High Estradiol Differentially Affects the Expression of the Glucose Transporter Type 4 in Pelvic Floor Muscles of Rats / 대한배뇨장애요실금학회지

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.

Establishment of an In Vivo Rat Model to Investigate Female Vaginal Arousal Response / 대한남성과학회지

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.

The Expression of p27(kip1) and p57(kip2) in Mouse Endometrium / 대한산부인과학회잡지

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.

The Expression of p57(kip2) in Mouse Endometrium During Estrus Cycle and Pre- and Peri-implantation Periods / 대한산부인과학회잡지

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.

Influence of the stage of the cycle on olfactory sensitivity in laboratory mice.

Females in estrus showed maximum olfactory sensitivity as judged by their ability in locating the buried bait. The results suggest that olfactory sensitivity in females varies during the stages of the estrous cycle. The findings further indicate that gonadal steroids play an important role in the expression of olfactory sensitivity in females.

Protein Expression of Matrix Metalloproteinases of Mouse Reproductive Organs During Estrous Cycle / 대한불임학회지

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.

Glycoconjugate Histochemistry of Rat Vaginal Epithelium During the Estrous Cycle / 체질인류학회지

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.

Aspectos morfológicos do epitélio uterino de ratas nude atímicas nas fases de proestro e de metaestro do ciclo estral / Morphological characteristics of the uterine epithelium of athimic nude rats, in the proestrus and metestrus phases of estrous cycle

O presente experimento compara a nível de microscopia de luz a morfologia do epitélio uterino de ratas congenitamente atímicas (rnu/rnu) com a de suas "littermtes" eutímicas (rnu/ +), da linhagen Rowett, nas fases de proestro e de metaestro do ciclo estral. Pudemos observar que a morfologia do epitélio uterino das ratas homozigotas (rnu/rnu) foi semelhante à das heterozigotas (rnu/ +), em ambas as fases estudadas e compatível com a morfologia uterina descrita na literatura referente a ratas comuns. No metaestro verificamos epitélio do tipo cilíndrico simples e áreas deste epitélio em desarranjo com algumas das células apresentando vacuolizaçäo citoplasmática e núcleo picnótico. No proestro, o epitélio cilíndrico simples apresentou-se constituído por células contendo núcleo alongado deslocado para a porçäo basal

Ultrastructural characteristics of the endometrial stroma of athymic nude rats in the proestrus and metestrus phases of the estrous cycle

En el presente estudio se observa la ultraestructura del estroma endometrial de ratas congenitamente atímicas (rnu/rnu), homozigotas para el gen nude recesivo y de sus camadas eutímicas (+/rnu), heterozigotas para el mismo gen, ambas de la línea Rowet, con el propósito de detectar posibles alteraciones en las ratas rnu/rnu. Estudios morfológicos del aparato reproductor de éstas, usando microscopio de luz, no muestran diferencias de las ratas +/rnu. Estudios ultraestructurales referentes a este asunto no fueron encontrados en la literatura consultada. La ultraestructura del estroma endometrial de rnu/rnu y +/rnu mostraron fibroblastos con una GER más dilatada en el proestro que el metaestro y una menor frecuencia de eosinófilos y macrófagos en las ratas rnu/rnu que en las +/rnu en proestro. La frecuencia de monocitos fue mas alta en el proestro mientras que la de los neutrófilos fue mas baja en el proestro y metaestro, en ambos grupos de ratas. Los resultados ultraestructurales sugieren que la menor frecuencia de eosinófilos y macrófagos encontrados en la ratas rnu/rnu en fase de proestro pueden estar relacionados con atmia congénita y/o alteraciones en los niveles de hormonas esteroidales encontradas en esas ratas

Alterations in feeding and sexual behaviour during reproductive cycle in female rats.

Alterations in feeding and sexual behaviour were studied in adult female rats. During estrous phase there was decrease in food intake and increased sexual behaviour, as manifested by increased lordosis quotient. During met-and diestrous phases, increase in food intake and depressed sexual behaviour were observed, indicating inverse relationship between these two behaviours. Multisensory hypothalamic neurons may explain interactions among regulatory systems.

Lipid pattern in female reproductive tissues during different phases of estrous cycle.

The normal pattern of lipids during various phases of estrous cycle in different reproductive tissues of female rats was analysed. The various lipid calsses undergo striking cyclical changes in harmony with the rhythm of endogenous gonadal and gonadotrophic hormones. A comparison of lipid pattern was made between various phases of estrous cycle. Ovary exhibited marked alterations in the distribution of lipids in different phases of the cycle compared to other tissues. Total lipids were maximum in diestrus phospholipids in estrus with a concomittant fall in triglycerides in all the tissues. The fluctuation in oviductal and vaginal lipids was much less compared to other tissues. Hormone-lipid interrelationship and tissue specificity may be attributed to these changes.