Weakley B S, Bowker S J, James J L
Cell Tissue Res. 1984;235(2):379-86. doi: 10.1007/BF00217863.
Ovaries of immature and adult hamsters were incubated in medium containing thiamine pyrophosphate (TPP) to determine the age at which TPPase-reactive cytoplasmic structures first appear in the germ cells, and at what age the structures cease to be present. The structures were found only in oocytes from animals 8-15 days of age. They occur in predictyate germ cells in polyovular follicles and in very early dictyate oocytes in unilaminar follicles. The TPPase-reactive structures were never observed in atretic oocytes, in unilaminar follicles of adult animals, nor in multilaminar follicles of animals at any age. Ovaries of 8-12-day-old animals and adults were then incubated in media in which one of the following substrates was substituted for TPP: uridine diphosphate (UDP), inosine diphosphate (IDP), and adenosine monophosphate (AMP). Half of the samples in each experiment were incubated in medium containing the inhibitor L-p-bromotetramisole. beta-Glycerophosphate was used in control incubations, or the substrate was omitted entirely. The cytoplasmic structures were found to be reactive after incubation in UDP-containing media, but not after incubation in media containing AMP. With IDP as substrate, reactions were atypical and confined to peripheral regions of the cytoplasm. Other sites of enzyme activity after incubation with the various substrates (cell membranes, zona pellucida, endoplasmic reticulum and Golgi apparatus) are also described and discussed.
将未成熟和成年仓鼠的卵巢置于含有硫胺素焦磷酸(TPP)的培养基中进行培养,以确定TPP酶反应性细胞质结构首次出现在生殖细胞中的年龄,以及该结构何时不再存在。这些结构仅在8至15日龄动物的卵母细胞中发现。它们出现在多卵卵泡中的前定形生殖细胞和单层卵泡中非常早期的双线期卵母细胞中。在闭锁卵母细胞、成年动物的单层卵泡或任何年龄动物的多层卵泡中均未观察到TPP酶反应性结构。然后将8至12日龄动物和成年动物的卵巢置于以下底物之一替代TPP的培养基中进行培养:尿苷二磷酸(UDP)、肌苷二磷酸(IDP)和腺苷一磷酸(AMP)。每个实验中的一半样本在含有抑制剂L-对溴四咪唑的培养基中培养。对照培养中使用β-甘油磷酸,或完全省略底物。发现细胞质结构在含有UDP的培养基中培养后具有反应性,但在含有AMP的培养基中培养后无反应性。以IDP为底物时,反应不典型且局限于细胞质的周边区域。还描述并讨论了与各种底物孵育后酶活性的其他位点(细胞膜、透明带、内质网和高尔基体)。