Shotgun Proteomics-based distinctive fingerprint of sperm-surface revealed over- representation of proteins driving spermatogenesis, motility, and fertility DOI Creative Commons
Vipul Batra,

Komal Dagar,

Maharana Pratap Diwakar

et al.

Research Square (Research Square), Journal Year: 2022, Volume and Issue: unknown

Published: Dec. 15, 2022

Abstract Background Numerous distinct secretagogues such as (glyco) proteins including the GPI-anchored are added to sperm surface during their transit through male reproductive tract (MRT). This remodelling of is critical for maturation, survival and function in female (FRT). study aimed characterize present on buffalo surface. Results A surface-specific proteomic fingerprint was generated using shotgun proteomics (LC-MS/MS). The protein informatics platform, Proteome Discoverer (v2.2) identified 1342, 678, 982 isoforms (P < 0.05, FDR 0.01) salt-extracted, PI-PLC treated capacitated samples, respectively. Overall, 1695 unique (minimum 2 peptides) with ≥ 1 high-quality PSM/s (proteoforms) were identified. Descriptive statistical analysis indicated that these buffalo-specific proteoforms exhibit remarkable heterogeneity molecular weight, pI, distribution expression from genome functional roles MRT FRT. Subsequent a thorough literature search revealed fertility-related, reproduction-specific constituted more than 50% (873) sperm-surface proteome (1695). Discussion These since database, NCBI reference (translated latest chromosome level assembly, (UOA_WB_1) used space. mapped 252 implicated regulation various aspects physiology across multiple species. Besides, 200 orphan, (undefined locus, uncharacterized, P also our computational strategy. allowed us consider novel considered mapping ontology terms. led elucidation biological functions hitherto unreported, by extrapolation sequence orthologs several ruminant non-ruminant (e.g. Primates Rodents) mammalian. uncharacterized constitute an extensive, yet unexplored, repertoire. Conclusions signature driving production, discovered this unparalleled vis-à-vis depth identification fertility-related cell-surface proteins. results would facilitate advances understanding mammalian function.

Language: Английский

Cryostress induces fragmentation and alters the abundance of sperm transcripts associated with fertilizing competence and reproductive processes in buffalo DOI
Sellappan Selvaraju,

Laxman Ramya,

Divakar Swathi

et al.

Cell and Tissue Research, Journal Year: 2023, Volume and Issue: 393(1), P. 181 - 199

Published: April 20, 2023

Language: Английский

Citations

3
Transcriptome and proteome reveal abnormal spermatozoa in precocious Chinese mitten crab, Eriocheir sinensis DOI Creative Commons
Huan Liu, Lei Guo, Weiwei Zhang

et al.

Aquaculture Reports, Journal Year: 2022, Volume and Issue: 27, P. 101392 - 101392

Published: Nov. 1, 2022

In juvenile Chinese mitten crabs, Eriocheir sinensis, the occurrence of sexual gland maturation in first year is known as precocity and significantly affects culture productivity profitability. The miniaturization individuals a key characteristic precocious crabs. Most related studies focus on underlying causes, nutritional quality, biochemical components differentially expressed mRNAs crab tissues. reproductive performance crabs evaluated based offspring quality. However, status parental gametes (especially spermatozoa) directly to has not been reported. To clarify differences spermatozoa between normal mature spermatozoal transcriptome proteome data were obtained via high-throughput sequencing technology. results revealed total 856 genes 150 proteins spermatozoa. Functional analysis showed that these identified E. sinensis enriched categories substance energy metabolism. And more, spermatogenic cells was also This indicates metabolic disorders might be main reason for males. molecular defects sperm closely imbalances essential These findings provide foundational information insights into hidden biological irregularities associated with phenotypes will contribute further study males even other crustaceans.

Language: Английский

Citations

5
New insights into semen separation techniques in buffaloes DOI Creative Commons

Crina Raluca Andrei,

Florin Petrișor Posastiuc, Nicolae Tiberiu Constantin

et al.

Frontiers in Veterinary Science, Journal Year: 2024, Volume and Issue: 10

Published: Jan. 10, 2024

Male infertility is frequently caused by idiopathic or unexplained reasons, resulting in an increase demand for assisted reproductive technologies. In buffaloes, more than other animals due to hardiness, successful fertilization needs spermatozoa effectively transit the female system reach oocyte. This mechanism naturally picks high-quality sperm cells conception, but when artificial technologies such as

Language: Английский

Citations

0
Effects of Tcte1 knockout on energy chain transportation and spermatogenesis: implications for male infertility DOI Creative Commons
Marta Olszewska, Agnieszka Malcher, Tomasz Stokowy

et al.

Human Reproduction Open, Journal Year: 2024, Volume and Issue: 2024(2)

Published: Jan. 1, 2024

Abstract STUDY QUESTION Is the Tcte1 mutation causative for male infertility? SUMMARY ANSWER Our collected data underline complex and devastating effect of single-gene on testicular molecular network, leading to reproductive failure. WHAT IS KNOWN ALREADY Recent have revealed mutations in genes related axonemal dynein arms as morphology motility abnormalities spermatozoa infertile males, including dysplasia fibrous sheath (DFS) multiple morphological sperm flagella (MMAF). The nexin–dynein regulatory (N-DRC) coordinates arm activity is built from DRC1–DRC7 proteins. DRC5 (TCTE1), one N-DRC elements, has already been reported a candidate abnormal beating; however, only restricted manner with no clear explanation respective observations. DESIGN, SIZE, DURATION Using CRISPR/Cas9 genome editing technique, mouse gene knockout line was created basis C57Bl/6J strain. potential, semen characteristics, expression levels, ATP, testis apoptosis level measurements were then assessed, followed by visualization proteins sperm, protein modeling silico. Also, pilot genomic sequencing study samples human males (n = 248) applied screening TCTE1 variants. PARTICIPANTS/MATERIALS, SETTING, METHODS To check potential KO mice, adult animals crossed delivery three litters per caged pair, but longer than 6 months, various combinations zygosity. All experiments performed wild-type (WT, control group), heterozygous Tcte1+/− homozygous Tcte1−/− mice. Gross anatomy epididymis samples, analysis. Sequencing RNA (RNAseq; Illumina) done mice tissues. STRING interactions checked protein–protein interactions, based changed levels corresponding identified RNAseq experiments. Immunofluorescence situ staining detect proteins: (Drc5), Drc7, Fbxl13 (Drc6), Eps8l1 (Drc3) spermatozoa. determine amount ATP spermatozoa, luminescence measured. In addition, immunofluorescence via caspase 3 samples. DNA whole blood 137 non-obstructive azoospermia or cryptozoospermia, n 111 spectrum oligoasthenoteratozoospermia, 47 asthenozoospermia) extracted perform (WGS, WES, Sanger). Protein prediction human-identified variants exon structure deleted also performed. MAIN RESULTS AND THE ROLE OF CHANCE No progeny at all found which while fertile manifested oligozoospermia, suggesting haploinsufficiency. RNA-sequencing tissue showed influence pattern 21 responsible mitochondrial processing linked spermatogenesis. residual amounts head nucleus not transported flagella, other components. Decreased (2.4-fold lower) together disturbed tail:midpiece ratios, tail beating. Casp3-positive signals (indicating apoptosis) observed spermatogonia only, similar genotypes. Mutation novel five ultra-rare heterogeneous (predicted disease-causing) 6.05% patients studied. changes surface charge disruption helix flexibility its dynamics, thus disrupted binding partners located within axoneme. LARGE SCALE DATA generated analyzed during this are included published article supplementary information files. available GEO database (https://www.ncbi.nlm.nih.gov/geo/) under accession number GSE207805. results described publication whole-genome exome includes sensitive form patient-specific germline Information regarding such must be shared publicly following European Union legislation, therefore access raw that support findings author upon reasonable request. LIMITATIONS, REASONS FOR CAUTION study, vitro fertilization performance checked. WIDER IMPLICATIONS FINDINGS This contains comprehensive concerning role infertility. next should added ‘male infertility list’ because crucial spermatogenesis proper functioning. FUNDING/COMPETING INTEREST(S) work supported National Science Centre Poland, grants no.: 2015/17/B/NZ2/01157 2020/37/B/NZ5/00549 (to M.K.), 2017/26/D/NZ5/00789 A.M.), HD096723, GM127569-03, NIH SAP #4100085736 PA DoH A.N.Y.). authors declare there conflict interest could perceived prejudicing impartiality research reported.

Language: Английский

Citations

0
The proteomic landscape of sperm surface deciphers its maturational and functional aspects in buffalo DOI Creative Commons
Vipul Batra,

Komal Dagar,

Maharana Pratap Diwakar

et al.

Frontiers in Physiology, Journal Year: 2024, Volume and Issue: 15

Published: June 28, 2024

Buffalo is a dominant dairy animal in many agriculture-based economies. However, the poor reproductive efficiency (low conception rate) of buffalo bulls constrains realization its full production potential. This turn leads to economic and welfare issues, especially for marginal farmers such The mammalian sperm surface proteins have been implicated regulation survival function spermatozoa female tract (FRT). Nonetheless, lack specific studies on makes it difficult researchers explore investigate role these mechanisms associated with protection, survival, function. study aimed generate surface-specific proteomic fingerprint (LC-MS/MS) predict functional roles identified proteins. three treatments used remove protein viz. Elevated salt, phosphoinositide phospholipase C (PI-PLC)

Language: Английский

Citations

0
Analysis of histomorphology and SERNINA5 gene expression in different regions of epididymis of cattleyak DOI
Haiyan Li,

Cheng Pan,

Yifei Wang

et al.

Journal of Molecular Histology, Journal Year: 2024, Volume and Issue: 55(5), P. 825 - 834

Published: Aug. 6, 2024

Language: Английский

Citations

0
Regulatory role of exosomal proteins MIF and PEBP1 in sperm storage in black rockfish (Sebastes schlegelii) ovary DOI
Ying Chen,

Yu Men,

Kai Yan

et al.

Aquaculture, Journal Year: 2024, Volume and Issue: unknown, P. 741985 - 741985

Published: Dec. 1, 2024

Language: Английский

Citations

0
Shotgun Proteomics-based distinctive fingerprint of buffalo sperm surface unravels over-representation of proteins driving spermatogenesis, motility, and fertility DOI Creative Commons
Vipul Batra,

Komal Dagar,

Maharana Pratap Diwakar

et al.

Research Square (Research Square), Journal Year: 2022, Volume and Issue: unknown

Published: Oct. 27, 2022

Abstract The mammalian sperm surface is radically modified in its biomolecular composition and structure during the transit of through male female reproductive tract (MRT, FRT). Numerous distinct secretagogues such as (glyco)proteins many GPI-anchored proteins (GPI-APs) are applied peripheral coats on MRT, which critical for maturation, survival function spermatozoa. This study aimed to characterize buffalo by extracting them either using elevated salt (NaCl) or Phosphatidylinositol-specific phospholipase-C (for GPI-APs) inducing vitro capacitation. A surface-specific proteomic fingerprint was subsequently generated protein informatics platform, Proteome Discoverer (v2.2) identification. Overall, 1695 unique isoforms (proteoforms) (≥ 1 high-quality PSM/s, minimum 2 peptides, P < 0.05, FDR 0.01) that exhibited remarkable heterogeneity their molecular weight, pI, distribution expression from genome functional roles MRT FRT were identified. We characterized identified including 200 orphan extrapolation sequence orthologs BLAST2GO software. extensive, yet unexplored, reproduction-specific sperm-surface repertoire discovered this unparalleled vis-à-vis depth identification fertility-related cell-surface proteins.

Language: Английский

Citations

0
Tcte1knockout influence on energy chain transportation, apoptosis and spermatogenesis – implications for male infertility DOI Creative Commons
Marta Olszewska, Agnieszka Malcher, Tomasz Stokowy

et al.

medRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Nov. 25, 2022

Abstract STUDY QUESTION: Is Tcte1 mutation causative for male infertility? SUMMARY ANSWER Collected data underline the complex and devastating effect of single-gene on testicular molecular network, leading to reproductive failure. WHAT IS KNOWN ALREADY Latest revealed mutations in genes related axonemal dynein arms as morphology motility abnormalities spermatozoa infertile males, including dysplasia fibrous sheath (DFS) multiple morphological sperm flagella (MMAF). The nexin-dynein regulatory (N-DRC) coordinates arm activity, is built from DRC1-DRC7 proteins. DRC5 (TCTE1) – one N-DRC element, has been already reported a candidate abnormal beating, however, only restricted manner with no clear explanation respective observations. DESIGN, SIZE, DURATION Using CRISPR/Cas9 genome editing technique, mouse knockout line gene was created basis C57Bl/6J strain. Then, potential, semen characteristics, expression level, ATP testis apoptosis level measurements have performed, followed by visualization proteins sperm, protein modeling silico . Also, pilot genomic sequencing study samples human males (n=248) applied screening TCTE1 variants. PARTICIPANTS/MATERIALS, SETTING, METHODS To check potential KO mice, adult animals were crossed delivery three litters per caged pair, but longer than 6 months, various combinations zygosity. All experiments performed wild type (WT control group), heterozygous +/- , homozygous -/- mice. Gross anatomy epididymis, analysis. Sequencing RNA (RNAseq; Illumina) done mice tissues. STRING interactions checked protein-protein interactions, based changed corresponding identified RNAseq experiments. Immunofluorescence situ staining detect proteins: (Drc5), Drc7, Fbxl13 (Drc6), Eps8l1 (Drc3) spermatozoa. determine amount spermatozoa, luminescence measured. immunofluorescent via caspase 3 samples. DNA whole blood (n=137 non-obstructive azoospermia or cryptozoospermia, n=111 spectrum oligoasthenoteratozoospermia, n=47 asthenozoospermia) extracted perform (WGS, WES Sanger). Protein prediction variants exon structure deleted also performed. MAIN RESULTS AND THE ROLE OF CHANCE No progeny at all found while (fertile) manifested oligozoospermia, suggesting haploinsufficiency. RNA-sequencing tissue showed influence pattern 21 responsible mitochondrial processing, linked apoptosis, spermatogenesis. In residual amounts head nucleus, not transported flagella, other components. Decreased (2.4-fold lower) together disturbed tail:midpiece ratio, tail beating. Casp3-positive signals (indicating apoptosis) observed spermatogonia only, similar genotypes. Mutation 1 novel 5 ultrarare heterogeneous (predicted disease causing) 6.05% patients studied. changes surface charge disruption helix flexibility its dynamics, thus, disrupted interaction binding partners located within axoneme. What does this mean patients? Abnormal parameters (sperm count, morphology) are known first symptoms that may be fertility problems. This aimed role infertility using model. structural elements building N-DRC, an axoneme coordination strictly motility. We model two phenotypes, dependent zygosity: oligoasthenoteratozoospermic homozygotes, fertile oligozoospermic heterozygotic haploinsufficiency mechanism. Pilot wide spectra quality (from azoospermia, severe oligoasthenozoospermia). Thus, next should added ‘male list’ because crucial spermatogenesis influencing variety networks (incl. energy machinery processing) proper function.

Language: Английский

Citations

0
Shotgun Proteomics-based distinctive fingerprint of sperm-surface revealed over- representation of proteins driving spermatogenesis, motility, and fertility DOI Creative Commons
Vipul Batra,

Komal Dagar,

Maharana Pratap Diwakar

et al.

Research Square (Research Square), Journal Year: 2022, Volume and Issue: unknown

Published: Dec. 15, 2022

Abstract Background Numerous distinct secretagogues such as (glyco) proteins including the GPI-anchored are added to sperm surface during their transit through male reproductive tract (MRT). This remodelling of is critical for maturation, survival and function in female (FRT). study aimed characterize present on buffalo surface. Results A surface-specific proteomic fingerprint was generated using shotgun proteomics (LC-MS/MS). The protein informatics platform, Proteome Discoverer (v2.2) identified 1342, 678, 982 isoforms (P < 0.05, FDR 0.01) salt-extracted, PI-PLC treated capacitated samples, respectively. Overall, 1695 unique (minimum 2 peptides) with ≥ 1 high-quality PSM/s (proteoforms) were identified. Descriptive statistical analysis indicated that these buffalo-specific proteoforms exhibit remarkable heterogeneity molecular weight, pI, distribution expression from genome functional roles MRT FRT. Subsequent a thorough literature search revealed fertility-related, reproduction-specific constituted more than 50% (873) sperm-surface proteome (1695). Discussion These since database, NCBI reference (translated latest chromosome level assembly, (UOA_WB_1) used space. mapped 252 implicated regulation various aspects physiology across multiple species. Besides, 200 orphan, (undefined locus, uncharacterized, P also our computational strategy. allowed us consider novel considered mapping ontology terms. led elucidation biological functions hitherto unreported, by extrapolation sequence orthologs several ruminant non-ruminant (e.g. Primates Rodents) mammalian. uncharacterized constitute an extensive, yet unexplored, repertoire. Conclusions signature driving production, discovered this unparalleled vis-à-vis depth identification fertility-related cell-surface proteins. results would facilitate advances understanding mammalian function.

Language: Английский

Citations

0