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Changing Proteins in Granulosa Cells during Follicular Development in Pig  

Chae, In-Soon (College of Animal Life Sciences, Kangwon National University)
Jang, Dong-Min (CHA Stem Cell Institute, CHA Hospital)
Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University)
Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University)
Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.33, no.3, 2009 , pp. 183-187 More about this Journal
Abstract
This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is $2{\sim}4\;mm$ and $6{\sim}10\;mm$, were collected from ovary of slaughtered pig that each follicle of diameter $1{\sim}4\;mm$ and $6{\sim}10\;mm$. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $200{\mu}l$. Immobilized pH gradient(IPG) strip used 18 cm, $3{\sim}10\;NL$. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of $6{\sim}10\;mm$ follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.
Keywords
Two-dimensional electrophoresis; MALDI-TOF; Granulosa cell; Protein profiling; Pig follicle;
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