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Identification and functional analysis of CG3526 in spermatogenesis of Drosophila melanogaster

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Insect Science 2024年第1期31卷 79-90页

作者： Qihao Hu Yanhong Xiao Runnan Wei Ting Tang Liang Wen Yuzhen Lu Xiao-Qiang Yu Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology Guangzhou Key Laboratory of Insect Development Regulation and Application ResearchInstitute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhou510631 China

spermatogenesis is a critical part of reproduction in insects;however,its molecular mechanism is still largely *** this study,we identified a testis-specific gene CG3526 in Drosophila *** analysis showed that CG3526 contains a zinc binding domain and 2 C2H2 type zinc fingers,and it is clustered to the vertebrate really interesting new gene(RING)family E3 ubiquitin-protein *** CG3526 was knocked down by RNA interference(RNAi),the testis became much smaller in size,and the apical tip exhibited a sharp and thin end instead of the blunt and round shape in the control *** importantly,compared to the control flies,only a few mature sperm were present in the seminal vesicle of C587-Gal4>CG3526 RNAi *** staining of the testis from CG3526 RNAi flies showed that the homeostasis of testis stem cell niche was disrupted,cell distribution in the apical tip was scattered,and the process of spermatogenesis was not ***,we found that the phenotype of CG3526 RNAi flies’testis was similar to that of testis of Stat92E RNAi flies,the expression level of CG3526 was significantly downregulated in the Stat92EF06346 mutant flies,and the promoter activity of CG3526 was upregulated by *** together,our results indicated that CG3526 is a downstream effector gene in the JAK-STAT signaling pathway that plays a key role in the spermatogenesis of Drosophila.

关键词： CG3526 JAK-STAT spermatogenesis STAT92E testis

Pyruvate kinase M in germ cells is essential for sperm motility and male fertility but not spermatogenesis

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Asian Journal of Andrology 2024年第2期26卷 212-219页

作者： Gao-Qing Qian Xiao-Chen Wang Xi Zhang Bin Shen Qiang Liu State Key Laboratory of Reproductive Medicine and Offspring Health Nanjing Medical UniversityNanjing 211166China Shanghai Key Laboratory of Reproductive Medicine Department of HistoembryologyGenetics and Developmental BiologyShanghai Jiao Tong University School of MedicineShanghai 200025China State Key Laboratory of Molecular Biology Shanghai Key Laboratory of Molecular AndrologyShanghai Institute of Biochemistry and Cell BiologyCenter for Excellence in Molecular Cell ScienceChinese Academy of SciencesUniversity of Chinese Academy of SciencesShanghai 200031China

Male germ cells employ specific metabolic pathways throughout their developmental *** a previous study,we discovered heightened expression of pyruvate kinase M(PKM),a pivotal glycolytic enzyme,in spermatogonia and *** gain deeper insights into PKM's roles in spermatogenesis,sperm function,and male fertility,we engineered a conditional-knockout mouse model(Pkm-vkO mice)to selectively disrupt the Pkm gene within germ *** maintaining regular testicular histology and sperm morphology,the male Pkm-vko mice were infertility,characterized by significant impairments in sperm motility and adenosine triphosphate(ATP)*** addition,Pkm-null spermatozoa exhibited similar deficits in protein tyrosine phosphorylation linked to capacitation,as well as compromised performance in in vitro fertilization *** conclude,PKM's presence is not obligatory for the entirety of spermatogenesis in male germ cells;however,it emerges as a critical factor influencing sperm motility and overall male fertility.

关键词： glycolysis male fertility pyruvate kinase M sperm motility spermatogenesis

Adenine nucleotide translocase 2(Ant2)is required for individualization of spermatogenesis of Drosophila melanogaster

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Insect Science 2024年第4期31卷 1055-1072页

作者： Zhen He Yang Fang Fengchao Zhang Yang Liu Xinkai Cheng Jiajia Wang Dechen Li Dengsong Chen Fan Wu Industrial Crops Institute Hubei Academy of Agriculture SciencesWuhanChina Institute of Evolution&Marine Biodiversity Ocean University of ChinaQingdaoShandong ProvinceChina

Successful completion of spermatogenesis is crucial for the perpetuation of the *** Drosophila,spermatid individualization,a process involving changes in mitochondrial structure and function is critical to produce functional mature ***2,encoding a mitochondrial adenine nucleotide translocase,is highly expressed in male testes and plays a role in energy metabolism in the ***,its molecular function remains ***,we identified an important role of Ant2 in spermatid *** Ant2 knockdown testes,spermatid individualization complexes composed of F-actin cones exhibited a diffuse distribution,and mature sperms were absent in the seminal vesicle,thus leading to male *** most striking effects in Ant2-knockdown spermatids were decrease in tubulin polyglycylation and disruption of proper mitochondria derivatives *** apoptotic cells were also observed in Ant2-knockdown *** further investigate the phenotype of Ant2 knockdown in testes at the molecular level,complementary transcriptome and proteome analyses were *** the mRNA level,868 differentially expressed genes were identified,of which 229 genes were upregulated and 639 were downregulated induced via Ant2 ***-labeling proteome analysis revealed 350 differentially expressed proteins,of which 117 proteins were upregulated and 233 were *** expression of glutathione transferase(GstD5,GstE5,GstE8,and GstD3),proteins involved in reproduction were significantly regulated at both the mRNA and protein *** results indicate that Ant2 is crucial for spermatid maturation by affecting mitochondrial morphogenesis.

关键词： adenine nucleotide translocase Drosophila melanogaster individualization complexes mitochondria spermatogenesis tubulin polyglycylation

Dynamic transcriptome profiles and novel markers in bovine spermatogenesis revealed by single-cell sequencing

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Journal of Integrative Agriculture 2024年第7期23卷 2362-2378页

作者： Yuan Gao Fuxia Bai Qi Zhang Xiaoya An Zhaofei Wang Chuzhao Lei Ruihua Dang Key Laboratory of Animal Genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100China College of Life Sciences Northwestern Polytechnical UniversityXi’an 710072China

Testicular development is an important biological process in male and requires interaction between the male germ cells and somatic ***,the mechanisms of testicular development in livestock,particularly in cattle,are poorly ***,cellular heterogeneity hinders the profiling of different cell types at different developmental *** this study,we first performed a single-cell transcriptomic study of the bovine testis development during puberty by using 10×genomics single-cell RNA sequencing(scRNA-seq).By collecting the scRNA-seq data from 11,083 cells from prepubertal and pubertal bovine testes,a high-resolution scRNA-seq atlas was described,identifying 9 somatic and 13 spermatogenic *** also distinguished several stage-specific marker genes for bovine germ cells and somatic cells,such as GRAF2 and MORC1 for SSC(spermatogonial stem cells),HJURP and TCF19 for differentiating spermatogonia,ARSE for immature Sertoli,CLEC12B for mature Sertoli,LOC112441470 for *** conclusion,we have examined the transcription levels and constructed the single-cell developmental maps of germ cells and somatic cells during testicular development in Angus *** datasets provided new insights into spermatogenesis and testicular somatic cell development in cattle.

关键词： scRNA-seq cattle spermatogenesis testis development

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Assessment of spermatogenesis in camels via seminiferous tubule staging and testosterone profiling

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Asian pacific Journal of Reproduction 2024年第3期13卷 133-142页

作者： Saif Ullah Abdul Quddus Quanwei Wei Wael Ennab Sheeraz Mustafa Jiahao Sun Rahmani Mohammad Malyar Abdul Baseer Dagan Mao Fangxiong Shi College of Animal Science and Technology Nanjing Agricultural UniversityNanjing 210095China Faculty of Veterinary and Animal Sciences Lasbela University of Agriculture Water and Marine SciencesBalochistanPakistan Faculty of Veterinary and Animal Sciences Ziaddin University Karachi Pakistan

Objective:To investigate the successive morphological stages of spermatogenesis,hormonal regulation,and testosterone profile in dromedary camel ***:Testicular tissue samples were obtained from 12 dromedary bull camels aged 7 to 8 at a local *** histological assessment involved tissue processing,hematoxylin and eosin(H&E)staining,and examination under a *** analysis,germ cell identification,and assessment of seminiferous tubules and maturation were *** assay was performed by radioimmunoassay using blood samples collected at regular ***:The study revealed 12 phases of the dromedary camel's seminiferous epithelium cycle,highlighting distinct morphological characteristics and cellular *** migration,maturation,cap formation,and the Golgi-mediated synthesis of proacrosomal vesicles were also explained in dimension,as were the steps of acrosome *** and mature sperm cells were present when spermatogenesis phases were *** analysis of the dimensions of seminiferous tubules revealed specific measures for diameter,area,and epithelial height about luminal ***,there were noticeable variations in the serum testosterone concentrations during the study period,indicating temporal ***:This study outlines the spermatogenesis process in dromedary camels across 12 stages,emphasizing cellular dynamics and acrosomal *** also provides seminiferous tubule measurements and observes seasonal testosterone fluctuations,offering insights into reproductive regulation and potential strategies for camel breeding conservation.

关键词： spermatogenesis Camel Testosterone profile Morphological staging

Pulsatile gonadotropin-releasing hormone therapy induces spermatogenesis in pituitary stalk interruption syndrome:A case report and review of the literature

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World Journal of Clinical Cases 2024年第20期12卷 4348-4356页

作者： Jin-Long Xie Hai-Ying Zhu Yang Dong Ping-Ping Sun Dan-Dan Qi Su-Xian Luan Yan Zhang Hua-Gang Ma The Reproductive Medicine Centre Weifang People's HospitalWeifang 261000Shandong ProvinceChina Department of Radiology Weifang People's HospitalWeifang 261000Shandong ProvinceChina

BACKGROUND Pituitary stalk interruption syndrome(PSIS)is a rare anatomical defect of the pituitary gland falling under the spectrum of holoprosencephaly *** is characterized by a deficiency in anterior pituitary hormones,such as growth hormone,gonadotropins,and thyroid *** to the syndrome's rarity and nonspecific manifestations,there is a lack of standardized treatment ***,early diagnosis through imaging and on-time intervention are crucial for improving patients’*** SUMMARY A 30-year-old man presented with absent secondary sexual characteristics and *** evaluation revealed a deficiency in gonadotropins,while thyroid function was mostly within normal *** resonance imaging of the pituitary gland showed pituitary stalk agenesis,hypoplasia of the anterior pituitary,and ectopic posterior pituitary,leading to the diagnosis of ***,the patient underwent 6 mo of gonadotropin therapy without significant changes in hormone levels and secondary sexual *** gonadotropin-releasing hormone therapy was then administered,resulting in the detection of sperm in the semen analysis within 3 *** 6 mo,routine semen tests showed normal semen *** couple faced challenges in conceiving due to abstinence and underwent three cycles of artificial insemination,which was *** also attempted in vitro fertilization,but unfortunately,the woman experienced a miscarriage 10 wk after the embryo *** Early detection,accurate diagnosis,and timely treatment are crucial in improving the quality of life and fertility of PSIS patients.

关键词： Pituitary stalk interruption syndrome Hypogonadotropic hypogonadism spermatogenesis Pulsatile gonadotropinreleasing hormone therapy Case report

Micro RNA profiles and potential regulatory pattern during the early stage of spermatogenesis in mice

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Science China(Life Sciences) 2015年第11期58卷 442-450页

作者： LUO MengMeng HAO LiLi HU Fen DONG YaNan GOU LiXia ZHANG WenDian WANG Xin ZHAO YuHui JIA MengChun HU SongNian ZHANG XiuJun College of Life Sciences College of Psychology Hebei United University CAS Key Laboratory of Genome Sciences and Information Beijing Institute of Genomics Chinese Academy of Sciences Hebei Normal University for Nationalities Department of Reproductive Endocrinology National Research Institute for Family Planning

spermatogenesis is a complicated and poorly understood process that relies on the precise regulation of the self-renewal and differentiation of spermatogonia. In many organisms, micro RNAs(mi RNAs) are involved in multiple developmental processes as critical regulators of transcriptional and post-transcriptional gene silencing. This study investigated the expression pattern of mi RNAs in type B spermatogonia cells(BSc) and primary spermatocytes(PSc) of mice, using a high-throughput small RNA sequencing system. The results revealed that the expression levels of Let-7 family mi RNAs were remarkably high in both cell types. Furthermore, the expression levels of mi R-21, mi R-140-3p, mi R-103, mi R-30 a, mi R-101 b and mi R-99 b were decreased during the transformation from BSc to PSc. These mi RNAs target vital genes that participate in apoptosis, cell proliferation and differentiation, junction assembly and cell cycle regulation. These results highlight the indispensable role of mi RNAs in spermatogenesis.

关键词： spermatogenesis miRNA profiling post-transcriptional regulation focal adhesion cytoskeleton dynamic Wnt signaling pathway

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Identification of a Novel Gene SRG4 Expressed at Specific Stages of Mouse spermatogenesis

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Acta Biochimica et Biophysica Sinica 2004年第5期36卷 351-359页

作者： Xiao-Wei XING, Lu-Yun LI, Gang LIU, Jun-Jiang FU, Xiao-Jun TAN, and Guang-Xiu LU* Human Reproduction and Stem Cell Engineering Institute, Central South University, Changsha 410078, China Human Reproduction and Stem Cell Engineering Institute Central South University Changsha 410078 China

spermatogenesis is a complex process. Two spermatocytes expression sequence tags (ESTs) BG101130 and BG100990 were found. Their putative amino acid sequences have high homology with rat Spag4 (sperm antigen 4). By electrical hybridization, a novel cDNA encoding polypeptide of 348 amino acid residues was identified from a mouse testis cDNA library. The new gene was designated as SRG4 (spermatogenesis related gene 4) (GenBank accession No. AY307077). Results of Northern blot and RT- PCR revealed that SRG4 expressed specifically in mouse testis. Changes of SRG4 expression in mouse dif- ferent development stages were observed by RT-PCR. The SRG4 mRNA was hardly detected in 2 weeks postpartum, and expressed abundantly from 3 weeks later, reaching top lever at 4–5 weeks, while slightly down in aging mouse testis. Results of in situ hybridization showed that SRG4 gene expressed abundantly in spermatocytes, round spermatids. This indicated SRG4 gene may play an important role in mouse meiotic divisions of spermatocytes.

关键词： gene cloning testis tissues expression pattern in situ hybridization spermatogenesis

Vitamin B12-induced spermatogenesis recovery in cimetidine-treated rats： effect on the spermatogonia number and sperm concentration

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Asian Journal of Andrology 2017年第5期19卷 567-572页

作者： Flavia L Beltrame Estela Sasso-Cerri Department of Morphology and Genetics Federal University of Sao Paulo (UNIFESP/EPM) Sao Paulo Brazil Department of Morphology Laboratory f Histology and Embryology Dental School - Sao Paulo State University (UNESP/FOAr) Araraquara Sao Paulo Brazil

The H2-receptor antagonist cimetidine is an antiulcer drug also used for the treatment of cancer due to its antiangiogenic effect. However, this drug has caused structural changes in the seminiferous tubules. Vitamin B12 has been used as a therapeutic agent for the treatment of male infertility. The supplementation of rats with vitamin B12 during cimetidine treatment has recovered the damaged seminiferous tubules, but how this vitamin restores the seminiferous epithelium has not been clarified. In this study, we evaluated whether vitamin B12 improves the number of spermatogonia, spermatocytes, and sperm concentration in cimetidine-treated rats. Adult male rats were treated for 50 days as follows： cimetidine group received 100 mg kg^-1 b.w. of cimetidine, cimetidine-B12 group received cimetidine and 3 μg of vitamin B12-hydroxocobalamin, B12 group received only 3 μg of vitamin, and control group received saline. Sperm concentration was calculated and historesin-embedded testes sections were used for the quantitative analyses of spermatogonia （A; In/B） and spermatocytes. TUNEL method and PCNA immunofluorescence were performed. Cimetidine caused a significant reduction in sperm concentration. TUNEL-positive spermatogonia and spermatocytes were correlated to a significant reduction in the number of these cells. In cimetidine-B12 group, sperm concentration was higher than cimetidine group and a significant increase in the number of spermatogonia （stages Ⅱ-Ⅵ） was correlated to a high incidence of PCNA-immunolabeled spermatogonia and spermatocytes. The results show that the supplementation of rats with vitamin B12 during cimetidine treatment increases sperm concentration and exerts a potential effect in the recovery of spermatogonia and spermatocytes.

关键词： cimetidine fertility spermatogenesis testis vitamin B12