Identification of metaphase II-specific gene transcripts in porcine oocytes and their expression in early stage embryos
Xiang-Shun Cui* A B , Hyuk Song* C and Nam-Hyung Kim A DA Department of Animal Science, Chungbuk National University, Cheongju, Chungbuk, Korea.
B Jilin Agricultural University, Changchun City, Jilin Province, China.
C Department of Animal Science, Kunkuk University, Choongju, Chungbuk, Korea.
D Corresponding author. Email: nhkim@chungbuk.ac.kr
Reproduction, Fertility and Development 17(6) 625-631 https://doi.org/10.1071/RD05019
Submitted: 17 February 2005 Accepted: 26 May 2005 Published: 1 July 2005
Abstract
Annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR) was used to identify the genes that are specifically or prominently expressed in porcine oocytes at the metaphase II (MII) and germinal vesicle (GV) stages. By using 60 ACPs, 13 differentially expressed genes (DEGs) were identified. The cloned genes or expressed sequence tags (ESTs) showed sequence similarity with known genes or ESTs of other species in GenBank. The mRNA expression during oocyte maturation and early embryonic development in both pigs and mice of four of these genes (namely transcription factor TZP, annexin A2, hypoxia-inducible protein 2, and ATPase 6) was further characterised by real-time quantitative reverse transcription–PCR. All four genes were markedly upregulated in pig and mouse MII oocytes compared with GV-stage oocytes. The expression levels of the four genes decreased gradually during early cleavage. Thus, these genes may play important roles during oocyte maturation and/or early cleavage in mammals. Although the detailed functions of these genes remain to be determined, their identification in the present study provides insights into meiotic maturation and fertilisation.
Extra keywords: annealing control primer, differentially expressed genes, maturation.
Acknowledgments
This research was supported by grants from the Agricultural Research R&D Center, by the Research Center for Bioresource and Health (RCBH), the Korea Science and Engineering Foundation and National Research Laboratory Program in Ministry of Science and Technology (to NHK).
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* X.-S. Cui and H. Song contributed equally to this paper.