The following experiments were designed to examine the effect of serum of spayed dogs on superovulation response in mice and rats. In Experiment 1, female mice at diestrus (n=30) were divided into three equal groups and superovulated with either administration of 5 IU pregnant mare serum gonadotropin (PMSG) or recombinant follicle stimulating hormone (rFSH) (reducing dose from 2.5 to 0.5 IU) and 5 IU human chorionic gonadotropin (hCG) administered 48h later. Serum of spayed dogs was administered intraperitoneally at a reduced dose from 0.1 to 0.025 mL in a 48 h period. In Experiment 2, female rats (n=30) at diestrus stage were divided into three equal groups. Superovulation was induced using either 30 IU PMSG, or a dose reduced from 5 to 1 IU rFSH and 25 IU hCG administered 48h later. Serum of spayed dogs was administered in a reduced dose from 0.6 to 0.1 mL in a 48 hour period. Female mice and rats were mated 24 h following hCG administration. On day 14 after mating, animals were euthanized and ovarian sections were fixed for histopathological evaluation and corpus luteum (CL) counting. No significant difference observed in mean (±SEM) number of CLs between the PMSG group and the mice that received serum of spayed dog (10.4±1.3 vs 9.2±1.0). Mean (±SEM) number of CLs tended to be lower in rats that received serum of spayed dog than those of rats which received either PMSG or rFSH (15.1±1.9 vs 23.6±3.1 and 23.1±2.9, P=0.06, respectively). In conclusion, serum of spayed dogs is able to induce a superovulatory response in mice and rats.
Animals ; Chorionic Gonadotropin ; Corpus Luteum ; Diestrus ; Dogs* ; Female* ; Follicle Stimulating Hormone ; Gonadotropins ; Humans ; Mice* ; Rats* ; Superovulation*

Opinions on ovariohysterectomy (OHE) of bitches vary depending on region and country. In this descriptive, prospective cross-sectional study, uterine tracts and ovaries exhibiting gross pathologic findings (n = 76) were collected post-surgery from a reference population of 3,600 bitches (2.11% incidence) that underwent elective OHE during September to November 2013 and evaluated by histopathology examination. Data were evaluated by using descriptive statistics and chi-squared tests. Bitches were of crossbred background with average age 5 years (range 0.6–8.0 years) and most were nulliparous (69.7%) with no anamnesis of reproductive diseases (81.6%). Frequencies of proestrus, estrus, and diestrus were 42.1%, 6.6%, and 19.7%, respectively. The presence of mammary gland masses (5.3%) significantly correlated with histopathologic findings in ovaries and age of the bitch (p < 0.05). Predominant uterine histopathologies included cystic endometrial hyperplasia, periglandular fibrosis, lymphoplasmocytary endometritis, and adenomyosis (19.7%, 14.5%, 4.0%, and 2.6%, respectively). In ovaries, hyperplasia of rete ovarii, follicular cysts, oophoritis, adenoma of the rete ovarii, cysts of superficial structures, and granulosa cell tumors (10.5%, 10.5%, 7.9%, 4.0%, 2.6%, and 2.6%, respectively) were observed. The results reveal the presence of subclinical pathologies in healthy bitches, suggesting that OHE at an early age is beneficial for prevention of reproductive pathologies.
Adenoma ; Adenomyosis ; Animals ; Cross-Sectional Studies* ; Diestrus ; Dogs* ; Endometrial Hyperplasia ; Endometritis ; Estrus ; Female ; Fibrosis ; Follicular Cyst ; Granulosa Cell Tumor ; Hyperplasia ; Mammary Glands, Human ; Oophoritis ; Ovary* ; Pathology ; Proestrus ; Prospective Studies ; Uterus*

A primary characteristic of autism, which is a neurodevelopmental disorder, is impaired social interaction and communication. Furthermore, patients with autism frequently show abnormal social recognition. In mouse models of autism, social recognition is usually assessed by examining same-sex social behavior using various tests, such as the three-chamber test. However, no studies have examined the ability of male mice with autism to recognize the estrous cycle of female partners. In this study, we investigated the sexual behaviors, especially mounting and ultrasonic vocal communication (USV), of BTBR T+ tf/J (BTBR) mice, which are used as a well-known mouse model of autism, when they encountered estrus or diestrus female mice. As expected, C57BL/6 mice mounted more female mice in the estrus stage compared with the diestrus stage. We found that BTBR mice also mounted more female mice in the estrus stage than female mice in the diestrus stage. Although the USV emission of male mice was not different between estrus and diestrus female mice in both strains, the mounting result implies that BTBR mice distinguish sexual receptivity of females.
Animals ; Autistic Disorder ; Diestrus ; Estrous Cycle* ; Estrus ; Female* ; Humans ; Interpersonal Relations ; Male* ; Mice* ; Neurodevelopmental Disorders ; Sexual Behavior* ; Social Behavior ; Ultrasonics*

PURPOSE: We established a rat model to investigate the female vaginal arousal response and the effects of the estrous cycle on vaginal blood flow in vivo. MATERIALS AND METHODS: Laser Doppler flowmetry was used to measure changes in vaginal blood flow induced by pelvic nerve stimulation(PNS) in 30 female Sprague-Dawley rats. Frequency response data were obtained in each animal. In addition, the rat's stage in the estrous cycle was determined according to the cell types observed in a vaginal smear. Changes in blood flow at 10 Hz frequency caused maximal response, and these responses were evaluated by comparing the relative peak flow, time to peak, duration of response, and relative area under the curve(AUC) according to the estrous cycle. RESULTS: Reproducible frequency dependent increases in vaginal blood flow were observed in response to PNS. Relative peak flow, time to peak, duration of response and relative AUC were slightly greater in proestrus and estrus groups(relatively higher estradiol level, n=17) than those in metestrus and diestrus groups(relatively lower estradiol level, n=13). However, these differences were not statistically significant. CONCLUSIONS: Our data suggest that the rat is a useful and reliable animal model for investigating the vaginal arousal response. In addition, the estrous cycle of the animal does not seem to be an important confounding factor in this animal model.
Animals ; Area Under Curve ; Arousal* ; Diestrus ; Estradiol ; Estrous Cycle ; Estrus ; Female* ; Humans ; Laser-Doppler Flowmetry ; Metestrus ; Models, Animal* ; Proestrus ; Rats* ; Rats, Sprague-Dawley ; Vagina ; Vaginal Smears

Animals ; Diestrus ; drug effects ; Female ; Hyperprolactinemia ; physiopathology ; Panax ; Prolactin ; blood ; Rats ; Rats, Wistar ; Saponins ; pharmacology

BACKGROUND: Pituitary adenylate cyclase-activating polypeptide (PACAP) plays the role of a hypophysiotropic factor, which regulates the synthesis and secretion of pituitary hormones through the hypothalamo-hypophysial portal system. No clear evidence has yet been reported regarding the regulation of prolactin (PRL) by PACAP. In the present study, we tested a hypothesis that PACAP regulates the synthetic machinery of PRL during the estrus cycle and pubertal process using intracerebroventricular (i.c.v.) injection of an antisense oligodeoxynucleotide (ODN) against type I PACAP receptor (PAC1). METHODS: An RNase protection assay (RPA) was used to determine the pattern of hypothalamic PACAP and PAC1 mRNA expressions during the estrus cycle. Antisense PAC1 ODN was administered via i.c.v. injection to the female rats in normal estrus cycle of pubertal process. Northern blot analysis was used to determine the mRNA ievel of PRL in the pituitary gland. RESULTS: 1) PACAP mRNA in the medial basal hypothalamus was significantly increased at the diestrus I, while PAC1 mRNA showed no significant change. 2) PRL mRNA level of pituitary was increased by an injection of antisense PAC1 ODN at the proestrus and estrus stages. 3) PRL mRNA level of pituitary was significantly decreased by antisense PAC1 ODN injection at stage of prepuberty and initiate puberty, while its level was increased at stage of puberty. CONCLUSION: These data suggest that PACAP suppresses PRL mRNA synthesis through the PAC1 signaling pathway in the certain estrus cycle environments. It may be also involved in the regulation of pituitary PRL gene expression during the pubertal process
Adolescent ; Animals ; Blotting, Northern ; Diestrus ; Estrus ; Female* ; Gene Expression* ; Humans ; Hypothalamus ; Pituitary Adenylate Cyclase-Activating Polypeptide* ; Pituitary Gland* ; Pituitary Hormones ; Portal System ; Proestrus ; Prolactin* ; Puberty ; Rats* ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide ; Ribonucleases ; RNA, Messenger

OBJECTIVE: This study was to investigate the localization of CDK inhibitor, p57(kip2) in mouse endometrium during the estrus cycle and pre- and peri-implantation periods. METHODS: The p57(kip2) protein was immunostained from endometrium of mouse sacrificed at diestrus, proestrus, estrus, and metestrus cycle, and at day 1-6 post-coitum (p.c.). RESULTS: The staining in the luminal epithelium was very weak in comparison with glandular and stromal cells. In diestrus stage, immunoreactivity of p57(kip2) was heterogeneously strong in parts of decidualized or degenerated stromal cells. In proestrus stage, strong immunoreactivity p57(kip2) was largely found in stromal cells. But, p57(kip2) was showed low immunoreactivity in estrus stage. In metestrus stage, immunoreactivity of p57(kip2) was heterogeneously strong in decidualized stromal cells. In day 1-2 p.c., immunoreactivity of p57(kip2) was low in some endometrial stromal cells. In day 3-4 p.c., immunoreactivity of p57(kip2) was strong in some endometrial stromal cells. In day 5-6 p.c., immunoreactivity of p57(kip2) was strong in decidual cells. CONCLUSION: These suggest that p57(kip2) may play an essential role in endometrial differentiation for maintenance of implantation, especially decidualization of endometrial stromal cells.
Animals ; Diestrus ; Endometrium* ; Epithelium ; Estrus* ; Female ; Metestrus ; Mice* ; Phenobarbital ; Proestrus ; Stromal Cells

OBJECTIVE: This study was to investigate the expression of CDK inhibitors, p27(kip1) and p57(kip2) in mouse endometrium during the estrus cycle and pregnant period. METHODS: Total RNA and protein were extracted from endometrium of mouse sacrificed at diestrus, proestrus, estrus, and metestrus cycle, and at day 1-6 post-coitum (p.c.), then semi-quantitative RT-PCR and western blotting of p27(kip1) and p57(kip2) was carried out. RESULTS: p27(kip1) and p57(kip2) mRNA was highly expressed in diestrus and proestrus stage than estrus and metestrus stage. In comparison with estrus cycle, p27(kip1) and p57(kip2) mRNA level was highly maintained in gestational endometrium (except p27(kip1) of day 5 p.c). p57(kip2) protein level was relatively low from day 1 p.c. to day 4 p.c. But it was significantly increased in day 5 p.c. and day 6 p.c. CONCLUSION: These results show that p27(kip1) and p57(kip2) may play a role in endometrial differentiation for regular estrus cycle and implantation, and especially p57(kip2) may play an essential role in endometrial differentiation for maintenance of implantation.
Animals ; Blotting, Western ; Diestrus ; Endometrium* ; Estrus ; Female ; Metestrus ; Mice* ; Proestrus ; RNA ; RNA, Messenger

OBJECTIVE: Bok, Bcl-2-related ovarian killer, is a proapoptotic Bcl-2 family protein identified in the ovary based on its dimerization with the antiapoptotic protein Mcl-1. The present study examined the hormonal regulation and localization of Bok messenger RNA levels in the rat ovary during the follicle development. METHODS: We used the female rats of SD strain. Bok mRNA levels in the ovary were determined by Northern blot analysis. In situ hybridization were performed to determine the specific ovarian cell type expressing Bok mRNA. RESULTS: Northern blot analysis of ovaries obtained from immature rats revealed increasing levels of Bok mRNA during postnatal development. The major cell types expressing Bok mRNA were the granulosa cells of preantral and atretic follicles. Treatment of immature rats with diethylstilbestrol (DES) for 24-48 h increased ovarian Bok mRNA levels. Bok mRNA was remained the same levels in rats removed DES for 24- 48 h to induce apoptosis. High signals of Bok mRNA after DES treatment were detected in granulosa cells of early antral follicles. Treatment of immature rats with pregnant mare's serum gonadotropin (PMSG) for 12 h increased markedly ovarian Bok mRNA expression which was detected mainly in preantral and atretic follicles. Interestingly, low levels of Bok mRNA were also expressed in granulosa cells of preovulatory follicles. Treatment of PMSG-primed rats with human chorionic gonadotropin (hCG) stimulated strongly ovarian Bok mRNA expression at 6-9 h. At that time, Bok mRNA was expressed in granulosa cells of atretic and small growing follicles. In adult estrus cyclic ovaries, Bok gene expression was higher on proestrus and estrus than metaestrus and diestrus. Moreover, the highly increased expression of Bok mRNA was found in rat ovaries at 48 h after hypophysectomy. CONCLUSION: These results demonstrate that Bok is one of proapoptotic Bcl-2 members expressed in early growing and atretic follicles during the ovarian follicular development. Gonadotropins induce a transient increase of Bok gene expression in granulosa cells of preantral and preovulatory follicles indicating some role in the ovulatory process.
Adult ; Animals ; Apoptosis* ; Blotting, Northern ; Chorionic Gonadotropin ; Diestrus ; Diethylstilbestrol ; Dimerization ; Estrus ; Female ; Gene Expression ; Gonadotropins ; Granulosa Cells ; Humans ; Hypophysectomy ; In Situ Hybridization ; Ovarian Follicle ; Ovary* ; Proestrus ; Rats* ; RNA, Messenger

Protein expression patterns of matrix metalloproteinases (MMPs) were examined in mouse reproductive organs during estrous cycle. Estrous cycle was classified into diestrus, proestrus, estrus or metestus and MMP expression was analyzed by zymography using gelatin as a substrate. Uterine fluid (UF) obtained both at diestrus and proestrus exhibited 4 major MMPs including 106kDa, 64kDa, 62kDa and 59kDa gelatinases. However, in UF at estrus, the gelatinolytic activity of 64kDa MMP disappeared and that of 106kDa and 62kDa MMPs dramatically decreased. At metestrus, 64kDa MMP activity reappeared and 106kDa and 62kDa MMP exhibited increased activities such that the band intensity of 106kDa was comparable to that in UF at diestrus. Gelatinolytic activity of 59kDa MMP was not changed throughout the cycle. Both ovarian and oviductal tissue homogenate revealed 4 MMPs which corresponded to the 4 MMPs of UF. However, unlike UF MMPs, gelatinolytic activity of these MMPs did not show distinct changes throughout the cycle. Either an inhibitor of MMP, 1, 10-phenanthroline, or a metal chelator, EDTA, abolished the appearance of the above MMP activities in gelatinated gel whereas a serine proteinase inhibitor, phenylmethylsulfonyl fluoride, failed to inhibit the appearance of MMP activities, proving that gelatinolytic activity of the above reproductive tissues were due to the enzymatic activity of MMP. When gelatinolytic activity of mouse serum was examined, it revealed 5 MMPs (131kDa, 106kDa, 89kDa, 64kDa and 62kDa bands) and one gelatinase (84kDa) band. From these results, it is concluded that the protein expression of MMPs of mouse reproductive organs, particularly uterus, is temporally regulated during estrous cycle and uterine 106kDa, 64kDa and 62kDa MMP,3 are suggested to play an important role in cyclic tissue remodeling of mouse uterus.
Animals ; Diestrus ; Edetic Acid ; Estrous Cycle* ; Estrus ; Female ; Gelatin ; Gelatinases ; Matrix Metalloproteinases* ; Metestrus ; Mice* ; Oviducts ; Phenylmethylsulfonyl Fluoride ; Proestrus ; Serine Proteases ; Uterus