Gene expression patterns in granulosa cells and oocytes at various stages of follicle development as well as in in vitro grown oocyte-and-granulosa cell complexes.
Follicle development is accompanied by proliferation of granulosa cells and increasing oocyte size. To obtain high-quality oocytes in vitro, it is important to understand the processes that occur in oocytes and granulosa cells during follicle development and the differences between in vivo and in vitro follicle development. In the present study, oocytes and granulosa cells were collected from early antral follicles (EAFs, 0.5-0.7 mm in diameter), small antral follicles (SAFs, 1-3 mm in diameter), large antral follicles (LAFs, 3-7 mm in diameter), and in vitro grown oocyte-and-granulosa cell complexes (OGCs), which were cultured for 14 days after collection from EAFs. Gene expression was analyzed comprehensively using the next-generation sequencing technology. We found top upstream regulators during the in vivo follicle development and compared them with those in in vitro developed OGCs. The comparison revealed that HIF1 is among the top regulators during both in vivo and in vitro development of OGCs. In addition, we found that HIF1-mediated upregulation of glycolysis in granulosa cells is important for the growth of OGCs, but the cellular metabolism differs between in vitro and in vivo grown OGCs. Furthermore, on the basis of comparison of upstream regulators between in vivo and in vitro development of OGCs, we believe that low expression levels of FLT1 (VEGFA receptor), SPP1, and PCSK6 can be considered causal factors of the suboptimal development under in vitro culture conditions.
Department of Animal Sciences, Tokyo University of Agriculture, Kanagawa 243-0034, Japan.
Yasuhisa
Y
Munakata
Ryoka
R
Kawahara-Miki
Shogo
S
Shiratsuki
Hidetaka
H
Tasaki
Nobuhiko
N
Itami
Koumei
K
Shirasuna
Takehito
T
Kuwayama
Hisataka
H
Iwata
D000818:Animals; D018929:Cell Culture Techniques; D005260:Female; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D006107:Granulosa Cells / Q000378:metabolism*; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism*; D006080:Ovarian Follicle / Q000254:growth & development / Q000378:metabolism*; D013552:Swine