Anomalies in sperm chromatin packaging: implications for assisted reproduction techniques

doi:10.1016/S1472-6483(10)60124-1

Reproductive BioMedicine Online

Volume 18, Issue 4, 2009, Pages 486-495

https://doi.org/10.1016/S1472-6483(10)60124-1 Get rights and content

Abstract

Sperm protamine deficiency and DNA damage were analysed employing chromomycin A₃ (CMA₃) staining and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling assay, respectively, in 132 patients (82 IVF, 50 intracytoplasmic sperm injection [ICSI]). The antioxidant ability of seminal plasma was analysed in 10 men, using the total oxidant scavenging capacity assay. A significant negative correlation was found between abnormal protamination and sperm parameters, including sperm DNA fragmentation (P < 0.01). A close relationship was found between sperm protamination and fertilization and pregnancy only in IVF (P = 0.004 and P < 0.04, respectively); in ICSI there was a correlation between DNA fragmentation and pregnancy (P = 0.031). Finally, there was a negative correlation between chromatin under-protamination and the antioxidant ability of seminal plasma (P < 0.01). Results of this study underline that, despite sperm abnormal protamination and DNA fragmentation being positively correlated, they affect the reproductive outcome in different ways: in particular there was good prognostic value for CMA₃ analysis only in IVF, whereas DNA fragmentation analysis was prognostic only for ICSI outcome. Data are also provided to support the idea of a relationship between defective antioxidant system activity and impairment of chromatin packaging.

Section snippets

Nicoletta Tarozzi obtained her PhD in 2004 at the University of Modena, Italy, working in the Genetics Laboratory of the Animal Biology Department. In 2004 she joined the team of Tecnobios Procreazione, Centre for Reproductive Health in Bologna. Dr Tarozzi has published several papers in the field of sperm DNA damage. Current research interests focus on male infertility, especially regarding molecular biology of the male germ cell and structure of sperm chromatin.

References (61)

RJ Aitken et al.
Origins and consequences of DNA damage in male germ cells
Reproductive BioMedicine Online
(2007)
RJ Aitken et al.
Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic disease
Reproductive BioMedicine Online
(2003)
G Balercia et al.
Placebo-controlled double-blind randomized trial on the use of l-carnitine, l-acetylcarnitine, or combined l-carnitine and l-acetylcarnitine in men with idiopathic asthenozoospermia
Fertility and Sterility
(2005)
F Bianchi et al.
Interaction of human P1 and P2 protamines with DNA
Biochemical and Biophysical Research Communications
(1994)
A Borini et al.
Pregnancies and births after oocyte cryopreservation
Fertility and Sterility
(2004)
A Borini et al.
Perifollicular vascularity monitoring and scoring: a clinical tool for selecting the best oocyte
European Journal of Obstetrics, Gynecology, and Reproductive Biology
(2004)
G D'Auria et al.
Structure and function of protamines: an ¹H nuclear magnetic resonance investigation of the interaction of clupeines with mononucleotides
Biochimica et Biophysica Acta
(1993)
JP Dadoune
The nuclear status of human sperm cells
Micron
(1995)
D Evenson et al.
Meta-analysis of sperm DNA fragmentation using the sperm chromatin structure assay
Reproduction BioMedicine Online
(2006)
JG Franco et al.
Significance of large nuclear vacuoles in human spermatozoa: implications for ICSI
Reproduction BioMedicine Online
(2008)

XL Gao et al.

Structure refinement of the chromomycin dimer-DNA oligomer complex in solution

Journal of Molecular Biology

(1992)

ME Hammadeh et al.

Comparison of reactive oxygen species concentration in seminal plasma and semen parameters in partners of pregnant and non-pregnant patients after IVF/ICSI

Reproductive BioMedicine Online

(2006)

A Hazout et al.

High-magnification ICSI overcomes paternal effect resistant to conventional ICSI

Reproductive BioMedicine Online

(2006)

CC Huang et al.

Sperm DNA fragmentation negatively correlates with velocity and fertilization rates but might not affect pregnancy rates

Fertility and Sterility

(2005)

NV Hud et al.

Identification of the elemental packing unit of DNA in mammalian sperm cells by atomic force microscopy

Biochemical and Biophysical Research Communications

(1993)

YJ Ménézo

Paternal and maternal factors in preimplantation embryogenesis: interaction with the biochemical environment

Reproductive BioMedicine Online

(2006)

MH Nasr-Esfahani et al.

Failed fertilization after ICSI and spermiogenic defects

Fertility and Sterility

(2008)

MH Nasr-Esfahani et al.

Effect of protamine-2 deficiency on ICSI outcome

Reproductive BioMedicine Online

(2004)

M O'Connell et al.

Differences in mitochondrial and nuclear DNA status of high-density and low-density sperm fractions after density centrifugation preparation

Fertility and Sterility

(2003)

AK Olsen et al.

How do male germ cells handle DNA damage?

Toxicology and Applied Pharmacology

(2005)

GC Pogany et al.

DNA and protein content of mouse sperm. Implications regarding sperm chromatin structure

Experimental Cell Research

(1981)

L Ramos et al.

Chromatin condensation and DNA damage of human epididymal spermatozoa in obstructive azoospermia

Reproductive BioMedicine Online

(2004)

RA Saleh et al.

Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility

Fertility and Sterility

(2003)

N Tarozzi et al.

Clinical relevance of sperm DNA damage in assisted reproduction

Reproductive BioMedicine Online

(2007)

FW van Overveld et al.

Tyrosine as important contributor to the antioxidant capacity of seminal plasma

Chemico-biological Interactions

(2000)

GW Winston et al.

A rapid gas chromatographic assay for determining oxyradical scavenging capacity of antioxidants and biological fluids

Free Radical Biology and Medicine

(1998)

A Zini et al.

Free thiols in human spermatozoa: correlation with sperm DNA integrity

Urology

(2001)

JG Alvarez et al.

Role of glutathione peroxidase in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation

Gamete Research

(1989)

VW Aoki et al.

Protamine levels vary between individual sperm cells of infertile human males and correlate with viability and DNA integrity

Journal of Andrology

(2006)

VW Aoki et al.

Identification and evaluation of a novel sperm protamine abnormality in a population of infertile males

Human Reproduction (Oxford, England)

(2005)

Cited by (60)

Proteomic and genetic dissection of testis-specific histone 2B in infertile men reveals its contribution to meiosis and sperm motility
2022, F and S Science
To investigate testis-specific histone 2B (TSH2B) and its gene anomalies in infertile men.
Case-control study.
Basic science laboratory.
Fertile and infertile men.
Not applicable.
The histone and protamine status of sperm was studied by aniline blue and chromomycin A3 staining, respectively. Testis-specific histone 2B, total H2B, and phosphorylated TSH2B (pTSH2B) were estimated by Western blot analysis. The frequency of genetic polymorphisms and rare variants in H2BC1 was studied by Sanger sequencing. Phosphosites on TSH2B in sperm were identified by reverse-phase high-performance liquid chromatography purification of TSH2B followed by mass spectrometric analysis.
Aniline blue and chromomycin A3 staining revealed significantly higher histone retention and low protamine in sperm of infertile men. Sperm TSH2B and total H2B levels were significantly lower in oligozoospermic and oligoasthenozoospermic men (in both groups). The TSH2B levels were comparable in asthenozoospermic men; however, the pTSH2B level was significantly low. The H2BC1 gene sequencing identified 6 variants, of which 2 are rare variants (rs368672899 and rs544942090) and 4 (rs4711096, rs4712959, rs4712960 and rs4712961) are single nucleotide polymorphisms. Minor allele frequency of 5′-untranslated region variant rs4711096 was significantly lower in infertile men (OR = 0.65). The rare nonsynonymous variant, rs368672899, p.Ser5Pro was seen in 1 oligoasthenoteratozoospermic individual. Interestingly, mass spectrometric analysis identified a site on TSH2B to bear a phosphate group in the sperm of fertile men.
Our study reveals a defect in the replacement of somatic histones with testis-specific variants in infertile men. Chromatin compaction positively correlates with sperm motility, which is suggestive of its utility in diagnostic semen analysis of infertile individuals. Our observations with TSH2B and its cognate gene in sperm of infertile men indicate an essential role for TSH2B in meiosis and its phosphorylation in sperm motility, respectively.
Glutathione S-transferase Mu 3 is associated to in vivo fertility, but not sperm quality, in bovine
2022, Animal
Citation Excerpt :
Regarding the putative relationship between GSTM3 levels and sperm chromatin status, our work did not find any significant relationship between both parameters, suggesting that GSTM3 is not involved in sperm chromatin (de)condensation. In contrast, Tarozzi et al. (2009) reported a negative relationship between the antioxidant capacity of seminal plasma and the protamination status of sperm chromatin in humans. These differences could be explained because sperm GSTM3 might not play an antioxidant role on sperm, which agrees with the fact that no relationship between its levels and the parameters concerning oxidative stress was observed.
In the dairy breeding industry, pregnancy of dairy cows is essential to initiate milk production, so that high fertility rates are required to increase their productivity. In this regard, sperm proteins that are indicative of sperm quality and/or fertility have become an important target of study. Glutathione S-transferase Mu 3 (GSTM3) has been established as a fertility and sperm quality parameter in humans and pigs and, consequently, it might be a potential biomarker in cattle. For this reason, the present work aimed to determine if GSTM3 could predict sperm quality and in vivo fertility in this species. Sperm quality was assessed with flow cytometry and computer-assisted sperm analysis. Immunoblotting and immunofluorescence analysis were performed to determine the presence and localisation pattern of sperm GSTM3. This enzyme was found to be present in bovine sperm and to be localised along the sperm tail and the equatorial segment of the head. No significant associations between sperm GSTM3 and sperm quality parameters were observed, except a negative association with morphologically abnormal sperm having a coiled tail. In addition, and more relevant, higher levels of GSTM3 in sperm were seen in bulls showing lower in vivo fertility rates. In conclusion, our data evidenced the presence of GSTM3 in bovine sperm. Moreover, we suggest that, despite not being associated with sperm quality, GSTM3 might be an in vivo subfertility biomarker in cattle sperm, and that high levels of this protein could be an indicative of defective spermatogenesis and/or epididymal maturation.
A systematic review identifying fertility biomarkers in semen: a clinical approach through Omics to diagnose male infertility
2022, Fertility and Sterility
To identify the most robust molecular biomarkers in sperm and seminal plasma for the diagnosis of male infertility, and to evaluate their clinical use.
Systematic review.
Not applicable.
Accessible studies reporting well-defined (in)fertile populations and semen molecular biomarkers were included in this review.
A systematic search of the literature published in MEDLINE-PubMed and EMBASE databases was performed, following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines.
The primary outcome was the content, expression, or activity of molecular biomarkers in human semen samples. Only studies reporting a receiver-operating characteristic (ROC) analysis values were included.
Eighty-nine studies were included. Direct evaluation of sperm DNA damage has high potential as a diagnostic biomarker of fertility and assisted reproductive technology outcomes (area under the curve [AUCs] median = 0.67). Regarding strand break-associated chromatin modifications, γH2AX levels show good predictive value for the diagnosis of male infertility (AUCs median = 0.93). Some noncoding ribonucleic acid (RNA) exhibit excellent predictive values; miR-34c-5p in semen is the most well-characterized and robust transcriptomic biomarker (AUCs median = 0.78). While many proteins in semen show fair diagnostic value for sperm quality and fertilizing capacity, the levels of some, such as TEX101, in seminal plasma have an excellent diagnostic potential (AUCs median = 0.69). Although individual metabolites and metabolomic profiles in seminal plasma present good predictive value, the latter seem to be better than the former when inferring sperm quality and fertilizing capacity.
The current review supports that some Omics (e.g., DNA structure and integrity, genomics and epigenomics, transcriptomics, metabolomics, and proteomics) could be considered relevant molecular biomarkers that may help identify infertility etiologies and fertilization prognosis with cost-effective, simple, and accurate diagnosis.
Revisión sistemática en la identificación de biomarcadores de fertilidad en el semen: un abordaje clínico a través de las ómicas para el diagnóstico de la infertilidad masculina.
Identificar los biomarcadores moleculares más robustos en el semen y plasma seminal para el diagnóstico de la infertilidad masculina y evaluar su utilidad clínica.
Revisión sistemática.
No aplica.
Se incluyeron en esta revisión estudios accesibles que reportan poblaciones (in)fértiles bien definidas y biomarcadores moleculares seminales.
Se realizó una búsqueda sistemática de la literatura publicada en las bases de datos de MEDLINE-PubMed y EMBASE, siguiendo las guías de Elementos de informe preferidos para revisiones sistemáticas y metanálisis (PRISMA).
Los resultados primarios fueron el contenido, la expresión o actividad de los biomarcadores moleculares en las muestras de semen humano. Solo se incluyeron estudios que reportan valores de análisis de características operativas del receptor (ROC).
Se incluyeron ochenta y nueve estudios. La evaluación directa del daño en el DNA de los espermatozoides tiene un alto potencial como biomarcador diagnóstico de fertilidad y los resultados de las tecnologías de reproducción asistida (área por debajo de la curva [AUCs] mediana = 0.67). Con respecto a la rotura de hebras asociadas a modificaciones de la cromatina, los niveles de ᵧH2AX muestran buenos valores predictivos para el diagnóstico de la infertilidad masculina (AUCs mediana = 0.93). Algunos ácidos ribonucleicos (ARN) no codificantes exhiben buenos valores predictivos; el miR-34c-5p en semen es el biomarcador transcriptómico mejor caracterizado y robusto (AUCs median = 0.78). Mientras que muchas proteínas en el semen muestran valores pobres para el diagnóstico de la calidad seminal y la capacidad de fecundación, tales como TEX101 en el plasma seminal tiene un excelente potencial de diagnóstico (AUCs mediana = 0.69). Aunque, metabolitos individuales y perfiles metabolómicos en el plasma seminal presentan buenos valores predictivos, los últimos parecen ser mejores que los primeros a la hora de inferir sobre la calidad seminal y la capacidad de fecundación.
La revisión actual apoya que algunas ómicas (por ejemplo, la integridad y estructura del ADN, genómica y epigenómica, transcriptómica, metabolómica y proteómica) podrían ser consideradas biomarcadores moleculares relevantes que ayuden a identificar etiologías de infertilidad y pronóstico de fecundación con diagnósticos costo-efectivos, simples y precisos.
Review: Diagnosis and impact of sperm DNA alterations in assisted reproduction
2017, Best Practice and Research: Clinical Obstetrics and Gynaecology
Sperm nuclear and chromatin abnormalities are common among infertile men and are known to influence natural reproduction. These abnormalities are also considered detrimental to normal fertilization, embryo development, and successful implantation and pregnancies following assisted reproductive treatment (ART). Abnormalities in the sperm nucleus can be broadly classified into sperm chromosomal abnormalities (aneuploidies) and sperm DNA abnormalities such as abnormal packing, DNA integrity, or DNA fragmentation. For the past 30 years, numerous tests have been developed to quantify these abnormalities in sperm. In this chapter, we review the causes of sperm DNA and chromosomal abnormalities, describe the commonly used tests to evaluate these abnormalities, and finally review the impact of these abnormalities on male fertility and ART outcomes. We also performed a comprehensive meta-analysis and systematic review from the existing literature to summarize the effect of sperm DNA fragmentation on ART outcomes such as fertilization rate, embryo quality, and clinical pregnancies. A review of the literature presented in this chapter suggests that sperm nuclear and chromatin abnormalities are associated with male infertility, and they reduce the probability of a successful pregnancy following ART.
Contribution of semen to early embryo development: fertilization and beyond
2023, Human Reproduction Update
Advanced Sperm Selection Strategies as a Treatment for Infertile Couples: A Systematic Review
2022, International Journal of Molecular Sciences

View all citing articles on Scopus

: Declaration: The authors report no financial or commercial conflicts of interest.

View full text

ArticleAnomalies in sperm chromatin packaging: implications for assisted reproduction techniques

Abstract

Section snippets

Reproductive BioMedicine Online

Reproductive BioMedicine Online

Fertility and Sterility

Biochemical and Biophysical Research Communications

Fertility and Sterility

European Journal of Obstetrics, Gynecology, and Reproductive Biology

Biochimica et Biophysica Acta

Micron

Reproduction BioMedicine Online

Reproduction BioMedicine Online

Journal of Molecular Biology

Reproductive BioMedicine Online

Reproductive BioMedicine Online

Fertility and Sterility

Biochemical and Biophysical Research Communications

Reproductive BioMedicine Online

Fertility and Sterility

Reproductive BioMedicine Online

Fertility and Sterility

Toxicology and Applied Pharmacology

Experimental Cell Research

Reproductive BioMedicine Online

Fertility and Sterility

Reproductive BioMedicine Online

Chemico-biological Interactions

Free Radical Biology and Medicine

Urology

Role of glutathione peroxidase in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation

Gamete Research

Protamine levels vary between individual sperm cells of infertile human males and correlate with viability and DNA integrity

Journal of Andrology

Identification and evaluation of a novel sperm protamine abnormality in a population of infertile males

Human Reproduction (Oxford, England)

Article
Anomalies in sperm chromatin packaging: implications for assisted reproduction techniques