Dynamics of male canine germ cell development

Autor(es)

de Souza, Aline F. ^{[1, 2]} ; Godoy Pieri, Naira C. ^[3] ; Roballo, Kelly C. S. ^[1] ; Bressan, Fabiana F. ^{[1, 2]} ; Casals, Juliana B. ^[2] ; Ambrosio, Carlos E. ^{[1, 2]} ; Perecin, Felipe ^{[1, 2]} ; Martins, Daniele S. ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Fac Anim Sci & Food Engn, Dept Vet Med, Pirassununga, SP - Brazil

^[2] Univ Sao Paulo, Fac Vet Med & Anim Sci, Dept Surg, Sao Paulo, SP - Brazil

^[3] Univ Sao Paulo, Fac Vet Med, Dept Reprod, Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 13, n. 2 FEB 28 2018.
• Citações Web of Science: 5

Resumo

Primordial germ cells (PGCs) are precursors of gametes that can generate new individuals throughout life in both males and females. Additionally, PGCs have been shown to differentiate into embryonic germ cells (EGCs) after in vitro culture. Most studies investigating germinative cells have been performed in rodents and humans but not dogs (Canis lupus familiaris). Here, we elucidated the dynamics of the expression of pluripotent (POU5F1 and NANOG), germline (DDX4, DAZL and DPPA3), and epigenetic (5mC, 5hmC, H3K27me3 and H3K9me2) markers that are important for the development of male canine germ cells during the early (22 +/- 30 days post-fertilization (dpf)), middle (35 +/- 40 dpf) and late (45 +/- 50 dpf) gestational periods. We performed sex genotype characterization, immunofluorescence, immunohistochemistry, and quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) analyses. Furthermore, in a preliminary study, we evaluated the capacity of canine embryo PGCs (30 dpf) to differentiate into EGCs. To confirm the canine EGCs phenotype, we performed alkaline phosphatase detection, immunohistochemistry, electron and transmission scanning microscopy and RT-qPCR analyses. The PGCs were positive for POU5F1 and H3K27me3 during all assessed developmental periods, including all periods between the gonadal tissue stage and foetal testes development. The number of NANOG, DDX4, DAZL, DPPA3 and 5mC-positive cells increased along with the developing cords from 35 +/- 50 dpf. Moreover, our results demonstrate the feasibility of inducing canine PGCs into putative EGCs that present pluripotent markers, such as POU5F1 and the NANOG gene, and exhibit reduced expression of germinative genes and increased expression of H3K27me3. This study provides new insight into male germ cell development mechanisms in dogs. (AU)

• Processo FAPESP: 13/10542-5 - Epigenética da reprogramação em células germinativas embrionárias caninas
• Beneficiário: Aline Fernanda de Souza
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 12/05609-0 - Espermatogênese xenogênica pós-transplante de células tronco caninas no testículo de camundongos
• Beneficiário: Naira Caroline Godoy Pieri
• Modalidade de apoio: Bolsas no Brasil - Doutorado