Proteomic analysis of human sperm proteins in relation to sperm motility, morphology and energy metabolism
Year: 2010
Type: dissertation
Abstract: Male infertility is often associated with impaired sperm motility and morphology (asthenoteratozoospermia) for which there is no specific therapeutic treatment.It has come to light that the modification and expression of human sperm proteins play a crucial role in sperm function.In the present study, we present proteomic data of human spermatozoa in the context of sperm dysfunction.Novel techniques have been used to successfully isolate and identify differences in protein expression on a cellular level associated with asthenoteratozoospermia.In the first part of the study, differences in protein expression within the total sperm proteome were investigated between immature and mature sperm populations.Semen was collected from healthy donors (n=23) and separated into mature and immature sperm populations by 3-layer Percoll gradient centrifugation.Cells were washed and motility and morphology were measured by computer assisted sperm analysis (CASA).For the proteomic investigation cells were lysed and proteins separated by means of two-dimensional gel electrophoresis (2D electrophoresis).PD-Quest was used to identify the differentially expressed proteins.The protein spots of interest were excised and subjected to in-gel digestion.Peptides were separated by High Pressure Liquid Chromatography (HPLC) analysis and amino acid sequences determined by mass spectrophotometry.Proteins were identified by Mascot, using the Swiss Prot database.
Author: Llewelen Rapuling
Language: English
Subfield: Reproductive Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W1795695370