Dataset Information

Chicken spermatozoa protein extract analyzed by Top-down HRMS

ABSTRACT: Currently, evaluation of quality semen is based on traditional in vitro sperm tests as measures of motility, viability and acrosomic reaction. However, the results are not always well correlated with real male fertility. In the present study, we evaluated Intact Cell MALDI-TOF Mass Spectrometry (ICM-MS) method as diagnostic molecular tool to substitute in vitro tests in order to phenotype semen with more robustness. For that, we investigated a large male population of known fertility that included two highly different genetic lines (meat and egg laying lines). Our optimized ICM-MS-based method through automation and the construction of fertility-predictive mathematical models was able to discriminate individuals on their reproductive capacity in the two lines studied. Furthermore, we showed a better diagnostic accuracy than traditional in vitro sperm quality tests and demonstrated that differential markers presenting a high discriminating power between fertile and subfertile sperm cells in sperm were implicated in fertility processes. To characterize m/z peaks observed in ICM-MS spectra, we performed high-throughput top-down protein identification of sperm cell extracts. Therefore, we identified more than 75% of the m/z peaks, revealing that these comprises mostly degradation products of testis-specific proteins implicated in the most important functional pathways in sperm cells such as energy metabolism and structure/movement.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Gallus Gallus (chicken)

TISSUE(S): Sperm

SUBMITTER: Labas Valerie

LAB HEAD: PAIB INRA platform, Mass spectrometry Laboratory, Valérie Labas

PROVIDER: PXD002768 | Pride | 2016-04-11

REPOSITORIES: Pride

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Publications

Intact Cell MALDI-TOF MS on Sperm: A Molecular Test For Male Fertility Diagnosis.

Soler Laura L Labas Valérie V Thélie Aurore A Grasseau Isabelle I Teixeira-Gomes Ana-Paula AP Blesbois Elisabeth E

Molecular & cellular proteomics : MCP 20160404 6

Currently, evaluation of sperm quality is primarily based on in vitro measures of sperm function such as motility, viability and/or acrosome reaction. However, results are often poorly correlated with fertility, and alternative diagnostic tools are therefore needed both in veterinary and human medicine. In a recent pilot study, we demonstrated that MS profiles from intact chicken sperm using MALDI-TOF profiles could detect significant differences between fertile/subfertile spermatozoa showing th ...[more]

PMID: 27044871

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Project description:In mammals, the capacity of the female germ cell, the oocyte, to develop into embryo is acquired throughout meiotic maturation. Immature oocyte cannot be fertilized while mature oocyte is apt to accept spermatozoa and to develop an embryo. In a follicle, the oocyte is surrounded by mural granulosa cells (GC) and is physically and metabolically coupled with specialized granulosa cumulus cells (CC) which play an important role in oocyte maturation and fertilization. Factors expressing in GC and CC during maturation may reflect the oocyte quality, i.e. its capacity to be fertilized and assure early embryo development. However, the modifications of the content and the amount of peptide/proteins in the oocyte and the surrounding CC during oocyte maturation are mostly unknown and so there is not an accurate way of evaluating/monitoring how different in vitro maturation (IVM) protocols being in use in assisted reproduction technologies, can affect the process. In this context, Intact Cell MALDI-TOF Mass Spectrometry (ICM-MS) was applied to bovine follicular cells (bovine single oocytes, cumulus cells and granulosa cells) in order to characterize proteomic changes that occur in the follicle during female gamete development. In order to characterize finely endogenous molecular species observed on ICM-MS profiles and to identify markers of interest with their post-translational modifications, we carried out top-down proteomic on the different follicular cells from oocyte, oocyte-cumulus complexes, cumulus cells and granulosa cells protein extracts. Prior to top-down MS using a dual linear ion trap Fourier Transform Mass Spectrometer LTQ Orbitrap Velos, depending on the amount of available biological material, we employed three analytic strategies as a direct infusion, a mono-dimensional liquid chromatography (µLC-1D-MS/MS) and an off-line multi-dimensional liquid chromatography combining four fractionations (based on reverse phase or gel filtration LC) to µLC-MS/MS. Here, we deposited our dataset from µLC-1D-MS/MS (analyses of oocytes, oocyte-cumulus complexes, cumulus cells and granulosa cells protein extracts) and MDLC-MS/MS (analyses of granulosa cells protein extract).

Project description:<h4><strong>BACKGROUND: </strong>Semen quality is negatively correlated with male age and is mainly quantified by a routine semen analysis, which is descriptive and inconclusive. Sperm proteins or semen metabolites are used as the intermediate or end-products, reflecting changes in semen quality, and hold much promise as a new biomarker to predict fertility in advanced-aged males.</h4><h4><strong>OBJECTIVES: </strong>In this study, we sought to assess whether the semen metabolome and proteome of aged males can affect semen quality and serve as biomarkers for predicting semen quality.</h4><p><strong>MATERIALS AND METHODS: </strong>We retrospectively analyzed 12825 males that underwent semen routine analysis to understand the age-dependent changes in sperm quality. To identify the difference between aged and young adults, metabolomics (n=60) analyses of semen and proteomics (n=12) analyses of sperm were conducted. Finally, integrated machine learning of metabolomics was conducted to screen biomarkers to identify aging semen.</p><p><strong>RESULTS:</strong> We discovered that male age was positively correlated with sperm concentration as well as DNA fragmentation index(DFI), and negatively with progressive motile sperm count, total sperm count, sperm volume and progressive sperm motility. The differential metabolites were significantly enriched in various metabolic pathways, and four of these differential metabolites (Pipamperone, 2,2-Bis(hydroxymethyl)-2,2',2''-nitrilotriethanol, Arg-Pro and Triethyl phosphate) were utilized to establish a biomarker panel to identify aging semen. Proteomic analysis showed that differential proteins were significantly enriched in protein digestion and absorption and some energy-related pathways. An integrated analysis of the metabolome and proteome identified differential energy metabolism and oxidative stress-related proteins, which could explain the decreased motility and the increased DFI of aging sperm.</p><p><strong>DISCUSSION AND CONCLUSION:</strong> We provide compelling evidence that the changes in semen metabolome and sperm proteome are related to the decline of semen quality in aged males. Moreover, a biomarker panel based on four metabolites was established to identify aging semen.</p>

Dataset Information

Chicken spermatozoa protein extract analyzed by Top-down HRMS

Publications

Intact Cell MALDI-TOF MS on Sperm: A Molecular Test For Male Fertility Diagnosis.

Similar Datasets

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