Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

Abstract

The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

Influence of caprine arthritis encephalitis on expression of ovulation related genes and activation of primordial follicles cultured in presence of phytohemagglutinin, epidermal growth factor or both

“…In infected animals, presence of PHA in culture medium led to an increase in primordial follicle diameter. Cunha et al (2013) observed that such a presence of PHA maintained the viability and ultrastructure of in vitro cultured follicles, and promoted the formation of an antral cavity.…”

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

“…In this context, Velasquez et al ., (2013) demonstrated that the lectin concanavalin A (Con A) inhibited the stimulatory effect of FSH and induces oocyte degeneration and death of rat granulosa cells. On the other hand, addition of 10 mg/ml phytohemagglutinin (PHA) during in vitro culture caprine secondary follicles stimulated antrum formation and expression of FSH-R and PCNA mRNAs, which helped to keep ultrastructural integrity of cultured follicles (Cunha et al ., 2013).…”

Transcriptome profiling reveals the immune response of goose T cells under selenium stimuli

“…; Cunha et al . ). There are two major sources of Se for poultry: inorganic Se, in the form of selenite or selenate, and organic Se, commonly in the form of selenomethionine, which are the principal chemical forms of Se in most poultry diets (Boostani et al .…”

“…Selenium (Se) is an essential trace element for poultry. Selenium plays a critical role in immune response, enhancing lymphocyte proliferation and T cell function in chickens (Boscolo et al 2005;Cunha et al 2013). There are two major sources of Se for poultry: inorganic Se, in the form of selenite or selenate, and organic Se, commonly in the form of selenomethionine, which are the principal chemical forms of Se in most poultry diets (Boostani et al 2015;Burk & Hill 2015).…”