Shim Yoo Jin, Hong Yeon Hee, Lee Jaewang, Jee Byung Chul
Department of Obstetrics and Gynecology, Dongguk University Ilsan Medical Center, Ilsan, Korea.
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea.
Clin Exp Reprod Med. 2021 Dec;48(4):347-351. doi: 10.5653/cerm.2021.04735. Epub 2021 Nov 23.
We investigated the impact of vitamin D3 (VD3) supplementation during mouse preantral follicle culture in vitro and the mRNA expression of 25-hydroxylase (CYP2R1), 1-alpha-hydroxylase (CYP27B1), and vitamin D receptor (VDR) in mouse ovarian follicles at different stages.
Preantral follicles were retrieved from 39 BDF1 mice (7-8 weeks old) and then cultured in vitro for 12 days under VD3 supplementation (0, 25, and 50 pg/mL). Follicular development and the final oocyte acquisition were assessed. Preantral follicles were retrieved from 15 other BDF1 mice (7-8 weeks old) and cultured without VD3 supplementation. Three stages of mouse ovarian follicles were obtained (preantral, antral, and ruptured follicles). Total RNA was extracted from the pooled cells (from 20 follicles at each stage), and then reverse transcriptase-polymerase chain reaction was performed to identify mRNA for CYP2R1, CYP27B1, and VDR.
The survival of preantral follicles, rates of antrum formation and ruptured follicles (per initiated follicle) and the number of total or mature oocytes were all comparable among the three groups. Both CYP2R1 and CYP27B1 were expressed in antral and ruptured follicles, but not in preantral follicles. VDR was expressed in all three follicular stages.
VD3 supplementation in vitro (25 or 50 pg/mL) did not enhance mouse follicular development or final oocyte acquisition. Follicular stage-specific expression of CYP2R1, CYP27B1, and VDR was observed.
我们研究了在小鼠体外窦前卵泡培养过程中补充维生素D3（VD3）的影响，以及不同阶段小鼠卵巢卵泡中25-羟化酶（CYP2R1）、1-α-羟化酶（CYP27B1）和维生素D受体（VDR）的mRNA表达。
从39只BDF1小鼠（7-8周龄）中获取窦前卵泡，然后在补充VD3（0、25和50 pg/mL）的条件下进行体外培养12天。评估卵泡发育和最终获得的卵母细胞情况。从另外15只BDF1小鼠（7-8周龄）中获取窦前卵泡，在不补充VD3的情况下进行培养。获得小鼠卵巢卵泡的三个阶段（窦前、窦状和破裂卵泡）。从汇集的细胞（每个阶段20个卵泡）中提取总RNA，然后进行逆转录聚合酶链反应以鉴定CYP2R1、CYP27B1和VDR的mRNA。
三组之间窦前卵泡的存活率、窦腔形成率和破裂卵泡率（每个起始卵泡）以及总卵母细胞或成熟卵母细胞的数量均相当。CYP2R1和CYP27B1在窦状和破裂卵泡中表达，但在窦前卵泡中不表达。VDR在所有三个卵泡阶段均有表达。
体外补充VD3（25或50 pg/mL）并未增强小鼠卵泡发育或最终卵母细胞的获得。观察到CYP2R1、CYP27B1和VDR在卵泡阶段特异性表达。
