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- Title
Knockdown of PRKAR2B Results in the Failure of Oocyte Maturation.
- Authors
Yoon, Hyemin; Jang, Hoon; Kim, Eun-Young; Moon, Sohyeon; Lee, Sangho; Cho, Minha; Cho, Hye Jung; Ko, Jung Jae; Chang, Eun Mi; Lee, Kyung-Ah; Choi, Youngsok
- Abstract
<bold><italic>Background/Aims:</italic></bold> Cyclic adenosine monophosphate (cAMP)-dependent type 2 regulatory subunit beta (<italic>Prkar2b</italic>) is a regulatory isoform of cAMP-dependent protein kinase (PKA), which is the primary target for cAMP actions. In oocytes, PKA and the pentose phosphate pathway (PPP) have important roles during the germinal vesicle (GV) stage arrest of development. Although the roles of the PKA signal pathway have been studied in the development of oocyte, there has been no report on the function of PRKAR2B, a key regulator of PKA. <bold><italic>Methods:</italic></bold> Using reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time PCR (qRT-PCR), immunohistochemistry, and immunofluorescence, we determined the relative expression of <italic>Prkar2b</italic> in various tissues, including ovarian follicles, during oocyte maturation. <italic>Prkar2b</italic>-interfering RNA (RNAi) microinjection was conducted to confirm the effect of <italic>Prkar2b</italic> knockdown, and immunofluorescence, qRT-PCR, and time-lapse video microscopy were used to analyze <italic>Prkar2b</italic>-deficient oocytes. <bold><italic>Results:</italic></bold><italic>Prkar2b</italic> is strongly expressed in the ovarian tissues, particularly in the growing follicle. During oocyte maturation, the highest expression of <italic>Prkar2b</italic> was during metaphase I (MI), with a significant decrease at metaphase II (MII). RNAi-mediated <italic>Prkar2b</italic> suppression resulted in MI-stage arrest during oocyte development, and these oocytes exhibited abnormal spindle formation and chromosome aggregation. Expression of other members of the PKA family (except for <italic>Prkaca</italic>) were decreased, and the majority of the PPP factors were also reduced in <italic>Prkar2b</italic>-deficient oocytes. <bold><italic>Conclusion:</italic></bold> These results suggest that <italic>Prkar2b</italic> is closely involved in the maturation of oocytes by controlling spindle formation and PPP-mediated metabolism.
- Subjects
CYCLIC-AMP-dependent protein kinase; OVUM; GERMINAL vesicles; PENTOSE phosphate pathway; RNA interference; DEVELOPMENTAL biology
- Publication
Cellular Physiology & Biochemistry (Karger AG), 2018, Vol 45, Issue 5, p2009
- ISSN
1015-8987
- Publication type
Article
- DOI
10.1159/000487978