Dataset Information

Binding partners to Testis Specific Antigen, TSA70

ABSTRACT: A testis specific antigen, TSA70 was identified in the lab which was homolgous to the sperm tail outer dense fiber protein (ODF2). ODFs are disulphide stabilised during epididymal maturation and are required for sperm motility. ODF2 is reported to bind macrophage migration inhibitory factor (MIF), a moonlighting oxidoreductase. It is not known whether such proteins exist in the female reproductive tract that ensure the disulphide stability. The aim was to identify full quorum of such redundant proteins in both male and female reproductive tract fluids. Using recombinant testis specific antigen, TSA70/ODF2, as bait we report the identification of binding partners from the epididymal fluid. Since MIF is a 12.5kDa protein, we focussed only on the low molecular weight proteins.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Epididymis, Epithelial Cell

SUBMITTER: Kaushiki Kadam

LAB HEAD: Kaushiki Kadam

PROVIDER: PXD002296 | Pride | 2020-01-23

REPOSITORIES: Pride

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Publications

Thiol based mechanism internalises interacting partners to outer dense fibers in sperm.

Kadam Kaushiki K Khole Vrinda V VV Ghosalkar Kanaka K Jagtap Dhanashree D Yarramala Deepthi S DS Ramachandran Bini B

Free radical biology & medicine 20200107

The sperm tail outer dense fibres (ODFs) contribute passive structural role in sperm motility. The level of disulphide cross-linking of ODFs and their structural thickness determines flagellar bending curvature and motility. During epididymal maturation, proteins are internalized to modify ODF disulphide cross-linking and enable motility. Sperm thiol status is further altered during capacitation in female tract. This suggests that components in female reproductive tract acting on thiol/disulphid ...[more]

PMID: 31923584

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Project description:Male-derived accessory gland proteins (Acps) that are transferred to females during mating have profound effects on female reproductive physiology including increased ovulation, mating inhibition, and effects on sperm utilization and storage. The extreme rates of evolution seen in Acps may be driven by sperm competition and sexual conflict, processes which may ultimately drive complex interactions between female- and male-derived molecules and sperm. However, little is known of how gene expression in female reproductive tissues changes in response to the presence of male molecules and sperm. To characterize this response, we conducted parallel genomic and proteomic analyses of gene expression in the reproductive tract of 3-day-old unmated and mated female Drosophila melanogaster. Using DNA microarrays, we identified 539 transcripts that are differentially expressed in unmated vs. mated females and revealed a striking peak in differential expression at 6 hrs postmating and a marked shift from primarily down-regulated to primarily up-regulated transcripts within 3 hrs after mating. Combining two-dimensional gel electrophoresis and liquid chromatography mass spectrometry analyses, we identified 84 differentially expressed proteins at 3 hrs postmating, including proteins which appeared to undergo post-translational modification. Together, our observations define transcriptional and translational response to mating within the female reproductive tract and suggest a bimodal model of postmating gene expression initially correlated with mating and the final stages of female reproductive tract maturation and later with the declining presence of male reproductive molecules and with sperm maintenance and utilization. Experiment Overall Design: Three-day-old mated and unmated females were dissected to remove the lower reproductive tract (upper uterus, sperm-storage organs, and accessory glands). Mated females were dissected either immediately following mating (0 hr) or at 3, 6, or 24 hrs following the termination of mating. Tracts of 12-40 females of like category were pooled and total RNA extracted via a TRIzol-based protocol. Processing and labeling of transcript was performed by the Molecular Biology Core Facility at the Medical College of Georgia. Arrays from mated females at the different timepoints were compared to unmated females.

Dataset Information

Binding partners to Testis Specific Antigen, TSA70

Publications

Thiol based mechanism internalises interacting partners to outer dense fibers in sperm.

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