Spatial and temporal expression of the myometrial mitogen-activated protein kinases p38 and ERK1/2 in the human uterus during pregnancy and labor.
OBJECTIVE
We have recently identified a novel putative spliced variant of the activating transcription factor 2 (ATF2) in the human myometrium during pregnancy and labor. This protein, termed ATF2-sm like full-length ATF2, acts as a potent transactivator of cyclic adenosine monophosphate response element (CRE)-containing promoter reporter genes. Similarly, employing microarray gene profiling in myometrial cells, we have shown ATF2-sm to affect the expression of several specific myometrial genes associated with regulating uterine activity during pregnancy and labor. At some point after conception this transcription factor becomes spatially expressed within the body of the uterus, with significantly higher levels detected in the upper (corpus) compared to the lower uterine segment. Because ATF2 species are the primary substrate for phosphorylation by the mitogen-activated protein kinases (MAPKs) p38 and ERK1/2, the purpose of the current investigation was to define the expression levels of these kinases in upper and lower segment myometrium during pregnancy and labor to see if they also correlated with expression of ATF2-sm.


METHODS
Paired myometrial samples were collected from the upper (corpus) and lower uterine segments from term nonlaboring and spontaneously laboring women undergoing elective and emergency cesarean deliveries, respectively. Non-pregnant myometrial samples were collected from premenopausal women having hysterectomies for benign gynecologic disorders. The MAPKs p38 and ERK1/2 present in individual myometrial homogenates were resolved using sodium dodecyl sulfate polacrylamide gel electropheresis (SDS-PAGE) with subsequent Western blotting with specific antibodies and scanning densitometry. Expression of the individual MAPKs in myometrial tissues was confirmed in situ using immunohistochemistry.


RESULTS
In non-pregnant tissues, p38 and ERK1/2 expression was uniform throughout the uterus. In term pregnant nonlaboring and spontaneously laboring samples expression of p38 and ERK1 was significantly elevated in the upper uterine segment compared to the lower segment, respectively. In contrast, there was no difference in ERK2 expression.


CONCLUSIONS
The data from this study indicate that both p38 and ERK1 are spatially regulated in different uterine regions during pregnancy/labor and suggest that they may be involved in regulating the activity of ATF2 isoforms and their subsequent effects on myometrial function.
Otun
Harry A
HA
School of Surgical and Reproductive Sciences, University of Newcastle-upon-Tyne, Newcastle-upon-Tyne, United Kingdom. H.A.Otun@Ncl.ac.uk
MacDougall
Malcolm W J
MW
Bailey
Jarrod
J
Europe-Finner
G Nicholas
GN
Robson
Stephen C
SC
D051698:Activating Transcription Factor 2; D000293:Adolescent; D000328:Adult; D015153:Blotting, Western; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D005260:Female; D006801:Humans; D007743:Labor, Obstetric / Q000378:metabolism*; D008875:Middle Aged; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009215:Myometrium / Q000201:enzymology; D011247:Pregnancy / Q000378:metabolism*; D012016:Reference Values; D014157:Transcription Factors / Q000378:metabolism; D014599:Uterus / Q000201:enzymology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
C497105:ATF2 protein, human; D051698:Activating Transcription Factor 2; D017362:Cyclic AMP Response Element-Binding Protein; D014157:Transcription Factors; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases