Studies on the protamine 1 gene in the sperm DNA of male albino rats treated with local gin using BseR I endonuclease
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Minari, J. B.
Salau, O. Y.
Asian Pacific Journal of Reproduction
Objective: To investigate the impact of local brewed gin (ogogoro) on the integrity of sperm DNA through the evaluation of the protamine 1 (PRM1) gene. Methods: Twenty four 9-week-old male albino rats were divided into ﬁve groups; the ﬁrst group which served as the control were fed with normal ﬁsh feed and water ad libitum while the rest were fed with different volumes of the local gin (1, 2, 3 and 4 mL respectively) in conjunction with the normal ﬁsh feed and water for 24 d. The caudal epididymis was macerated in saline water and the sperm solution collected in an Eppendorf tube. The extraction of DNA was done using Jena Bioscience DNA preparation kit and the protocol was based on the spin column based genomic DNA puriﬁcation from blood, animal and plant cells. The primer was designed to detect mutation in Protamine 1 gene associated with male infertility. BseR I, a restriction endonuclease was used to digest the amplicon at 37 oC for 10 min and then inactivated at 80 oC for 20 min. The digest was separated on 2% agarose gel. Results: There was variation in the thickness of bands and the thinnest band was observed in the group that consumed the highest volume of local gin. There was also a close similarity between the amplicons from the group fed with 1 mL of ogogoro and the control group that were given distilled water. There was no visible digestion by BseRI. There was a signiﬁcant increase in the body weight across the entire experimental group and the highest weight was recorded in the 4 mL group. Multiple tumors were also seen in the livers of all the experimental groups. Conclusion: This study has shown that local gin can affect the sperm DNA by making it susceptible to damage because of the reduced viability of the PRM1 gene whose product i.e., PRM1 protein is needed for the proper compaction of the sperm DNA.
Male infertility, Protamine 1, Local gin (ogogoro), BseR I endonuclease