Laboratory of Theriogenology, Department of Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Present: Laboratory of Reproductive Physiology, Faculty of Environmental, Life, Natural Science and Technology, Okayama University, Okayama 700-8530, Japan.
J Reprod Dev. 2024 Jun 1;70(3):184-191. doi: 10.1262/jrd.2023-096. Epub 2024 Apr 18.
Heat stress reduces the developmental competence of bovine oocytes during the growth phase; however, the detailed mechanisms remain unclear. Amino acids play various critical roles in follicular development, including protein synthesis and as energy sources. We performed in vitro growth (IVG) culture of oocyte-cumulus-granulosa complexes (OCGCs) to assess the amino acid metabolism of small follicles at high temperatures. We isolated OCGCs from early antral follicles (0.5-1.0 mm) and subjected them to IVG culture for 12 days. OCGCs in the heat shock group were cultured under a temperature cycle of (38.5°C: 5 h, 39.5°C: 5 h, 40.5°C: 5 h, and 39.5°C: 9 h) to reproduce the body temperature of lactating cows under a hot environment. OCGCs in the control group were cultured at a constant temperature of 38.5°C for 24 h. Of the surviving OCGCs, those showing similar morphology and size between the groups were selected for amino acid analysis. We analyzed the free amino acids and their metabolites in the culture medium and calculated the depletion or appearance of molecular species. The depletion of three essential amino acids (isoleucine, leucine, and valine), two non-essential amino acids (aspartic acid and glycine), and ornithine was higher in the heat shock group (P < 0.05). Alanine depletion was lower in the heat shock group (P < 0.05). We concluded that heat exposure alters the amino acid metabolism of OCGCs isolated from early antral follicles, which might be involved with the diminished developmental potential of oocytes during summer.
热应激会降低牛卵母细胞在生长阶段的发育能力;然而,其详细机制尚不清楚。氨基酸在卵泡发育中发挥着各种关键作用,包括蛋白质合成和作为能量来源。我们进行了卵母细胞-颗粒细胞复合体(OCGCs)的体外生长(IVG)培养,以评估高温下小卵泡的氨基酸代谢。我们从早期腔前卵泡(0.5-1.0mm)中分离出 OCGCs,并将其进行 IVG 培养 12 天。热休克组的 OCGCs 在温度循环(38.5°C:5h,39.5°C:5h,40.5°C:5h 和 39.5°C:9h)下进行培养,以模拟热环境下泌乳奶牛的体温。对照组的 OCGCs 在 38.5°C 的恒定温度下培养 24h。在存活的 OCGCs 中,选择那些在组间具有相似形态和大小的 OCGCs 进行氨基酸分析。我们分析了培养基中的游离氨基酸及其代谢物,并计算了分子物种的消耗或出现。热休克组中三种必需氨基酸(异亮氨酸、亮氨酸和缬氨酸)、两种非必需氨基酸(天冬氨酸和甘氨酸)和鸟氨酸的消耗更高(P<0.05)。热休克组中丙氨酸的消耗较低(P<0.05)。我们得出结论,热暴露改变了从早期腔前卵泡中分离出的 OCGCs 的氨基酸代谢,这可能与夏季卵母细胞发育潜力下降有关。