Top

Journal of Assisted Reproduction and Genetics

Published in:

01-11-2015 | Technological Innovations

A single-cell assay for telomere DNA content shows increasing telomere length heterogeneity, as well as increasing mean telomere length in human spermatozoa with advancing age

Authors: Danielle M. F. Antunes, Keri H. Kalmbach, Fang Wang, Roberta C. Dracxler, Michelle L. Seth-Smith, Yael Kramer, Julia Buldo-Licciardi, Fabiana B. Kohlrausch, David L. Keefe

Published in: Journal of Assisted Reproduction and Genetics | Issue 11/2015

Abstract

Purpose

The effect of age on telomere length heterogeneity in men has not been studied previously. Our aims were to determine the relationship between variation in sperm telomere length (STL), men’s age, and semen parameters in spermatozoa from men undergoing in vitro fertilization (IVF) treatment.

Methods

To perform this prospective cross-sectional pilot study, telomere length was estimated in 200 individual spermatozoa from men undergoing IVF treatment at the NYU Fertility Center. A novel single-cell telomere content assay (SCT-pqPCR) measured telomere length in individual spermatozoa.

Results

Telomere length among individual spermatozoa within an ejaculate varies markedly and increases with age. Older men not only have longer STL but also have more variable STL compared to younger men. STL from samples with normal semen parameters was significantly longer than that from samples with abnormal parameters, but STL did not differ between spermatozoa with normal versus abnormal morphology.

Conclusion

The marked increase in STL heterogeneity as men age is consistent with a role for ALT during spermatogenesis. No data have yet reported the effect of age on STL heterogeneity. Based on these results, future studies should expand this modest sample size to search for molecular evidence of ALT in human testes during spermatogenesis.

Blackburn EH. Telomere states and cell fates. Nature. 2000;408:53–6.CrossRefPubMed

Liu L et al. An essential role for functional telomeres in mouse germ cells during fertilization and early development. Dev Biol. 2002;249:74–84.CrossRefPubMed

Keefe DL et al. Telomere length predicts embryo fragmentation after in vitro fertilization in women—toward a telomere theory of reproductive aging in women. Am J Obst Gynecol. 2005;192:1256–60.CrossRef

Sahin E et al. Telomere dysfunction induces metabolic and mitochondrial compromise. Nature. 2011;470:359–65.PubMedCentralCrossRefPubMed

Schaetzlein S et al. Telomere length is reset during early mammalian embryogenesis. Proc Natl Acad Sci U S A. 2004;101:8034–8.PubMedCentralCrossRefPubMed

Kalmbach KH et al. Telomeres and human reproduction. Fertil Steril. 2013;99:23–9.CrossRefPubMed

Krawetz SA. Paternal contribution: new insights and future challenges. Nat Rev Genet. 2005;6:633–42.CrossRefPubMed

Moskovtsev SI et al. Disruption of telomere-telomere interactions associated with DNA damage in human spermatozoa. Syst Biol Reprod Med. 2010;56:407–12.CrossRefPubMed

Kimura M et al. Offspring’s leukocyte telomere length, paternal age, and telomere elongation in sperm. PLoS Genet. 2008;4, e37.PubMedCentralCrossRefPubMed

10.

Ioannou D, Griffin DK. Male fertility, chromosome abnormalities, and nuclear organization. Cytogenet Genome Res. 2011;133:269–79.CrossRefPubMed

11.

Lee HW et al. Essential role of mouse telomerase in highly proliferative organs. Nature. 1998;392:569–74.CrossRefPubMed

12.

Reig-Viader R et al. Telomeric repeat-containing RNA (TERRA) and telomerase are components of telomeres during mammalian gametogenesis. Biol Reprod. 2014;90(5):103.CrossRefPubMed

13.

Reig-Viader R et al. Telomere homeostasis is compromised in spermatocytes from patients with idiopathic infertility. Fertil Steril. 2014;102(3):728–38.CrossRefPubMed

14.

Allsopp RC et al. Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci U S A. 1992;89:10114–8.PubMedCentralCrossRefPubMed

15.

Aston KI et al. Divergence of sperm and leukocyte age-dependent telomere dynamics: implications for male-driven evolution of telomere length in humans. Mol Hum Reprod. 2012;18:517–22.PubMedCentralCrossRefPubMed

16.

Baird DM et al. Telomere instability in the male germline. Hum Mol Genet. 2006;15:45–51.CrossRefPubMed

17.

Sartorius GA, Nieschlag E. Paternal age and reproduction. Hum Reprod Update. 2010;16:65–79.CrossRefPubMed

18.

Bryan TM et al. The telomere lengthening mechanism in telomerase-negative immortal human cells does not involve the telomerase RNA subunit. Hum Mol Genet. 1997;6:921–6.CrossRefPubMed

19.

Cesare AJ, Reddel RR. Alternative lengthening of telomeres: models, mechanisms and implications. Nat Rev Genet. 2010;11:319–30.CrossRefPubMed

20.

Dolcetti R, De Rossi A. Telomere/telomerase interplay in virus-driven and virus-independent lymphomagenesis: pathogenic and clinical implications. Med Res Rev. 2012;32:233–53.CrossRefPubMed

21.

Bryan TM et al. Telomere elongation in immortal human cells without detectable telomerase activity. EMBO J. 1995;14:4240–8.PubMedCentralPubMed

22.

Liu L et al. Telomere lengthening early in development. Nat Cell Biol. 2007;9:1436–41.CrossRefPubMed

23.

Li B, Lustig AJ. A novel mechanism for telomere size control in Saccharomyces cerevisiae. Genes Dev. 1996;10:1310–26.CrossRefPubMed

24.

Bucholc M, Park Y, Lustig AJ. Intrachromatid excision of telomeric DNA as a mechanism for telomere size control in Saccharomyces cerevisiae. Mol Cell Biol. 2001;21:6559–73.PubMedCentralCrossRefPubMed

25.

von Zglinicki T, Saretzki G, Docke W, Lotze C. Mild hyperoxia shortens telomeres and inhibits proliferation of fibroblasts: a model for senescence? Exp Cell Res. 1995;220:186–93.CrossRef

26.

Aitken RJ et al. Oxidative stress and male reproductive health. Asian J Androl. 2014;16:31–8.PubMedCentralCrossRefPubMed

27.

Thilagavathi J et al. Analysis of sperm telomere length in men with idiopathic infertility. Arch Gynecol Obstet. 2013;287:803–7.CrossRefPubMed

28.

Ferlin A et al. In young men sperm telomere length is related to sperm number and parental age. Hum Reprod. 2013;28:3370–6.CrossRefPubMed

29.

World Health Organization. WHO laboratory manual for the examination and processing of human semen. 2010; 5th edition

30.

Wang F et al. Robust measurement of telomere length in single cells. Proc Natl Acad Sci U S A. 2013;110:E1906–12.PubMedCentralCrossRefPubMed

31.

Nordstokke DW, Zumbo BD. A new nonparametric Levene test for equal variances. Psicológica. 2010;31:401–30.

32.

Turner S, Hartshorne GM. Telomere lengths in human pronuclei, oocytes and spermatozoa. Mol Hum Reprod. 2013;19:510–8.CrossRefPubMed

33.

Blackburn EH, Greider CW, Szostak JW. Telomeres and telomerase: the path from maize, Tetrahymena and yeast to human cancer and aging. Nat Med. 2006;12:1133–8.CrossRefPubMed

Title: A single-cell assay for telomere DNA content shows increasing telomere length heterogeneity, as well as increasing mean telomere length in human spermatozoa with advancing age
Authors: Danielle M. F. Antunes
Keri H. Kalmbach
Fang Wang
Roberta C. Dracxler
Michelle L. Seth-Smith
Yael Kramer
Julia Buldo-Licciardi
Fabiana B. Kohlrausch
David L. Keefe
Publication date: 01-11-2015
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 11/2015
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-015-0574-3

Keynote webinar | Spotlight on medication adherence

Springer Medicine

A single-cell assay for telomere DNA content shows increasing telomere length heterogeneity, as well as increasing mean telomere length in human spermatozoa with advancing age

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 11/2015

Marijuana, phytocannabinoids, the endocannabinoid system, and male fertility

Live birth following IVF/ICSI using oocytes from donor who was conceived via IVF: a case report

Case reports to suggest an algorithm for management of total fertilisation failure prior to use of donor gametes

Macrozoospermia: screening for the homozygous c.144delC mutation in AURKC gene in infertile men and estimation of its heterozygosity frequency in the Tunisian population

Evidence of an age-related correlation of ovarian reserve and FMR1 repeat number among women with “normal” CGG repeat status

Diagnosis of abnormal human fertilization status based on pronuclear origin and/or centrosome number