Dataset Information

Differential protein expression profiles of testes in Tmprss12-/- and wild-type mice

ABSTRACT: Serine protease is considered a relative upstream regulator in the physiological processes including male reproduction. Transmembrane serine protease 12 (TMPRSS12) has been shown to regulate sperm motility and uterotubal junction migration in mice, but its role in the testis remains unknown.In this study, we confirmed the important role of TMPRSS12 in spermatogenesis. To further investigate the mechanism of TMPRSS12 in male reproduction, we used 2-D electrophoresis combined with Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry(MALDI-TOF/TOF) to construct differential protein expression profiles of testes in Tmprss12-/- and WT mice. A total of 64 spots with 2-fold or more differential expression between two groups were identified using MALDI-TOF/TOF. As a result, 54 unique proteins were identified successfully, including 39 proteins with decreased expression and 15 proteins with increased expression in Tmprss12-/- mice.

INSTRUMENT(S): ultraflex

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Testis

SUBMITTER: Xinli Zhou

LAB HEAD: Hui Zhu

PROVIDER: PXD028014 | Pride | 2022-05-20

REPOSITORIES: Pride

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Publications

TMPRSS12 Functions in Meiosis and Spermiogenesis and Is Required for Male Fertility in Mice.

Zhang Jingjing J Zhou Xinli X Wan Danyang D Yu Li L Chen Xu X Yan Tong T Wu Zhu Z Zheng Meimei M Zhu Feng F Zhu Hui H

Frontiers in cell and developmental biology 20220425

Serine proteases are involved in many physiological activities as initiators of proteolytic cascades, and some members have been reported to play roles in male reproduction. Transmembrane serine protease 12 (TMPRSS12) has been shown to regulate sperm motility and uterotubal junction migration in mice, but its role in the testis remains unknown. In this study, we verified that TMPRSS12 was expressed in the spermatocytes and spermatids of testis and the acrosome of sperm. Mice deficient in <i>Tmpr ...[more]

PMID: 35547804

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Project description:Data analysis. Spot detection and matching were performed with a comparative cross analysis of all the gels using DeCyder software v.6.5 (GE Healthcare). 178 spots were selected based on 1.15-fold for protein ratio cut-off, allowing for the appearance of the spots in 23 out of 28 gels (69 out of 84 total images). Data from 95 spots were submitted. 93 spots were identified with high confidence. Spot picking and Trypsin digestion. The spots of interest were picked up by Ettan Spot Picker (GE Healthcare) based on the in-gel analysis and spot picking design by DeCyder software. The gel spots were washed a few times then digested in-gel with modified porcine trypsin protease (Promega, Fitchburg, WI). The digested tryptic peptides were desalted using a Zip-tip C18 (Millipore, Billerica, MA). Peptides were eluted from the Zip-tip with 0.5 uL of matrix solution (alpha-cyano-4-hydroxycinnamic acid, 5 mg/mL in 50% acetonitrile, 0.1% trifluoroacetic acid, 25mM ammonium bicarbonate) and spotted on a MALDI plate. Mass Spectrometry. MALDI-TOF MS and TOF/TOF tandem MS/MS were performed on AB SCIEX TOF/TOF 5800 System (AB SCIEX). MALDI-TOF mass spectra were acquired in reflectron positive ion mode, averaging 4000 laser shots per spectrum. TOF/TOF tandem MS fragmentation spectra were acquired for each sample, averaging 4000 laser shots per fragmentation spectrum on each of the 7-10 most abundant ions present in each sample (excluding trypsin autolytic peptides and other known background ions). Database search. Both the resulting peptide mass and the associated fragmentation spectra were submitted to GPS Explorer workstation equipped with MASCOT search engine (Matrix Science, Boston, MA) to search the Swiss-Prot database. Searches were performed without constraining protein molecular weight or isoelectric point, with variable carbamidomethylation of cysteine and oxidation of methionine residues, and with one missed cleavage also allowed in the search parameters. Candidates with either protein score C.I.% or Ion C.I.% greater than 95 were considered significant. When multiple IDs were significant for a given spot, the selection was made by evaluating apparent molecular weight, isoelectric point, the location of the spot in the gel, and the presence of strips of multiple protein isoforms in the adjacent spots.

Dataset Information

Differential protein expression profiles of testes in Tmprss12-/- and wild-type mice

Publications

TMPRSS12 Functions in Meiosis and Spermiogenesis and Is Required for Male Fertility in Mice.

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