Effects of jacalin and follicle-stimulating hormone onin vitrogoat primordial follicle activation, survival and gene expression

Abstract

This study aims to investigate the effects of jacalin and follicle-stimulating hormone (FSH) on activation and survival of goat primordial follicles, as well as on gene expression in cultured ovarian tissue. Ovarian fragments were cultured for 6 days in minimum essential medium (MEM) supplemented with jacalin (10, 25, 50 or 100 μg/ml - Experiment 1) or in MEM supplemented with jacalin (50 μg/ml), FSH (50 ng/ml) or both (Experiment 2). Non-cultured and cultured tissues were processed for histological and ultrastructural analysis. Cultured tissues from Experiment 2 were also stored to evaluate the expression of BMP-15, KL (Kit ligand), c-kit, GDF-9 and proliferating cell nuclear antigen (PCNA) by real-time polymerase chain reaction (PCR). The results of Experiment 1 showed that, compared with tissue that was cultured in control medium, the presence of 50 μg/ml of jacalin increased both the percentages of developing follicles and viability. In Experiment 2, after 6 days, higher percentages of normal follicles were observed in tissue cultured in presence of FSH, jacalin or both, but no synergistic interaction between FSH and jacalin was observed. These substances had no significant effect on the levels of mRNA for BMP-15 and KL, but FSH increased significantly the levels of mRNA for PCNA and c-kit. On the other hand, jacalin reduced the levels of mRNA for GDF-9. In conclusion, jacalin and FSH are able to improve primordial follicle activation and survival after 6 days of culture. Furthermore, presence of FSH increases the expression of mRNA for PCNA and c-kit, but jacalin resulted in lower GDF-9 mRNA expression.

Supplementation of in vitro culture medium with FSH to grow follicles and mature oocytes can be replaced by extracts of Justicia insularis

“…Several genes have been identified in follicular walls as biomarkers to understand the dynamics of follicular development. Among them GPx related to oxidative stress [64], KL indicative of follicular growth [65]; cyclin B1 and HAS2 indicative of progression of oocyte maturation [66]. We observed that after the IVM of COCs from secondary follicles, the follicular walls (granulosa and theca cells) expressed similar transcript levels.…”

Kit ligand promotes the transition from primordial to primary follicles afterin vitroculture of ovine ovarian tissue

“…The tissues were cut into 5-μm sections, stained with hematoxylin–eosin and then examined by microscopy (Nikon, Tokyo, Japan; ×400 magnification). Follicles were classified as primordial (one layer of flattened or flattened and cuboidal granulosa cells around the oocyte), primary (one layer of cuboidal granulosa cells) or secondary (two or more layers of cuboidal granulosa cells around the oocyte) (Ribeiro et al , 2014), and as morphologically normal or atretic, according to the presence or absence of pyknotic nucleus, retracted oocyte, and/or disorganized granulosa cells detached from the basement membrane. To evaluate follicular activation, only morphologically normal follicles with a visible oocyte nucleus were recorded, and the proportion of primordial and growing follicles was calculated at day 0 (fresh control) and after 7 days of culture.…”

A utilização de GnRH ou Galectina-1 no momento da IA aumenta a taxa de concepção em vacas de corte submetidas a IATF?

“…Assim, os autores demonstraram a importância fundamental da GAL-1 na tolerância materno-fetal e na sinergia entre GAL-1 e progesterona na manutenção da gestação. Ribeiro et al, (2015) investigaram o efeito de diferentes concentrações de uma lectina vegetal no desenvolvimento, maturação e fertilização de folículos, e verificaram que esta molécula estimula a ativação dos folículos e contribui para a manutenção da viabilidade embrionária, bem como exerce efeito na expressão de mRNA.…”