Increases in phosphorylation of SAPK/JNK and p38MAPK correlate negatively with mouse embryo development after culture in different media.
OBJECTIVE
To test whether signal transduction proteins that mediate stress may be used to detect responses of embryos to different media in a prospective randomized study.


METHODS
Controlled laboratory study.


METHODS
None.


METHODS
None.


METHODS
Mouse embryos isolated at E3.5 (3.5 days after fertilization) or E1.5 were cultured in different media for 24 hours or 72 hours, respectively. Expression of p38 mitogen activated protein kinases (MAPKs) and stress-activated protein kinase/Jun kinase (SAPK/JNK) phosphoproteins in the mouse embryo and their correlation with preimplantation development were studied.


METHODS
[1] In E3.5 embryos, SAPK/JNK and p38MAPK are phosphorylated at different levels in different media after 24 hours, with Ham's F10+BSA and M-16 having the highest intensity of both SAPK/JNK and p38MAPK phosphorylation and Quinn's cleavage medium and potassium simplex optimized medium supplemented with amino acids (KSOM+AA) the lowest intensity. [2] The stress-induced increase in phosphorylation of SAPK/JNK and p38MAPK appears to be post-translational in embryos. [3] The intensity of SAPK/JNK phosphorylation measured at E1.5+72 hours culture is inversely correlated with 4-cell/compaction rate, morula formation rate, blastocyst formation rate, and hatching rate.


RESULTS
SAPK/JNK and p38MAPK phosphoprotein levels, but not all forms of protein, are affected during culture of preimplantation embryos in seven different media. During culture, the rate of progress to four developmental events was assayed and each rate was inversely proportional to the level of SAPK/JNK phosphorylation measured by immunocytochemical means or Western blot analysis at the end of culture.


CONCLUSIONS
Culture stresses embryos; different media exert different levels of stress on the embryos. There is a negative correlation between the amount of stress and the development rate. Taken together, the data suggest that SAPK/JNK phosphorylation may constitute a measure of homeostatic response to negative stimuli of media.
Wang
Yingchun
Y
C.S. Mott Center for Human Growth and Development, Department of Obstetrics and Gynecology, Hutzel Hospital, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI 48201, USA.
Puscheck
Elizabeth E
EE
Lewis
Jennifer J
JJ
Trostinskaia
Anna B
AB
Wang
Fangfei
F
Rappolee
Daniel A
DA
D000818:Animals; D015153:Blotting, Western; D002021:Buffers; D003470:Culture Media / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D007552:Isotonic Solutions / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D011247:Pregnancy; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D002021:Buffers; D003470:Culture Media; D007552:Isotonic Solutions; C083923:nutrient mixture F-10 Ham; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases