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; H1R, H2R, H3R, and H4R 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 in vitro 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 H1R, H2R, and H3R on mouse oocytes in IF and RT-PCR analyses. The treatment of IVF with histamine receptor antagonists (H1R: pyrilamine; H2R: cimetidine; H3R: 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 H1R, H2R, and H3R on mouse oocytes possibly will suggest the involvement of histamine in fertilization.
Keywords: Fertilization; Gene expression; Histamine receptor; Mouse; RT-PCR.
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