Follicular growth and atresia in the ovaries of hens (Gallus domesticus) with diminished egg production rates.

Diets containing 3.5, 1.0 and 0.1% calcium were fed from the age of 42 weeks to individually caged laying hens. Ovaries were examined at 46-49 and 70 weeks of age for changes in the follicular population corresponding to the lowered egg production rates of birds given calcium-deficient diets (1.0% and 0.1%) and older birds given a normal diet (3.5%). Growth rates of follicles from 3.5 mm diameter to ovulation were not changed by the level of dietary calcium in 46-49-week-old birds. The number of atretic small follicles (less than or equal to 8 mm diam.) increased in old and calcium-deprived birds, resulting in lower numbers of viable follicles in the intermediate stages of growth (3-8 mm diam.). There was also an increase in the number of small follicles (1-2 mm diam.) starting to grow in 70-week-old birds which may have partly compensated for the increased loss by atresia. Birds of all ages on all diets were able to produce large follicles up to ovulable size. The main feature of poor laying birds was a reduction in the ovulation rate due to the loss of large follicles (greater than 8 mm diam.) by atresia, an event seen rarely in the birds with good laying performance. As atresia is the normal fate of most of the small follicles, the mechanisms controlling atresia in the small follicles and the large follicles appear to be independent.

Role of atresia in establishing the follicular hierarchy in the ovary of the domestic hen (Gallus domesticus).

At peak laying periods the ovary of the domestic hen contained 30-100 small yolky follicles with diameters varying between 1 and 8 mm. In general, the number of these healthy follicles decreased with increasing size in that there were about 20 follicles with a diameter of 1-2 mm and 1 follicle (mean less than 1) with a diameter of 7-8 mm. The number of follicles with diameters greater than 8 mm (the hierarchy of large, yolky follicles) varied between 4 and 7. By using a dye-marker, growth from 3 to 5 mm was estimated to take 3 days, from 5 to 8 mm, 2 days and from 8 mm to ovulation, 6 days. No information was obtained for growth between 1 and 3 mm because the dye did not enter these smaller follicles. Between 5 and 25 small yolky follicles were atretic. The reduction in the number of follicles with time and the high incidence of atresia suggests that this is a normal fate of small yolky follicles in birds with a high rate of lay. In marked contrast, only one large yolky follicle was observed to be atretic throughout the whole experiment. These two very different levels of atresia serve to define two distinct groups of follicles in the size range studied. Ovulation rate appears to be the product of two complementary mechanisms, one for the initiation of growth and the other controlling the rate at which the small yolky follicles are lost through atresia.

Variations in the number of nucleoli in the granulosa cells of the domestic fowl during follicular growth.

1. The proportions of granulosa cells containing one, two, or more nucleoli were determined in ovarian follicles of the domestic fowl (Gallus domesticus). Follicles ranged in size from 5 to 37 mm diameter. 2. The greatest proportion of cells (65%) contained one nucleolus. Cells containing two nucleoli were fewer (29%) and those containing more than two were rare (6%). 3. There was a tendency for the proportion of multinuclear cells to decrease with increasing size of the follicle. Conversely there was a similar tendency for the proportion of cells with single nucleoli to increase with size of the follicle. These relationships are curvilinear. 4. Suitable linear models confirmed these observations (P less than 0.005). 5. These results may provide an explanation for the previously reported progressive decrease in cellular DNA as the follicle increases in size.

Cellular changes in the granulosa layer of the maturing ovarian follicle of the domestic fowl.

1. The relationships between length, width and height of the maturing ovarian follicle of the domestic fowl (Gallus domesticus) are shown. 2. During the rapid-growth phase (yolk deposition) of the oocyte, i.e. an increase in size from about 9 mm diameter to 35 mm diameter, there is about a 15-fold increase in its surface area. During this time the number of granulosa cells adjacent to the oocyte increases by about 5-fold. 3. The granulosa cells become flatter during the growth period of the follicle, consequently increasing their surface area adjacent to the oocyte by about 3-fold. 4. Together the change in shape of the granulosa cells and the increase in their number account for the increase in area of the granulosa layer during follicular growth. 5. Measurement of the DNA content of the granulosa layer indicated a progressive decrease in the cellular content of DNA as the follicle matures.

Effect of ovine LH on the progesterone content of the granulosa cells in preovulatory follicles of the domestic fowl (Gallus Domesticus).

Laying hens were injected i.v. with a single dose of ovine LH (100 microgram) 6-8 hr after a known ovulation, i.e. when plasma LH concentrations are low. Groups of 4 birds were killed at 15 min intervals up to 1 h after injection and the complete granulosa was obtained from 4 or 5 of the largest preovulatory follicles of each bird for progesterone measurement by g.l.c. Compared with the saline-injected controls which had negligible levels of progesterone, granulosa progesterone increased in all the LH-treated birds. The maximum response (mean progesterone content 1966 ng/follicle) occurred 45 min after injection but there was a significant decline by 60 min. It is concluded that the granulosa cells provide most of the preovulatory plasma progesterone increase.