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Department and Section of Physiology, College of Veterinary Medicine and Division of Biological Sciences, Cornell University, Ithaca, New York 14853
Address all correspondence and requests for reprints to: Joanne E. Fortune, Ph.D., Cornell University, 923 VRT, Ithaca, New York 14853. E-mail: jf11{at}cornell.edu
Mechanisms that allow selection of a dominant ovarian follicle from a
cohort of growing follicles are unknown. Large healthy, estrogen-active
follicles contain more LH receptors than atretic estrogen-inactive
follicles, and levels of messenger RNA (mRNA) for LH receptor increase
in the granulosa cells of dominant follicles as growth progresses. The
aim of the present study was to test the hypothesis that changes in the
temporal pattern of expression of mRNA for LH and FSH receptors are
associated with selection of dominant follicles in cattle. Based on
size, the dominant and two largest subordinate follicles were collected
from the ovaries of heifers on days 2 (n = 3) or 3 (n = 3) of
a follicular wave. On day 2, the dominant follicle was 1 mm larger than
the largest subordinate follicle, but by day 3 of the wave the dominant
follicle was 2–4 mm larger than the largest subordinate. Follicular
fluid concentrations of estradiol and estradiol secretion in
vitro by pieces of follicle wall (granulosa and theca cells)
were greatest by the dominant compared with the subordinate follicles
(P < 0.05). These data indicate that selection of
a dominant follicle had occurred by the second day of the follicular
wave. By in situ hybridization, mRNAs for LH and FSH
receptors, P450 aromatase and P450 17
-hydroxylase (17-OH) were
localized in frozen sections from each follicle. The expression of mRNA
for LH receptor in granulosa cells was always at or near background and
was not different between days or follicle types (P
= 0.63). In contrast, the expression of mRNA for LH receptor in theca
cells of the same sections was readily detectable; there was no
difference between follicle types on the second day of the follicular
wave, but by the third day expression in the subordinate follicles had
decreased (P < 0.05). The expression of mRNA for
FSH receptor was highest in granulosa cells of dominant follicles
collected on day 3 of the follicular wave (P <
0.05) and was not different between dominant and subordinate follicles
on day 2 of the wave (P > 0.05). The expression of
mRNA for aromatase in granulosa cells was similar
(P > 0.05) between the dominant follicles on days
2 and 3 and the largest subordinate follicle on day 2 of the follicular
wave and was much lower in the remaining follicles
(P < 0.01). On day 2 of the wave, the expression
of mRNA for 17-OH was not different between the dominant and
subordinate follicles, but by day 3 the dominant follicles had more
mRNA for 17-OH than the subordinate follicles (P
< 0.05).
These data show that the dominant follicle had been selected by the second day of the follicular wave (based on diameter and estradiol secretion) and that selection occurred in the absence of detectable levels of mRNA for LH receptor in the granulosa cells or differences between dominant and subordinate follicles in mRNA for LH receptor in theca cells or FSH receptor in granulosa cells. However, the divergent pattern of growth between dominant and subordinate follicles (after follicle selection) was associated with higher levels of mRNA for gonadotropin receptors and steroidogenic enzymes in dominant compared with subordinate follicles. Therefore, selection of the dominant follicle in cattle does not appear to involve the regulation of expression of mRNA for gonadotropin receptors, although such regulation may be important at other stages of differentiation of the dominant follicle.
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