|Kayser, Jean Patrick|
|Cerny, Ronald - UNIV NEBRASKA, LINCOLN|
Submitted to: Biology of Reproduction Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: May 1, 2004
Publication Date: August 1, 2004
Citation: Kayser, J.R., Kim, J.G., Cerny, R.L., Vallet, J.L. 2004. Characterization of the intrauterine proteome in pigs [abstract]. Biology of Reproduction. 70(Supplement):93. Technical Abstract: The proteins present within the intrauterine environment change dramatically at the time of maternal recognition of pregnancy in gilts (d 10-13). The objective of this experiment was to identify these proteins. Sixteen mature gilts were either mated (day 0) or remained cyclic and were slaughtered at either d 10 or 13 (n=4 per status by day). At slaughter, each uterine horn was flushed with 20 ml of medium. Flushings were dialysed extensively against distilled water. A .5 ml aliquot of each was lyophilized, and the lyophilized sample was subjected to 2 dimensional PAGE, followed by coomassie staining. An image of each gel was used to match proteins between treatments and to obtain the relative concentration of each protein (2D advanced software). After statistical analysis, spots that differed due to either the effect of day, status or their interaction were excised and digested in-gel with trypsin. Resulting peptides were analyzed using tandem mass spectrometry (MS/MS) using a QTOF Ultima mass spectrometer. The MS/MS data for each spot was used to identify the corresponding protein. There were 280 matching spots, 131 were significantly (p<.05 or .01) affected by either pregnancy status or day. Most (71%) increased on day 13 with no effect of pregnancy. Several of these were identified as proteases (matrix metalloproteinase, cathepsin-L) or their inhibitors (tissue inhibitor of metalloproteinase-2, serpin, alpha-2 macroglobulin). Others potentially modify glycolipids (acid ceramidase, saposin) and/or glycoproteins (fucosidase, N-acetylglucosamine-6-sulfatase). These results suggest that the concentrations of many proteins within the intrauterine environment are independent of the conceptus. Results also suggest that many could play roles in either elongation or superficial attachment of the conceptus to the endometrium through regulation of the endometrial or conceptus glycocalyx.