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免疫着丝粒蛋白 A 和弗氏完全佐剂诱导的抗着丝粒抗体可能会干扰小鼠卵母细胞减数分裂。

Anticentromere antibody induced by immunization with centromere protein a and Freund's complete adjuvant may interfere with mouse oocyte meiosis.

机构信息

Department of Obstetrics and Gynecology, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Reproductive Medicine Center, The Third Affiliated Hospital of Guangzhou Medical University, 63 Duobao Road, Liwan District, Guangzhou, China.

出版信息

Reprod Biol Endocrinol. 2021 Apr 1;19(1):50. doi: 10.1186/s12958-021-00737-w.

Abstract

BACKGROUND

Anticentromere antibody (ACA) is a member of the antinuclear antibody (ANA) family, and recent studies have found that ACA may be associated with oocyte maturation disorders; however, the possible mechanism behind this phenomenon remains unknown. We conducted this study to investigate whether ACA could penetrate into the living oocytes and interfere with oocyte meiosis in a mouse model.

METHODS

We divided mice into three groups: human recombinant centromere protein-A (human CENP-A, HA) and complete Freund's adjuvant (CFA) were used to immunize mice for the study group (HA + CFA), and mice injected with CFA (CFA group) or saline (Saline group), respectively, served as controls. After immunization, serum anti-CENP-A antibody was detected by indirect immunofluorescence assay (IIFT) and enzyme-linked immunosorbent assay (ELISA). Chromosome alignment and intracellular IgG localization in MI- and MII-stage oocytes were investigated by immunofluorescence analysis.

RESULTS

Positive ACAs were successfully induced by immunization with CENP-A and CFA, and results showed that the serum level of anti-CENP-A antibody was significantly higher in the HA + CFA group compared with the control groups. There was marked increase of chromosome misalignments in MI and MII oocytes in the HA + CFA group compared to the control groups. However, no oocytes from any of the three groups showed intracellular antibody immunofluorescence.

CONCLUSIONS

The development and maturation of oocytes were impaired in peripheral ACA positive mice, which exhibited severe chromosomal misalignments in metaphase meiosis; however, no evidence of ACAs entering the oocytes was observed, thus the underlying mechanism needs further exploration.

摘要

背景

抗着丝点抗体(ACA)是抗核抗体(ANA)家族的一员,最近的研究发现 ACA 可能与卵母细胞成熟障碍有关;然而,这种现象背后的可能机制尚不清楚。我们进行这项研究是为了探讨 ACA 是否可以穿透活卵母细胞并在小鼠模型中干扰卵母细胞减数分裂。

方法

我们将小鼠分为三组:用人重组着丝粒蛋白-A(human CENP-A,HA)和完全弗氏佐剂(CFA)免疫小鼠作为研究组(HA+CFA),分别用 CFA(CFA 组)或生理盐水(Saline 组)注射的小鼠作为对照组。免疫后,通过间接免疫荧光法(IIFT)和酶联免疫吸附试验(ELISA)检测血清抗-CENP-A 抗体。通过免疫荧光分析研究 MI 和 MII 期卵母细胞中的染色体排列和细胞内 IgG 定位。

结果

用 CENP-A 和 CFA 免疫成功诱导出阳性 ACA,结果显示,HA+CFA 组血清抗-CENP-A 抗体水平明显高于对照组。与对照组相比,HA+CFA 组 MI 和 MII 期卵母细胞中的染色体错位明显增加。然而,三组中的任何一组卵母细胞均未显示细胞内抗体免疫荧光。

结论

外周血 ACA 阳性小鼠的卵母细胞发育和成熟受到损害,中期减数分裂中出现严重的染色体错位;然而,没有观察到 ACA 进入卵母细胞的证据,因此需要进一步探索其潜在机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cc4/8015055/4a1027c6e15b/12958_2021_737_Fig1_HTML.jpg

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