Evaluation of protamine 2 genes in spermatozoa of teratospermic and oligospermic men in Nigeria

Emmanuel Sunday Oni, Ojo Moses Oke, Josephine Kpalap, Samuel Kehinde Wojuade, Adewale Oke, Emmanuel Ayomide Oni
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Abstract

There have been several reports of single nucleotide polymorphisms (SNPs) linked to different kinds of male infertility. The aim of the study was to evaluate single nucleotide polymorphisms in protamine 2 gene (PRM2) in spermatozoa of teratospermic and oligospermic infertile men in Nigeria. At some fertility clinics in Lagos, Nigeria, twenty-two (22) teratospermic and thirty-five (35) oligospermic infertile men as well as thirty (35) normospermic fertile men (control) who volunteered and gave consent were recruited for the cross-section study after meeting the inclusion criteria and confirmation of their fertility statuses by the use of computer-Assisted Sperm Analyzer (CASA). Semen was collected from the participants under the WHO guideline for semen collection and processing. Spermatozoa’s DNA was extracted with the use of Proteinase K Storage Buffer. Nanodrop 1000 spectrophotometer was used to quantify the isolated genomic DNA. PRM II F: 5-AGGGCCCTGCTAGTTGTGA-3' and PRM II R: 3'- CAGATCTTGTGGGCTTCTCG -5, were used as primers. Sequencing of sperm DNA was done using the BigDye Terminator kit on a 3510 ABI sequencer by Inqaba Biotechnological, Pretoria South Africa. Agarose gel electrophoresis was used to show the amplified PRM2. In the study, 7 SNPs in the teratospermic infertile men, 8 SNPs in the oligospermic infertile men and 7 SNPs in the normospermic fertile men were discovered. The SNPs between the teratospermic infertile men and the oligospermic infertile men are significantly different. We propose that considerably larger genome-wide investigations are required to confidently validate these SNPs and find new SNPs linked with male infertility.
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尼日利亚畸形精子症和少精子症男性精子中原胺 2 基因的评估
关于单核苷酸多态性(SNPs)与不同类型的男性不育症有关的报道已有数篇。本研究旨在评估尼日利亚畸形精子症和少精子症不育男性精子中原胺 2 基因(PRM2)的单核苷酸多态性。在尼日利亚拉各斯的一些不孕不育诊所招募了 22 名畸形精子症和 35 名少精子症不育男性,以及 30 名精子正常的可育男性(对照组),这些男性在自愿并同意的情况下符合纳入标准,并通过计算机辅助精子分析仪(CASA)确认其生育状况后,被纳入横断面研究。研究人员按照世界卫生组织的精液采集和处理指南采集精液。使用蛋白酶 K 储存缓冲液提取精子的 DNA。使用 Nanodrop 1000 分光光度计对分离的基因组 DNA 进行定量。引物为 PRM II F: 5-AGGGCCCTGCTAGTTGTGA-3' 和 PRM II R: 3'- CAGATCTTGTGGGCTTCTCG -5。在南非比勒陀利亚 Inqaba Biotechnological 公司的 3510 ABI 测序仪上使用 BigDye Terminator 试剂盒对精子 DNA 进行测序。研究发现,畸精不育男性中有 7 个 SNPs,少精不育男性中有 8 个 SNPs,正常精子可育男性中有 7 个 SNPs。畸精不育男性和少精不育男性的 SNPs 有显著差异。我们建议,需要进行更大规模的全基因组调查,才能有把握地验证这些 SNPs,并找到与男性不育有关的新 SNPs。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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