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仓鼠卵母细胞成熟过程中凝集素与透明带结合亲和力的变化

Appearance of lectin binding affinity to the zona pellucida during hamster oocyte maturation.

作者信息

Kimura J, Sato K, Tsukise A, Okano M

机构信息

Department of Veterinary Anatomy, College of Agriculture and Veterinary Medicine, Nihon University, Kanagawa, Japan.

出版信息

Cell Mol Biol. 1991;37(6):623-9.

PMID:1747897
Abstract

In our previous study, a drastic change in terminal saccharides of glycoconjugates of the hamster zona pellucida associated with oocyte maturation was observed using light microscopic methods of lectin cytochemistry. To understand the mechanism of this change, in the present study, the correlation between the cytochemical appearance of saccharide residues in the zona pellucida and nuclear maturation was examined. Immature hamsters were treated with PMSG and hCG to induce follicular development and ovulation. The animals were euthanized 0 to 26 hrs. after the injection of PMSG or 0,1,2,3,4,5,7,9 or 11 hrs. after the injection of hCG, and ovaries were dissected out, fixed, paraffin embedded and sectioned serially. Every other paraffin section was stained with hematoxylin to observe the status of nuclei and to classify follicular growth and only the fully developed preovulatory follicles were examined in experiments. The peroxidase-labelled lectin-diaminobenzidine procedure was applied to sections. The lectins employed were WGA, SBA, MPA, UEA-I, LotusA and AAA. Germinal vesicle breakdown was observed within 3 hrs. after the administration of hCG. A positive reaction of WGA, SBA or MPA for zonae pellucidae in the fully developed preovulatory follicles appeared 1 hr. after hCG injection, and remained so for the next 10 hrs. UEA-I, Lotus A and AAA reactions were negative for all of the zonae pellucidae observed. The data indicate that the synthesis of saccharide residues such as GlcNAc and GalNAc forming zona components in the follicles is not triggered by germinal vesicle breakdown.

摘要

在我们之前的研究中,使用凝集素细胞化学的光学显微镜方法观察到,仓鼠透明带糖缀合物的末端糖类在卵母细胞成熟过程中发生了剧烈变化。为了解这种变化的机制,在本研究中,我们检测了透明带中糖残基的细胞化学表现与核成熟之间的相关性。对未成熟的仓鼠注射孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(hCG)以诱导卵泡发育和排卵。在注射PMSG后的0至26小时或注射hCG后的0、1、2、3、4、5、7、9或11小时对动物实施安乐死,然后取出卵巢,进行固定、石蜡包埋并连续切片。每隔一张石蜡切片用苏木精染色以观察细胞核的状态并对卵泡生长进行分类,实验中仅检查完全发育的排卵前卵泡。将过氧化物酶标记的凝集素-二氨基联苯胺方法应用于切片。所使用的凝集素为小麦胚芽凝集素(WGA)、大豆凝集素(SBA)、麦芽凝集素(MPA)、荆豆凝集素I(UEA-I)、百脉根凝集素(LotusA)和花生凝集素(AAA)。在注射hCG后3小时内观察到生发泡破裂。在完全发育的排卵前卵泡中,WGA、SBA或MPA对透明带的阳性反应在注射hCG后1小时出现,并在接下来的10小时内保持。在所观察的所有透明带中,UEA-I、Lotus A和AAA反应均为阴性。数据表明,卵泡中形成透明带成分的糖残基(如N-乙酰葡糖胺和N-乙酰半乳糖胺)的合成不是由生发泡破裂触发的。

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