brain Development and in Cultured Neurons, Astrocytes, and Oligodendrocytes

Abstract: The development and distribution of cytosolic creatine kinase (CK) activity was studied in rat brain and in cell culture. The activity of CK in whole brain increased almost fivefold during the period from birth to day 40 when adult levels of 18–19 U/mg of protein were attained. The distribution of CK activity was examined in dissected regions of the adult brain and was nonuniform; the cerebellum, the striatum, and the pyramidal tracts contained significantly higher CK activity than did whole brain. The cellular compartmentation of CK was investigated using primary cultures of purified neurons, astrocytes, and oligodendrocytes. The CK activity in neurons increased fourfold greater than that measured at the time of isolation to 4 U/mg of protein. The CK activity in astrocytes cultured for 20 days was 3.5 U/mg of protein and was 1.5‐fold greater than that measured at the time of isolation. In contrast, the CK activity in cultured oligodendrocytes (day 20) was three‐ to fourfold higher than that determined in astrocytes and almost sevenfold higher than the activity measured at the time the cells were isolated. The high levels of CK in cultured oligodendrocytes suggest a role for this enzyme in oligodendrocyte function and/or myelinogenesis.

[1]  G. Pfleiderer,et al.  Quantitation of creatine kinase isoenzymes in human tissues and sera by an immunological method. , 1975, Clinica chimica acta; international journal of clinical chemistry.

[2]  K. Ikeda,et al.  The presence of creatine kinase (CK)-immunoreactive neurons in the zona incerta and lateral hypothalamic area of the mouse brain , 1987, Brain Research.

[3]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[4]  H. Maker,et al.  CHANGES DURING DEVELOPMENT OF MOUSE BRAIN IN THE ACTIVITIES AND SUBCELLULAR DISTRIBUTIONS OF CREATINE AND ADENYLATE KINASES , 1974, Journal of neurochemistry.

[5]  K. McCarthy,et al.  Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue , 1980, The Journal of cell biology.

[6]  B. Shapiro,et al.  Metabolite channeling: A phosphorylcreatine shuttle to mediate high energy phosphate transport between sperm mitochondrion and tail , 1985, Cell.

[7]  N. Sims,et al.  2′,3′-Cyclic Nucleotide 3′-Phosphodiesterase , 1978 .

[8]  C. Merril,et al.  [30] Gel protein stains: Silver stain , 1984 .

[9]  F. Suzuki,et al.  Highly Sensitive Immunoassay for Rat Brain‐Type Creatine Kinase: Determination in Isolated Purkinje Cells , 1986, Journal of neurochemistry.

[10]  J. de Vellis,et al.  Developmental expression of glial-specific mRNAs in primary cultures of rat brain visualized by in situ hybridization. , 1988, Journal of neuroscience research.

[11]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[12]  S. Schneider,et al.  A Sensitive Fluorometric Assay for 2′,3′‐Cyclic Nucleotide 3′‐Phosphohydrolase , 1980, Journal of neurochemistry.

[13]  J. Gershoni,et al.  Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to a positively charged membrane filter. , 1982, Analytical biochemistry.

[14]  I. T. Oliver A spectrophotometric method for the determination of creatine phosphokinase and myokinase. , 1955, The Biochemical journal.

[15]  J. de Vellis,et al.  Capacity for substrate utilization in oxidative metabolism by neurons, astrocytes, and oligodendrocytes from developing brain in primary culture , 1987, Journal of neuroscience research.

[16]  W. E. Jacobus,et al.  Creatine kinase of heart mitochondria. Control of oxidative phosphorylation by the extramitochondrial concentrations of creatine and phosphocreatine. , 1986, The Journal of biological chemistry.

[17]  H. Maker,et al.  Regional changes in cerebellar creatine phosphate metabolism during late maturation. , 1973, Experimental neurology.

[18]  S. Bessman The creatine phosphate energy shuttle--the molecular asymmetry of a "pool". , 1987, Analytical biochemistry.

[19]  G. Mckhann,et al.  ACTIVITY OF 2′,3′‐CYCLIC‐NUCLEOTIDE 3′‐PHOSPHODIESTERASE IN REGIONS OF RAT BRAIN DURING DEVELOPMENT: QUANTITATIVE RELATIONSHIP TO MYELIN BASIC PROTEIN , 1978, Journal of neurochemistry.

[20]  R. Thompson,et al.  Immunohistochemical localization of creatine kinase-BB isoenzyme to astrocytes in human brain , 1980, Brain Research.

[21]  R. Wevers,et al.  Mitochondrial creatine kinase (EC 2.7.3.2) in the brain. , 1982, Clinica chimica acta; international journal of clinical chemistry.

[22]  K. Ikeda,et al.  Creatine kinase immunoreactivity: Localization in nerve terminals in the hypothalamic area and superior colliculus of the mouse brain , 1988, Neuroscience Letters.

[23]  T. Yoshimine,et al.  Immunohistochemical localization of creatine kinase BB-isoenzyme in human brain: Comparison with tubulin and astroprotein , 1983, Brain Research.

[24]  S E Poduslo,et al.  MYELINATION IN RAT BRAIN: CHANGES IN MYELIN COMPOSITION DURING BRAIN MATURATION 1 , 1973, Journal of neurochemistry.

[25]  H. Eppenberger,et al.  THE ONTOGENY OF CREATINE KINASE ISOZYMES. , 1964, Developmental biology.