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Histochemical identification of primordial germ cells and differentiation of the gonads in homozygous tetraploid mouse embryos.

机译:纯合四倍体小鼠胚胎中原始生殖细胞的组织化学鉴定和性腺分化。

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摘要

This study was undertaken to establish whether primordial germ cells are differentiated by homozygous tetraploid mouse embryos produced by the technique of electrofusion, and to study the morphological features of their gonads. Tetraploid embryos were transferred to the oviducts of pseudopregnant recipients, and these were autopsied either on day 11 or days 15 or 16 of gestation. In the developmentally less advanced group, embryos in which cytogenetic analysis of their extraembryonic membranes confirmed that they had a tetraploid chromosome constitution were analysed histochemically in order to demonstrate the presence of intracellular alkaline phosphatase enzyme activity. This enabled the presence or absence of germ cells to be established. Out of a total of 9 early limb-bud stage embryos studied, all contained primordial germ cells. The latter were mostly located in association with the hindgut, though some germ cells were still present at the base of the allantois. The sex ratio in this group was close to unity. In the 2nd group in which recipients were autopsied on either day 15 or 16 of gestation, a total of 7 healthy tetraploid embryos were recovered. All displayed the characteristic craniofacial features seen in tetraploid embryos. Four of these embryos had a normal postcranial axial morphology, and their crown-rump lengths were only slightly less (81-91%) than those of developmentally matched control diploid embryos. Three of the tetraploid embryos had an abnormal postcranial axis associated with a body wall closure defect involving the anterior abdominal and lower thoracic region. In all 7 of these embryos, gonadal differentiation was consistent with their developmental age.
机译:进行这项研究以确定是否通过电融合技术产生的纯合四倍体小鼠胚胎分化原始生殖细胞,并研究其性腺的形态特征。将四倍体胚胎转移到假妊娠受体的输卵管,并在妊娠第11天或第15天或第16天进行尸检。在发育较不先进的组中,对其组织进行了细胞外遗传学分析,证实了它们具有四倍体染色体组成,对这些胚胎进行了组织化学分析,以证明细胞内碱性磷酸酶活性的存在。这使得能够确定是否存在生殖细胞。在总共研究的9个早期肢芽芽早期胚胎中,所有胚胎都含有原始生殖细胞。后者大多位于后肠附近,尽管尿囊底部仍存在一些生殖细胞。该组中的性别比接近统一。在第二组中,在妊娠的第15天或第16天对接受者进行了尸检,共回收了7个健康的四倍体胚胎。全部显示出在四倍体胚胎中看到的特征性颅面特征。这些胚胎中有四个具有正常的颅后轴形态,其冠臀长度仅比发育匹配的对照二倍体胚胎略短(81-91%)。四倍体胚胎中的三个具有颅后轴异常,与涉及前腹和下胸部区域的体壁闭合缺陷相关。在所有这些胚胎的7个中,性腺分化与它们的发育年龄一致。

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