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小鼠卵母细胞上组胺受体的检测及其与受精潜能的关系。

Detection of histamine receptors on mouse oocytes and their involvement in fertilization potential.

作者信息

Yusoff Maisarah, Hashim Noor Hashida, Mohd-Yusuf Yusmin

机构信息

Institute for Advanced Studies, University of Malaya, Kuala Lumpur, Malaysia.

Centre for Foundation Studies in Science, University of Malaya, Kuala Lumpur, Malaysia.

出版信息

Vet Res Forum. 2022 Sep;13(3):331-337. doi: 10.30466/vrf.2021.136028.3050. Epub 2022 Sep 15.

Abstract

Histamine widely involves in local immune responses, physiological function in the gut, and acting as a neurotransmitter in the brain. Scientist also found the importance of histamine in the reproductive systems. The present study aimed to determine the existence of histamine receptor subtypes; , , , and on mouse oocytes through immunofluorescence (IF) staining and reverse transcription- polymerase chain reaction (RT-PCR). These further confirmed by the involvement of histamine receptor antagonists in fertilization (IVF). In IF staining, mouse oocytes were incubated with primary antibody against histamine receptor, followed by incubation with fluorescence conjugated secondary antibody. Then RT-PCR analysis was carried out for the undetected receptors during IF for confirmation. The RT-PCR used RNA extracted from mice COCs and cumulus free oocytes. In IVF, sperm was cultured in a group of treated histamine receptor antagonists oocytes. This investigation revealed the existance of , , and on mouse oocytes in IF and RT-PCR analyses. The treatment of IVF with histamine receptor antagonists (: pyrilamine; : cimetidine; : thioperamide) led to a significant reduction quantity of 2-cell embryos (4.61 ± 2.44%; 5.83 ± 4.65%; 3.83 ± 1.82%, respectively) as compared with the control group (22.50 ± 6.44%). Therefore, according to the results of this study, the presence of , , and on mouse oocytes possibly will suggest the involvement of histamine in fertilization.

摘要

组胺广泛参与局部免疫反应、肠道生理功能,并在大脑中作为神经递质发挥作用。科学家还发现了组胺在生殖系统中的重要性。本研究旨在通过免疫荧光(IF)染色和逆转录聚合酶链反应(RT-PCR)确定小鼠卵母细胞上组胺受体亚型H1、H2、H3和H4的存在情况。组胺受体拮抗剂参与体外受精(IVF)进一步证实了这些结果。在IF染色中,将小鼠卵母细胞与抗组胺受体的一抗孵育,随后与荧光偶联二抗孵育。然后对IF过程中未检测到的受体进行RT-PCR分析以进行确认。RT-PCR使用从小鼠卵丘卵母细胞复合体(COCs)和裸卵中提取的RNA。在IVF中,将精子与一组经组胺受体拮抗剂处理的卵母细胞共同培养。这项研究在IF和RT-PCR分析中揭示了小鼠卵母细胞上H1、H2和H3的存在。用组胺受体拮抗剂(H1:吡苄明;H2:西咪替丁;H3:硫代乙酰胺)处理IVF导致2细胞胚胎数量显著减少(分别为4.61±2.44%;5.83±4.65%;3.83±1.82%),与对照组(22.50±6.44%)相比。因此,根据本研究结果,小鼠卵母细胞上H1、H2和H3的存在可能表明组胺参与了受精过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5da9/9548215/2ad990763f59/vrf-13-331-g001.jpg

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