Male infertility caused by spermiogenic defects: Lessons from gene knockouts

doi:10.1016/j.mce.2009.03.003

Molecular and Cellular Endocrinology

Volume 306, Issues 1–2, 10 July 2009, Pages 24-32

https://doi.org/10.1016/j.mce.2009.03.003 Get rights and content

Abstract

Spermiogenesis refers to the process by which postmeiotic spermatids differentiate into elongated spermatids and eventually spermatozoa. During spermiogenesis, round spermatids undergo dynamic morphologic changes, which include nuclear condensation and elongation, formation of flagella and acrosome, reorganization of organelles and elimination of cytoplasm upon spermiation. This cellular differentiation process is unique to male haploid germ cells, which may explain why ∼half of the testis-specific genes are exclusively expressed in spermiogenesis. The spermiogenesis-specific expression implies that these genes contribute to either structural or functional aspects of future sperm. Many such genes have been inactivated in mice and some of these gene knockout mice display male infertility due to nonfunctional sperm which display no or various degrees of structural abnormalities. Since the majority of these spermiogenesis-specific genes are highly conserved between mice and humans, findings from knockout mouse studies may be applicable to human infertility. Here, I briefly review some of these spermatid-specific gene knockouts. The mouse studies strongly suggest that sperm quality rather than quantity is a better indicator of male fertility and novel assays should be developed to determine sperm functionality.

Section snippets

Spermiogenesis is unique to male germ cell development and many of the testis-specific genes are expressed in spermiogenesis

Spermatogenesis is a complex differentiation process through which highly specialized haploid spermatozoa are differentiated from diploid germline stem cells (Clermont, 1972). The developmental process begins in the basal compartment of the seminiferous epithelium with spermatogonia proliferating by mitosis and differentiating into spermatocytes. Since mitotic multiplication of spermatogonia is the main event in this phase, it has been termed the mitotic phase. Primary spermatocytes then

Genes required for acrosome formation

The acrosome is a structure attached to the nucleus of a mature spermatozoon (Fig. 2). The acrosome contains proteolytic enzymes which are released to dissolve zona pellucida proteins during fertilization. Acrosome formation involves three consecutive phases: the Golgi phase, also called the granule phase, starts when numerous pro-acrosomic granules are formed in trans-Golgi stacks followed by a fusion into a single, large acrosomic granule that associates with the nuclear envelope (Abou-Haila

What do these knockout mice tell us?

Since virtually all spermiogenic genes are dedicated for transforming round spermatids into spermatozoa, it is not surprising that structural and functional defects arise in the absence of these genes. The knockout mouse lines discussed above show many commonalities, which are of importance for us to understand the mechanisms underlying the regulation of haploid male germ cell development. Some of the findings suggest that more sensitive assays should be developed to efficiently detect subtle

Acknowledgements

David Young is acknowledged for editing the text and comments. I apologize to colleagues whose work is not cited here due to publisher's space limit. The research in the Yan Laboratory is supported by NIH grants HD048855 and HD050281, as well as a startup grant from the University of Nevada, Reno.

References (140)

A. Abou-Haila et al.
Mammalian sperm acrosome: formation, contents, and function
Arch. Biochem. Biophys.
(2000)
R.F. Burk et al.
Selenoprotein metabolism and function: evidence for more than one function for selenoprotein P
J. Nutr.
(2003)
A.E. Carlson et al.
Identical phenotypes of CatSper1 and CatSper2 null sperm
J. Biol. Chem.
(2005)
M.A. Carmell et al.
MIWI2 is essential for spermatogenesis and repression of transposons in the mouse male germline
Dev. Cell
(2007)
A.H. Dam et al.
Homozygous mutation in SPATA16 is associated with male infertility in human globozoospermia
Am. J. Hum. Genet.
(2007)
W. Deng et al.
Miwi, a murine homolog of piwi, encodes a cytoplasmic protein essential for spermatogenesis
Dev. Cell
(2002)
A.S. Edwards et al.
A-kinase anchoring proteins: protein kinase A and beyond
Curr. Opin. Cell Biol.
(2000)
D.W. Fawcett
The mammalian spermatozoon
Dev. Biol.
(1975)
M. Hampsey et al.
RNA polymerase II as a control panel for multiple coactivator complexes
Curr. Opin. Genet. Dev.
(1999)
K.D. Hinsch et al.
Voltage-dependent anion-selective channels VDAC2 and VDAC3 are abundant proteins in bovine outer dense fibers, a cytoskeletal component of the sperm flagellum
J. Biol. Chem.
(2004)

N. Iguchi et al.

Molecular cloning of haploid germ cell-specific tektin cDNA and analysis of the protein in mouse testis

FEBS Lett.

(1999)

S. Kashiwabara et al.

Identification of a novel isoform of poly(A) polymerase, TPAP, specifically present in the cytoplasm of spermatogenic cells

Dev. Biol.

(2000)

S.M. King

The dynein microtubule motor

Biochim. Biophys. Acta

(2000)

S. Kuramochi-Miyagawa et al.

Two mouse piwi-related genes: miwi and mili

Mech. Dev.

(2001)

L. Lalonde et al.

Male infertility associated with round-headed acrosomeless spermatozoa

Fertil. Steril.

(1988)

M. Larsson et al.

The spatial and temporal expression of Tekt1, a mouse tektin C homologue, during spermatogenesis suggest that it is involved in the development of the sperm tail basal body and axoneme

Eur. J. Cell Biol.

(2000)

I. Martianov et al.

Late arrest of spermiogenesis and germ cell apoptosis in mice lacking the TBP-like TLF/TRF2 gene

Mol. Cell

(2001)

K. Miki et al.

Targeted disruption of the Akap4 gene causes defects in sperm flagellum and motility

Dev. Biol.

(2002)

M.A. Motsenbocker et al.

Effect of dietary selenium on plasma selenoprotein P, selenoprotein P1 and glutathione peroxidase in the rat

J. Nutr.

(1984)

L.I. Neilson et al.

cDNA cloning and characterization of a human sperm antigen (SPAG6) with homology to the product of the Chlamydomonas PF16 locus

Genomics

(1999)

J. O’Brien et al.

Sperm DNA integrity and male infertility

Urology

(2005)

R.J. Aitken et al.

Analysis of sperm function in globozoospermia: implications for the mechanism of sperm–zona interaction

Fertil. Steril.

(1990)

A. Aravin et al.

A novel class of small RNAs bind to MILI protein in mouse testes

Nature

(2006)

C.P. Austin et al.

The knockout mouse project

Nat. Genet.

(2004)

S.A. Beyler et al.

Binding of antibodies against antigenic domains of murine lactate dehydrogenase-C4 to human and mouse spermatozoa

Biol. Reprod.

(1985)

J.A. Blendy et al.

Severe impairment of spermatogenesis in mice lacking the CREM gene

Nature

(1996)

P.H. Boer et al.

The testis-specific phosphoglycerate kinase gene pgk-2 is a recruited retroposon

Mol. Cell Biol.

(1987)

P.R. Brown et al.

A-kinase anchoring protein 4 binding proteins in the fibrous sheath of the sperm flagellum

Biol. Reprod.

(2003)

D.O. Bunch et al.

Glyceraldehyde 3-phosphate dehydrogenase-S protein distribution during mouse spermatogenesis

Biol. Reprod.

(1998)

R.F. Burk et al.

Selenoprotein P: an extracellular protein with unique physical characteristics and a role in selenium homeostasis

Annu. Rev. Nutr.

(2005)

C. Cho et al.

Protamine 2 deficiency leads to sperm DNA damage and embryo death in mice

Biol. Reprod.

(2003)

C. Cho et al.

Haploinsufficiency of protamine-1 or -2 causes infertility in mice

Nat. Genet.

(2001)

Y. Clermont

Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal

Physiol. Rev.

(1972)

A. Cochrane

Controlling HIV-1 Rev function

Curr. Drug Targets Immune Endocr. Metabol. Disord.

(2004)

T.G. Cooper

Cytoplasmic droplets: the good, the bad or just confusing?

Hum. Reprod.

(2005)

D.N. Cox et al.

A novel class of evolutionarily conserved genes defined by piwi are essential for stem cell self-renewal

Genes Dev.

(1998)

D.N. Cox et al.

Piwi encodes a nucleoplasmic factor whose activity modulates the number and division rate of germline stem cells

Development

(2000)

J.P. Dadoune

The cellular biology of mammalian spermatids: a review

Bull. Assoc. Anat. (Nancy)

(1994)

J. Drews

Drug discovery: a historical perspective

Science

(2000)

S.K. Dutcher et al.

Genetic dissection of the central pair microtubules of the flagella of Chlamydomonas reinhardtii

J. Cell Biol.

(1984)

E.M. Eddy

Male germ cell gene expression

Recent Prog. Horm. Res.

(2002)

E.M. Eddy et al.

Fibrous sheath of mammalian spermatozoa

Microsc. Res. Tech.

(2003)

D. Escalier et al.

Spermatogenesis of mice lacking CK2alpha’: failure of germ cell survival and characteristic modifications of the spermatid nucleus

Mol. Reprod. Dev.

(2003)

G. Esposito et al.

Mice deficient for soluble adenylyl cyclase are infertile because of a severe sperm-motility defect

Proc. Natl. Acad. Sci. U.S.A.

(2004)

N.S. Foulkes et al.

Developmental switch of CREM function during spermatogenesis: from antagonist to activator

Nature

(1992)

L.R. Fraser

Dibutyryl cyclic AMP decreases capacitation time in vitro in mouse spermatozoa

J. Reprod. Fertil.

(1981)

A. Girard et al.

A germline-specific class of small RNAs binds mammalian Piwi proteins

Nature

(2006)

U.W. Goodenough et al.

Substructure of inner dynein arms, radial spokes, and the central pair/projection complex of cilia and flagella

J. Cell Biol.

(1985)

S.T. Grivna et al.

A novel class of small RNAs in mouse spermatogenic cells

Genes Dev.

(2006)

S.T. Grivna et al.

MIWI associates with translational machinery and PIWI-interacting RNAs (piRNAs) in regulating spermatogenesis

Proc. Natl. Acad. Sci. U.S.A.

(2006)

Cited by (160)

Trace and essential elements as vital components to improve the performance of the male reproductive system: Implications in cell signaling pathways
2024, Journal of Trace Elements in Medicine and Biology
Successful male fertilization requires the main processes such as normal spermatogenesis, sperm capacitation, hyperactivation, and acrosome reaction. The progress of these processes depends on some endogenous and exogenous factors. So, the optimal level of ions and essential and rare elements such as selenium, zinc, copper, iron, manganese, calcium, and so on in various types of cells of the reproductive system could affect conception and male fertility rates. The function of trace elements in the male reproductive system could be exerted through some cellular and molecular processes, such as the management of active oxygen species, involvement in the action of membrane channels, regulation of enzyme activity, regulation of gene expression and hormone levels, and modulation of signaling cascades. In this review, we aim to summarize the available evidence on the role of trace elements in improving male reproductive performance. Also, special attention is paid to the cellular aspects and the involved molecular signaling cascades.
Molecular characterization of SPATA6 and association of its SNPs with testicular size in sheep
2024, Theriogenology
The testis is an important organ for maintaining fertility in males, and testis size is positively correlated with ejaculate volume, sperm motility, thus fertility. Spermatogenesis-associated 6 (SPATA6) is an evolutionarily conserved testis-specific gene reported in many species. However, the effect of SPATA6 expression levels on testicular development and the effect of single nucleotide polymorphisms (SNPs) on testis and epididymis phenotype in sheep have not been studied. The purpose of the research was to investigate the expression profile of SPATA6 and its effect on testicular development and to confirm the effect of SNPs on the testis and epididymis phenotype. In this study, we detected a 1245bp coding sequence (CDS) of SPATA6 and encoded 414 amino acids. The expression levels of SPATA6 were significantly higher in the testis than in other tissues and gradually increased with testis development. Moreover, the expression level in the large testis was significantly higher than that in the small testis at six months. A total of 11 SNPs were detected in the coding region of SPATA6 by cDNA-pooling sequencing and improved multiplex ligation detection reaction (iMLDR) methods. Correlation analysis showed that SNP2 (c. 3631C > G) significantly affected left epididymis weight (LEW) and right epididymis weight (REW), and SNP10 (c. 937 A > G) significantly affected REW. And the combined genotype of SNP1 (c. 4245 G > A) and SNP2 significantly affected REW. The current study concluded that SPATA6 plays an important role in testicular development and the SNPs significantly associated with the epididymis phenotype can provide molecular markers for the early selection of high-fertility Hu sheep.
Calicin is a key sperm head-shaping factor essential for male fertility
2022, Science Bulletin
Flurochloridone induced abnormal spermatogenesis by damaging testicular Sertoli cells in mice
2022, Ecotoxicology and Environmental Safety
Citation Excerpt :
In mice, it is specifically expressed from leptotene to zygotene, and its deletion causes meiotic stagnation (Pittman et al., 1998; Laan et al., 2005). In the late stage of spermatogenesis, the morphology of round spermatids will undergo significant changes (Yan, 2009), whereas abnormal sperm morphology is an important factor affecting the occurrence of infertility (WHO, 2010). Concentration, motility, and kinematic parameters can effectively reflect sperm fertilization ability.
Flurochloridone (FLC), a selective herbicide used on a global scale, has been reported to have male reproductive toxicity whose evidence is limited, but its mechanism remains unclear. The present study was conducted to systematically explore the male reproductive toxicity of FLC, including sperm quality, spermatogenesis, toxicity targets, and potential mechanisms.
Male C57BL/6 mice aged 6–7 weeks received gavage administration of FLC (365/730 mg/kg/day) for 28 consecutive days. Then, the tissue and sperm of mice were collected for analysis. We measured the gonadosomatic index and analyzed sperm concentration, motility, malformation rate, and mitochondrial membrane potential (MMP). Spermatocyte immunofluorescence staining was performed to analyze meiosis. We also performed pathological staining on the testis and epididymis tissue and TUNEL staining, immunohistochemical analysis, and ultrastructural observation on the testicular tissue.
Results showed that FLC caused testicular weight reduction, dysfunction, and architectural damage in mice, but no significant adverse effect was found in the epididymis. The exposure interfered with spermatogonial proliferation and meiosis, affecting sperm concentration, motility, kinematic parameters, morphology, and MMP, decreasing sperm quality. Furthermore, mitochondrial damage and apoptosis of testicular Sertoli cells were observed in mice treated with FLC.
We found that FLC has significant adverse effects on spermatogonial proliferation and meiosis. Meanwhile, apoptosis and mitochondrial damage may be the potential mechanism of Sertoli cell damage. Our study demonstrated that FLC could induce testicular Sertoli cell damage, leading to abnormal spermatogenesis, which decreased sperm quality. The data provided references for the toxicity risk and research methods of FLC application in the environment.
Homeodomain-interacting protein kinase HIPK4 regulates phosphorylation of manchette protein RIMBP3 during spermiogenesis
2022, Journal of Biological Chemistry
Nonobstructive azoospermia (NOA) is the most serious form of spermatogenesis abnormalities in male infertility. Genetic factors are important to consider as elements leading to NOA. Although many pathogenic genes have been reported, the causative genes of NOA for many patients are still unknown. In this study, we found ten point mutations in the gene encoding homeodomain-interacting protein kinase 4 (HIPK4) in patients with NOA, and using in vitro studies, we determined a premature termination point mutation (p. Lys490∗, c.1468A>T) that can cause decreased expression of HIPK4. Our phosphoproteomic analysis of Hipk4^−/− testes revealed phosphorylation of multiple proteins regulated by HIPK4 during spermiogenesis. We also confirmed that a substrate of HIPK4 with four downregulated phosphorylation sites matching the xSPx motif is the known manchette-related protein RIMS-binding protein 3, which is required for sperm head morphogenesis. Therefore, we conclude HIPK4 regulates the phosphorylation of manchette protein RIMS-binding protein 3 and plays essential roles in sperm head shaping and male fertility.
GATA-1 mutation alters the spermatogonial phase and steroidogenesis in adult mouse testis
2022, Molecular and Cellular Endocrinology
GATA-1 is a transcription factor from the GATA family, which features zinc fingers for DNA binding. This protein was initially identified as a crucial regulator of blood cell differentiation, but it is currently known that the Gata-1 gene expression is not limited to this system. Although the testis is also a site of significant GATA-1 expression, its role in testicular cells remains considerably unexplored. In the present study, we evaluated the testicular morphophysiology of adult ΔdblGATA mice with a mutation in the GATA-1 protein. Regarding testicular histology, GATA-1 mutant mice exhibited few changes in the seminiferous tubules, particularly in germ cells. A high proportion of differentiated spermatogonia, an increased number of apoptotic pre-leptotene spermatocytes (Caspase-3-positive), and a high frequency of sperm head defects were observed in ΔdblGATA mice. The main differences were observed in the intertubular compartment, as ΔdblGATA mice showed several morphofunctional changes in Leydig cells. Reduced volume, increased number and down-regulation of steroidogenic enzymes were observed in ΔdblGATA Leydig cells. Moreover, the mutant animal showed lower serum testosterone concentration and high LH levels. These results are consistent with the phenotypic and biometric data of mutant mice, i.e., shorter anogenital index and reduced accessory sexual gland weight. In conclusion, our findings suggest that GATA-1 protein is an important factor for germ cell differentiation as well as for the steroidogenic activity in the testis.

View all citing articles on Scopus

View full text

ReviewMale infertility caused by spermiogenic defects: Lessons from gene knockouts

Abstract

Section snippets

Spermiogenesis is unique to male germ cell development and many of the testis-specific genes are expressed in spermiogenesis

Genes required for acrosome formation

What do these knockout mice tell us?

Acknowledgements

Arch. Biochem. Biophys.

J. Nutr.

J. Biol. Chem.

Dev. Cell

Am. J. Hum. Genet.

Dev. Cell

Curr. Opin. Cell Biol.

Dev. Biol.

Curr. Opin. Genet. Dev.

J. Biol. Chem.

FEBS Lett.

Dev. Biol.

Biochim. Biophys. Acta

Mech. Dev.

Fertil. Steril.

Eur. J. Cell Biol.

Mol. Cell

Dev. Biol.

J. Nutr.

Genomics

Urology

Analysis of sperm function in globozoospermia: implications for the mechanism of sperm–zona interaction

Fertil. Steril.

A novel class of small RNAs bind to MILI protein in mouse testes

Nature

The knockout mouse project

Nat. Genet.

Binding of antibodies against antigenic domains of murine lactate dehydrogenase-C4 to human and mouse spermatozoa

Biol. Reprod.

Severe impairment of spermatogenesis in mice lacking the CREM gene

Nature

The testis-specific phosphoglycerate kinase gene pgk-2 is a recruited retroposon

Mol. Cell Biol.

A-kinase anchoring protein 4 binding proteins in the fibrous sheath of the sperm flagellum

Biol. Reprod.

Glyceraldehyde 3-phosphate dehydrogenase-S protein distribution during mouse spermatogenesis

Biol. Reprod.

Selenoprotein P: an extracellular protein with unique physical characteristics and a role in selenium homeostasis

Annu. Rev. Nutr.

Protamine 2 deficiency leads to sperm DNA damage and embryo death in mice

Biol. Reprod.

Haploinsufficiency of protamine-1 or -2 causes infertility in mice

Nat. Genet.

Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal

Physiol. Rev.

Controlling HIV-1 Rev function

Curr. Drug Targets Immune Endocr. Metabol. Disord.

Cytoplasmic droplets: the good, the bad or just confusing?

Hum. Reprod.

A novel class of evolutionarily conserved genes defined by piwi are essential for stem cell self-renewal

Genes Dev.

Piwi encodes a nucleoplasmic factor whose activity modulates the number and division rate of germline stem cells

Development

The cellular biology of mammalian spermatids: a review

Bull. Assoc. Anat. (Nancy)

Drug discovery: a historical perspective

Science

Genetic dissection of the central pair microtubules of the flagella of Chlamydomonas reinhardtii

J. Cell Biol.

Male germ cell gene expression

Recent Prog. Horm. Res.

Fibrous sheath of mammalian spermatozoa

Microsc. Res. Tech.

Spermatogenesis of mice lacking CK2alpha’: failure of germ cell survival and characteristic modifications of the spermatid nucleus

Mol. Reprod. Dev.

Mice deficient for soluble adenylyl cyclase are infertile because of a severe sperm-motility defect

Proc. Natl. Acad. Sci. U.S.A.

Developmental switch of CREM function during spermatogenesis: from antagonist to activator

Nature

Dibutyryl cyclic AMP decreases capacitation time in vitro in mouse spermatozoa

J. Reprod. Fertil.

A germline-specific class of small RNAs binds mammalian Piwi proteins

Nature

Review
Male infertility caused by spermiogenic defects: Lessons from gene knockouts