Relationship between cyclic follicle recruitment and ovulation in the hen ovary

Determining whether follicle recruitment in the domestic hen is functionally linked to ovulation would inform investigations on the exact time of cyclic recruitment and subsequently, the earliest cellular event mediating recruitment. The objective of the present studies was to determine if the absence of ovulation results in the failure of cyclic recruitment. 3 groups of Hy-Line W-36 hens were studied: no-ovulation, first-ovulation, or late-ovulation within the sequence. Time and occurrence of oviposition and ovulation were documented by video recording and cloacal palpation. To determine the presence of a recently recruited follicle, ovaries were collected during the last hour of photoperiod, and follicles weighing 280 to 900 mg, with yellow yolk incorporated, and diameter of 9 to 12 mm, were considered recently recruited. To compare the amount of yolk uptake in recruited follicles, hens were fed Sudan IV dye 6 h before ovary collection. Percent dyed yolk was quantified from cross-sections of the recruited follicles. To confirm follicle viability, granulosa cell (GC) mitotic activity was assessed via flow cytometry. Initial results indicated the presence of a recently recruited follicle in 10 of 11, 7 of 10, and 9 of 10, no-ovulation, first-ovulation, and late-ovulation ovaries, respectively, with no significant differences in weight (543 ± 62 mg, 456 ± 71 mg, 620 ± 75 mg, respectively). There were no significant differences in yolk incorporation among the most recently recruited follicles with 19.0 ± 2.0%, 18.0 ± 3.7%, and 19.8 ± 3.2% dyed yolk in no-ovulation, first-ovulation, and late-ovulation ovaries, respectively. Finally, there were no significant differences in % GC in the S/G2-M phase of mitosis among recruited follicles (19.0 ± 5.0%, 22.0 ± 4.2%, 15.67 ± 1.0%, in no-ovulation, first-ovulation, and late-ovulation ovaries, respectively), confirming viability of all recruited follicles. We conclude that cyclic recruitment occurs independently of ovulation and propose that the order and timing of cyclic recruitment is predetermined at an earlier stage of follicle development.