DNA methylation and histone post-translational modifications in the mouse germline following in-vitro maturation of fresh or cryopreserved prepubertal testicular tissue

Autor: Justine Saulnier, Antoine Oblette, Julie Rondeaux, Ludovic Dumont, Nathalie Rives, Marion Delessard, Aurélie Rives, Christine Rondanino
Přispěvatelé: Gamétogenèse et Qualité du Gamète - ULR 4308 (GQG), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université de Lille, CHU Rouen, Normandie Université (NU)
Rok vydání: 2019
Předmět:
Zdroj: 2019, 39 (3), pp.383-401. ⟨10.1016/j.rbmo.2019.05.007⟩
Reproductive BioMedicine
Reproductive BioMedicine Online, Elsevier, 2019, 39 (3), pp.383-401. ⟨10.1016/j.rbmo.2019.05.007⟩
ISSN: 1472-6483
1472-6491
DOI: 10.1016/j.rbmo.2019.05.007
Popis: Research question Do cryopreservation and in-vitro culture procedures affect the expression of DNA methyltransferases (DNMT) and histone-modifying enzymes, as well as the establishment of DNA methylation and histone post-translational modifications (PTM) in germ cells in prepubertal mouse testicular tissue? Design This study investigated the expression of epigenetic modification enzymes, DNA methylation and histone PTM, and the spermatogenic progression after in-vitro maturation of fresh or cryopreserved mouse prepubertal testicular tissue. Fresh or cryopreserved testicular fragments from 6–7 days post-partum mice were cultured for 30 days in the presence of retinol with or without FSH. Results The in-vitro maturation of fresh or cryopreserved tissue allowed the differentiation of spermatogonia into spermatozoa. Differences in the levels of transcripts encoding epigenetic modification enzymes (Dnmt1, Dnmt3a, Jarid1b, Src1, Sirt1, Hdac1) were found between 30-day tissue cultures and age-matched in-vivo controls. DNMT1/DNMT3a expression and the presence of 5-methylcytosine (5mC) were detected in spermatogonia and leptotene/zygotene spermatocytes in cultures. The relative 5mC fluorescence intensity was similar in spermatozoa produced in cultures of cryopreserved tissues or in vivo. H3K4me3, H3K9ac and H4K8ac were present in all germ cell types but differences in the proportion of germ cells containing these epigenetic marks were found after cultures. Conclusions Despite differences with the in-vivo situation, DNA methylation and histone methylation and acetylation occur in the mouse germline in in-vitro matured fresh or cryopreserved mouse prepubertal testicular tissue, and the expression of the enzymes catalysing these epigenetic modifications are maintained in vitro.
Databáze: OpenAIRE