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sertoli cell and spermatogonial development in pigs

Journal of Animal Science and Biotechnology 2022年第4期13卷 1028-1044页

作者： Yi Zheng Qiang Gao Tianjiao Li Ruifang Liu Zechao Cheng Ming Guo Jinhong Xiao De Wu Wenxian Zeng Key Laboratory for Animal Genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100ShaanxiChina Zhumei Porcine Breeding Corporation ZhengzhouHenanChina Key Laboratory for Animal Disease Resistance Nutrition of the Ministry of Education of China Institute of Animal NutritionSichuan Agricultural UniversityChengdu 611100SichuanChina.

Background:Spermatogenesis is an intricate developmental process during which undifferentiated spermatogonia,containing spermatogonial stem cells(SSCs),undergo self-renewal and differentiation to generate eventually mature spermatozoa.Spermatogenesis occurs in seminiferous tubules within the testis,and the seminiferous tubules harbor sertoli and germ cells.sertoli cells are an essential somatic cell type within the microenvironment that support and steer male germ cell development,whereas spermatogonia are the primitive male germ cells at the onset of spermatogenesis.While the developmental progression of sertoli cells and spermatogonia has been well established in mice,much less is known in other mammalian species including pigs.Results:To acquire knowledge of sertoli cell and spermatogonial development in pigs,here we collected as many as nine ages of Duroc porcine testes from the neonate to sexual maturity,i.e.,testes from 7-,30-,50-,70-,90-,110-,130-,150-and 210-day-old boars,and performed histological and immunohistochemical analyses on testis sections.We first examined the development of spermatogenic cells and seminiferous tubules in porcine testes.Then,by immunofluorescence staining for marker proteins(AMH,SOX9,DBA,UCHL1,VASA,KIT,Ki67 and/or PCNA),we delved into the proliferative activity and development of sertoli cells and of spermatogonial subtypes(pro-,undifferentiated and differentiating spermatogonia).Besides,by immunostaining forβ-catenin and ZO-1,we studied the establishment of the blood-testis barrier in porcine testes.Conclusions:In this longitudinal study,we have systematically investigated the elaborate sertoli cell and spermatogonial developmental patterns in pigs from the neonate to sexual maturity that have so far remained largely unknown.The findings not only extend the knowledge about spermatogenesis and testicular development in pigs,but also lay the theoretical groundwork for porcine breeding and rearing.

关键词： Pig sertoli cell Spermatogenesis Spermatogonia Testis

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Applications of single-cell RNA sequencing in spermatogenesis and molecular evolution

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Zoological Research 2024年第3期45卷 575-585页

作者： Wen-Bo Chen Meng-Fei Zhang Fan Yang Jin-Lian Hua College of Veterinary Medicine Shaanxi Centre of Stem Cells Engineering&TechnologyNorthwest A&F UniversityYanglingShaanxi 712100China Key Laboratory of Livestock Biology Northwest A&F UniversityYanglingShaanxi 712100China

Spermatogenic cell heterogeneity is determined by the complex process of spermatogenesis differentiation.However,effectively revealing the regulatory mechanisms underlying mammalian spermatogenic cell development and differentiation via traditional methods is difficult.Advances in technology have led to the emergence of many single-cell transcriptome sequencing protocols,which have partially addressed these challenges.In this review,we detail the principles of 10x Genomics technology and summarize the methods for downstream analysis of single-cell transcriptome sequencing data.Furthermore,we explore the role of single-cell transcriptome sequencing in revealing the heterogeneity of testicular ecological niche cells,delineating the establishment and disruption of testicular immune homeostasis during human spermatogenesis,investigating abnormal spermatogenesis in humans,and,ultimately,elucidating the molecular evolution of mammalian spermatogenesis.

关键词： Single-cell RNA sequencing(scRNA-seq) Spermatogenesis Molecular evolution sertoli cell

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Expression of IL-1α, IL-6, TGF-β, FasL and ZNF265 During sertoli cell Infection by Ureaplasma Urealyticum

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cellular & Molecular Immunology 2009年第3期6卷 215-221页

作者： Rongping Li Yebin Xi Xiuzhi Liu Guangjie Chen Baoguo Wang Lihua Jiang Weiyi Li Shanghai Jiao Tong University School of Medicine Department of ImmunoLogy Shanghai Institute of Immunology Shanghai 200025 China

To investigate immunoregulatory mechanisms of sertoli cells in the testis in vitro and in vivo, we utilized our well-characterized Ureaplasma Urealyticum （UU）-induced model. We investigated the expressions of IL-1α, IL-6, TGF-β, FasL and ZNF265 at the first, second and third weeks post-infection. During recovery from inflammation and with the help of negative regulators TGF-β and FasL, the high levels of IL-1α and IL-6 expressions were observed in the early stages of the infection, and decreased gradually in the later weeks both in vitro and in vivo. The trend of varied expression of ZNF265 was similar to those of TGF-β and FasL in vitro and in vivo for sertoli cells infected with UU.

关键词： ZNF265 TGF-β FasL sertoli cell immunoregulation

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17β-Estradiol Regulates Cultured Immature Boar sertoli cell Proliferation via the cAMP-ERK1/2 Pathway and the Estrogen Receptor β

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Agricultural Sciences in China 2010年第8期9卷 1201-1210页

作者： WANG Xian-zhong ZHAO Bo-chuan ZHOU Yu-lan ZHOU Yin-tao MA Kai-ge ZHANG Jia-hua Chongqing Key Laboratory of Forage ＆ Herbivore College of Animal Science and Technology Southwest University Chongqing 400716P.R. China

Estrogen plays an important role in regulating sertoli cell number in the testis. The objective of the study was to identify whether 17β-estradiol affected the proliferation of cultured, immature boar sertoli cells via the estrogen receptor β （ERβ） and the cAMP-extracellular signal-regulated kinase （ERK1/2） pathway. Low levels （10-10-10-8 mol L-1） of 17β-estradiol increased cell number, but high levels （10-7-10-6 mol L-1） decreased it （P〈0.05）. sertoli cell number began to recover for an additional 24 h in the medium without 17β-estradiol （10-6 mol L-l）（P〉0.05）. The effects of 17β-estradiol （10-9 mol L-1） peaked at the first 24 h （P〈0.05）. 17β-estradiol activated ERK1/2 from 5 min to 24 h, but the activiy of ERK1/2 began to decrease after 4 h. Both PD98059 and U0126, two ERK inhibitors, blocked cell division （P〈0.05）. 17β-estradiol （10-10-10-6 mol L-1） dose-dependently increased cAMP production （P 〈 0.05）, and both 17β-estradiol （10-9 mol L-1） and forskolin, which increases cAMP levels, induced cell proliferation and activated ERK1/2 （P〈 0.05）. Rp-cAMP, an antagonist of cAMP, blocked this 17β-estradiol activity （P〈 0.05）. Two estrogen receptor antagonists, ICI 182780 and ERβ antagonist （ERβAnt）, reduced sertoli cell number, cAMP production and ERK1/2 activation （P〈 0.05）, but ERaAnt did not （P〉 0.05）. Therefore, 17β- estradiol mainly promotes pig sertoli cell proliferation via ERβ to induce cAMP production and ERK activation to promote cell proliferation.

关键词： 17β-estradiol sertoli cell cell proliferation estrogen receptor ERK1/2

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Effect of FasL High Expression on Immune Regulative Function of sertoli cell in Transgenic Mouse

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Journal of Reproduction and Contraception 2004年第4期15卷 209-218页

作者： Wei-yi LI Zhao-ping LIU Ye-bin XI Bo LIN Ying-hua MA Jia-hua HU Guang-jie CHEN Bao-guo WANG Shi-san BAO Li-he GUO Shanghai Institute of Immunology Shanghai Second Medical University Shanghai 200025 China Institute of Biochemistry and cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Department of Veterinary Anatomy ＆ Pathology B14 University of Sydney NSW 2006 Australia

Objective To study the immune regulative function of sertoli cell on testis local infection Methods Ureaplasma urealyticum （UU） was directly injected into bladders of FasL transgenic mice and wild-type mice, which mimicked an ascending infectious way. At week 1, 2 and 3 after injection respectively, the mice were killed to observe the pathological alterations in testis section. And at the same time cytokines was tested by immunohistochemistry. Comparison of levels of FasL, TGF-β, IL-1α and IL-6 between UU-infected and control groups of wild mice and FasL transgenic mice was made respectively. Then the capability of sertoli cell （FasL^＋） to mediate apoptosis of Fas^＋ cells between wild control and wild UU-infected groups was analyzed. Results The pathological changes of testis in FasL transgenic mice were more seriously compared with wild counterpart and the changing mode of cytokines secreted by sertoli cells were different between the two kinds of mice. The UU-infected sertoli cells increased Fas^＋ Jurkat cell apoptosis. Conclusions High expression of FasL in FasL transgenic mice can influence the cytokines secretion during anti-infection, thus affecting the testis immune response to infection and immune balance. The high expression of FasL is not beneficial for body＇s anti-inflection immune response.

关键词： sertoli cell immune regulative function FasL TGF-β IL-1α IL-6

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Effects of Ureaplasma Urealyticum on Fas Ligand Expression in Rat sertoli cell

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Journal of Reproduction and Contraception 2000年第3期11卷 128-134页

作者：陈广洁李伟毅席晔斌王保国周葵 ShanghaiInstituteofImmunologyofShanghaiSecondMedicalUniversity Shanghai200025China BasicMedicalCollegeofShanghaiSecondMedicalUniversity Shanghai2000025China

To investigate the effect of ureaplasma urealyticum (UU) on the expression of Fas ligand (FasL) on rat sertoli cell Materials & Method Isolated rat sertoli cells were infected by living UU, UU super- natants, inactivated UU, then Fluorescence Activated cell Sorter and observed fluores- cence microscopy were used to assay for the FasL expression on the surface of sertoli cells. Results UU infection could increase the expression of FasL in sertoli cell. Conclusion The functional expression of FasL is related to the immune privilege and can give the immune regulation on the testis.

关键词： sertoli cell Fas Ligand (FasL) Ureaplasma urealyticum (UU) Immune privilege Immune regulation

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硼对大鼠睾丸sertoli cell及种公猪精液品质的影响

硼对大鼠睾丸Sertoli Cell及种公猪精液品质的影响

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作者：张倩安徽科技学院

学位级别：硕士

本文研究硼对大鼠睾丸sertoli cell及种公猪精液品质的影响。试验首先选用18-22日龄雄性SD大鼠,分离sertoli cell,经过免疫荧光进行细胞鉴定,通过HE染色、细胞活性试验、流式细胞术研究不同浓度硼对sertoli cell形态、活性、增殖与调亡... 详细信息

本文研究硼对大鼠睾丸sertoli cell及种公猪精液品质的影响。试验首先选用18-22日龄雄性SD大鼠,分离sertoli cell,经过免疫荧光进行细胞鉴定,通过HE染色、细胞活性试验、流式细胞术研究不同浓度硼对sertoli cell形态、活性、增殖与调亡的影响,试验分为对照组、0.25、0.5、1.0、5.0、10.0、40.0、80.0 mmoL/L 8个梯度硼试验组。进而研究雌激素阻断剂特异性阻断雌激素受体,探究硼是否通过ER来影响sertoli cell的增殖、凋亡及相关蛋白、基因的表达量,以及sertoli cell分泌的细胞因子含量,主要采用流式细胞术、Western Blot、荧光定量PCR、Elisa等方法检测相关指标。然后试验再选用30头健康性成熟的种公猪,随机分为3组,每组10个重复,试验组分别给其饮水中添加5 mg/L和10 mg/L硼浓度,对照组常规饲料饲喂,饲喂30 d后进行采集精液和采血,观察并检测不同硼水平下猪精液品质的影响及血清中激素水平含量。试验结果如下:1、硼对sertoli cell凋亡、增殖的影响通过改进sertoli cell的分离、培养方法,采用免疫荧光方法对sertoli cell进行鉴定,结果显示,所分离培养的细胞为sertoli cell,经显微镜下计数其纯度达95%以上。此方法简便了分离培养sertoli cell的步骤,同时提高了 sertoli cell的纯度和细胞数量。硼对sertoli cell凋亡、增殖的影响,与对照组相比,硼处理浓度低于5.0 mmol/L,可促进sertoli cell增殖并抑制其凋亡,增加细胞周期的S、G2/M增殖期细胞比例,硼处理浓度高于5 mmol/L,对sertoli cell有明显毒性作用,细胞凋亡率升高,细胞周期的增殖被完全阻滞在G0/G1期,细胞周期中增殖期的细胞比例明显减少。2、硼对ER阻断前后sertoli cell凋亡、增殖的影响试验组分为空白对照组、0.5、40 mmol/L硼处理组和加ER阻断剂的对照组、加ER阻断剂的0.5 mmol/L和加ER阻断剂的40 mmol/L硼处理组。分别采用流式细胞仪、Western Blot、荧光定量PCR方法检测。结果显示,硼对ER阻断前后sertoli cell凋亡、增殖的影响,进一步证实了低浓度硼（0.5 mmol/L）有利于减少sertoli cell的凋亡,提高sertoli cell的增殖,高浓度硼（40 mmol/L）促进sertoli cell的凋亡;硼对添加MPP阻断剂的sertoli cell凋亡有一定的影响,在一定程度上阻断了硼对ERα通路上sertoli cell凋亡作用,但对细胞的增殖作用不显著;硼对添加PHTPP阻断剂处理的sertoli cell凋亡、增殖有显著性作用,硼可能通过ERβ通路对sertoli cell的凋亡、增殖起到调控作用;硼对添加G15阻断剂的sertoli cell的增殖有一定的影响,但是对GPR30通路上的凋亡、增殖基因和蛋白并没有完全阻断,硼可能通过GPR30通路调节sertoli cell增殖、凋亡,尤其是对sertoli cell的增殖。3、硼对ER阻断前后sertoli cell细胞因子的影响与空白对照组相比,低浓度硼（0.5 mmol/L）有利于提高sertoli cell细胞因子的含量,高浓度硼（40 mmol/L）可以显著降低sertoli cell的细胞因子的表达量。硼对ER阻断后sertoli cell细胞因子的影响结果是,硼对加了 PHTPP和G15阻断剂的ERβ和GPR30通路的sertoli cell细胞因子有一定的影响,分别对sertoli cell中与生殖相关的ABP、ER、TRF细胞因子的表达量有显著的调控作用。4、硼对种公猪精液品质的影响添加0.08和0.16 mmoL/L的硼可以显著增加种公猪精子活力,有效精子数和精子密度,在一定程度上提高了种公猪A、B级精液的水平;也提高了种公猪血清促黄体素（LH）含量,分别显著提高了 53.82%、330.83%,其中0.16 mmoL/L可以显著提高睾酮（T）含量,提高了 207.62%（P＜0.05）。

关键词： SD大鼠种公猪精液品质硼 sertoli cell 雌激素通路增殖与凋亡细胞因子

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Low XIST expression in sertoli cells of Klinefelter syndrome patients causes high susceptibility of these cells to an extra X chromosome

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Asian Journal of Andrology 2023年第6期25卷 662-673页

作者： Liang-Yu Zhao Peng Li Chen-Cheng Yao Ru-Hui Tian Yu-Xin Tang Yu-Zhuo Chen Zhi Zhou Zheng Li Department of Andrology The Center for Men’s HealthUrologic Medical CenterShanghai Key Laboratory of Reproductive MedicineShanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China Department of Urology The Fifth Affiliated Hospital of Sun Yat-sen UniversityZhuhai 519000China Department of Interventional Medicine Guangdong Provincial Key Laboratory of Biomedical ImagingThe Fifth Affiliated Hospital of Sun Yat-sen UniversityZhuhai 519000China School of Life Science and Technology ShanghaiTech UniversityShanghai 200120China

Klinefelter syndrome(KS)is the most common genetic cause of human male infertility.However,the effect of the extra X chromosome on different testicular cell types remains poorly understood.Here,we profiled testicular single-cell transcriptomes from three KS patients and normal karyotype control individuals.Among the different somatic cells,sertoli cells showed the greatest transcriptome changes in KS patients.Further analysis showed that X-inactive-specific transcript(XIST),a key factor that inactivates one X chromosome in female mammals,was widely expressed in each testicular somatic cell type but not in sertoli cells.The loss of XIST in sertoli cells leads to an increased level of X chromosome genes,and further disrupts their transcription pattern and cellular function.This phenomenon was not detected in other somatic cells such as Leydig cells and vascular endothelial cells.These results proposed a new mechanism to explain why testicular atrophy in KS patients is heterogeneous with loss of seminiferous tubules but interstitial hyperplasia.Our study provides a theoretical basis for subsequent research and related treatment of KS by identifying sertoli cell-specific X chromosome inactivation failure.

关键词： Klinefelter syndrome nonobstructive azoospermia sertoli cell spermatogenesis X chromosome inactivation

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Infection of SARS-CoV-2 causes severe pathological changes in mouse testis

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Journal of Genetics and Genomics 2023年第2期50卷 99-107页

作者： Min Chen Shihua Li Shujun Liu Yuhang Zhang Xiuhong Cui Limin Lv Bowen Liu Aihua Zheng Qihui Wang Shuguang Duo Fei Gao State Key Laboratory of Stem cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China Institute for Stem cell and Regeneration Chinese Academy of SciencesBeijing 100101China Beijing Institute for Stem cell and Regenerative Medicine Beijing 100101China University of Chinese Academy of Sciences Beijing 100101China CAS Key Laboratory of Pathogen Microbiology and Immunology Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China Institute of Zoology Chinese Academy of SciencesBeijing 100101China State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of ZoologyChinese Academy of SciencesBeijing 100101China CAS Center for Excellence in Biotic Interactions University of Chinese Academy of SciencesBeijing 100101China

Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARSCoV-2),has affected more than 600 million people worldwide.Several organs including lung,intestine,and brain are infected by SARS-CoV-2.It has been reported that SARS-CoV-2 receptor angiotensin-converting enzyme-2(ACE2)is expressed in human testis.However,whether testis is also affected by SARS-CoV-2 is still unclear.In this study,we generate a human ACE2(hACE2)transgenic mouse model in which the expression of hACE2 gene is regulated by hACE2 promoter.sertoli and Leydig cells from hACE2 transgenic mice can be infected by SARS-CoV-2 pseudovirus in vitro,and severe pathological changes are observed after injecting the SARS-CoV-2 pseudovirus into the seminiferous tubules.Further studies reveal that sertoli and Leydig cells from hACE2 transgenic mice are also infected by authentic SARS-CoV-2 virus in vitro.After testis interstitium injection,authentic SARS-CoV-2 viruses are first disseminated to the interstitial cells,and then detected inside the seminiferous tubules which in turn cause germ cell loss and disruption of seminiferous tubules.Our study demonstrates that testis is most likely a target of SARS-CoV-2 virus.Attention should be paid to the reproductive function in SARS-CoV-2 patients.

关键词： SARS-CoV-2 ACE2 sertoli cell Leydig cell Testis

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支柱细胞(sertoli cell)——间质细胞(Leydigcell)在睾丸功能调节中的相互作用

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西安交通大学学报(医学版) 1983年第1期 119-120页

作者：张德馨

间质细胞是睾酮的主要来源,并局部定位于管间组织,保证睾酮的局部性高浓度,这是完成精子发生所必需的条件之一。另一方面,尽管支柱细胞与生殖细胞有着紧密联系,但是这种细胞影响精子发生过程的方式尚知之甚少。现已确认支柱细胞对FSH起... 详细信息

间质细胞是睾酮的主要来源,并局部定位于管间组织,保证睾酮的局部性高浓度,这是完成精子发生所必需的条件之一。另一方面,尽管支柱细胞与生殖细胞有着紧密联系,但是这种细胞影响精子发生过程的方式尚知之甚少。现已确认支柱细胞对FSH起反应,产生雄激素结合蛋白(ABP)、抑制素(inhibin)和液体,并且组成血一睾屏障。

关键词：间质细胞功能 sertoli cell Leydigcell 支柱

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