Effects of Oxidative Stress on the Autophagy and Apoptosis of Granulosa Cells in Broody Geese.

Broodiness is an unfavorable trait associated with the cessation of egg laying. Studies have found that excessive granulosa cell apoptosis and autophagy occur during goose broodiness. Other studies have also confirmed that oxidative stress is an important cause of apoptosis and autophagy. However, whether oxidative stress occurs during goose broodiness and whether this oxidative stress causes apoptosis and autophagy have not been fully elucidated. In this study, we investigated the effects of oxidative stress on the autophagy and apoptosis of granulosa cells in broody geese. The results showed higher mRNA expression of genes related to antioxidative stress responses (GPX, SOD-1, SOD-2, COX-2, CAT and hsp70) in pre-broody and broody geese than in laying birds. In addition, increased levels of granulosa cell apoptosis and autophagy were observed in pre-broody geese than in laying geese. Additionally, granulosa cells treated with H2O2 exhibited increased apoptosis and autophagy in vitro, and these effects were responsible for goose granulosa cell death. Moreover, vitamin E treatment effectively protected granulosa cells from H2O2-induced oxidative stress by inhibiting ROS production. Correspondingly, granulosa cell apoptosis and autophagy were greatly alleviated by vitamin E treatment. Together, our results demonstrated serious oxidative stress and granulosa cell apoptosis and autophagy in broody geese, and oxidative stress promoted apoptosis and autophagy. Vitamin E alleviated the autophagy and apoptosis of granulosa cells by inhibiting oxidative stress.

MiR-34c-5p promotes granulosa cells apoptosis by targeting Bcl2 in broody goose (Anser cygnoides).

Goose (Anser cygnoides) are highly broody with low egg production, and large number of granulosa cells undergo apoptosis during broodiness. Our previous study has found that miR-34c-5p was highly abundant in the ovary of goose with broodiness phenotype. However, the mechanism that miR-34c-5p regulates granulosa cells function remains unclear. Here, we demonstrated that broody goose had higher levels of miR-34c-5p than that in laying goose by qRT-PCR. The dual luciferase reporter assay showed that Bcl2 was identified as a direct target of miR-34c-5p, which could be negatively regulated by miR-34c-5p. Furthermore, over-expression of miR-34c-5p significantly increased the rate of apoptosis and slowed down the proliferation of granulosa cells by inhibiting the Bcl2 expression, whereas the opposite trend was obtained when granulosa cells were supplemented with miR-34c-5p inhibitors. In addition, Bcl2 mRNA level was lower in goose with a brooding phenotype than that in goose with an egg-laying phenotype. Taken together, the data suggested that miR-34c-5p regulated granulosa cells apoptosis and brooding behavior by targeting Bcl2, which not only contribute to reveal the potential mechanism of miR-34c-5p underlying granulosa cells apoptosis in goose, but also provides an effective strategy to reduce the incidence of broodiness and improve the egg production.

Identifying molecular pathways and candidate genes associated with knob traits by transcriptome analysis in the goose (Anser cygnoides).

Anser cygnoides has a spherical crest on the beak roof, which is described as knob. However, the mechanisms affecting knob morphology are unclear. Here, we investigated the phenotypic characteristics and molecular basis of knob-size differences in Yangzhou geese. Anatomically, the knob was identified as frontal hump in the frontal area of the skull, rather than hump of upper beak. Although the frontal hump length, and height varied greatly in geese with different knob phenotypes, little was changed in the width. Histologically, knob skin in large-size knobs geese have a greater length in the stratum corneum, stratum spinosum, and stratum reticular than that in small-size knobs geese. Moveover, the 415 differentially expressed genes were found between the large knobs and small ones through transcriptome profiling. In addition, GO enrichment and KEGG pathway analysis revealed 455 significant GO terms and 210 KEGG pathways were enriched, respectively. Among these, TGF-ß signaling and thyroid hormone synthesis-signaling pathways were identified to determine knob-size phenotype. Furthermore, BMP5, DCN, TSHR and ADCY3 were recognized to involve in the growth and development of knob. Our data provide comprehensive molecular determinants of knob size phenotype, which can potentially promote the genetic improvement of goose knobs.

Effects of marketable ages on meat quality through fiber characteristics in the goose.

Goose meat is increasingly popular among consumers because of its good quality. The fiber characteristics have been well demonstrated to be key contributing factors of meat quality, and the marketable ages are also closely related to meat quality. However, little is known about the effect of different marketable ages on the quality of goose meat through its fiber characteristics. Here, fiber characteristics of Yangzhou geese of different marketable ages (70, 90, and 120 d) and their effect on meat quality were investigated. The results showed that only fast-twitch fibers were present in breast muscle, irrespective of age, and that few slow-twitch fibers could be identified in leg muscle, especially in gastrocnemius and extensor digitorum longus. Fiber diameter in breast muscle increased rapidly from age 70 d to 90 d, from 19.88 to 26.27 µm, and remained stable for 90 d thereafter. The diameter and cross-sectional area of muscle fiber continue to grow with day increasing in leg muscle. In addition, we measured the proximate composition and physical properties at different ages. Among the 3 marketable ages investigated, the 120-day-old geese had higher intramuscular fat and protein content, as well as lower moisture content, both in breast and leg meat. Greater lightness and pressing loss, with lower redness and shear force, were observed in the breast and leg meat of 70-day-old geese when compared with 90- or 120-day-old geese. Taken together, although older marketable age hardly affected muscle fiber type in geese, it would contribute to larger muscle fiber area, higher intramuscular fat and protein content, as well as redder and chewier meat. As a result, the reasonable marketable age should be taken into account to improve quality in goose meat production, and the marketable age of 90 or 120 d was recommended and it could potentially improve meat quality in goose meat production.

The effects of FAR1 and TGFBRAP1 on the proliferation and apoptosis of follicular granulosa cells in goose (Anser cygnoides).

The low laying performance of geese seriously damages the growth of the poultry industry, and is related to the development of pre- hierarchical follicles. Our previous studies have revealed that FAR1 and TGFBRAP1 were involved in follicular development, but the exact regulation mechanism still kept unclear. In recent studies, the expression of FAR1 and TGFBRAP1 mRNA were detected, and we found that their expression levels were relatively higher in hierarchical follicles than in pre-hierarchical follicles (P < 0.05). Moreover, generally the level of FAR1 and TGFBRAP1 mRNA gradually increased in hierarchical follicles. In addition, the proliferation and apoptosis of granulosa cells were assayed with overexpression or knockdown technology. The results showed that by the knockdown of FAR1 mRNA level, the proliferation rate of follicular granulosa cells increased significantly, the apoptosis rate decreased (P < 0.05), and the apoptosis rate also reduced obviously by transfecting TGFBRAP1-siRNA (P < 0.05). Finally, the overexpression of FAR1 or TGFBRAP1 resulted in the inhabitation to the secretion of E2 and P4 in granulosa cells, while the knockdown of FAR1 or TGFBRAP1 enhanced the secretion of E2 and P4. In conclusion, the results indicated that FAR1 and TGFBRAP1 regulated the apoptosis of follicular granulosa cells and cut the secretion of E2 and P4 in geese, which provided basic data for the understanding of the regulating process of goose reproduction.