Microtubule and chromatin behavior follow MAP kinase activity but not MPF activity during meiosis in mouse oocytes

Oocyte meiotic maturation is triggered by different stimuli (hormones, unknown signals through cell interactions) in different species. These stimuli indirectly lead to the activation of a major cell cycle regulating activity, the maturation promoting factor (MPF). Other factors, such as the product of the proto-oncogene c-mos or enzymes of the MAP kinase family, are also involved in the process of maturation. MAP kinase activation occurs during meiotic maturation in oocytes from different species with different kinetics. The relationships between MPF activation and MAP kinase activation have been well studied in species such as clam and Xenopus. In this paper, we study the precise timing of MAP kinase activation (as measured by phosphorylation of exogenous myelin basic protein and shifts in mobility of ERK 1 and ERK 2) versus MPF activation (as measured by phosphorylation of exogenous histone H1) during mouse oocyte maturation and, in parallel, morphological events such as changes in microtubule organization and chromatin condensation. We observed that MAP kinase activation was delayed after MPF activation and that this activity persisted throughout maturation whereas MPF activity dropped between the two meiotic metaphases. After parthenogenetic activation of ovulated eggs, MAP kinase inactivation was very slow compared to MPF inactivation. During the first mitotic cell cycle, a rise in myelin basic protein kinase activity at M-phase was observed but it was not related to MAP kinase activation. Furthermore, microtubules and chromatin remained in a metaphase-like state during the complete period of maturation (including the period between the two meiotic metaphases) and a few hours after activation.(ABSTRACT TRUNCATED AT 250 WORDS)

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Tyrosine phosphorylation and activation of homologous protein kinases during oocyte maturation and mitogenic activation of fibroblasts.

Molecular and Cellular Biology ◽

10.1128/mcb.11.5.2517 ◽

1991 ◽

Vol 11 (5) ◽

pp. 2517-2528 ◽

Cited By ~ 118

Author(s):

J Posada ◽

J Sanghera ◽

S Pelech ◽

R Aebersold ◽

J A Cooper

Keyword(s):

Cell Cycle ◽

Map Kinase ◽

Tyrosine Phosphorylation ◽

Oocyte Maturation ◽

Mammalian Cells ◽

Map Kinases ◽

Kinase Activity ◽

Mitotic Cell ◽

Meiotic Maturation ◽

Sea Star

Meiotic maturation of Xenopus and sea star oocytes involves the activation of a number of protein-serine/threonine kinase activities, including a myelin basic protein (MBP) kinase. A 44-kDa MBP kinase (p44mpk) purified from mature sea star oocytes is shown here to be phosphorylated at tyrosine. Antiserum to purified sea star p44mpk was used to identify antigenically related proteins in Xenopus oocytes. Two tyrosine-phosphorylated 42-kDa proteins (p42) were detected with this antiserum in Xenopus eggs. Xenopus p42 chromatographs with MBP kinase activity on a Mono Q ion-exchange column. Tyrosine phosphorylation of Xenopus p42 approximately parallels MBP kinase activity during meiotic maturation. These results suggest that related MBP kinases are activated during meiotic maturation of Xenopus and sea star oocytes. Previous studies have suggested that Xenopus p42 is related to the mitogen-activated protein (MAP) kinases of culture mammalian cells. We have cloned a MAP kinase relative from a Xenopus ovary cDNA library and demonstrate that this clone encodes the Xenopus p42 that is tyrosine phosphorylated during oocyte maturation. Comparison of the sequences of Xenopus p42 and a rat MAP kinase (ERK1) and peptide sequences from sea star p44mpk indicates that these proteins are close relatives. The family members appear to be tyrosine phosphorylated, and activated, in different contexts, with the murine MAP kinase active during the transition from quiescence to the G1 stage of the mitotic cell cycle and the sea star and Xenopus kinases being active during M phase of the meiotic cell cycle.

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Mitogen-activated protein kinase activity and microtuble organisation are altered by protein synthesis inhibition in maturing porcine oocytes

Zygote ◽

10.1017/s0967199400003105 ◽

1996 ◽

Vol 4 (3) ◽

pp. 191-198 ◽

Cited By ~ 13

Author(s):

Maki Inoue ◽

Kunihiko Naito ◽

Taisuke Nakayama ◽

Eimei Sato

Keyword(s):

Protein Synthesis ◽

Map Kinase ◽

Kinase Activity ◽

Germinal Vesicle ◽

Protein Kinase Activity ◽

Protein Synthesis Inhibition ◽

Germinal Vesicle Breakdown ◽

Kinase Activation ◽

Synthesis Inhibition ◽

Mitogen Activated Protein

SummaryPreviously we have shown that mitogen-activated protein (MAP) kinase activity abruptly increases at the first metaphase (M1) and remains significantly higher than that at the germinal vesicle (GV) stage until the second metaphase (M2) in porcine oocytes cultured in vitro. The present paper describes how the mechanism of the blockage of meiotic maturation by protein sythesis inhibition involves MAP kinase regulation. Cycloheximide arrested both germinal vesicle breakdown (GVBD) and the normal transition from M1 to M2. MAP kinase activation was also reduced in these maturation-inhibited oocytes. By using immunofluorescence microscopy with the monoclonal antibody raised against rat α-tubulin, we showed that cycloheximide caused morphological abnormality in a spindle at M1, but not at M2. All these results indicate that in porcine oocytes: (1) GV blockage by protein synthesis inhibition involves the suppression of both histone H1 kinase and MAP kinase activation, (2) during the transition from M1 to M2, maintenance of a normal metaphasic spindle and high MAP kinase activity require protein synthesis, and (3) once the M2 cytoskeletal structures have been completed, and/or after the ‘critical period’, cytostatic factor activity is independent of protein synthesis.

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Endothelin stimulates mitogen-activated protein kinase activity in mesangial cells through ETA.

Journal of the American Society of Nephrology ◽

10.1681/asn.v541074 ◽

1994 ◽

Vol 5 (4) ◽

pp. 1074-1080

Author(s):

Y Wang ◽

J Pouysségur ◽

M J Dunn

Keyword(s):

Map Kinase ◽

Map Kinases ◽

Kinase Activity ◽

Mesangial Cells ◽

Chinese Hamster ◽

Lung Fibroblasts ◽

Protein Kinase Activity ◽

Glomerular Mesangial Cells ◽

Kinase Activation ◽

Mitogen Activated Protein

Accumulating evidence suggests that endothelin (ET) contributes to the pathophysiology of such disorders as acute renal failure, cyclosporine-mediated renal and vascular toxicity, and perhaps even glomerular inflammation. The postreceptor signaling pathways that mediate the actions of ET in these pathophysiologic conditions may include activation of kinase cascades. Thus, the effects of ET isopeptides on p42 and p44 mitogen-activated protein (MAP) kinase activity in rat glomerular mesangial cells were examined. ET-1 activated both p42 and p44 MAP kinases with similar dose responses and different kinetics. The threshold for kinase activation was 10(-9) M ET-1. ET-1 stimulated p42 and p44 MAP kinases with similar rapid (5 min) but different sustained activation of p42 (3 to 6 h) and p44 (1 to 2 h). Endothelin-3 (ET-3) also activated both isoforms of MAP kinase but with a threshold at 10(-7) M. Compared with ET-1, ET-3 stimulated only a rapid increase of p42 MAP kinase activity. We further investigated which ET receptors are coupled to MAP kinase activation. BQ-123, an ETA blocker, completely blocked the responsiveness of the MAP kinase to either ET-1 or ET-3. In Chinese hamster lung fibroblasts transfected with ETA or ETB cDNA, both receptors showed a rapid stimulation of MAP kinase in response to ET-1. These results suggest that ET can activate MAP kinases through both ET receptors but act exclusively through ETA in glomerular mesangial cells.

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Dissociation of MAP kinase activation and MPF activation in hormone-stimulated maturation of Xenopus oocytes

Development ◽

10.1242/dev.126.20.4537 ◽

1999 ◽

Vol 126 (20) ◽

pp. 4537-4546 ◽

Cited By ~ 4

Author(s):

D.L. Fisher ◽

T. Brassac ◽

S. Galas ◽

M. Doree

Keyword(s):

Map Kinase ◽

Xenopus Oocytes ◽

Kinase Activity ◽

Early Response ◽

Meiotic Maturation ◽

Cdc2 Kinase ◽

Kinase Activation ◽

Cell Cycle Activation ◽

Map Kinase Phosphatase ◽

Universal Mechanism

MAP kinase activation occurs during meiotic maturation of oocytes from all animals, but the requirement for MAP kinase activation in reinitiation of meiosis appears to vary between different classes. In particular, it has become accepted that MAP kinase activation is necessary for progesterone-stimulated meiotic maturation of Xenopus oocytes, while this is clearly not the case in other systems. In this paper, we demonstrate that MAP kinase activation in Xenopus oocytes is an early response to progesterone and can be temporally dissociated from MPF activation. We show that MAP kinase activation can be suppressed by treatment with geldanamycin or by overexpression of the MAP kinase phosphatase Pyst1. A transient and low-level early activation of MAP kinase increases the efficiency of cell cycle activation later on, when MAP kinase activity is no longer essential. Many oocytes can still undergo reinitiation of meiosis in the absence of active MAP kinase. Suppression of MAP kinase activation does not affect the formation or activation of Cdc2-cyclin B complexes, but reduces the level of active Cdc2 kinase. We discuss these findings in the context of a universal mechanism for meiotic maturation in oocytes throughout the animal kingdom.

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Mos is required for MAP kinase activation and is involved in microtubule organization during meiotic maturation in the mouse

Development ◽

10.1242/dev.122.3.815 ◽

1996 ◽

Vol 122 (3) ◽

pp. 815-822 ◽

Cited By ~ 20

Author(s):

M.H. Verlhac ◽

J.Z. Kubiak ◽

M. Weber ◽

G. Geraud ◽

W.H. Colledge ◽

...

Keyword(s):

Map Kinase ◽

Meiotic Maturation ◽

Microtubule Organization ◽

Kinase Activation ◽

Mouse Oocytes ◽

M Phase ◽

In The Wild ◽

Cytostatic Factor ◽

Map Kinase Kinase Kinase ◽

Map Kinase Kinase

Mos is normally expressed during oocyte meiotic maturation in vertebrates. However, apart from its cytostatic factor (CSF) activity, its precise role during mouse meiosis is still unknown. First, we analyzed its role as a MAP kinase kinase kinase. Mos is synthesized concomitantly with the activation of MAP kinase in mouse oocytes. Moreover, MAP kinase is not activated during meiosis in oocytes from mos −/− mice. This result implies that Mos is necessary for MAP kinase activation in mouse oocytes. Raf-1, another MAP kinase kinase kinase, is already present in immature oocytes, but does not seem to be active when MAP kinase is activated. Moreover, the absence of MAP kinase activation in mos −/− oocytes demonstrates that Raf-1 cannot compensate for the lack of Mos. These results suggest that Raf-1 is not involved in MAP kinase activation. Second, we analyzed the organization of the microtubules and chromosomes in oocytes from mos −/− mice. We observed that during the transition between two meiotic M-phases, the microtubules and chromosomes evolve towards an interphase-like state in mos −/− oocytes, while in the control mos +/− oocytes they remain in an M-phase configuration, as in the wild type. Moreover, after spontaneous activation, the majority of mos −/− oocytes are arrested for at least 10 hours in a third meiotic M-phase where they exhibit monopolar half-spindles. These observations present the first evidence, in intact oocytes, of a role for the Mos/…/MAP kinase cascade in the control of microtubule and chromatin organization during meiosis.

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Activation of ribosomal S6 kinase (RSK) during porcine oocyte maturation

Zygote ◽

10.1017/s0967199402002058 ◽

2002 ◽

Vol 10 (1) ◽

pp. 31-36 ◽

Cited By ~ 14

Author(s):

K. Sugiura ◽

K. Naito ◽

N. Iwamori ◽

H. Kagii ◽

S. Goto ◽

...

Keyword(s):

Map Kinase ◽

Kinase Activity ◽

Meiotic Maturation ◽

Protein Kinase Activity ◽

Specific Substrate ◽

Dependent Manner ◽

Germinal Vesicle Breakdown ◽

S6 Kinase ◽

Substrate Peptide ◽

Ribosomal S6 Kinase

The normal kinetics of ribosomal S6 kinase (RSK) during the meiotic maturation of porcine oocytes were examined. The phosphorylation states of RSK and extracellular signal-regulated kinase (ERK), major mitogen-activated protein (MAP) kinases in maturating porcine oocytes, were detected by Western blotting analysis. The S6 protein kinase activity was assayed using a specific substrate peptide which contained the major phosphorylation sites of S6 kinase. Full phosphorylation of RSK was correlated with ERK phosphorylation and was observed before germinal vesicle breakdown. S6 kinase activity was low in both freshly isolated and 20 h cultured oocytes. S6 kinase activity was significantly elevated in matured oocytes to a level about 6 times higher than that in freshly isolated oocytes. Furthermore, full phosphorylation of RSK was inhibited when oocytes were treated with U0126, a specific MAP kinase kinase inhibitor, in dose-dependent manner, indicating that RSK is one of the substrates of MAP kinase. These results suggest that the activation of RSK is involved in the regulation of meiotic maturation of porcine oocytes.

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Rapid stimulation of mitogen-activated protein kinase of rat liver by prolactin

Biochemical Journal ◽

10.1042/bj3030429 ◽

1994 ◽

Vol 303 (2) ◽

pp. 429-433 ◽

Cited By ~ 37

Author(s):

R Piccoletti ◽

P Maroni ◽

P Bendinelli ◽

A Bernelli-Zazzera

Keyword(s):

Protein Kinase ◽

Map Kinase ◽

Tyrosine Kinases ◽

Kinase Activity ◽

Mitogen Activated Protein Kinase ◽

Female Rats ◽

Protein Kinase Activity ◽

Kinase Activation ◽

Stimulation Of

Intraperitoneal prolactin administration to female rats caused a rapid and transient stimulation of hepatic mitogen-activated kinase (MAP kinase) activity measured in vitro as cytosolic phosphotransferase capacity towards two specific substrates. Myelin basic protein kinase activity of MAP kinase immunoprecipitates confirmed the specificity and magnified the prolactin effect. Immunoblot experiments with anti-(MAP kinase) and anti-phosphotyrosine antibodies showed changes in both electrophoretic mobility and phosphotyrosine content of 40 and 44 kDa isoenzymes suggesting that prolactin affects these isoforms. Concomitant with the increase in MAP kinase activity, prolactin induced tyrosine phosphorylation in a number of liver proteins, suggesting a rapid involvement of tyrosine kinases which might be correlated in some way with MAP kinase activation. Protein kinase C activity, which has been implicated in the regulation of MAP kinase and in mediating the prolactin effect, does not seem to participate in MAP kinase activation.

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Myelin basic protein kinase activity in tomato leaves is induced systemically by wounding and increases in response to systemin and oligosaccharide elicitors

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.94.20.11085 ◽

1997 ◽

Vol 94 (20) ◽

pp. 11085-11089 ◽

Cited By ~ 132

Author(s):

J. W. Stratmann ◽

C. A. Ryan

Keyword(s):

Protein Kinase ◽

Myelin Basic Protein ◽

Kinase Activity ◽

Basic Protein ◽

Protein Kinase Activity ◽

Oligosaccharide Elicitors

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Tyrosine phosphorylation and activation of homologous protein kinases during oocyte maturation and mitogenic activation of fibroblasts

Molecular and Cellular Biology ◽

10.1128/mcb.11.5.2517-2528.1991 ◽

1991 ◽

Vol 11 (5) ◽

pp. 2517-2528 ◽

Cited By ~ 1

Author(s):

J Posada ◽

J Sanghera ◽

S Pelech ◽

R Aebersold ◽

J A Cooper

Keyword(s):

Cell Cycle ◽

Map Kinase ◽

Tyrosine Phosphorylation ◽

Oocyte Maturation ◽

Mammalian Cells ◽

Map Kinases ◽

Kinase Activity ◽

Mitotic Cell ◽

Meiotic Maturation ◽

Sea Star

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Evidence that protein kinase C and mitogen activated protein kinase are not involved in the mechanism by which insulin stimulates translation in L6 myoblasts

Bioscience Reports ◽

10.1007/bf01200213 ◽

1995 ◽

Vol 15 (1) ◽

pp. 37-46 ◽

Cited By ~ 9

Author(s):

Michael G. Thompson ◽

Monique Pascal ◽

Steven C. Mackie ◽

Amanda Thom ◽

Kenneth S. Morrison ◽

...

Keyword(s):

Protein Synthesis ◽

Protein Kinase C ◽

Protein Kinase ◽

Myelin Basic Protein ◽

Map Kinase ◽

Basic Protein ◽

Protein Kinase Activity ◽

S6 Kinase ◽

Kinase C ◽

Mitogen Activated Protein

Insulin stimulated a concentration-dependent increase in protein synthesis in L6 myoblasts which was significant at 1 nM. This response was not prevented by the transcription inhibitor, actinomycin D. The protein kinase C (PKC) inhibitor, Ro-31-8220, and downregulation of PKC by prolonged incubation of cells with 12-O-tetradecanoylphorbol-13-acetate (TPA), had no effect on the ability of insulin to stimulate protein synthesis whilst completely blocking the response to TPA. In contrast, insulin failed to enhance protein synthesis significantly in the presence of either ibuprofen, a selective cyclooxygenase inhibitor or rapamycin, an inhibitor of the 70 kDa S6 kinase. When cell extracts were prepared and assayed for total myelin basic protein kinase activity, a stimulatory effect of insulin was not observed until the concentration approached 100-fold (i.e. 100 nM) that required to elicit increases in protein synthesis. Upon fractionation on a Mono-Q column, 100 nM insulin increased the activity of 3 peaks which phosphorylated myelin basic protein. Two of these peaks were identified as the 42 and 44 kDa forms of Mitogen Activated Protein (MAP) kinase by immunoblotting. In contrast, 1 nM insulin had no effect on the activity of these peaks. The data suggest that physiologically relevant concentrations of insulin do not stimulate translation in L6 cells through either PKC or the 42/44 kDa isoforms of MAP kinase and that this response is, at least in part, mediated through the activation of the 70 kDa S6 kinase by cyclooxygenase metabolites.

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