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Ovaries were collected from prepubertal ( 10 months, n = 12 ovaries) bitches after routine ovariohysterectomies and fixed in formalin. Ovaries were bisected, embedded in paraffin wax and 20 serial sections were made at intervals of 10 microns. Sections were stained with haematoxylin and eosin to examine follicles and oocytes in a cross-section of cortex of known size. Counts were made on all sections, resulting in examination of the entire cortical area present in the sections. Oocytes were counted and classified as nucleate or anucleate. Follicles were counted and classified as large (> 100 microns in diameter) or small (

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The relationships between activities of matrix metalloproteinases (MMPs) in the canine uterus and the occurrence of degeneration of the luminal epithelium, cystic endometrial hyperplasia, pyometra and uterine remodelling post partum were determined. Mature bitches (n = 10) were ovariectomized, treated with hormones (oestradiol benzoate, progestagen) and investigated at stages simulating pro-oestrus (n = 2), oestrus (n = 2), dioestrus (n = 2), and mid- (n = 2) and late (n = 2) anoestrus (3 and 9 weeks, respectively, after cessation of treatment with progestagen). Untreated bitches (n = 1 per group) served as controls (Expt 1). An additional 10 ovariectomized bitches, at the end of treatment-induced simulated dioestrus, were treated each day for a further 3 weeks either with the same dose (standard dose, n = 3), a decreased dose (n = 3) or an increased dose (n = 3) of progestagen, or no treatment (withdrawal dose, n = 1). These bitches were then investigated (Expt 2). A suture was placed in the lumen of one uterine horn of another five bitches at ovariectomy. Three of these bitches were treated to induce simulated dioestrus and two bitches served as untreated controls. In the hormone-treated bitches, the suture resulted in cystic endometrial hyperplasia in one bitch and in cystic endometrial hyperplasia with pyometra in two bitches. The control bitches showed no cystic endometrial hyperplasia or pyometra (Expt 3). Four intact bitches were studied at 2 (n = 1), 3 (n = 2) and 11 (n = 1) weeks post partum. Uterine tissues were also collected from two bitches with naturally occurring cystic endometrial hyperplasia with pyometra (Expt 4). All uteri were examined histologically and the activities of MMP-2, -7 and -9 (latent and active forms) were assessed using zymography of extracts of endometrium. In Expts 1 and 2, marked degeneration of the luminal epithelium in six of 25 bitches (simulated mid-anoestrus, withdrawal dose and decreased dose groups) was not associated with changes in MMP activities. Markedly increased activities of MMP-2 (active form), -7 (latent form) and -9 (active and latent forms) were observed in the bitches with cystic endometrial hyperplasia with pyometra (but not with cystic endometrial hyperplasia alone) and in the bitches at 2 and 3 weeks post partum (but not at 11 weeks post partum). These results indicate that MMPs are not involved with degeneration of the luminal epithelium, but are involved with uterine remodelling in the postpartum bitch and with cystic endometrial hyperplasia with pyometra.

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Introduction - Although the hormonal changes during estrus cycle of bitches are well- known, knowledge in luteal phase and anestrus are incomplete. Information on phenomenon perform in cytosol during estrous cycle, especially in bitch are extremely rare. The crosstalk among the different signals determines the fate of the ovarian follicle. Because there are multiple paracrine and autocrine signals, it is still not completely understood which are the critical factors that discriminate between the follicles destined for elimination by apoptosis (the major population and the follicles) and between few of the follicles that will continue to develop to reach the final stage of a Graafian preovulatory follicle. Progesterone formation and release during the estrous cycle may play an important role in the fertilization of the oocyte during ovulation. It has been suggested that early progesterone production in the preovulatory follicle impairs the quality of the mature egg during fertilization (Lindheim et al 1998, Fanchin et al 1997, Urman et al 1999). In contrast, proper timing of progesterone production and the duration of its secretion seems to be critical for maintaining functional granulosa-lutein cells, subsequent to the LH surge and maintenance of the corpus luteum during pregnancy. Progesterone interaction with its cytosolic receptor may play a part in the survival activity of the granulosa-lutein cells (Eva et al 2001). 041b061a72