Environmental Public Health Commons^™

Sperm Epigenetics Mediates The Effects Of Paternal Preconception Phthalate Exposures And Aging On Reproductive Outcomes And Embryonic Programming, Oladele Amos Oluwayiose

Doctoral Dissertations

Infertility, which is the inability to achieve clinical pregnancy within 12 months of unprotected sexual intercourse, affects 15% of all couples globally. Emerging evidence shows, just as females, males are equally likely to contribute to couple infertility, suggesting the relevance of male factor in couple reproductive success. The potential factors thought to drive male infertility include male preconception environment and aging. Thus, this dissertation focused on two (DNA methylation and small non-coding RNAs (sncRNAs)) of the three major potential epigenetic mediators that occur during spermatogenesis and sperm maturation prior to conception using human and mice model. First, to explore the …

Phthalates, Embryo Development, And Sperm Dna Methylation, Haotian Wu

Doctoral Dissertations

Infertility affects 15% of all couples and presents a major public health issue. Animal and human data indicate that epigenetic dysregulation in sperm is associated with poor male reproductive health and extensive epigenetic reprogramming during spermatogenesis presents a window of vulnerability for environmentally-induced epigenetic dysregulation. Phthalates is a class of ubiquitous environmental contaminant and global health concern. Phthalate exposure in humans has been associated with diminished male fertility, adverse birth outcomes, and altered offspring development. In order to facilitate additional research on this topic, we aimed to developing a higher throughput method of sperm nucleic acid extraction. In addition, this …

Rapid Method Of Processing Sperm For Nucleic Acid Extraction In Clinical Research, Matthew K. De Gannes

Masters Theses

Background: Sperm contain highly compact nuclei, inhibiting DNA extraction using traditional techniques. Current methods extracting sperm DNA involve lengthy lysis and no means of stabilizing DNA, hindering clinical research.

Objective: We sought to optimize an efficient method of extracting high quality human sperm DNA.

Methods: Sperm from three volunteers were isolated using PureCeption. We tested 1) proteinase K with DNA/RNA Shield, 2) DTT and TCEP as reducing agents, 3) QIAshredder homogenization, and 4) stability of sperm DNA fresh (baseline) or after 4 weeks of storage at 4^OC in DNA/RNA Shield using modified Quick-gDNA MiniPrep. DNA was PCR amplified …