Anti-Müllerian hormone type II receptor in avian follicle development.

Anti-Müllerian hormone (AMH) helps maintain the ovarian reserve by regulating primordial follicle activation and follicular selection in mammals, although its role within the avian ovary is unknown. In mammals, AMH is primarily produced in granulosa cells of preantral and early antral follicles. Similarly, in the hen, the granulosa cells of smaller follicles are the predominant source of AMH. The importance of AMH in mammalian ovarian dynamics suggests the protein and its specific Type II receptor, AMHRII, may have conserved functions in the hen. AMHRII mRNA expression is highest (P < 0.01) in small follicles of the hen and decreases as follicle size increases. Similarly, expression of AMHRII and AMH is highest in granulosa cells from small follicles as compared to larger follicles. Dissection of 3-5 mm follicles into ooplasm and granulosa components shows that AMHRII mRNA levels are greater in ooplasm than granulosa cells. Furthermore, immunohistochemistry also revealed AMHRII staining in the oocyte and granulosa cells. AMH expression in mammals is elevated during periods of reproductive dormancy, possibly protecting the ovarian reserve. AMHRII and AMH mRNA were significantly higher (P < 0.05) in nonlaying ovaries of broiler hens. In molting layer hens, AMHRII mRNA was significantly greater (P < 0.05) compared to nonmolting hen ovaries. These results suggest that AMH may have a direct effect on the oocyte and, thereby, contribute to bidirectional communication between oocyte and granulosa cells. Enhanced expression of AMHRII and AMH during reproductive quiescence supports a potential role of AMH in protecting the ovarian reserve in hens.

Occludin expression and regulation in small follicles of the layer and broiler breeder hen.

Synchronized yolk accumulation and follicle development are essential for egg production in oviparous species. In birds, yolk is incorporated into the oocyte by an avian specific yolk receptor (LR8), and it has been suggested that occludin (OCLN), a tight junction protein, mediates transfer of yolk material to the oocyte surface. OCLN may be a key regulator of yolk accumulation and follicle growth, however, the expression and regulation of OCLN in granulosa cells during various stages of follicle development is unknown. In the first experiment, we found that LR8 and OCLN mRNA were highest in small follicles within the ovary. In addition, OCLN decreased with increasing follicle size. OCLN mRNA was more abundant in the germinal disc region of the granulosa cell layer than the non-germinal disc region. In addition, we found epidermal growth factor (EGF) and activin B, decreased OCLN mRNA, while activin A increased OCLN. In the second experiment, restricted fed (RF) broiler breeder hens were randomly divided into two groups and one group remained on RF and the other was fed ad libitum (FF). OCLN expression in granulosa cells of 3-5mm follicles of FF hens was lower compared to RF hens and yolk weights were higher in the FF group, however, LR8 mRNA in small whole follicles (<3mm) did not differ between groups. In conclusion, the level of feed intake is related to or may directly regulate OCLN mRNA expression or may have an indirect effect through paracrine or autocrine factors in the ovary.

Bone morphogenetic protein 15 may promote follicle selection in the hen.

In the hen, optimal ovulation rate depends on selection of a single follicle into the pre-ovulatory hierarchy. Follicle selection is associated with increased oocyte growth and changes in gene expression in granulosa cells surrounding the oocyte, in preparation for ovulation. This study investigated the expression, function and regulation of bone morphogenetic protein-15 (BMP15) during follicle development in the hen. BMP15 mRNA expression was analyzed in the ooplasm and granulosa cells of 3mm follicles and was confirmed to be primarily in the ooplasm. BMP15 was detected by immunoblotting in 6 and 8mm follicles near the time of follicle selection. Expression of mRNA for BMP15 receptors (BMPR1B and BMPR2) in granulosa cells increased with follicle size, indicating that BMP15 may play an important role around follicle selection. The function of BMP15 was examined by culturing granulosa cells from 3-5mm and 6-8mm follicles with recombinant human BMP15 (rhBMP15). BMP15 increased expression of follicle stimulating hormone receptor (FSHR) mRNA and decreased anti-Müllerian hormone (AMH) mRNA and occludin (OCLN), factors associated with follicle maturation and growth in the hen. Hormonal regulation of BMP15 was assessed by whole follicle culture with estradiol (E2) which increased BMP15 mRNA expression. The distinct expression pattern of BMP15 and its receptors, coupled with the effects of BMP15 to increase FSHR mRNA and decrease AMH mRNA and OCLN mRNA and protein expression suggest that the oocyte may have a role in follicle selection in the chicken.

The domestic chicken: Causes and consequences of an egg a day.

The domestic laying chicken has been intensely selected to be a persistent ovulator. That is, the tendency for broodiness has been nearly eliminated and, given the appropriate lighting and nutrition, many strains of laying hens produce an egg on almost every day. The regulatory mechanisms involved in coordination of neuroendocrine and ovarian events have been well studied and described. In spite of this, there has been little attention focused on the oocyte itself. Recent findings in mammals have indicated that the oocyte produces several oocyte-specific factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic factor 15 (BMP15), which influence the surrounding cells and follicular development. Our studies indicate that GDF9 is present in the hen oocyte and influences granulosa cell proliferation. Additionally, Bmp15 mRNA is most abundant in oocytes of small follicles and stimulates an increase in follicle stimulating hormone (FSH) receptor mRNA in granulosa cells. BMP15 also enhances yolk uptake in growing follicles by decreasing tight junctions between granulosa cells. These studies indicate that the oocyte likely contributes to follicle development. Commercial laying hens also spontaneously develop ovarian cancer at a high rate, and susceptibility to this disease has been associated with ovulatory events in women. Studies have shown that ovulation, or events associated with ovulation, increase the prevalence of ovarian cancer in hens. Inhibition of ovulation in hens through a hormonal strategy mimicking oral contraceptives results in a decrease of ovarian cancer incidence. Recent studies in women have suggested that some ovarian tumors may arise from the distal oviduct. Gene expression profiles in very early stage tumors from hens show a high expression of oviduct-related genes, supporting the possibility of oviduct origin for some ovarian tumors. Genetic selection for high productivity in commercial laying hens has generated an efficient and valuable food source as well as an important animal model for human ovarian cancer.