Conservação de material genético de espécies silvestres do bioma caatinga utilizando a manipulação oócitos inclusos em folículos ovarianos pré antrais (MOIFOPA)
The objective of the present thesis was to use the manipulation of oocytes enclosed in preantral follicles (MOEPF) as a tool for the female gametes rescue and optimization, from wild species of Caatinga biome. The thesis was divided into 4 experiments. At first experiment, it was performed the es...
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| Formato: | doctoralThesis |
| Idioma: | por |
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Universidade Federal do Rio Grande do Norte
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| Endereço do item: | https://repositorio.ufrn.br/jspui/handle/123456789/19810 |
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| Resumo: | The objective of the present thesis was to use the manipulation of oocytes
enclosed in preantral follicles (MOEPF) as a tool for the female gametes rescue and
optimization, from wild species of Caatinga biome. The thesis was divided into 4
experiments. At first experiment, it was performed the estimative and description of the
agouti (Dasyprocta leporina) preantral follicles (PF) histologic and ultrastructural
features, in which it was estimated 4419.8 ± 532.26 and 5397.52 ± 574.91 follicles for
the right and left ovary, respectively, and the majority (86,63%) belonged to the
primordial follicles category (P<0.05). Most of the population consists of
morphologically normal follicles (70.78%), presenting a large and central nuclei and
uniform cytoplasm. At ultrastructural evaluation it was verified the presence of a great
number of round mitochondrias associated to lipid droplets. In the second experiment, it
was performed the estimative and description of yellow-toothed cavies (Galea spixii) PF
characteristics, also, the evaluation of the effect of solid surface vitrification (SSV) on the
in situ PF morphology. The total of 416.0 ± 342.8 PF was estimated for the ovary pair
and the presence of a large quantity of primary follicles (P<0.05) was evidenced. Most of
the PF was morphologically normal (94.6%), in which the oocyte nuclei presented
condensed granules of heterochromatin. Round or elongated shaped mitochondria
constituted the most abundant organelles. In regard of the SSV, the protocol using the
dimethylsulfoxide (DMSO) 3M possibility the preservation of 69.5% of morphologically
normal PF, which was evidenced by the light and transmission electronic microscopy. At
third experiment, the evaluation of the SSV procedure on the morphology and viability
in situ PF form collared peccaries (Pecari tajacu) was performed. No differences were
observed among treatments, in which the use of DMSO, ethylene glycol (EG) and
dimethylformamide (DMF) as cryoprotectants, regardless its concentration, promoted the
morphology preservation of much than 70% of PF. Concerning the PF viability, the
DMSO and EG promoted the best preservation. The fourth experiment aimed to evaluate
the effect of α MEM+ or TCM199 associated or not to 50 ng of FSHr on the morphology,
activation and growth of collared peccaries PF, in vitro cultured (IVC) during 1 or 7 days
and the effect on the extracellular matrix (ECM). After 7 days of IVC only the use of
TCM199/FSH maintained the proportion of intact PF, similar to day 1(63.2%), however,
no differences were observed among treatments (P>0.05). Also, an improvement of the
proportion of intact growing PF was verified (P>0.05). By the Ag-NOR analysis it was
observed that only the treatment using TCM199/FSH promoted the maintenance of cell
proliferation similar to day 1 (P>0.05). The picrosirius red stain revealed that ECM
remained intact in all treatments (P>0.05). Thus, as the general conclusion, the use of
MOEPF in the refereed species allowed the knowledge of aspects related to its
reproductive morphology and physiology, enabling the germplasm conservation, with the
possibility of germplasm bank formation, as the elucidation of mechanisms related to the
PF survive and in vitro development. |
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