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MKK7-mediated phosphorylation of JNKs regulates the proliferation and apoptosis of human spermatogonial stem cells

World Journal of Stem Cells 2021年第11期13卷 1797-1812页

作者： Zeng-Hui Huang Chuan Huang Xi-Ren Ji Wen-Jun Zhou Xue-Feng Luo Qian Liu Yu-Lin Tang Fei Gong Wen-Bing Zhu Institute of reproductive and Stem Cell Engineering Central South UniversityChangsha 410008Hunan ProvinceChina Department of reproductive Center Reproductive and Genetic Hospital of CITIC-XiangyaChangsha 410008Hunan ProvinceChina Department of sperm bank Reproductive and Genetic Hospital of CITICXiangyaChangsha 410008Hunan ProvinceChina

BACKGROUND Human spermatogonial stem cells(SSCs)are the basis of ***,little is known about the developmental regulatory mechanisms of SSC due to sample origin and species *** To investigates the mechanisms involved in the proliferation of human *** The expression of mitogen-activated protein kinase kinase 7(MKK7)in human testis was identified using immunohistochemistry and western blotting(WB).MKK7 was knocked down using small interfering RNA,and cell proliferation and apoptosis were detected by WB,EdU,cell counting kit-8 and fluorescenceactivated cell *** bioinformatic analysis,the interaction of MKK7 with c-Jun N-terminal kinases(JNKs)was verified by protein co-immunoprecipitation and *** phosphorylation of JNKs was inhibited by SP600125,and the phenotypic changes were detected by WB,cell counting kit-8 and fluorescenceactivated cell *** MKK7 is mainly expressed in human SSCs,and MKK7 knockdown inhibits SSC proliferation and promotes their ***7 mediated the phosphorylation of JNKs,and after inhibiting the phosphorylation of JNKs,the phenotypic changes of the cells were similar to those after MKK7 *** expression of MKK7 was significantly downregulated in patients with abnormal spermatogenesis,suggesting that abnormal MKK7 may be associated with spermatogenesis *** MKK7 regulates the proliferation and apoptosis of human SSC by mediating the phosphorylation of JNKs.

关键词： MKK7 spermatogonial stem cell Proliferation Apoptosis JNKs

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Exploring the impact of pericentric inversion of chromosome 9 on fertility in sperm donors

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Asian Journal of Andrology 2024年第3期26卷 302-307页

作者： Qian Liu Wen-Jun Zhou Zeng-Hui Huang Xiu-Hai Huang Jian Wu Xi-Ren Ji Xue-Feng Luo Yu-Ling Tang Rui-Jun Wang Li-Qing Fan Chuan Huang Wen-Bing Zhu Institute of reproductive and Stem Cell Engineering Basic Medicine CollegeCentral South UniversityChangsha 410013China Clinical Research Center for Reproduction and genetics in Hunan Province Reproductive and Genetic Hospital of CITIC-XiangyaChangsha 410005China

Pericentric inversion of chromosome 9(inv[9])is a common chromosomal structural variant,but its impact on clinical outcomesremains *** screening criteria of sperm banks are rarely mentioned to individuals with inv(9).In this study,we evaluatedthe fertility of sperm donors with inv(9)who met eligibility criteria for sperm banks(inv[9]-eligible donors).From March 2004 toMay 2022,chromosomal analysis of 16124 sperm donors at citic-xiangya Human sperm bank in Hunan Province(Changsha,China)found that 251(1.6%)had chromosome variations,with inv(9)being the most prevalent at 1.1%.All 169 inv(9)-eligibledonors were contacted to collect fertility outcome data,along with 206 eligible donors without inv(9)as *** addition,semen samples from inv(9)-eligible donors and eligible donors underwent assessments of sperm fluorescence in situ hybridization(FISH),mitochondrial membrane potential,DNA fragmentation index,acrosome integrity,reactive oxygen species(ROS),andsperm *** showed that inv(9)did not significantly increase reproductive risks *** detecting ROSlevel differences,the clinical impact may be *** study provides new data on the inv(9)population that can serveas a valuable reference for decision-making by sperm banks as well as for genetic counseling and clinical guidance for individualscarrying inv(9)variant.

关键词： male fertility pericentric inversion of chromosome 9 sperm bank sperm functional parameters

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sperm flagellar 2(SPEF2)is essential for sperm flagellar assembly in humans

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Asian Journal of Andrology 2022年第4期24卷 359-366页

作者： Dong-Yan Li Xiao-Xuan Yang Chao-Feng Tu Wei-Li Wang Lan-Lan Meng Guang-Xiu Lu Yue-Qiu Tan Qian-Jun Zhang Juan Du Institute of reproductive and Stem Cell Engineering School of Basic Medical ScienceCentral South UniversityChangsha 410078China Clinical Research Center for Reproduction and genetics in Hunan Province Reproductive and Genetic Hospital of CITIC-XiangyaChangsha 410078China

spermiogenesis is a complex and tightly regulated process,consisting of acrosomal biogenesis,condensation of chromatin,flagellar assembly,and disposal of extra *** studies have reported that sperm flagellar 2(SPEF2)deficiency causes severe asthenoteratozoospermia owing to spermiogenesis failure,but the underlying molecular mechanism in humans remains ***,we performed proteomic analysis on spermatozoa from three SPEF2 mutant patients to study the functional role of SPEF2 during sperm tail development.A total of 1262 differentially expressed proteins were detected,including 486 upregulated and 776 *** constructed heat map of the differentially expressed proteins showed similar *** these,the expression of proteins related to flagellar assembly,including SPEF2,sperm associated antigen 6(SPAG6),dynein light chain tctex-type 1(DYNLT1),radial spoke head component 1(RSPH1),translocase of outer mitochondrial membrane 20(TOM20),EF-hand domain containing 1(EFHC1),meiosis-specific nuclear structural 1(MNS1)and intraflagellar transport 20(IFT20),was verified by western *** clustering analysis indicated that these differentially expressed proteins were specifically enriched for terms such as spermatid development and flagellar ***,we showed that SPEF2 interacts with radial spoke head component 9(RSPH9)and IFT20 in vitro,which are well-studied components of radial spokes or intra-flagellar transport and are essential for flagellar *** results provide a rich resource for further investigation into the molecular mechanism underlying the role that SPEF2 plays in sperm tail development and could provide a theoretical basis for gene therapy in SPEF2 mutant patients in the future.

关键词： SPEF2 flagellar assembly male infertility protein interaction proteomics

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Update on techniques for cryopreservation of human spermatozoa

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Asian Journal of Andrology 2022年第6期24卷 563-569页

作者： Chuan Huang Yu-Lin Tang Jian-Ling Hu Wen-Jun Zhou Zeng-Hui Huang Xue-Feng Luo Zheng Li Wen-Bing Zhu Institute of reproductive and Stem Cell Engineering Basic Medicine CollegeCentral South UniversityChangsha 410000China reproductive and genetic hospital of citic-xiangya Changsha 410000China Department of Andrology Center for Men's HealthShanghai General HospitalShanghai Key Lab of Reproductive MedicineShanghai 200025China

In the 1960s,sperm cryopreservation was developed as a method to preserve ***,techniques for the cryopreservation of human spermatozoa have been widely used in assisted ***,although sperm cryobiology has made notable achievements,the optimal method for the recovery of viable spermatozoa after cryopreservation remains *** sperm quality can be affected by cryoprotectants,ice formation,storage conditions,and osmotic stress during the freezing *** review discusses recent advances in different cryopreservation techniques,cryoprotectants,and freezing and thawing methods during cryopreservation and new indications for the use of cryopreserved spermatozoa.

关键词： cryodamage cryoprotectant dry preservation freezing and thawing sperm cryopreservation

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Influence of sperm morphology on pregnancy outcome and offspring in in vitro fertilization and intracytoplasmic sperm injection:a matched case-control study

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Asian Journal of Andrology 2021年第4期23卷 421-428页

作者： Wen-Jun Zhou Chuan Huang Su-Hua Jiang Xi-Ren Ji Fei Gong Li-Qing Fan Wen-Bing Zhu Institute of reproductive and Stem Cell Engineering Basic Medicine CollegeCentral South UniversityChangsha 410008China reproductive and genetic hospital of citic-xiangya Changsha 410008China Clinical Research Center for Reproduction and genetics in Hunan Province Changsha 410008China

sperm morphology was once believed as one of the most predictive indicators of pregnancy outcome in assisted reproductive technology(ART).However,the impact of teratozoospermia on in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)outcomes and its offspring remains *** order to evaluate the influence of teratozoospermia on pregnancy outcome and newborn status after IVF and ICSI,a retrospective study was *** was a matched case-control study that included 2202 IVF cycles and 2574 ICSI cycles and was conducted at the reproductive and genetic hospital of citic-xiangya in Changsha,China,from June 2013 to June *** were divided into two groups based on sperm morphology:teratozoospermia and normal sperm *** pregnancy outcome and newborn outcome were *** results indicated that couples with teratozoospermia had a significantly lower optimal embryo rate compared to those with normal sperm morphology in IVF(P=0.007),while there were no statistically significant differences between the two groups in terms of the fertilization rate,cleavage rate,implantation rate,and pregnancy rate(all P>0.05).Additionally,teratozoospermia was associated with lower infant birth weight in multiple births after *** regard to ICSI,there was no significant difference in both pregnancy outcome and newborn outcome between the teratozoospermia and normal groups(both P>0.05).Furthermore,no increase in the risk of birth defects occurred in the teratozoospermia group after IVF/***,we believe that teratozoospermia has limited predictive value for pregnancy outcomes in IVF/ICSI,and has little impact on the resulting offspring if multiple pregnancy is avoided.

关键词： in vitro fertilization intracytoplasmic sperm injection newborn outcome pregnancy outcome teratozoospermia

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In sight on multiple morphological abno rmalities of sperm flagella in male infertility:what is new?

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Asian Journal of Andrology 2020年第3期22卷 236-245页

作者： Wei-Li Wang Chao-Feng Tu Yue-Qiu Tan Institute of reproductive and Stem Cell Engineering School of Basic Medical ScienceCentral South UniversityChangsha 410078China reproductive and genetic hospital of citic-xiangya Changsha 410078China National Engineering and Research Center of Human Stem Cell Changsha 410078China

The syndrome of multiple morphological abnormalities of the sperm flagella(MMAF)is a specific kind of asthenoteratozoospermia with a mosaic of flagellar morphological abnormalities(absent,short,bent,coiled,and irregular flagella).MMAF was proposed in 2014 and has attracted increasing attention;however,it has not been clearly *** this review,we elucidate the definition of MMAF from a systematical view,the difference between MMAF and other conditions with asthenoteratozoospermia or asthenozoospermia(such as primary mitochondrial sheath defects and primary ciliary dyskinesia),the knowledge regarding its etiological mechanism and related genetic findings,and the clinical significance of MMAF for intracytoplasmic sperm injection and genetic coun sell *** review provides the basic kno wledge for MMAF and puts forward some suggestions for further investigations.

关键词： asthe no teratozoospermia disease-causing ge nes in tracytoplasmic sperm in jecti on male in fertility multiple morphological abnormalities of the sperm flagella

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Novel deletion mutations of the PIH1D3 gene in an infertile young man with primary ciliary dyskinesia and his cousin with Kartagener’s syndrome

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Asian Journal of Andrology 2021年第3期23卷 330-332页

作者： Chuan Huang Na-Chuan Liu Xiao-Bo Wang Ben-Hong Gu Jian-Xiong Zhang Li-Zhang Zheng Li Department of Andrology Center for Men's HealthShanghaiChina Department of ART Institute of Urologic Medical CenterShanghai General HospitalShanghaiKey Lab of Reproductive MedicineShanghai Jiao Tong UniversityShanghai 200080China The Human sperm bank Reproductive and Genetic Hospital of CITIC-XiangyaChangsha 410000China Department of Urology Tongren HospitalShanghai Jiao Tong UniversitySchool of MedicineShanghai 200080China.

Dear Editor,Male infertility is defined as the inability to achieve pregnancy in a fertile female partner even after 12 months of unprotected *** humans,it accounts for 40%-50%of cases of infertility,1 and it occurs most often due to problems with sperm production or sperm *** infertility is commonly due to deficiencies in semen,and semen quality is used as a surrogate measure of male *** ciliary dyskinesia(PCD)is a rare cause of male infertility.2,3 It is an autosomal recessive genetic condition that causes abnormal function of *** are microscopic finger-like projections on the surface of the cells.

关键词： partner protected cilia

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A novel homozygous frameshift mutation in MNS1 associated with severe oligoasthenoteratozoospermia in humans

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Asian Journal of Andrology 2021年第2期23卷 197-204页

作者： Yong Li Wei-Li Wang Chao-Feng Tu Lan-Lan Meng Tong-Yao Hu Juan Du Ge Lin Hong-Chuan Nie Yue-Qiu Tan Institute of reproductive and Stem Cell Engineering School of Basic Medical ScienceCentral South UniversityChangsha 410078China reproductive and genetic hospital of citic-xiangya Changsha 410078China Clinical Research Center for Reproduction and genetics in Hunan Province Changsha 410078China

Oligoasthenoteratozoospermia(OAT)refers to the combination of various sperm abnormalities,including a decreased sperm count,reduced motility,and abnormal sperm *** a few genetic causes have been shown to be associated with ***,we identified a novel homozygous frameshift mutation in meiosis-specific nuclear structural 1(MNS1;NM_018365:c.603_604insG:***202Glufs*6)by whole-exome sequencing in an OAT proband from a consanguineous Chinese *** variant screening identified four additional heterozygous MNS1 variants in 6/219 infertile individuals with oligoasthenospermia,but no MNS1 variants were observed among 223 fertile *** analysis showed MNS1 to be normally located in the whole-sperm flagella,but was absent in the proband's *** analysis by Western blot also confirmed that MNS1 was absent in the proband's *** flagellum morphology and ultrastructural disturbances in outer doublet microtubules were observed in the proband's sperm.A total of three intracytoplasmic sperm injection cycles were carried out for the proband's wife,but they all failed to lead to a successful ***,this is the first study to report a loss-of-function mutation in MNS1 causing OAT in a Han Chinese patient.

关键词： intracytoplasmic sperm injection male infertility meiosis-specific nuclear structural 1(MNS1) oligoasthenoteratozoospermia whole-exome sequencing

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Improving native human sperm freezing protection by using a modified vitrification method

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Asian Journal of Andrology 2021年第1期23卷 91-96页

作者： Dai Zhou Xing-Ming Wang Rui-Xue Li Yi-Ze Wang Yuan-Chi Chao Zhi-Zhong Liu Zeng-Hui Huang Hong-Chuan Nie Wen-Bing Zhu Yue-Qiu Tan Li-Qing Fan Hnstitute of Reproduction and Stem Cell Engineering School of Basic Medicine ScienceCentral South UniversityChangsha 410000China reproductive and genetic hospital of citic-xiangya Changsha 410000China Department of Urology Hunan Cancer HospitalChangsha 410000China

Slow freezing is the most commonly used technique for the cryopreservation of spermatozoa in clinical ***,it has been shown to have a negative impact on sperm function and *** as a successful alternative method has been proved to have better protective effects on human embryos,but vitrification of spermatozoa is still subject to low recovery *** this study,a modified vitrification method for native spermatozoa was developed.A total of 28 semen samples were included;each sample was divided into three equal parts and assigned to fresh,slow freezing,and vitrification *** vitality,motility,morphology,DNA integrity,and acrosome reaction were assessed for each of the *** results showed that vitrification achieves better results for several sperm protection parameters than slow freezing;vitrification achieves a higher recovery rate(Pspermatozoa after *** component analysis of all sperm parameters revealed that the vitrification cluster was closer to the fresh cluster,indicating that spermatozoa are better preserved through *** conclusion,while both slow freezing and vitrification have negative effects on sperm function and structure,the vitrification protocol described here had a relatively better recovery rate(65.8%)and showed improved preservation of several sperm quality parameters compared with slow freezing.

关键词： cryopreservation cryoprotectant slow freezing sperm vitrification

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Defects in phospholipase C zeta cause polyspermy and low fertilization after conventional IVF:not just ICSI failure

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Asian Journal of Andrology 2024年第2期26卷 175-182页

作者： Jian-Fang Che Hui-Xia Wu Si-Cong Zeng Yue-Ren Wu Jing Dai De-Hua Cheng Fei Gong Guang-Xiu Lu Ge Lin Can Dai School of Medicine Hunan Normal UniversityChangsha 410013China reproductive and genetic hospital of citic-xiangya Changsha 410008China Clinical Research Center for Reproduction and genetics in Hunan Province Changsha 410078China National Engineering and Research Center of Human Stem Cell Changsha 410205China

Phospholipase C zeta(PLC)is a key sperm-borne oocyte-activating factor that triggers Ca^(2+)oscillations and the subsequent block to polyspermy following gamete *** in PLCZ1,the gene encoding PLCζ,cause male infertility and intracytoplasmic sperm injection(ICSI)fertilization failure;and PLCζ expression and localization patterns are significantly correlated with ICSI fertilization rate(FR).However,in conventional in vitro fertilization(cIVF),whether and how sperm PLCζ affects fertilization remain ***,we identified one previously reported and two novel PLCZ1 mutations associated with polyspermy in vitro that are characterized by excessive sperm-zona binding and a delay in pronuclei(PN)*** staining and oocyte activation testing revealed that virtually all spermatozoa from patients lacked functional PLCζ and were thus unable to evoke Ca^(2+) *** with an artificial oocyte activation treatment successfully rescued the polyspermic phenotype and resulted in a live ***,we analyzed PLCζ in an additional 58 males after cIVF treatment in the reproductive and genetic hospital of citic-xiangya(Changsha,China)between February 2019 and January *** found that the proportion of spermatozoa that expressed PLCζ was positively correlated with both 2PN rate and total *** optimal cutoff value below which males were likely to experience low FR(total FR≤30%)after clVF was 56.7%for the proportion of spermatozoa expressing *** study expands the mutation and the phenotypic spectrum of PLCZ1 and further suggests that PLCζ constitutes a promising biomarker for identifying low FRs cases in cIVF due to sperm-related oocyte activation deficiency and that sperm PLCζ analysis may benefit the widermale population and not onlymen with IcsI failure.

关键词： conventional IVF gene variants PLCζ polyspermy

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