貉早期胚胎发育内环境的研究

Study on Inner Environment of Early Embryos Development in Racoon Dog

为研究貉早期胚胎发育体内微环境变化,试验选用年龄相同、体重相似的成年健康母貉23只,在繁殖季节自然交配,并受配1～2次,以第1次交配为零点开始计时,分别于初配后29～99h(n=7)、100～126h(n=7)、169～268h(n=9)随机处死貉,用免疫组化、透射及扫描电镜的方法研究貉早期胚胎发育过程中输卵管和子宫的形态学和超微结构变化;用X射线能谱技术对貉输卵管液和子宫液中的钾、钙、铁、锌等9种元素进行测定。结果显示:①随着貉早期胚胎的发育,其后期输卵管长度、黏膜厚度、皱襞高度和上皮高度均显著减小(P<0.05),输卵管直径有所降低,但差异不显著(P>0.05);子宫黏膜厚度、皱襞高度及子宫腺密度均显著增加(P<0.05),子宫的直径和长度有所增加,但差异不显著(P>0.05)。②随着貉早期胚胎的发育,貉子宫黏膜上皮微绒毛、脂滴和溶酶体含量增多。③随着貉早期胚胎的发育,其后期输卵管液中硫、钙、铁、铜和锌元素含量均显著升高(P<0.05),磷含量显著降低(P<0.05),而钾、氯和钠含量呈波动性变化;子宫液中硫、氯、钾的相对含量逐渐降低(P>0.05),锌的相对含量显著降低(P<0.05),磷、钙、铁和铜的含量呈波动性变化。本试验初步揭示了貉早期胚胎发育内环境的变化,为貉胚胎体外培养体系的建立提供参考。

To investigate the micro-environment in vivo of preimplantation embryos in the raccoon dog,23 adult healthy females with the same age and similar body weight were selected.After 1-2 times of copulation during breeding season,starting with the first mating(0 h),the females were killed randomly at 29-99 h(n=7),100-126 h(n=7),169-268 h(n=9),respectively.The morphological and ultrastructural changes of the oviduct and uterus during early embryonic development were analyzed using immunohistochemistry,transmission and scanning electron microscopy.9 elements such as potassium,calcium,iron,zinc of sputum oviduct fluid and uterus fluid during early embryo development were determined by X-ray energy spectroscopy.The results showed that:(1)With the development of early embryos of raccoon dog,at the later stage,the length,mucosal thickness,duplicatures’height and epithelial height of oviduct were significantly reduced(P<0.05);Although the diameter of oviduct was also decreased,but there was no significant difference(P >0.05).On the other hand,the mucosal thickness,duplicatures’height and density of uterine gland were significantly increased(P<0.05);Although the diameter and length of uterus were enhanced,but the difference was not significant(P >0.05).(2)With the development of early embryos of raccoon dog,the numbers of microvilli,lipid droplets and lysosomes in the uterine mucosa epithelium were magically increased.(3)With the development of early embryos of raccoon dog,at the later stage,the contents of S,Ca,Fe,Cu and Zn in the oviduct fluids were significantly improved(P<0.05),while the content of P in the oviduct fluids was significantly decreased(P<0.05),the contents of K,Cl and Na in the oviduct fluids showed the fluctuation change with the development of embryos.The contents of S,Cl,K in the uterus fluids were decreased(P >0.05),the content of Zn in the uterus fluids was significantly decreased(P<0.05),the contents of P,Ca,Fe and Cu presented the fluctuation change with the development of embryos.This study preliminarily investigated the physical and chemical micro-environment in vivo of preimplantation embryos in raccoon dogs,which would provide reference for the establishment of culture system of preimplantation embryos in vitro in raccoon dogs.