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Expression and activation of mitogen-activated protein kinases in matured porcine oocytes under thermal stress.

Yen SY, Tseng JK, Chuang SM, Chen SE, Ju JC - J. Reprod. Dev. (2014)

Bottom Line: When MII oocytes were heated, only the phosphorylated p38 levels relative to the total p38 levels decreased (P < 0.01) after HS, but no clear relationship with HS treatments was observed in the ERK, JNK and p90(rsk) expressions of matured oocytes.To confirm p38 activation in matured oocytes, immunocytochemical staining was performed to localize its expression and distribution in the ooplasm, and the results were largely consistent with previous Western blot analyses.These alterations of p38 and activation of MAPKAPK2 may be associated with porcine oocyte viability under HS conditions, and a potential cross-talk between p38 MAPK and other signaling cascades may exist, which warrants additional investigation.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan, ROC.

ABSTRACT
In this study, we determined the expression and activation of p38 MAPK in matured porcine oocytes subjected to heat shock (HS). When MII oocytes were heated, only the phosphorylated p38 levels relative to the total p38 levels decreased (P < 0.01) after HS, but no clear relationship with HS treatments was observed in the ERK, JNK and p90(rsk) expressions of matured oocytes. To confirm p38 activation in matured oocytes, immunocytochemical staining was performed to localize its expression and distribution in the ooplasm, and the results were largely consistent with previous Western blot analyses. Moreover, when matured oocytes were co-cultured with a P38 MAPK inhibitor, SB203580, for 4 h at 41.5 C, the activation of its immediate downstream substrate MAPKAPK-2 was not inhibited within any of the treatment groups. It appears that the MAPKAPK2 levels increased only under prolonged culture (HS4h and C4h) compared with the control group. In conclusion, p38 activity in porcine oocytes was decreased after exposure to HS and prolonged culture. These alterations of p38 and activation of MAPKAPK2 may be associated with porcine oocyte viability under HS conditions, and a potential cross-talk between p38 MAPK and other signaling cascades may exist, which warrants additional investigation.

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Expression of JNK in matured porcine oocytes after in vitro heat shock at 41.5 C for 0, 1, 2 or 4h. (a) A representative immunoblot of JNK (upper panel) and total JNK (lower panel, 54 and 46 kDa,respectively). There were no significant differences among treatment groups in either panel. Data are expressed asfolds of the C0h group. Each lane of the SDS-PAGE gel contained 100 oocytes (four replicates). Bars indicate means ±SEM. M, marker.
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fig_002: Expression of JNK in matured porcine oocytes after in vitro heat shock at 41.5 C for 0, 1, 2 or 4h. (a) A representative immunoblot of JNK (upper panel) and total JNK (lower panel, 54 and 46 kDa,respectively). There were no significant differences among treatment groups in either panel. Data are expressed asfolds of the C0h group. Each lane of the SDS-PAGE gel contained 100 oocytes (four replicates). Bars indicate means ±SEM. M, marker.

Mentions: With various durations of HS treatment, activation of ERK1/2 and its downstream target molecules, p90rsk and JNK, in maturedoocytes was not significantly different among treatment groups (Figs. 1 and 2Fig. 1.


Expression and activation of mitogen-activated protein kinases in matured porcine oocytes under thermal stress.

Yen SY, Tseng JK, Chuang SM, Chen SE, Ju JC - J. Reprod. Dev. (2014)

Expression of JNK in matured porcine oocytes after in vitro heat shock at 41.5 C for 0, 1, 2 or 4h. (a) A representative immunoblot of JNK (upper panel) and total JNK (lower panel, 54 and 46 kDa,respectively). There were no significant differences among treatment groups in either panel. Data are expressed asfolds of the C0h group. Each lane of the SDS-PAGE gel contained 100 oocytes (four replicates). Bars indicate means ±SEM. M, marker.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219997&req=5

fig_002: Expression of JNK in matured porcine oocytes after in vitro heat shock at 41.5 C for 0, 1, 2 or 4h. (a) A representative immunoblot of JNK (upper panel) and total JNK (lower panel, 54 and 46 kDa,respectively). There were no significant differences among treatment groups in either panel. Data are expressed asfolds of the C0h group. Each lane of the SDS-PAGE gel contained 100 oocytes (four replicates). Bars indicate means ±SEM. M, marker.
Mentions: With various durations of HS treatment, activation of ERK1/2 and its downstream target molecules, p90rsk and JNK, in maturedoocytes was not significantly different among treatment groups (Figs. 1 and 2Fig. 1.

Bottom Line: When MII oocytes were heated, only the phosphorylated p38 levels relative to the total p38 levels decreased (P < 0.01) after HS, but no clear relationship with HS treatments was observed in the ERK, JNK and p90(rsk) expressions of matured oocytes.To confirm p38 activation in matured oocytes, immunocytochemical staining was performed to localize its expression and distribution in the ooplasm, and the results were largely consistent with previous Western blot analyses.These alterations of p38 and activation of MAPKAPK2 may be associated with porcine oocyte viability under HS conditions, and a potential cross-talk between p38 MAPK and other signaling cascades may exist, which warrants additional investigation.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan, ROC.

ABSTRACT
In this study, we determined the expression and activation of p38 MAPK in matured porcine oocytes subjected to heat shock (HS). When MII oocytes were heated, only the phosphorylated p38 levels relative to the total p38 levels decreased (P < 0.01) after HS, but no clear relationship with HS treatments was observed in the ERK, JNK and p90(rsk) expressions of matured oocytes. To confirm p38 activation in matured oocytes, immunocytochemical staining was performed to localize its expression and distribution in the ooplasm, and the results were largely consistent with previous Western blot analyses. Moreover, when matured oocytes were co-cultured with a P38 MAPK inhibitor, SB203580, for 4 h at 41.5 C, the activation of its immediate downstream substrate MAPKAPK-2 was not inhibited within any of the treatment groups. It appears that the MAPKAPK2 levels increased only under prolonged culture (HS4h and C4h) compared with the control group. In conclusion, p38 activity in porcine oocytes was decreased after exposure to HS and prolonged culture. These alterations of p38 and activation of MAPKAPK2 may be associated with porcine oocyte viability under HS conditions, and a potential cross-talk between p38 MAPK and other signaling cascades may exist, which warrants additional investigation.

Show MeSH
Related in: MedlinePlus