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Abnormal spermatogenesis in mice unable to synthesize ascorbic acid

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Abstract

Although exposure to environmental toxicants, including endocrine-disrupting chemicals, is thought to be a possible cause of male infertility, the pathogenesis of male reproductive disorders remains unclear. In the present study, we used Gulo-/- mutant mice, which are unable to synthesize ascorbic acid, to study the importance of dietary vitamin C (VC) on spermatogenesis. Regular chow containing approximately 110 mg/ kg VC is unable to support the growth of these mutant mice, but a VC supplement in their drinking water (330 mg/L) is able to ameliorate the VC deficiency. Testes of Gulo-/- mutants born from heterozygous mothers without VC supplement (VC-deficient mice) and those born from mothers given a VC supplement (VC-sufficient mice) were examined by morphological and biochemical analyses. Morphological analysis revealed that apoptosis of spermatocytes occurred frequently in VC-deficient mice at 20 days of age. Two-dimensional electrophoresis analysis revealed the specific disappearance of heat-shock protein (Hsp) 70 in the testes of 20-day-old VC-deficient mice. In the present study, the relationship between the apoptosis of spermatocytes and Hsp70 in VC-deficient mice is discussed.

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References

  • Anthony CT, Skinner MK (1989) Actions of extracellular matrix on Sertoli cell morphology and function.Biol Reprod 40, 691–702.

    Article  PubMed  CAS  Google Scholar 

  • Bauche F, Fouchard MH, Jegou B (1994) Antioxidant system in rat testicular cells.FEBS Lett 349, 392–6.

    Article  PubMed  CAS  Google Scholar 

  • Bellve AR, Cavicchia JC, Millette CFet al. (1977) Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization.J Cell Biol 74, 68–85.

    Article  PubMed  CAS  Google Scholar 

  • Borland K, Muffy KE, Hall PF (1986) Production of components of extracellular matrix by cultured rat Sertoli cell.Biol Reprod 35, 997–1008.

    Article  PubMed  CAS  Google Scholar 

  • Buettner GR (1993) The pecking order of free radicals and antioxidants: Lipid peroxidation, α-tocopherol, and ascorbate.Arch Biochem Biophys 300, 535–43.

    Article  PubMed  CAS  Google Scholar 

  • Chinoy NJ, Mehta RR, Seethalakshmi L, Sharma JD, Chinoy MR (1986) Effects of vitamin C deficiency on physiology of male reproductive organs of guinea pigs.Int J Fertil 31, 232–9.

    PubMed  CAS  Google Scholar 

  • Cleveland DW, Fisher SG, Kirschner MW, Laemmli UK (1977) Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis.J Biol Chem 252, 1102–6.

    PubMed  CAS  Google Scholar 

  • Cooke HJ, Saunders PT (2002) Mouse models of male infertility.Nat Rev Genet 3, 790–801.

    Article  PubMed  CAS  Google Scholar 

  • Davis CM, Papadopoulos V, Sommers CL, Kleinman HD, Dym M (1990) Differential expression of extracellular matrix components in rat Sertoli cells.Biol Reprod 43, 860–9.

    Article  PubMed  CAS  Google Scholar 

  • De Rooij DG (2001) Proliferation and differentiation of spermatogonial stem cells.Reproduction 121, 347–54.

    Article  PubMed  Google Scholar 

  • De Rooij DG, Grootegoed A (1998) Spermatogonial stem cells.Curr Opin Cell Biol 10, 694–701.

    Article  PubMed  Google Scholar 

  • Diaz ES, Pellizzari E, Meroni S, Cigorraga S, Lustig L, Denduchis B (2002) Effect of extracellular matrix proteins onin vitro testosterone production by rat Leydig cells.Mol Reprod Dev 61, 493–503.

    Article  PubMed  CAS  Google Scholar 

  • Dix DJ, Allen JW, Collins BWet al. (1996) Targeted gene disruption of Hsp 70-2 results in failed meiosis, germ cell apoptosis, and male infertility.Proc Natl Acad Sci USA 93, 3264–8.

    Article  PubMed  CAS  Google Scholar 

  • Dunkel L, Hirvonen V, Erkkila K (1997) Clinical aspects of male germ cell apoptosis during testis development and spermatogenesis.Cell Death Differ 4, 171–9.

    Article  PubMed  CAS  Google Scholar 

  • Dym M, Fawcett DW (1970) The blood-testis barrier in the rat and the physiological compartmentation of the seminiferous epithelium.Biol Reprod 3, 308–26.

    PubMed  CAS  Google Scholar 

  • Eddy EM (1999) Role of heat shock protein HSP70-2 in spermatogenesis.Rev Reprod 4, 23–30.

    Article  PubMed  CAS  Google Scholar 

  • Erkkila K, Aito H, Aalto K, Pentikainen V, Dunkel L (2002) Lactate inhibits germ apoptosis in the human testis.Mol Hum Reprod 8, 109–17.

    Article  PubMed  CAS  Google Scholar 

  • Escalier D (2001) Impact of genetic engineering on the understanding of spermatogenesis.Hum Reprod Update 7, 191–210.

    Article  PubMed  CAS  Google Scholar 

  • Fawcett DW (1975) Ultrastructure and function of the Sertoli cell. In: Hamilton DW, Greep RO, eds.Handbook of Physiology, Section 7: Endocrinology. American Physiological Society, Washington DC, 21–55.

    Google Scholar 

  • Fukuda H, Ito Y, Hirota R, Tsuji M, Mori H (1991) Production of corticosterone and testosterone in scorbutic mutant rats: Difference ofin vivo production between adrenal gland and testis.Acta Endocrinol 124, 425–33.

    PubMed  CAS  Google Scholar 

  • Griswold MD (1998) The central role of Sertoli cells in spermatogenesis.Semin Cell Dev Biol 9, 411–16.

    Article  PubMed  CAS  Google Scholar 

  • Hadley MK, Dym M (1987) Immunocytochemistry of extracellular matrix in the lamina propria of the rat testis: Electron microscopic localization.Biol Reprod 37, 1283–9.

    Article  PubMed  CAS  Google Scholar 

  • Hasegawa M, Wilson G, Russell LD, Meistrich ML (1997) Radiation- induced cell death in the mouse testis: Relationship to apoptosis.Radiat Res 147, 457–67.

    Article  PubMed  CAS  Google Scholar 

  • Hirano H (1982) Varietal differences of leaf protein profiles in mulberry.Phytochemistry 21, 1513–18.

    CAS  Google Scholar 

  • Hirano H, W-Liebold B (1989) Protein microsequence analysis with dansyl-amino-PITC. In: W-Liebold B, ed.Methods in Protein Sequence Analysis. Springer, Berlin, 42–51.

    Google Scholar 

  • Ito Y, Tsuji M, Terada N, Miyano M, Mori H (1992) Testosterone production in mature scorbutic mutant rats unable to synthesize ascorbic acid.Int J Androl 15, 160–9.

    Article  PubMed  CAS  Google Scholar 

  • Kasahara E, Sato EF, Miyoshi Met al. (2002) Role of oxidative stress in germ cell apoptosis induced by di(2-ethylhexyl) phthalate.Biochem J 365, 849–56.

    PubMed  CAS  Google Scholar 

  • Khassaf M, McArdle A, Esanu Cet al. (2003) Effect of vitamin C supplements on antioxidant defect and stress proteins in human lymphocytes and skeletal muscle.J Physiol 549, 645–52.

    Article  PubMed  CAS  Google Scholar 

  • Legare C, Thabet M, Sullivan R (2004) Expression of heat shock protein 70 in normal and cryptorchid human excurrent duct.Mol Hum Reprod 10, 197–202.

    Article  PubMed  CAS  Google Scholar 

  • Leu YH, Hikim AP, Swerdloff RSet al. (1999) Signal exposure to heat induces stage-specific germ cell apoptosis in rats: Role of intratesticular testosterone on stage specificity.Endocrinology 140, 1709–17.

    Article  Google Scholar 

  • Lewis SE, Sterling ES, Young IS, Thompson W (1997) Comparison of individual antioxidants of sperm and seminal plasma in fertile and infertile men.Fertil Steril 67, 142–7.

    Article  PubMed  CAS  Google Scholar 

  • Maeda N, Hagihara H, Nakata Y, Hiller S, Wilder J, Reddick R (2000) Aortic wall damage in mice unable to synthesize ascorbic acid.Proc Natl Acad Sci USA 97, 841–6.

    Article  PubMed  CAS  Google Scholar 

  • Mauduit C, Hamamah S, Benahmed M (1999) Stem cell factor/ c-kit system in spermatogenesis.Hum Reprod Update 5, 535–45.

    Article  PubMed  CAS  Google Scholar 

  • McLachlan JA, Newbold RR, Burow ME, Li SF (2001) From malformations to molecular mechanisms in the male: Three decades of research onendocrine disrupters.APMIS 109, 236–72.

    Article  Google Scholar 

  • McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy.J Histochem Cytochem 22, 1077–83.

    PubMed  CAS  Google Scholar 

  • Mori C, Nakamura N, Dix DJet al. (1997) Morphological analysis of germ cell apoptosis during postnatal testis development in normal and Hsp 70-2 knockout mice.Dev Dyn 208, 125–36.

    Article  PubMed  CAS  Google Scholar 

  • Mori C, Hamamatsu A, Fukata Het al. (2002) Temporal changes in testis weight during the past 50 years in Japan.Anat Sci Int 77, 109–16.

    Article  PubMed  Google Scholar 

  • Nakano M, Kishida R, Funakoshi Ket al. (2003) Central projections of thoracic splanchnic and somatic nerves and the location of sympathetic preganglionic neurons inXenopus levis. J Comp Neurol 456, 321–37.

    Article  Google Scholar 

  • O’Bryan MK, Schlatt S, Phillips DJ, de Kretser DM, Hedger MP (2000) Bacterial ipopolysaccharide-induced inflammation compromises testicular function at multiple levelsin vivo.Endocrinology 141, 238–46.

    Article  PubMed  CAS  Google Scholar 

  • O’Farrell PH (1975) High resolution two-dimensional electro- phoresis of proteins.J Biol Chem 250, 4007–21.

    PubMed  CAS  Google Scholar 

  • Paranko J, Seitz J, Meinhardt A (1996) Developmental expression of heat shock protein 60 (HSP60) in the rat testis and ovary.Differentiation 60, 159–67.

    Article  PubMed  CAS  Google Scholar 

  • Rao AV, Shaha C (2000) Role of glutathione S-transferases in oxidative stress-induced male germ cell apoptosis.Free Radic Biol Med 29, 1015–27.

    Article  PubMed  CAS  Google Scholar 

  • Richburg JH (2000) The relevance of spontaneous- and chemically- induced alterations in testicular germ cell apoptosis to toxicology.Toxicol Lett 112-113, 79–86.

    Article  PubMed  CAS  Google Scholar 

  • Rockett JC, Mapp FL, Garges JB, Luft JC, Mori C, Dix DJ (2001) Effects of hyperthermia on spermatogenesis, apoptosis, gene expression, and fertility in adult male mice.Biol Reprod 65, 229–39.

    Article  PubMed  CAS  Google Scholar 

  • Rodriguez I, Ody C, Araki K, Garcia I, Vassalli P (1997) An early and massive wave of germinal cell apoptosis is required for the development of functional spermatogenesis.EMBO J 16, 2262–70.

    Article  PubMed  CAS  Google Scholar 

  • Setchell BP, Waites GMH (1975) The blood-testis barrier. In: Hamilton DW, Greep RO, eds.Handbook of Physiology, Section 7: Endocrinology. American Physiological Society, Washington DC, 143–72.

    Google Scholar 

  • Shaban SF (2003) Male Infertility Overview. Assessment, Diagnosis, and Treatment. IVF Com., Atlanta, GA. Available on-line: http://www.ivf.com/shaban.html (accessed April 2005).

  • Sharma RK, Agarwal A (1996) Role of reactive oxygen species in male infertility.Urology 48, 835–50.

    Article  PubMed  CAS  Google Scholar 

  • Suzaki E, Kataoka K (1992) Lectin cytochemistry in the gastrointestinal tract with special reference to glycosylation in the golgi apparatus of Brunner’s gland cells.J Histochem Cytochem 43, 379–85.

    Google Scholar 

  • Swan S, Elkin E, Fenster L (1997) Reanalysis of international data finds sharp decline in sperm density.Environ Health Perspect 105, 1228–32.

    Article  PubMed  CAS  Google Scholar 

  • Toshimori K, Ito C, Maekawa M, Toyama Y, Suzuki-Toyota F, Saxena DK (2004) Impairment of spermatogenesis leading to infertility.Anat Sci Int 79, 101–11.

    Article  PubMed  CAS  Google Scholar 

  • Triendl R (2001) Genes may solve hormone-disrupter debate.Nature 409, 274.

    Article  PubMed  CAS  Google Scholar 

  • Tung PS, Fritz IB (1986) Extracellular matrix components and testicular peritubular cells influence the rate and pattern of Sertoli cell migrationin vitro. Dev Biol 113, 119- 34.

    CAS  Google Scholar 

  • Tung PS, Fritz IB (1987) Morpholgenetic restructuring and formation of basement membranes by Sertoli cells and testis peritubular cells in coculture: Inhibition of morphogenetic dascade by cyclic AMP derivative and by blocking direct cell contact.Dev Biol 120, 139–53.

    Article  PubMed  CAS  Google Scholar 

  • Verma A, Kanwar KC (1998) Human sperm motility and lipid peroxidation in different ascorbic acid concentrations: Anin vitro analysis.Andrologia 30, 325–9.

    PubMed  CAS  Google Scholar 

  • Vernon RB, Lane TF, Angello JC, Sage H (1991) Adhesion, shape, proliferation, and gene expression of mouse Leydig cells are influenced by extracellular matrixin vivo.Biol Reprod 44, 157–70.

    Article  PubMed  CAS  Google Scholar 

  • Wang RA, Nakane PK, Koji T (1998) Autonomous cell death of mouse male germ cells during fetal and postnatal periods.Biol Reprod 58, 1250–6.

    Article  PubMed  CAS  Google Scholar 

  • Yang Y, Cheng JZ, Singhal SS, Saini M, Pandya U, Awasthi YC (2001) Role of glutathione S-transferases in protection against lipid peroxidation. Overexpression of hGSTA2-2 in K562 cells protects against hydrogen peroxide-induced apoptosis and inhibits JNK and caspase 3 activation.J Biol Chem 276, 19220–30.

    Article  PubMed  CAS  Google Scholar 

  • Yazama F, Nishida T, Kurohmaru M, Hayashi Y (1988) The fine structure of the blood-testis barrier in the boar.Jpn J Vet Sci 50, 1259–61.

    CAS  Google Scholar 

  • Yazama F, Esaki M, Sawada H (1997) Immunocytochemistry of extracellular matrix components in the rat seminiferous tubule: Electron microscopic localization with improved methodology.Anat Rec 248, 51–62.

    Article  PubMed  CAS  Google Scholar 

  • Yoganathan T, Eskild W, Hansson V (1989) Investigation of detoxification capacity of rat testicular germ cells and Sertoli cells.Free Radic Biol Med 7, 355–9.

    Article  PubMed  CAS  Google Scholar 

  • Zhou XC, Han XB, Hu ZY, Zhou RJ, Liu YX (2001) Expression of Hsp70-2 in unilateral cryptorchid testis of rhesus monkey during germ cell apoptosis.Endocrine 16, 89–95.

    Article  PubMed  CAS  Google Scholar 

  • Zhu D, Dix DJ, Eddy EM (1997) HSP70-2 is required for CDC2 kinase activity in meiosis I of mouse spermatocytes.Development 134, 3007–14.

    Google Scholar 

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Authors and Affiliations

  1. Department of Life Sciences, Laboratory of Cell Biology and Morphology, Prefectural University of Hiroshima, 562 Nanatsuka, 727-0023, Shobara City, Hiroshima, Japan

    Futoshi Yazama, Kazuko Furuta, Miwa Fujimoto & Hiroko Shigetomi

  2. Department of Life Sciences, Laboratory of Gamete Biotechnology, Shobara

    Toshitaka Horiuchi

  3. Department of Life Sciences, Laboratory of Molecular Physiology, Shobara

    Manabu Yamada

  4. Department of Life Sciences, Laboratory of Cell Biochemistry, Prefectural University of Hiroshima, Shobara

    Norio Nagao

  5. Department of Molecular Biodefense Research, Yokohama City University School of Medicine, Yokohama, Japan

    Tomoko Sonoda

  6. Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, North Carolina, USA

    Nobuyo Maeda

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  1. Futoshi Yazama
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  2. Kazuko Furuta
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  3. Miwa Fujimoto
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Correspondence to Futoshi Yazama.

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Yazama, F., Furuta, K., Fujimoto, M. et al. Abnormal spermatogenesis in mice unable to synthesize ascorbic acid. Anato Sci Int 81, 115–125 (2006). https://doi.org/10.1111/j.1447-073X.2006.00139.x

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  • DOI: https://doi.org/10.1111/j.1447-073X.2006.00139.x

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