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Human testicular biopsies iTRAQ proteomics

ABSTRACT: We collected testes biopsies from NOA (severe spermatogenic defects, no spermatids or sperm) and OA (normal spermatogenesis, tubular lumen obstruction) patients for iTRAQ proteomics analyses, to screen the genes for which the altered expression is involved in spermatogenic defects

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Testis

SUBMITTER: Zhenguo Chen

LAB HEAD: Zhenguo Chen

PROVIDER: PXD060811 | Pride | 2025-06-11

REPOSITORIES: Pride

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			Action	DRS
	checksum.txt	Txt
	iTRAQ_testes_test1.group	Other
	iTRAQ_testes_test1_1.raw	Raw
	iTRAQ_testes_test1_10.raw	Raw
	iTRAQ_testes_test1_11.raw	Raw

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Publications

Adenylate kinase 1 deficiency disrupts mouse sperm motility under conditions of energy stress†.

Xie Minyu M Zhang Guofei G Zhang Hanbin H Chen Feilong F Chen Yan Y Zhuang Yuge Y Huang Zicong Z Zou Feng F Liu Min M An Geng G Kang Xiangjin X Chen Zhenguo Z

Biology of reproduction 20201001 5

Mammalian spermatozoa are highly polarized cells characterized by compartmentalized cellular structures and energy metabolism. Adenylate kinase (AK), which interconverts two ADP molecules into stoichiometric amounts of ATP and AMP, plays a critical role in buffering adenine nucleotides throughout the tail to support flagellar motility. Yet the role of the major AK isoform, AK1, is still not well characterized. Here, by using a proteomic analysis of testis biopsy samples, we found that AK1 levels ...[more]

PMID: 32744313

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Project description:Actin-related proteins (Arp) are classified according to their similarity to actin and are involved in diverse cellular processes. ACTL7B is a testis-specific Arp and highly conserved in rodents and primates. ACTL7B is specifically expressed in round and elongating spermatids during spermiogenesis. Here, we have generated an Actl7b-null allele in mice to unravel the role of ACTL7B in sperm formation. Male mice homozygous for the Actl7b-null allele (Actl7b-/-) were infertile, while heterozygous males (Actl7b+/-) were fertile. Severe spermatid defects such as detached acrosomes, disrupted membranes and failed elongation of the axoneme start to appear at spermiogenesis step 9 in Actl7b-/- mice, finally resulting in spermatogenic arrest. Abnormal spermatids, were degraded. Co-immunoprecipitation experiments identified interaction between ACTL7B and the LC8 dynein light chains DYNLL1 and DYNLL2, which are first detected in step 9 spermatids and mislocalized when ACTL7B is absent. Our data unequivocally establishes that mutations in ACTL7B are directly related to male infertility, pressing for additional research in men.

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A non-canonical bivalent gene Wfdc15a controls spermatogenic protease and immune homeostasis

Project description:Male infertility can be caused by chromosomal abnormalities, mutations, and epigenetic defects. Epigenetic modifiers pre-program hundreds of spermatogenic genes in spermatogonial stem cells (SSCs) for expression later in spermatids, but it remains mostly unclear whether and how those genes are involved in fertility. Here, we report that Wfdc15a, a WFDC family protease inhibitor pre-programmed by KMT2B, is essential for spermatogenesis. We found that Wfdc15a is a non-canonical bivalent gene carrying both H3K4me3 and facultative H3K9me3 in SSCs but is later activated along with the loss of H3K9me3 and acquisition of H3K27ac during meiosis. We show that WFDC15A deficiency causes defective spermiogenesis at the beginning of spermatid elongation. Notably, depletion of WFDC15A causes substantial disturbance of the testicular protease-antiprotease network and leads to an orchitis-like inflammatory response associated with TNFa expression in round spermatids. Together, our results reveal a unique epigenetic program regulating innate immunity crucial for fertility.

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A non-canonical bivalent gene Wfdc15a controls spermatogenic protease and immune homeostasis

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A non-canonical bivalent gene Wfdc15a controls spermatogenic protease and immune homeostasis [P25]

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10x Genonics Drop-seq single-cell RNA-seq of isolated Adult ID4-EGFP mouse spermatogonia, spermatocytes, spermatids & steady-state spermatogenic cells

Project description:To reveal distinct transcriptomes associated with various spermatogenic cells, including spermatogonial stem cells and all of their subsequent progeny, single-cell transcriptomes from Adult ID4-EGFP+ spermatogonia (sorted for brightest or dimmest), StaPut-enriched spermatocytes and spermatids, or unselected steady-state spermatogenic cells were used for 10x Genomics analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (Drop-Seq) to perform single-cell RNA-seq

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Phenotyping spermatogenic defects by single-cell expression profiling

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The insufficient activation of RIG-I-like signaling pathway in IBRS-2 cells contributes to highly efficient replication of porcine picornaviruses

Project description:Seneca Valley Virus (SVV) or commonly known as Senecavirus A (SVA), is one of picornavirus that is associated with vesicular disease and neonatal mortality in swine herds. However, the pathogenesis of SVV remains largely elusive. Our previous study found that SVV replicates extremely faster in porcine IBRS-2 cells than that in porcine PK-15 cells. However, the underlying mechanism remains unknown. In this study, we comprehensively compared the expression features between IBRS-2 cells and PK-15 cells in response to SVV infection by an unbiased high-throughput quantitative proteomic analysis. We found that the innate immune response-realted pathways were efficiently activated in PK-15 cells but not in IBRS-2 cells during SVV infection. A large amount of interferon-stimulated genes (ISGs) were induced in PK-15 cells. In contrast, no ISGs were induced in IBRS-2 cells after SVV infection. This different expression features in the two cell lines was verified by qPCR analysis. Besides, we determined similar results in the two cell lines during another porcine picornavirus foot-and-mouth disease virus (FMDV) infection. Further study demonstrated that the Janus kinase signal transducer and activator of transcription (JAK-STAT) signaling pathway was functioning properly in both IBRS-2 and PK-15 cells. A systematic screening study revealed that the aberrant signal transduction from TBK1 to IRF3 in the RIG-I-like receptor signaling pathway in IBRS-2 cells was the fundamental cause of the different innate immune response manifestation and different viral replication rate in the two cell lines. Together, our findings determined the different feature of IBRS-2 and PK-15 cell lines which will provide useful guidance for choosing right cell line in porcine picornaviruses-mediated host immune signaling research and could be easily extended to other porcine viruses.

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The insufficient activation of RIG-I-like signaling pathway in IBRS-2 cells contributes to highly efficient replication of porcine picornaviruses

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A specific CBP/p300-dependent gene expression program drives the metabolic remodelling in late stages of spermatogenesis.

Project description:Histone hyperacetylation is thought to drive the replacement of histones by transition proteins that occurs in elongating spermatids after a general shut-down of transcription. The molecular machineries underlying this histone hyperacetylation remain still undefined. Here we focused our attention on the role of Cbp and p300 in histone hyperacetylation and in the preceding late gene transcriptional activity in elongating spermatids. A strategy was designed to partially deplete Cbp and p300 in elongating spermatids. These cells progressed normally through spermiogenesis and showed normal histone hyperacetylation and removal. However, a genome-wide transcriptomic analysis, performed in the round and elongating spermatids, revealed the existence of a gene regulatory circuit encompassing genes presenting high expression levels in pre-meiotic cells, undergoing a repressed state in spermatocytes and early post-meiotic cells, but becoming reactivated in elongating spermatids, just prior to the global shut-down of transcription. Interestingly, this group of genes was over-represented within the genes affected by Cbp/p300 knock-down and were all involved in metabolic remodelling. This study revealed the occurrence of a tightly regulated Cbp/p300-dependent gene expression program that drives a specific metabolic state both in progenitor spermatogenic cells and in late transcriptionally active spermatids and confirmed a special link between Cpb/p300 and cell metabolism programming previously shown in somatic cells. Total RNA were obtained from post meiotic male germ cells (round spermatids and condensing spermatids) of adult mice either wild-type or with a double knock down of Cbp/P300 in post-meiotic cells. In each experiments, 6 replicates of each genotype and cell type were used.

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