Head and flagella subcompartmental proteomic analysis of human spermatozoa

Abstract

Subcellular proteomics not only deepens our knowledge of what proteins are present within cells, but also opens our understanding as to where those proteins reside. Given the highly differentiated, cross-linked state of spermatozoa, such studies have proven difficult to perform. In this study we have fractionated spermatozoa into two components, consisting of either the head or flagellar region. Following SDS-PAGE, 1 mm slices were digested and used for LC-MS/MS analysis. In total, 1429 proteins were identified with 721 proteins being exclusively found in the tail and 521 exclusively in the head. Not only is this the largest reported proteomic analysis of human spermatozoa, but also it has provided novel insights into the compartmentalization of proteins, particularly receptors, never previously reported to be present in this cell type.

Compartmentalization of Distinct cAMP Signaling Pathways in Mammalian Sperm

“…As mentioned, immunofluorescence localization of G␣ s subunit suggests that the tmAC signaling pathway is present in the sperm plasma membrane surrounding the anterior acrosome. Our results also indicate that the PKA catalytic subunit is only present in the sperm flagellum, in agreement with a recent proteomic analysis by Aitken and co-workers (72). Consistent with this differential localization, FK did not activate PKA-dependent phosphorylation in mouse sperm.…”

A proteomic analysis on human sperm tail: comparison between normozoospermia and asthenozoospermia

“…In their recent proteomic studies of sperm, Baker et al (2013) and Amaral et al(2013) identified the proteins belonging to normal sperm head and tail domains [14,19].…”

“…SPANX B has been detected in both head and tail domains of sperm [19]. The function and exact location of this protein in flagella has not yet been clarified.…”

“…Then the sample was loaded on 5 ml of 80 % Percoll and centrifuged (650×g, 35 min, 4°C). The top 1.5 mL was discarded, and the following 1 ml of enriched sperm tail was collected [19] and centrifuged at 16,000×g, for 70 min at 4°C. It was then washed two times with TrisHCl 50 mM.…”

Identification of endogenous metabolites in human sperm cells using proton nuclear magnetic resonance (1H-NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS)

“…The identification of the metabolites and proteins as main determinants of the expression of the genomes and transcriptomes as also influenced by environmental cues, represents a first step to understand the normal composition and physiology of cells and tissues and their dysfunction associated with pathological states (Jodar et al, 2012;Wishart et al, 2013;Castillo et al, 2014a,b). In the case of the human spermatozoon, over 6000 proteins have been identified using mass spectrometry (MS), representing about 80% of the estimated proteome of this cell (Oliva et al, , 2009Amaral et al, 2013Amaral et al, , 2014aBaker et al, 2013;Azpiazu et al, 2014). However, and in *Correction added on 26 May 2015, after first online publication: article title corrected.…”