Hypoxia-mediated carbohydrate metabolism and transport promote early-stage murine follicle growth and survival

Yogeshwar Makanji, David J Tagler, Jennifer E Pahnke, Lonnie D Shea, Teresa K Woodruff

Research output: Contribution to journalArticleOther

28 Citations (Scopus)


Oxygen tension is critical for follicle growth and metabolism, especially for early-stage follicles, where vascularity is limited. Its role and underlying mechanism in the in vitro activation and maturation of immature to ovulatory follicles is largely unknown. In this study, early secondary (110 mum) murine follicles were isolated and encapsulated in alginate hydrogels to replicate the in vivo environment of the growing/maturing follicle. Encapsulated follicles were cultured for 8 days at either 2.5 or 20 O2. Survival (2.6-fold) and growth (1.2-fold) were significantly higher for follicles cultured at 2.5 compared with 20 O2. Using a mouse hypoxia-signaling pathway qRT-PCR array and GeneGo Metacore analysis, we found that direct target genes of the hypoxia-activated HIF1-complex were significantly upregulated in follicles cultured for 8 days at 2.5 compared with 20 O2, including the carbohydrate transport and metabolism genes Slc2a3, Vegfa, Slc2a1, Edn1, Pgk1, Ldha, and Hmox1. Other upregulated genes included carbohydrate transporters (Slc2a1, Slc2a3, and Slc16a3) and enzymes essential for glycolysis (Pgk1, Hmox1, Hk2, Gpi1, Pfkl, Pfkp, Aldoa, Gapdh, Pgam1, Eno1, Pkm2, and Ldha). For follicles cultured at 2.5 O2, a 7.2-fold upregulation of Vegfa correlated to an 18-fold increase in VEGFA levels, and a 3.2-fold upregulation of Ldha correlated to a 4.8-fold increase in lactate levels. Both VEGFA and lactate levels were significantly higher in follicles cultured at 2.5 compared with 20 O2. Therefore, enhanced hypoxia-mediated glycolysis is essential for growth and survival of early secondary follicles and provides vital insights into improving in vitro culture conditions.
Original languageEnglish
Pages (from-to)893 - 903
Number of pages11
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Issue number8
Publication statusPublished - 2014
Externally publishedYes

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