Division of Reproductive Biology, Department of Reproductive Science, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal, 576104, India.
Manipal Center for Biotherapeutic Research, Manipal Academy of Higher Education, Manipal, 576104, India.
Reprod Sci. 2023 Jul;30(7):2137-2151. doi: 10.1007/s43032-023-01166-3. Epub 2023 Jan 23.
Preimplantation-stage embryos are susceptible to various types of stress when cultured in vitro. Parthenogenetic embryos that lack spermatozoa contribution exhibit aberrant developmental dynamics due to their uniparental origin. Herein, we assessed whether the absence of paternal genome affects the susceptibility of the embryos to pH, osmotic and oxidative stress. Haploid parthenogenetic embryos (HPE) (activated oocytes with 1 pronucleus and 2 polar bodies) were generated by incubating cumulus oocyte complexes of Swiss albino mice with 10 mM strontium chloride for 3 h. Normally fertilized embryos (NFE) (fertilized oocytes with 2 pronuclei and 2 polar bodies) were derived using in vitro fertilization. At 2-cell stage, both HPE and NFE were exposed to various stressors including pH (6.8 to 8.2), osmotic (isotonic, hypotonic, and hypertonic), and peroxidatic oxidative (HO, 25 µM) stress. Endoplasmic reticulum stress response, mitochondrial membrane potential, and the rate of blastocyst development were assessed. HPE were susceptible to alteration in the pH that was well tolerated by NFE. Similarly, HPE displayed remarkable difference in sensitivity to hypertonic stress and oxidative stress compared to NFE. The results clearly indicate that the oocytes that develop into embryos in the absence of paternal contribution are more vulnerable to environmental stressors, further highlighting the importance of spermatozoa contribution and/or the ploidy status in mitigating these stressors and towards healthy early embryo development.
体外培养的胚胎在植入前阶段易受到各种类型的压力。由于单倍体起源,缺乏精子贡献的孤雌胚胎表现出异常的发育动力学。在此,我们评估了父本基因组的缺失是否会影响胚胎对 pH 值、渗透压和氧化应激的敏感性。通过用 10mM 氯化锶孵育瑞士白化病小鼠的卵丘卵母细胞复合物 3 小时,产生单倍体孤雌胚胎(HPE)(具有 1 个原核和 2 个极体的激活卵母细胞)。正常受精胚胎(NFE)(具有 2 个原核和 2 个极体的受精卵)通过体外受精获得。在 2 细胞阶段,将 HPE 和 NFE 暴露于各种应激源,包括 pH(6.8 至 8.2)、渗透压(等渗、低渗和高渗)和过氧化物氧化应激(HO,25µM)。评估内质网应激反应、线粒体膜电位和囊胚发育率。HPE 容易受到 pH 值变化的影响,而 NFE 则能很好地耐受 pH 值变化。同样,与 NFE 相比,HPE 对高渗应激和氧化应激的敏感性存在显著差异。结果清楚地表明,在没有父本贡献的情况下发育成胚胎的卵母细胞对环境应激源更敏感,进一步强调了精子贡献和/或倍性状态在减轻这些应激源和促进健康早期胚胎发育方面的重要性。