Kempisty Bartosz, Jackowska Marta, Woźna Magdalena, Antosik Paweł, Piotrowska Hanna, Zawierucha Piotr, Bukowska Dorota, Jaśkowski Jędrzej M, Nowicki Michał, Brüssow Klaus P
Department of Histology and Embryology, Poznan University of Medical Sciences, 6 Święcickiego St., 60-781 Poznań, Poland.
J Biomed Biotechnol. 2012;2012:742829. doi: 10.1155/2012/742829. Epub 2012 Nov 20.
Cumulus-oocyte-complexes (COCs) were collected from small (<3 mm), medium (3-5 mm), and large (>5 mm) porcine follicles, and the INHA and INHB expression and cellular localization were studied. Developmentally competent (BCB+) COCs were cultured for 44 h. Samples of mRNA were isolated before and after in vitro maturation (IVM) from oocytes collected from follicles of different size for RQ-PCR assay. The INHA and INHB protein distribution within the oocytes was observed by confocal microscopy. INHA mRNA expression was increased in oocytes from large compared to medium and small follicles before IVM (P < 0.001), and to oocytes of small follicles after IVM (P < 0.001). The INHB expression was not different before IVM, but the IHNB mRNA level was gradually higher in oocytes from large follicles after IVM (P < 0.01). INHA was not differently expressed before IVM; however, in large follicle oocytes the protein was distributed in the peripheral area of the cytoplasm; in oocytes from small follicles it was in the entire cytoplasm. After IVM, INHA was strongly expressed in oocytes from small follicles and distributed particularly in the zona pellucida (ZP). Similarly and both before and after IVM, INHB protein was highly expressed in small follicle oocytes and within the cytoplasm. In summary, INHs can be recognized as a marker of porcine oocyte quality.
从直径小于3毫米的小卵泡、直径3至5毫米的中卵泡和直径大于5毫米的大卵泡中采集卵丘-卵母细胞复合体（COCs），并研究抑制素α（INHA）和抑制素β（INHB）的表达及细胞定位。对发育能力良好（BCB+）的COCs进行44小时培养。从不同大小卵泡采集的卵母细胞在体外成熟（IVM）前后分离mRNA样本用于相对定量聚合酶链反应（RQ-PCR）检测。通过共聚焦显微镜观察卵母细胞内INHA和INHB蛋白的分布。IVM前，与中、小卵泡的卵母细胞相比，大卵泡卵母细胞中INHA mRNA表达增加（P<0.001），IVM后与小卵泡卵母细胞相比也增加（P<0.001）。IVM前INHB表达无差异，但IVM后大卵泡卵母细胞中INHB mRNA水平逐渐升高（P<0.01）。IVM前INHA表达无差异；然而，在大卵泡卵母细胞中，该蛋白分布于细胞质周边区域；在小卵泡卵母细胞中则分布于整个细胞质。IVM后，INHA在小卵泡卵母细胞中强烈表达，尤其分布在透明带（ZP）中。同样，无论IVM前后，INHB蛋白在小卵泡卵母细胞及细胞质中均高表达。总之，抑制素可被视为猪卵母细胞质量的标志物。
