RESUMO

Ovarian puncture has been widely used in assisted reproduction, but there are still gaps about its effects on ovarian morphophysiology, as well as the relationship between inflammation caused by this procedure and the follicular growth and fertility. The aim of this study was to investigate the effects of ovarian puncture on folliculogenesis and fertility. Mice (n = 24) were divided into two groups: (1) SHAM-both ovaries were exposed and repositioned and (2) Punctured-ovaries were exposed, punctured, and repositioned. After 96 h of surgery, ovaries were collected for morphofunctional analysis. New females were used for the superovulation (n = 10) and fertility assays (n = 10). Increased volumetric density of inflammatory cells-p = 0.0005, p = 0.0013; hemorrhagic foci-p < 0.0001; and inflammatory exudate-p < 0.0001 could be noticed on the punctured group, compared to SHAM. The percentage of primordial follicles was lower on the punctured ovaries (p = 0.00294). Ovarian puncture has also induced an increase in the proliferation of granulosa cells of primary (p = 0.0321) and antral follicles (p = 0.0395), and an increased apoptotic index of antral follicles (p = 0.0100). There was no influence on expression of some genes related to inflammation, collagen deposition and folliculogenesis progression. The reproductive aspects (oocyte retrieval and number of fetuses per female) were not altered (p > 0.05). Taken together, our findings strongly suggest that ovarian puncture results in a local inflammation that affects follicular growth and atresia. However, it does not affect female fertility, which strengthens the safety of this procedure.

RESUMO

Naproxen reduces the production of prostaglandins via inhibition of the cyclooxygenase. Studies have shown that its administration in women can be related to failed ovulation. Therefore, preclinical investigations must be performed in order to investigate its effects in experimental models. Thus, the aim of this study was to evaluate the effects of naproxen on murine folliculogenesis, ovulation, and female fertility. Female C57BL/6 mice (n = 128 - 6 weeks old) were divided into Control, low (10 mg/kg), and high naproxen (50 mg/kg) groups, who were treated for 8 days and directed to morphofunctional analyses. Follicular quantification showed a reduced percentage of antral follicles in naproxen-treated animals. These treated animals also showed smaller oocytes included in secondary and antral follicles, and the diameter of secondary and antral follicles was also reduced. A reduction in the percentage of Ki67-positive granulosa cells was observed in treated animals that also showed down-regulation of Igf1r compared to control. After an ovarian stimulation protocol, naproxen-treated animals showed a reduction in the percentage of secondary and antral follicles, a reduced number of ovulated oocytes and, corpora lutea, and an increased number of failed ovulations. Finally, naproxen-treated animals also showed a reduction in mating index and pregnancy rate. Our findings suggested that, in mice, naproxen administration (eight days treatment) negatively affects molecular and morphological aspects related to late folliculogenesis, ovulation, and fertility.

Assuntos

RESUMO

Conjugated linoleic acid (CLA) is a mixture of positional isomers of linoleic acid found in meat and dairy products from ruminants. It is a trans fat widely used by athletes as a food supplement, due to a supposed effect of maximizing the use of body fat reserves. The interest in diet and culture media supplementation with CLA is an emerging area, demanding studies in order to elucidate its benefits in the reproductive parameters, as well as in cryopreservation. Therefore, the aim of this review was to discuss the effects of CLA on the oocytes, sperm and embryos cryotolerance. Some studies have already demonstrated its use in cryopreservation of germline. Among those, it was observed that CLA supplementation during oocyte in vitro maturation can increase their viability post-freezing and developmental capacity. Regarding the use of CLA on sperm, there are few studies and their results are still inconclusive. Finally, studies about CLA supplementation on embryo culture media have shown promising results, indicating that this bioactive molecule is able to modulate lipid uptake on blastomeres. Altogether, these findings demonstrate the potential use of CLA as a bioactive molecule to improve germline and embryo cryotolerance and open new perspectives on human and animal reproduction field.(AU)

O ácido linoleico conjugado (CLA) é uma mistura de isômeros posicionais do ácido linoleico encontrado em carne e laticínios de ruminantes. É um tipo de gordura trans muito utilizada por atletas para como suplemento alimentar devido a um suposto efeito de maximizar a utilização das reservas de gordura corporal. O interesse na suplementação de dietas e meios de cultura com o CLA é uma área emergente, exigindo estudos para elucidar seus benefícios nos parâmetros reprodutivos e na criopreservação. Dessa forma, o objetivo dessa revisão foi discutir os efeitos do CLA na criotolerância de oócitos, espermatozóides e embriões. Alguns estudos já demonstraram seu uso na criopreservação da linhagem germinativa. Entre esses, foi observado que a suplementação de CLA durante a maturação in vitro de oócitos pode aumentar sua viabilidade pós-congelamento e capacidade de desenvolvimento. Em relação ao uso de CLA no esperma, existem poucos estudos e seus resultados ainda são inconclusivos. Por último, estudos sobre a suplementação de CLA em meios de cultura de embriões mostraram resultados promissores, indicando que essa molécula bioativa é capaz de modular a captação de lipídios em blastômeros. No total, essas descobertas demonstram o potencial uso do CLA como uma molécula bioativa para melhorar a linha germinativa e a criotolerância ao embrião e abrir novas perspectivas no campo da reprodução humana e animal.(AU)

Assuntos

RESUMO

Conjugated linoleic acid (CLA) is a mixture of positional isomers of linoleic acid found in meat and dairy products from ruminants. It is a trans fat widely used by athletes as a food supplement, due to a supposed effect of maximizing the use of body fat reserves. The interest in diet and culture media supplementation with CLA is an emerging area, demanding studies in order to elucidate its benefits in the reproductive parameters, as well as in cryopreservation. Therefore, the aim of this review was to discuss the effects of CLA on the oocytes, sperm and embryos cryotolerance. Some studies have already demonstrated its use in cryopreservation of germline. Among those, it was observed that CLA supplementation during oocyte in vitro maturation can increase their viability post-freezing and developmental capacity. Regarding the use of CLA on sperm, there are few studies and their results are still inconclusive. Finally, studies about CLA supplementation on embryo culture media have shown promising results, indicating that this bioactive molecule is able to modulate lipid uptake on blastomeres. Altogether, these findings demonstrate the potential use of CLA as a bioactive molecule to improve germline and embryo cryotolerance and open new perspectives on human and animal reproduction field.

O ácido linoleico conjugado (CLA) é uma mistura de isômeros posicionais do ácido linoleico encontrado em carne e laticínios de ruminantes. É um tipo de gordura trans muito utilizada por atletas para como suplemento alimentar devido a um suposto efeito de maximizar a utilização das reservas de gordura corporal. O interesse na suplementação de dietas e meios de cultura com o CLA é uma área emergente, exigindo estudos para elucidar seus benefícios nos parâmetros reprodutivos e na criopreservação. Dessa forma, o objetivo dessa revisão foi discutir os efeitos do CLA na criotolerância de oócitos, espermatozóides e embriões. Alguns estudos já demonstraram seu uso na criopreservação da linhagem germinativa. Entre esses, foi observado que a suplementação de CLA durante a maturação in vitro de oócitos pode aumentar sua viabilidade pós-congelamento e capacidade de desenvolvimento. Em relação ao uso de CLA no esperma, existem poucos estudos e seus resultados ainda são inconclusivos. Por último, estudos sobre a suplementação de CLA em meios de cultura de embriões mostraram resultados promissores, indicando que essa molécula bioativa é capaz de modular a captação de lipídios em blastômeros. No total, essas descobertas demonstram o potencial uso do CLA como uma molécula bioativa para melhorar a linha germinativa e a criotolerância ao embrião e abrir novas perspectivas no campo da reprodução humana e animal.

Assuntos

RESUMO

Ovarian tissue cryopreservation is emerging as a promising alternative for fertility preservation of cancer survivors. To date, more than a hundred couples have successfully had babies using this procedure, although it is still considered experimental and demands further investigation. In this work, we evaluated the effects of vitrification, warming and autotransplantation procedures on the morphology and gene expression of murine ovaries. Ovaries were removed from adult female C57BL6 mice (n = 15), vitrified, warmed and autotransplanted (vitrified group), additionally, ovaries were autotransplanted without vitrification (control group, n = 15). After twenty days, grafted ovaries were harvested and used for histological and ultrastructural analysis, germinal vesicle (GV) oocyte collection, RNA sequencing, and Transmission Electron Microscopy (TEM). All classes of follicles and GV were observed in both control and vitrified/warmed transplanted ovaries, and the numbers of primordial, antral and atretic follicles were not different (p > 0.05). Using RNA-seq, we detected 16,602 vs 13,527 expressed genes in vitrified and control ovaries, respectively; and 623 significantly dysregulated genes (fold change >1.5; 332 up-regulated and 291 down-regulated). Cellular membranes, cytoskeletons, and extracellular matrices were found as the main functions of the differentially expressed genes. Moreover, vitrified samples also presented ultrastructural alterations in the cytoskeleton, cell junctions, and endoplasmic reticulum. Taken together, this work showed for the first time that ovarian cells might trigger a compensatory gene regulation mechanism to maintain cellular structure and folliculogenesis progression after vitrification and autotransplantation.

Assuntos