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

Estrous cycle -related histological and histochemical changes in the vaginal epithelium of mature female rats were studied with PAS (periodic acid Schiff) alcian blue pH 2.5 and biotinylated lectins (DBA, SBA, PNA, BSL -1, sWGA, UEA -1, RCA -1, Con A and LCA).The prominent characteristic changes that occured during the estrous cycle were mucinous transformation in proestrus and cornification in estrus. In proestrus, the superficial mucinous cells of the epithelium were increased in number and enlarged in size, and the amount of acid and neutral mucosubstances was more increase in proestrus than in diestrus and metestrus. About the binding pattern of all lectins examined to the superficial mucinous cells, in diestrus, the binding pattern of these cells showed a similar affinity as in metestrus with intense DBA and UEA -1 reactivity. In proestrus, however, these cells were reactive with seven lectins examined except LCA and PNA, and DBA, SBA, BSL -1, RCA -1 and UEA -1 reacted more strongly than in diestrus and metestrus. In estrus, the superficial cornified cell layers showed a weak reactivity of SBA, BSL -1 and PNA. In diestrus and metestrus, the mucinous cells in the intermediate layers of the basal portion of vaginal fold stained with eight lectins examined except LCA and showed the same binding pattern to the superficial mucinous cells. About the distribution of glycoconjugates in the intermediate layer, the upper spindle cells showed different binding pattern according to the estrous stages. In diestrus, estrus, and metestrus, these cells showed a affinity for all lectins examined. In proestrus, however, DBA and PNA staining were not observed, and stained more intensely with sWGA, SBA and UEA -1, and less intensely with BSL -1 and RCA - 1. In estrus, DBA and PNA reactivity reappeared as trace, and RCA -1 and sWGA reactivity increased. In metestrus, sWGA reactivity reduced and BSL -1 and UEA -1 increased continually. The lower rounded cells of the intermediate layers stained with all lectins examined in estrus, with six lectins examined except Con A, DBA and UEA -1 in proestrus and with five lectins examined except DBA, UEA -1, sWGA and BSL -1 in diestrus and metestrus. BSL -1 reactivity for the layers increased in proestrus, estrus and metestrus, and PNA reactivity increased in estrus and reduced in metestrus. The basal layer of the vaginal epithelium showed different binding pattern to the different portion of vagina, and showed faint staining of BSL -1, SBA and RCA -1, and moderately staining of BSL -1 in proestrus and estrus. In conclusion, alpha /-N -acetyl -D -galactosamine, alpha /-D -galactose and alpha -L -fucose participate in the mucinous transformation of the vaginal epithelium, and beta -N -acetyl -D -glucosamine participates in the cornification of the vaginal epithelium.
Alcian Blue ; Animals ; Diestrus ; Epithelium* ; Estrous Cycle* ; Estrus ; Female ; Glycoconjugates ; Humans ; Hydrogen-Ion Concentration ; Lectins ; Metestrus ; Mucins ; Proestrus ; Rats* ; Vagina

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

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

PURPOSE: To characterize the relationship between serum estradiol levels and the expression of glucose transporter type 4 (Glut4) in the pubococcygeus and iliococcygeus muscles in female rats. METHODS: The muscles were excised from virgin rats during the metestrus and proestrus stages of the estrous cycle, and from sham and ovariectomized rats implanted with empty or estradiol benzoate–filled capsules. The expression of estrogen receptors (ERs) was inspected in the muscles at metestrus and proestrus. Relative Glut4 expression, glycogen content, and serum glucose levels were measured. Appropriate statistical tests were done to identify significant differences (P≤0.05). RESULTS: The pubococcygeus and iliococcygeus muscles expressed ERα and ERβ. Glut4 expression and glycogen content in the pubococcygeus muscle were higher at proestrus than at metestrus. No significant changes were observed in the iliococcygeus muscle. In ovariectomized rats, the administration of estradiol benzoate increased Glut4 expression and glycogen content in the pubococcygeus muscle alone. CONCLUSIONS: High serum estradiol levels increased Glut4 expression and glycogen content in the pubococcygeus muscle, but not in the iliococcygeus muscle.
Animals ; Benzoates ; Blood Glucose ; Capsules ; Estradiol* ; Estrous Cycle ; Female ; Glucose Transport Proteins, Facilitative* ; Glucose Transporter Type 4* ; Glucose* ; Glycogen ; Humans ; Metabolism ; Metestrus ; Muscles* ; Ovariectomy ; Pelvic Floor* ; Proestrus ; Rats* ; Receptors, Estrogen