The nuclear form of glutathione peroxidase 4 is associated with sperm nuclear matrix and is required for proper paternal chromatin decondensation at fertilization

Autor: Carla Boitani, Marcus Conrad, Franco Mangia, Rossella Puglisi, Irene Maccari, Simona Pipolo
Rok vydání: 2012
Předmět:
Male
metabolism [Recombinant Proteins]
Physiology
Clinical Biochemistry
GPX4
physiology [Oocytes]
Mice
chemistry.chemical_compound
Human fertilization
cytology [Epididymis]
Chlorocebus aethiops
deficiency [Glutathione Peroxidase]
Phospholipid-hydroperoxide glutathione peroxidase
enzymology [Nuclear Matrix]
Epididymis
Mice
Knockout
physiology [Chromatin Assembly and Disassembly]
Spermatozoa
physiology [Embryonic Development]
physiology [Fertilization]
enzymology [Spermatozoa]
Recombinant Proteins
Chromatin
genetics [Recombinant Proteins]
medicine.anatomical_structure
physiology [Spermatogenesis]
COS Cells
Female
Germ cell
endocrine system
genetics [Glutathione Peroxidase]
Embryonic Development
Fertilization in Vitro
Biology
Transfection
physiology [Spermatozoa]
Chromosomal Instability
ddc:570
medicine
Animals
Nuclear Matrix
Spermatogenesis
Glutathione Peroxidase
Embryogenesis
Cell Biology
Chromatin Assembly and Disassembly
Phospholipid Hydroperoxide Glutathione Peroxidase
Nuclear matrix
Molecular biology
Sperm
Mice
Inbred C57BL
metabolism [Glutathione Peroxidase]
chemistry
Fertilization
Oocytes
physiology [Chromosomal Instability]
enzymology [Epididymis]
Zdroj: Journal of cellular physiology / Supplement 227(4), 1420-1427 (2012). doi:10.1002/jcp.22857
ISSN: 0021-9541
DOI: 10.1002/jcp.22857
Popis: The nuclear isoform of the selenoprotein Phospholipid Hydroperoxide Glutathione Peroxidase (nGPx4) is expressed in haploid male germ cells, contains several cysteines and is able to oxidize protein thiols, besides glutathione. In this study we have investigated the subnuclear localization of this isoform in isolated mouse male germ cells at different steps of maturation. Immunoblotting and confocal microscopy analyses of subnuclear fractions showed that nGPx4 is localized to the nuclear matrix together with well known markers of this subnuclear compartment like lamin B and topoisomerase IIβ at all stages of germ cell differentiation. The peculiar nGPx4 distribution was confirmed by both biochemical and morphological analyses of COS-1 cells overexpressing Flag-tagged nGPx4. To test the functional role of nGPx4 in the process of chromatin assembly, sperm isolated from the caput and the cauda epididymides of wild-type (WT) and genetically deficient in nGPx4 (nGPx4-KO) mice were analyzed in an in vitro chromatin decondensation assay. Results showed that sperm from nGPx4-KO mice were more prone to decondense than those from WT mice at all stages of epididymal maturation, providing conclusive evidence that nGPx4 is required for a correct sperm chromatin compaction. We next addressed the issue of whether the lack of nGPx4 impacts on early events occurring at fertilization. Indeed, in vitro fertilization experiments showed an acceleration of sperm chromatin dispersion in oocytes fertilized by nGpx4-KO sperm compared with control. Overall these data indicate that the absence of nGPx4 leads to sperm nuclear matrix/chromatin instability that may negatively affect the embryo development.
Databáze: OpenAIRE
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