Department of Physiology, Facultad de Veterinaria, Campus Internacional de Excelencia de Educación Superior (Campus Mare Nostrum). Universidad de Murcia, Murcia, Spain.
Instituto Murciano de Investigación Biosanitaria Virgen de la Arrixaca (IMIB-Arrixaca), Murcia, Spain.
Mol Hum Reprod. 2020 Oct 1;26(10):748-759. doi: 10.1093/molehr/gaaa053.
This work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21-168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.
本研究旨在确定母猪生殖道内的温度条件,并研究其对辅助生殖技术(ARTs)的影响。使用腹腔镜单部位手术辅助方法和微型探头记录温度。分别使用母猪和小母猪来解决自然周期和卵巢刺激治疗的问题。根据体内值,在三种温度条件(37.0°C、38.5°C 和 39.5°C)下进行体外受精(IVF),并将假定的受精卵在这些条件下培养 20 小时,而进一步的胚胎培养(EC)(受精后 21-168 小时)则保持在 38.5°C。20 小时后,评估了不同的生育参数。在 EC 期间,评估了卵裂和囊胚阶段。使用差示扫描量热法和荧光恢复后光漂白技术研究了实验温度下精子膜的流动性。发现母猪输卵管和子宫之间存在 1.5°C 的温度梯度(P<0.05),当将该梯度转移到猪体外培养时,IVF 后的多核受精卵数量减少,囊胚百分比增加。此外,公猪精子膜的温度过渡阶段(37.0°C)与母猪输卵管中记录的温度相吻合,与在较高温度(38.5°C 和 39.5°C)下孵育的精子相比,37.0°C 下精子膜更具流动性。这些数据表明,生理温度梯度可能对人类胚胎发育有影响。