Expression of growth and differentiation factor 9 (GDF-9) and its effect on the in vitro culture of caprine preantral ovarian follicles
| Autor: | J.R.V. Silva, Ana Beatriz Graça Duarte, Valdevane Rocha Araújo, D.M. Magalhães, Cláudio Cabral Campello, Márcia Viviane Alves Saraiva, Isana Mara Aragão Frota, A.P. Almeida, C.A.P. Lopes, José Ricardo de Figueiredo |
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| Rok vydání: | 2011 |
| Předmět: |
Oocyte
endocrine system medicine.medical_specialty Basal medium Total rna mRNA expression Biology Caprine In vitro In vitro maturation Recombinant fsh Endocrinology medicine.anatomical_structure Food Animals Theca Internal medicine Follicular growth and viability embryonic structures medicine Animal Science and Zoology GDF-9 Antrum |
| Zdroj: | Small Ruminant Research. 100:169-176 |
| ISSN: | 0921-4488 |
| DOI: | 10.1016/j.smallrumres.2011.06.001 |
| Popis: | This study examined the expression of growth and differentiation factor 9 (GDF-9) in caprine ovarian follicles, and the effect of GDF-9 with or without FSH on the in vitro culture of preantral follicles. To evaluate the expression of GDF-9 in Experiment 1, follicles were recovered from 32 goat ovaries and the total RNA isolated and transcribed for real-time polymerase chain reaction (PCR). Experiments 2 and 3 each used a further 32 goat ovaries to provide preantral follicles of ≥150μm. These follicles were isolated and cultured individually in 100μL drops. In each experiment at least 45 follicles were used per treatment. Every 6 days, follicles were evaluated for viability, antrum formation and growth rate. At the end of the culture period, oocytes were submitted to in vitro maturation (IVM), viability tests and chromatin evaluation. In Experiment 2, follicles were cultured in a basal medium (control) or this medium supplemented with GDF-9 at a concentration of 100ng/mL (GDF-9 100) or 200ng/mL (GDF-9 200). The same media were used in Experiment 3, supplemented with recombinant FSH at a level of 100ng/mL from day 0, 500ng/mL from day 6 to 12 and 1000ng/mL from day 12 to 18 of culture to form the three treatments: control FSH, GDF-9 (100) plus FSH and GDF-9 (200) plus FSH. Relative GDF-9 expression (Experiment 1) was greater in the secondary (18units) than the primordial (1unit) and the primary (1unit) preantral follicles (P3mm diameter (1unit; P3mm (319.2 and 200.1units, respectively), compared to their respective granulosa and theca cells (1unit for each category, P |
| Databáze: | OpenAIRE |
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