In porcine production, porcine litter size is a quantitative trait and its heritability is especially low. So it is necessary to identify porcine reproductive gene and protein. The establishment of pregnancy requires performance of a receptive endometrium and ovary. The endometrium and ovary go through transformations in response to physiological changes initiated by local factors including ovarian hormones and uterine environment that make it for possible pregnancy. The endometrium and ovary secrete a wide array of growth factors, cytokines and proteins. Based on these background, we analyzed the endometrial tissue protein of porcine and would find out biomarker proteins related to porcine litter size.
We sorted the two groups according to litter size of porcine: a small litter size group (SLSG) (n=2) and a large litter size group (LLSG) (n=2). The porcine endometrial tissue and ovary samples were preprocessed for proteomic analysis. In order to comparison, samples of each 2mg endometrium protein and ovary protein were separated form pI and molecular weight in the same conditions by applying a pH 3.0-10.0 IPG gels for the first dimension and then 8-16% SDS-PAGE gel for the second dimension. After proteins were visualized by staining with Commassie brilliant blue (CBB), image analysis was performed with Image Master detect variations in protein spots between large litter size group and small litter size group endometrium. And then differential proteins were identified using MALDI-TOF analysis.
The master images of 2-DE gel images obtained from 2mg samples of large litter size group and small litter size group endometrial proteins at pH 3.0-10.0 revealed more than 400 protein spots in pH 3.0-10.0 range. When we analyzed the levels of expression of proteins that protein spots appeared more than 1.5-fold difference in endometrial tissue from porcine.
In comparison of SLSG(small litter size group) with LLSG(large litter size group), a total of 18 protein spots differentially expressed on porcine endometrial tissue 2-DE gels, among which 9 spots were up-regulated proteins as retinol dehydrogenase 16-like isoform 1, Acrosin-binding protein, alpha-N-acetylgalactosaminidase. phosphoglycerate kinase 2, Acrosin-binding protein in LLSG. And 8 spots were up-regulated proteins as phosphoglycerate kinase 2, prenylcysteine oxidase in SLSG.
포유류 난포의 폐쇄 과정은 매우 정교한 내분비적 조절작용에 의해 일어나며, 이 과정중에 발생하는 난포 내 과립세포의 퇴화는 핵응축 현상을 동반하는 것으로 알려져 있다. 본 연구는 핵응축 현상과 관련하여 돼지 난소 내 폐쇄난포의 과립세포가 퇴화 시 동반되는 세포 사멸이 아포토시스의 과정에 의해서 일어나는지의 여부와 아포토시스 관련 주요 단백질 분해 효소인 캐스파제-3과 연관된 세포 사멸 기전과 관련이 있는지에 대해서 조사하고자 하였다. 돼지 난소로부터 정