Nuclear status of immature and mature stallion spermatozoa.
Abstract: 'The highly packed chromatin of mature spermatozoa results from replacement of somatic-like histones by highly basic arginine- and cysteine-rich protamines during spermatogenesis, with additional conformational changes in chromatin structure during epididymal transit. The objective of the present study was to compare the nuclear characteristics of immature and mature epididymal stallion spermatozoa, using a variety of experimental approaches. Resistance to in vitro decondensation of chromatin, following exposure to SDS-DTT and alkaline thioglycolate, increased significantly in mature spermatozoa. Evaluation of the thiol-disulfide status (monobromobimane labeling) demonstrated that immature cells obtained from ductulli efferentes contained mostly thiol groups, whereas these groups were oxidized in mature cells collected from the cauda epididymidis. Based on atomic absorption spectrophotometry, maturation of stallion spermatozoa was accompanied by a 60% reduction in the Zn(2+) content of sperm cells, concomitant with increased concentrations of this ion in epididymal fluid. Furthermore, the degree of disulfide bonding was inversely correlated with susceptibility of chromatin to acid denaturation (SCSA). Collectively, these data were consistent with the hypothesis that maturation of stallion spermatozoa involves oxidation of sulphydryl groups to form intra- and intermolecular disulfide links between adjacent protamines, with loss of zinc as an integral feature. These changes endow mechanical and chemical resistance to the nucleus, ensuring efficient transmission of the paternal genome at fertilization.
Publication Date: 2006-01-19 PubMed ID: 16423383DOI: 10.1016/j.theriogenology.2005.10.024Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study focuses on the comparison of nuclear characteristics between immature and mature sperms in stallions, detailing how the differences in their structures facilitate the transformation of the sperm during maturation and play a vital role in successful fertilization.
Research Objective and Methodology
- The research aimed to compare the nuclear characteristics of immature and mature stallion sperm cells with a focus on the changes in chromatin structure, which are closely related to fertility.
- Resistance to chromatin decondensation was examined by exposing the cells to Sodium Dodecyl Sulfate-Dithiothreitol (SDS-DTT) and alkaline thioglycolate, two substances that help to unravel chromatin, and comparing the responses of mature and immature cells.
- Another method involved evaluating the status of thiol-disulfide in the immature and mature cells using monobromobimane labeling. Immature cells were harvested from ductulli efferentes and mature cells from the cauda epididymidis for this test.
- Lastly, atomic absorption spectrophotometry was used to note the decrease in Zn(2+) content accompanying the maturation of sperm cells. This corresponded with increased concentrations of this ion in the surrounding epididymal fluid.
Findings
- The results showed a significant increase in resistance to in vitro decondensation of chromatin in mature sperm cells.
- Thiol-disulfide status evaluation showed that immature cells contain mostly thiol groups, while these were oxidized in mature cells.
- The maturation process is marked by a 60% reduction in the Zn(2+) content in sperm cells.
- The levels of disulfide bonding were negatively correlated with the susceptibility of chromatin to acid denaturation.
Interpretation and Conclusion
- The study concludes that the maturation of stallion sperm cells involves oxidation of sulphydryl groups, forming intra- and intermolecular disulfide links between adjacent protamines.
- The loss of zinc ions during this process appears to be an intrinsic quality of maturation.
- These changes in the sperm cell’s structure and chemistry enhance the resistance of the cell nucleus, allowing it to effectively transmit the paternal genome during fertilization.
Cite This Article
APA
Dias GM, Retamal CA, Tobella L, Arnholdt AC, López ML.
(2006).
Nuclear status of immature and mature stallion spermatozoa.
Theriogenology, 66(2), 354-365.
https://doi.org/10.1016/j.theriogenology.2005.10.024 Publication
Researcher Affiliations
- Centro de Biociências e Biotecnologia, LBCT, Setor Biologia da Reprodução, Universidade Estadual do Norte Fluminense, Av. Alberto Lamêgo 2000, Horto, Campos dos Goytacazes CEP: 28013600, RJ, Brazil.
MeSH Terms
- Animals
- Chromatin / metabolism
- Chromatin / ultrastructure
- Ejaculation / physiology
- Epididymis / physiology
- Horses / physiology
- Male
- Oxidation-Reduction
- Sperm Maturation / physiology
- Spermatogenesis / physiology
- Spermatozoa / metabolism
- Spermatozoa / physiology
- Sulfhydryl Compounds / analysis
- Zinc / analysis
- Zinc / metabolism
Citations
This article has been cited 4 times.- Schneider S, Shakeri F, Trötschel C, Arévalo L, Kruse A, Buness A, Poetsch A, Steger K, Schorle H. Protamine-2 Deficiency Initiates a Reactive Oxygen Species (ROS)-Mediated Destruction Cascade during Epididymal Sperm Maturation in Mice.. Cells 2020 Jul 27;9(8).
- Mochida K, Hasegawa A, Ogonuki N, Inoue K, Ogura A. Early production of offspring by in vitro fertilization using first-wave spermatozoa from prepubertal male mice.. J Reprod Dev 2019 Oct 23;65(5):467-473.
- Yamauchi Y, Riel JM, Stoytcheva Z, Burgoyne PS, Ward MA. Deficiency in mouse Y chromosome long arm gene complement is associated with sperm DNA damage.. Genome Biol 2010;11(6):R66.
- Balhorn R. The protamine family of sperm nuclear proteins.. Genome Biol 2007;8(9):227.
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