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Spermatogenesis, lc-ms

Sellappan Selvaraju, Sivashanmugam Parthipan, Lakshminarayana Somashekar, B Krishnan Binsila, Atul P Kolte, Arunachalam Arangasamy, Janivara Parameshwaraiah Ravindra, Stephen A Krawetz
With artificial insemination (AI) and other precision dependent assisted reproductive technologies (ART) being followed in large scale in human and animal reproduction, assessing semen quality and fertilizability is under continuous scrutiny. Various tests have been developed to predict semen quality, but so far no single, highly reliable test is available. In this regard, transcriptomic profiling of spermatozoa assumes significance as it carries the information about spermatogenesis, sperm function, and paternal roles in post-fertilization events...
March 14, 2018: Systems Biology in Reproductive Medicine
Chung-Hsing Chen, Shih Sheng Jiang, I-Shou Chang, Hui-Ju Wen, Chien-Wen Sun, Shu-Li Wang
BACKGROUND: Phthalic acid esters are ubiquitous and antiandrogenic, and may cause systemic effects in humans, particularly with in utero exposure. Epigenetic modification, such as DNA methylation, has been hypothesized to be an important mechanism that mediates certain biological processes and pathogenic effects of in utero phthalate exposure. OBJECTIVE: The aim of this study was to examine the association between genome-wide DNA methylation at birth and prenatal exposure to phthalate...
April 2018: Environmental Research
Qianying Liu, Zhixin Lei, Anxiong Huang, Qirong Lu, Xu Wang, Saeed Ahmed, Ihsan Awais, Zonghui Yuan
Mequindox (MEQ) is a synthetic antimicrobial agent widely used in China since the 1980s. Although the toxicity of MEQ is well recognized, its testis toxicity has not been adequately investigated. In the present study, we provide evidence that MEQ triggers oxidative stress, mitochondrion dysfunction and spermatogenesis deficiency in mice after exposure to MEQ (0, 25, 55, and 110 mg/kg in the diet) for up to 18 months. The genotoxicity and adrenal toxicity may contribute to sperm abnormalities caused by MEQ. Moreover, using LC/MS-IT-TOF analysis, two metabolites, 3-methyl-2-(1-hydroxyethyl) quinoxaline- N 4-monoxide (M4) and 3-methyl-2-(1-hydroxyethyl) quinoxaline- N 1-monoxide (M8), were detected in the serum of mice, which directly confirms the relationship between the N →O group reduction metabolism of MEQ and oxidative stress...
2017: Frontiers in Pharmacology
Heike Laue, Susanne Kern, Remo P Badertscher, Graham Ellis, Andreas Natsch
Several aromatic aldehydes such as 3-(4-tert-butylphenyl)-2-methylpropanal were shown to adversely affect the reproductive system in male rats following oral gavage dose of ≥ 25 mg/kg bw/d. It was hypothesized that these aldehydes are metabolized to benzoic acids such as p-tert-butylbenzoic acid as key toxic principle and that Coenzyme A (CoA) conjugates may be formed from such acids. Here we performed a detailed structure activity relationship study on the formation of benzoic acids from p-alkyl-phenylpropanals and related chemicals in rat hepatocytes in suspension...
December 1, 2017: Toxicological Sciences: An Official Journal of the Society of Toxicology
Ying-Hung Lin, Chih-Chun Ke, Ya-Yun Wang, Mei-Feng Chen, Tsung-Ming Chen, Wei-Chi Ku, Han-Sun Chiang, Chung-Hsin Yeh
According to recent estimates, 2%-15% of couples are sterile, and approximately half of the infertility cases are attributed to male reproductive factors. However, the reasons remain undefined in approximately 25% of male infertility cases, and most infertility cases exhibit spermatogenic defects. Numerous genes involved in spermatogenesis still remain unknown. We previously identified Male Germ Cells Rab GTPase-Activating Proteins (MGCRABGAPs) through cDNA microarray analysis of human testicular tissues with spermatogenic defects...
January 5, 2017: International Journal of Molecular Sciences
Arafa I Hamed, Ridha Ben Said, Abdullah S Al-Ayed, Jaroslaw Moldoch, Usama A Mahalel, Ayman M Mahmoud, Hassan A Elgebaly, Andy J Perez, Anna Stochmal
A fingerprint of steroid saponins, the major constituent in 80% methanolic fraction from the male flowers of Phoenix dactylifera has been established. Under ESI-MS/MS conditions, the fragmentation patterns of [M - H](-) ions exclusively displayed signals corresponding to the cleavage of the glycosidic bonds, thus allowing a rapid identification of 21 steroidal saponins. Moreover, two unique among them conjugated with histidine were detected by LC-ESI (-)-MS and DFT and were given tentative names of 3-o-histidine-26-o-hexosyl-dioscin and 3-o-histidine-26-o-dihexosyl-hydroxydioscin...
September 2017: Natural Product Research
Ho-Geun Kwak, Naoshi Dohmae
Various histones, including testis-specific histones, exist during spermatogenesis and some of them have been reported to play a key role in chromatin remodeling. Mass spectrometry (MS)-based characterization has become the important step to understand histone structures. Although individual histones or partial histone variant groups have been characterized, the comprehensive analysis of histone variants has not yet been conducted in the mouse testis. Here, we present the comprehensive separation and characterization of histone variants from mouse testes by a top-down approach using MS...
November 15, 2016: Bioscience Trends
Lacey J Luense, Xiaoshi Wang, Samantha B Schon, Angela H Weller, Enrique Lin Shiao, Jessica M Bryant, Marisa S Bartolomei, Christos Coutifaris, Benjamin A Garcia, Shelley L Berger
BACKGROUND: During the process of spermatogenesis, male germ cells undergo dramatic chromatin reorganization, whereby most histones are replaced by protamines, as part of the pathway to compact the genome into the small nuclear volume of the sperm head. Remarkably, approximately 90 % (human) to 95 % (mouse) of histones are evicted during the process. An intriguing hypothesis is that post-translational modifications (PTMs) decorating histones play a critical role in epigenetic regulation of spermatogenesis and embryonic development following fertilization...
2016: Epigenetics & Chromatin
Yu-Lin Huang, Qiang Fu, Hong Pan, Fu-Mei Chen, Xiu-Ling Zhao, Huan-Jing Wang, Peng-Fei Zhang, Feng-Ling Huang, Yang-Qing Lu, Ming Zhang
The testicular seminiferous tubules contain Sertoli cells and different types of spermatogenic cells. They provide the microenvironment for spermatogenesis, but the precise molecular mechanism of spermatogenesis is still not well known. Here, we have employed tandem mass tag coupled to LC-MS/MS with the high-throughput quantitative proteomics technology to explore the protein expression from buffalo testicular seminiferous tubules at three different developmental stages (prepuberty, puberty, and postpuberty)...
July 2016: Proteomics
J Vitku, L Sosvorova, T Chlupacova, R Hampl, M Hill, V Sobotka, J Heracek, M Bicikova, L Starka
The general population is potentially exposed to many chemicals that can affect the endocrine system. These substances are called endocrine disruptors (EDs), and among them bisphenol A (BPA) is one of the most widely used and well studied. Nonetheless, there are still no data on simultaneous measurements of various EDs along with steroids directly in the seminal fluid, where deleterious effects of EDs on spermatogenesis and steroidogenesis are assumed. We determined levels of BPA and 3 estrogens using LC-MS/MS in the plasma and seminal plasma of 174 men with different degrees of infertility...
2015: Physiological Research
Cheng Chen, Mamiko Maekawa, Kenji Yamatoya, Masami Nozaki, Chizuru Ito, Toshihiko Iwanaga, Kiyotaka Toshimori
Basigin is a member of the immunoglobulin superfamily and plays various important roles in biological events including spermatogenesis. To examine the basigin molecular variants during spermatogenesis and sperm maturation in the mouse, immunoprecipitated basigin samples from testis and epididymal spermatozoa were analyzed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The results demonstrated that basigin molecules from the testis and spermatozoa were separable into two major bands and that the differences in the molecular sizes were possibly because of an endoproteolytic cleavage...
July 2016: Asian Journal of Andrology
Sergo Kasvandik, Gerly Sillaste, Agne Velthut-Meikas, Aavo-Valdur Mikelsaar, Triin Hallap, Peeter Padrik, Tanel Tenson, Ülle Jaakma, Sulev Kõks, Andres Salumets
A significant proportion of mammalian fertilization is mediated through the proteomic composition of the sperm surface. These protein constituents can present as biomarkers to control and regulate breeding of agricultural animals. Previous studies have addressed the bovine sperm cell apical plasma membrane (PM) proteome with nitrogen cavitation enrichment. Alternative workflows would enable to expand the compositional data more globally around the entire sperm's surface. We used a cell surface biotin-labeling in combination with differential centrifugation to enrich sperm surface proteins...
June 2015: Proteomics
Yan Zhang, Zhaoming Dong, Peiming Gu, Weiwei Zhang, Dandan Wang, Xiaomeng Guo, Ping Zhao, Qingyou Xia
The development of the testis involves a large number of tissue-specific proteins, possibly because the sperms in it are the most divergent of all cell types. In this study, LC-MS/MS was employed to investigate the protein compositions of the adult testis of silkworm. A total of 14,431 peptides were identified in the adult testis of Bombyx mori, which were matched to 2292 proteins. Thirty-two HSPs constitute a group of most abundant proteins in the adult testis, suggesting that they are critical for the development, differentiation, and survival of germ cells...
October 2014: Proteomics
Rubén Azpiazu, Alexandra Amaral, Judit Castillo, Josep Maria Estanyol, Marta Guimerà, Josep Lluís Ballescà, Juan Balasch, Rafael Oliva
STUDY QUESTION: Are there quantitative alterations in the proteome of normozoospermic sperm samples that are able to complete IVF but whose female partner does not achieve pregnancy? SUMMARY ANSWER: Normozoospermic sperm samples with different IVF outcomes (pregnancy versus no pregnancy) differed in the levels of at least 66 proteins. WHAT IS KNOWN ALREADY: The analysis of the proteome of sperm samples with distinct fertilization capacity using low-throughput proteomic techniques resulted in the detection of a few differential proteins...
June 2014: Human Reproduction
F M L Sousa, C H Lobo, E S B Menezes, J P A Rego, R V Oliveira, A C Lima-Souza, M Fioramonte, F C Gozzo, R C F F Pompeu, M J D Cândido, J T Oliveira, A A Moura
This study describes the reproductive parameters of Morada Nova rams, a breed of hair sheep from Brazil and with unique adaption to tropical environments. At 42 weeks of age, 15 rams were subjected to semen collection and, 1 week later, animals were slaughtered for collection of testes, epididymis and accessory sex glands. We conducted 2-D electrophoresis of seminal plasma proteins and major spots of stained gels were identified by LC-MS/MS. Total RNA was isolated from testis, epididymis and vesicular glands and subjected to qPCR...
June 2014: Reproduction in Domestic Animals, Zuchthygiene
John K Amory, Samuel Arnold, María C Lardone, Antonio Piottante, Mauricio Ebensperger, Nina Isoherranen, Charles H Muller, Thomas Walsh, Andrea Castro
OBJECTIVE: To determine whether decreased testicular levels of enzymes necessary for retinoic acid biosynthesis were associated with male infertility, as retinoic acid is known to be necessary for spermatogenesis. DESIGN: Observational analysis of testicular tissue samples, sperm indices, and serum hormone concentrations. SETTING: Two infertility centers in Chile. PATIENT(S): 32 infertile men and 11 control men. INTERVENTION(S): Measurement of the three enzymes necessary for retinoic acid biosynthesis, aldehyde dehydrogenase (ALDH) 1A1, 1A2, and 1A3, in testicular tissue by a novel liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) peptide assay...
April 2014: Fertility and Sterility
Florian Flenkenthaler, Stefanie Windschüttl, Thomas Fröhlich, J Ullrich Schwarzer, Artur Mayerhofer, Georg J Arnold
Spermatogonial stem cells (SSCs) are vital for lifelong spermatogenesis in man. In their niches, a special growth factor milieu and structural support by surrounding cells are thought to ensure their maintenance. In man, the cells of the wall of seminiferous tubules, human testicular peritubular cells (HTPCs), are considered to contribute to this microenvironment and the overall testicular microenvironment via secreted proteins. Therefore, the secretome of cultured HTPCs from five individual men was analyzed by LC-MS/MS...
March 7, 2014: Journal of Proteome Research
Zhimin Shi, Junjie Hou, Xuejiang Guo, Hongxia Zhang, Fuquan Yang, Jiayin Dai
Perfluorododecanoic acid (PFDoA) is a common environmental pollutant, which has been detected in human sera and has adverse effects on testicular function in animal models. Exploring phosphorylation events in testes helps elucidate the specific phosphorylation signals involved in testicular toxicity of PFDoA. Combining efficient prefractionation of tryptic peptide mixtures using self-packed reversed phase C18 columns with TiO2 and IMAC phosphopeptide enrichment techniques followed by 2D-LC-MS/MS, we identified 4077 unique phosphopeptides from 1777 proteins with a false discovery rate below 1...
August 14, 2013: Toxicology Letters
J J Nya-Ngatchou, S L M Arnold, T J Walsh, C H Muller, S T Page, N Isoherranen, J K Amory
Intratesticular retinoic acid is necessary for spermatogenesis, but the relationship between intratesticular retinoic acid and sperm quality in man has not been studied. We hypothesized that intratesticular concentrations of retinoic acid would be lower in men with abnormal semen analyses compared to men with normal semen analyses. We recruited men requiring scrotal or penile surgery in a pilot observational study examining the relationship between sperm quality and intratesticular and serum retinoic acid. Twenty-four men provided two pre-operative blood and semen samples, and underwent a testicular biopsy during surgery...
March 2013: Andrology
Rahul D Upadhyay, N H Balasinor, Anita V Kumar, Geetanjali Sachdeva, Priyanka Parte, Kushaan Dumasia
Proteomics, an interface of rapidly evolving advances in physics and biology, is rapidly developing and expanding its potential applications to molecular and cellular biology. Application of proteomics tools has contributed towards identification of relevant protein biomarkers that can potentially change the strategies for early diagnosis and treatment of several diseases. The emergence of powerful mass spectrometry-based proteomics technique has added a new dimension to the field of medical research in liver, heart diseases and certain forms of cancer...
January 2013: Biochimica et Biophysica Acta
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