Abstract
Purpose: Our purpose was to determine the association of calmodulin-dependent protein kinase II (CaMK II ) with oocyte activation and to explore the network of protein kinases during mammalian fertilization.
Methods: Mouse M-II oocytes were collected after superovulation induced by PMSG-hCG injection. The oocytes were inseminated or artificially activated by Ca ionophore (A23187) or 12-O-tetradecanoyl phorbol 13-acetate (TPA). The effects of KN-62, a specific and selective inhibitor of calmodulin-dependent protein kinase II, on second polar body emission (2PBE), pronuclearformation (PF), and cortical granule exocytosis (CGE) during fertilization or after artificial oocyte activation were investigated.
Results: KN-62 inhibited 2PBE and PF after sperm or Ca ionophore inducing activation. Additionally, PF was inhibited by KN-62 after TPA activation, whereas KN-62 did not inhibit CGE in any case. KN-04, an inactive form of KN-62, did not inhibit significantly 2PBE, CGE, or PF. When oocytes were exposed to KN-62 after Ca ionophore or TPA activation, no inhibitory effects on 2PBE or PF were observed.
Conclusions: The CaMK II activation that occurs after fertilization or artificial activation of mouse oocytes is presumably secondary to increases in the intracellular free calcium concentration. As determined by the use of inhibitor, CaMK II activity is associated with 2PBE and PF but not with CGE.
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REFERENCES
Cuthbertson KSR, Whittingham DG, Cobbold PH: Free Ca2+ increases in exponential phases during mouse oocyte activation. Nature 1981;294:754–757
Cuthbertson KSR, Cobbold PH: Phorbol ester and sperm activate mouse oocytes by inducing sustained oscillations in cell Ca2+. Nature 1985;316;541–542
Miyazaki S: Inositol 1,4,5-trisphosphate-induced calcium release and guanine nucleotide-binding protein-mediated periodic calcium rises in golden hamster eggs. J Cell Biol 1988;106:345–353
Durham ACH, Walton JM: Calcium ions and the control of proliferation in normal and cancer cells. Biosci Rep 1982;2:15–30
Rink TJ, Sanchez A, Hallam TJ: Diacylglycerol and phorbol ester stimulate secretion without raising cytoplasmic free calcium in human platelets. Nature 1983;305:317–319
Sha'afi RI, White JR, Molski TF, Shefcyk J, Volpi M, Naccache PH, Feinstein MB: Phorbol 12-myristate 13-acetate activates rabbit neutrophils without an apparent rise in the level of intracellular free calcium. Biochem Biophys Res Commum 1983;114:638–645
Steinhardt RA, Epel D, Carroll EJ, Yanagimachi R: Is calcium ionophore an universal activator for unfertilized eggs? Nature 1974;252:41–43
Fulton BP, Whitingham DG: Activation of mammalian oocytes by intracellular injection of calcium. Nature 1978;273:149–151
Kline D: Calcium-dependent events at fertilization of the frog: Injection of calcium buffer blocks ion channel opening, exocytosis, and formation of pronuclei. Dev Biol 1988;126:346–361
Kline D, Kline JT: Repetitive calcium transients and the role of calcium in exocytosis and cell cycle activation in the mouse egg. Dev Biol 1992;149:80–89
Xu Z, Lefevre L, Ducibella T, Schultz RM, Kopf GS: Effects of calcium-BAPTA buffers and calmodulin antagonist W-7 on mouse egg activation. Dev Biol 1996;180:594–604
Lorca T, Cruzalegui FH, Fesquet D, Cavadore J, Méry J, Means A, Doré M: Calmodulin-dependent protein kinase II mediates inactivation of MPF and CSF upon fertilization of Xenopus eggs. Nature 1993;366:270–273
Winston NJ, Maro B: Calmodulin-dependent kinase II is activated transiently in ethanol stimulated mouse oocytes. J Cell Sci 1995;170:350–352
Hidaka H, Inagaki M, Kawamoto S, Sasaki Y: Isoquinolinesulfonamides, novel and potent inhibitor of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry 1984;23:5036–5041
Tohda M, Nakamura J, Hidaka H, Nomura Y: Inhibitory effect of KN-62, a specific inhibitor of Ca/calmodulin-dependent protein kinase II, on serotonin-evoked Cl-current and 36C1− efflux in Xenopus oocytes. Neurosci Lett 1991;129:47–50
Tokumitsu H, Chijiwa T, Hagiwara M, Mizutani A, Terasawa M, Hidaka H: KN-62, 1-[N,O-bis (1,5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, a specific inhibitor of Ca2+-calmodulin-dependent protein kinase II. J Biol Chem 1990;265:4315–4320
Noronha KF, Mercier AJ: A role for calcium/calmodulin-dependent protein kinase in mediating synaptic modulation by a neuropeptide. Brain Res 1995;673:70–74
Maurer JA, Wenger BW, McKay DB: Effects of protein kinase inhibitors on morphology and function of cultured bovine adrenal chromaffin cells. J Neurochem 1996;66:105–113
Tsutsui M, Yanagihara N, Fukunaga K, Minami K, Nakashima Y, Kuroiwa A, Miyamoto E, Izumi F: Ca2+/calmodulin-dependent protein kinase II inhibitor KN-62 inhibits adrenal medullary chromaffin cell functions independent of its action on the kinase. J Neurochem 1996;66:2517–1522
Toyoda Y, Chang MC: Fertilization of rat eggs in vitro by epididymal spermatozoa and the development of eggs following transfer. J Reprod Fertil 1974;36:9–22
Kono T, Carroll J, Swann KS, Whittingham D: Nuclei from fertilized mouse embryos have calcium-releasing activity. Development 1995;121:1123–1128
Kono T, Carroll J, Jones K, Whittingham D, Nakahara T: How calcium oscillations at fertilization stop in the mouse egg. J Mammal Ova Res 1995;12:S68
Igusa Y, Miyazaki S: Effects of altered extracellular and intracellular calcium concentration on hyperpolarizing responses of the hamster egg. J Physiol 1986;340:611–632
Ducibella T, Anderson E, Albertini DF, Aalberg J, Rangarajan S: Quantitive studies of changes in cortical granule number and distribution in the mouse oocyte during meiotic maturation. Dev Biol 1988;130:184–197
Cherr GN, Drobnis EZ, Katz DF: Localization of cortical granule constituents before and after exocytosis in the hamster egg. J Exp Zool 1988;246:81–93
Inagaki N, Suzuki S, Kuji N, Kitai H, Nakatogawa N, Nozawa S: Egg activation induced by osmotic pressure change and the effects of amiloride on the cryopreservation of mouse oocytes. Mol Hum Reprod 1996;2:835–843
Meyer T, Hanson PI, Stryer L, Schulman H: Calmodulin trapping by calcium-calmodulin-dependent protein kinase. Science 1992;256:1199–1202
Moy GW, Kopf GS, Cache C, Vacquier VD: Calcium-mediated release of glucanase activity from cortical granules of sea urchin eggs. Dev Biol 1983;100:267–274
Masumoto N, Sasaki T, Tahara M, Mammoto A, Ikebuchi Y, Tasaka K, Tokunaga M, Takai Y, Miyake A: Involvement of Rabphilin-3A in cortical granule exocytosis in mouse eggs. J Cell Biol 1996;135:1741–1747
Nuoffer C, Balch WE: GTPases: Multifunctional molecular swithes regulating vesicular traffic. Annu Rev Biochem 1994;63:949–990
Tahara M, Tasaka K, Masumoto N, Mammoto A, Ikebuchi Y, Miyake A: Dynamics of cortical granule exocytosis at fertilization in living mouse eggs. Am J Physiol 1996;270:C1354–C1361
Geppert M, Bolshakov VY, Siegelbaum SA, Takei K, De Camilli P, Hammer RE, Sudhof TC: The role of Rab3A in neurotransmitter release. Nature 1994;369:493–497
Winston NJ, McGuinness O, Johnson MH, Maro B: The exit of mouse oocytes from meiotic M-phase requires an intact spindle during intracellular calcium release. J Cell Sci 1995;108:143–151
Ohta Y, Ohba T, Miyamoto E: Ca2+/calmodulin-dependent protein kinase II: Localization in the interphase nucleus and the mitotic apparatus of mammalian cells. Proc Natl Acad Sci USA 1990;87:5341–5345
Blankson H, Holen I, Seglen PO: Disruption of the cytokeratin cytoskeleton and inhibition of hepatocytic autophagy by okadaic acid. Exp Cell Res 1995;218:522–530
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Inagaki, N., Suzuki, S., Kitai, H. et al. Effect of KN-62, a Selective Inhibitor of Calmodulin-Dependent Kinase II, On Mouse Oocyte Activation. J Assist Reprod Genet 14, 609–616 (1997). https://doi.org/10.1023/A:1022541020458
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DOI: https://doi.org/10.1023/A:1022541020458