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Activation of the cannabinoid receptor type 2 by the agonist JWH133 promotes the first wave of in vitro spermatogenesis
Author(s) -
Dumont Ludovic,
RivesFeraille Aurélie,
Delessard Marion,
Saulnier Justine,
Rondanino Christine,
Rives Nathalie
Publication year - 2021
Publication title -
andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.947
H-Index - 43
eISSN - 2047-2927
pISSN - 2047-2919
DOI - 10.1111/andr.12928
Subject(s) - spermatogenesis , meiosis , germ cell , biology , andrology , gametogenesis , medicine , endocrinology , in vitro , microbiology and biotechnology , immunology , genetics , embryo , embryogenesis , gene
Background Oncological procedures have irreversible side effects on germ cells for childhood cancer survival boys. In vitro culture of prepubertal testicular tissue has been proposed to restore fertility; however, recent data on animal models showed that meiotic and post‐meiotic progression was impaired. Objectives As potential key inducers of the mitosis‐meiosis switch, type 2 cannabinoid receptor (CB 2 ) has been proposed to play a central role in the meiotic entry of male germ cells. Herein, the in vitro first spermatogenesis wave in mice was used to understand the impact of CB 2 activation on the differentiation of spermatogonia until elongated spermatids. Materials and methods A first set of cultured testicular explants of 6.5 days post‐partum (d pp ) mice was performed to assess the impact of a range of JWH133 supplementation (10 n m , 100 n m , 1 µ m , 10 µ m ). Then, the progressive development of germ cells at key timepoints of spermatogenesis was evaluated throughout (i) in vitro culture (day 2 [D2], D3, D6, D10, D18, and D30) coupled with (ii) in vivo counterparts (8.5, 9.5, 12.5, 16.5, 24.5, and 36.5 d pp ). Results CB 2 was detected at the plasma membrane of cells, and a successful completion of spermatogenesis was obtained in vitro . One day after the activation of CB 2 by 1 μ m of the agonist JWH133, percentage of zygotene spermatocyte I increased. Conclusion After 30 days of culture, (i) an enrichment of haploid germ cells detected by flow cytometry, (ii) a reduced necrotic area, and (iii) an increase in the density of post‐meiotic germ cells were observed. We showed that the activation of CB 2 improves in vitro entry into meiosis and differentiation of spermatogonia, mimicking physiological meiotic transition.

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