Mechanism of the development of permanent estrus in rats after transplantation of the ovaries into a low temperature medium

Autotransplantation of the ovaries to the ears of adult rats induces permanent estrus following 5-7 days. Autotransplantation of the ovaries beneath the renal capsules makes the sexual cycle return to normal after the same period. Autotransplantation of the ovaries to the ears of infantile rats brings about pubertas precox followed by normal sexual cycle. Permanent estrus ensues only after 4 months. Transplantation of the ovaries from infantile rats to adult ones and vice versa has shown that age-related differences in the alterations seen in the sexual cycle in response to gonadal transplantation to the ears are caused by age-related differences in the recipients, precisely by those in the cyclic center.     

The effects of continuous light and darkness on the activity of monoamine oxidase A and B in the hypothalamus, ovaries and uterus of rats

For assessing monoamine oxidase (MAO-A and -B) activities in the hypothalamus, ovaries and uterus, mature female rats were exposed to either continuous light or dark over 6 weeks. Confirming previous studies, continuous light induced constant estrus in all animals. The majority of animals kept under continuous dark during the six weeks remained mostly in diestrus with estrus appearing sporadically. The endocrinological function of the ovaries was disturbed by continuous light, which resulted in the development of polycystic ovaries, their morphological appearance being not significantly affected by continuous dark. Hypothalamic MAO-A activity was markedly reduced under the influence of both light (p < 0.05) and dark (p < 0.01). The activity of hypothalamic MAO-B was reduced only under the influence of dark (p < 0.01). In the ovaries, no significant differences were detected in either MAO-A or -B activity as a result of these environmental manipulations. In the uterus of rats exposed to continuous light which developed polycystic ovaries, MAO-A activity was significantly lower (p < 0.05) in comparison to either intact controls or rats exposed to continuous dark. These results demonstrate that chronic changes in photoperiodicity may considerably influence MAO-A activity and to a lesser extent MAO-B activity dependent on the tissue studied.     

In vitro sensitivity of rat ovarian follicles to the direct action of various hormones applied in particular stages of sexual cycle

The direct action of various hormones on the ovary of adult rat ovaries has been investigated. The ovaries were surgically exposed at particular stages of sexual cycle i.e. in estrus, metestrus, diestrus and proestrus, and were injected with various doses of gonadotropic hormones. Injected ovaries were excised 56 hrs after injection and examined histologically. The reactivity of the ovaries to hormone applied was dependent on stage of the cycle. The considerable increase of the amount of t1 and t2 follicles was observed after injection of gonadotropins in estrus stage. In metestrus stage the influence of gonadotropins on the follicles more advanced in development was pronounced. Numerous follicles t3 appeared in diestrus under the influence of all used hormones.     

Antiovulatory effect of a single injection of pure antiestrogen ZK 191703 at early stage of rat estrus cycle

The effects of ZK 191703 (ZK), a pure antiestrogen, on ovulation, follicle development and peripheral hormone levels were investigated in rats with 4-day estrus cycle and gonadotropin-primed immature rats in comparison to tamoxifen (TAM)-treatment. In adult rats, a single s.c. injection of ZK (5 mg/kg) or TAM (5 mg/kg) at an early stage of the estrus cycle (diestrus 9:00) inhibited ovulation, and was associated with suppression of the surge of preovulatory LH, FSH and progesterone. In rats treated with ZK or TAM at a late stage of the estrus cycle (proestrus 9:00), no inhibitory effects on ovulation, the gonadotropin and progesterone surge were detected. ZK treatment at diestrus 9:00, in contrast to TAM, increased the baseline LH level. When immature rats were treated with antiestrogens in the earlier stage of follicular development, 6 and 30 h but not 48 h or later after injection of gonadotropin (PMSG), ovulation was attenuated, associated with a lowered progesterone level. Unruptured preovulatory follicles were found in most of the ovaries from anovulatory animals treated with ZK or TAM. Antiestrogens, ZK and TAM administered at an early phase of the estrus cycle delay the follicular development functionally and inhibit ovulation in rats and suppression of the preovulatory progesterone surge.     

Pinealectomy and sexual rhythm in female rats.

The role of the pineal gland in inducing and maintaining the persistent estrus of rats exposed to continuous illumination was examined. Under the cyclic illumination, the pinealectomy or sham-operation revealed no effect on the estrous cycle, although weights of the ovaries, adrenals and hypophysis were slightly but definitely greater in the pinealectomized animals. In rats exposed to the continous illumination immediately after pinealectomy or sham-operation, both groups exhibited the persistent estrous states soon after the change of lighting condition. In these rats, neither the degree of persistent estrus nor the organ weights a autopsy showed any significant difference between the groups. Moreover, the pinealectomy could not alter the incidence of estrous in persistent estrous rats which had been established already by the continuous illumination.     

Light modes and ovulation function in rats in ontogenesis

Enhancement of illumination was shown to lead to occurrence of early and more obvious age-dependent changes of ovulation cycle in rats as compared with the same changes in animals staying in conditions of light deprivation or standard illumination. The effect of 24-hour illumination during a year induced more obvious and still earlier changes of the estrus cycle that the effect of natural illumination in the North-West of Russia with its peculiar yearly photo-periodicity (short light day in the autumn-winter period and "white nights" in the spring-summer period).     

Ovarian hormone secretory response to gonadotropins and nitric oxide following chronic nitric oxide deficiency in the rat

Ovarian hormone secretion is regulated by gonadotropins, and it has been demonstrated that this response is modulated by nitric oxide (NO). The focus of this study was to determine the effect of chronic NO deficiency on the secretion of ovarian steroids. Female rats were given N-nitro-L-arginine (L-NNA; 0.6 g/L) in their drinking water, and vaginal smears were obtained daily. By 4 wk of treatment, all the rats were in constant estrus or proestrus. At 6-8 wk the animals were killed; the ovaries were removed and incubated in the presence of eCG (1 IU/ml) and hCG (1 IU/ml) and/or S-nitroso-L-acetyl penicillamine (an NO donor, S-NAP; 0.1 mM) for 4 h. Medium was collected at 30-min intervals, and estradiol, progesterone, and androstenedione were measured. Ovaries from proestrous rats served as controls. Ovaries from L-NNA-treated animals had a greater basal and gonadotropin-stimulated release of estradiol but not of androstenedione or progesterone in comparison to ovaries from untreated controls. S-NAP decreased the gonadotropin-stimulated estradiol, progesterone, and androstenedione in ovaries from NO-deficient rats. Steroid secretion in controls was not responsive to S-NAP. We conclude that chronic NO inhibition produces constant estrus due to increased estradiol production and that NO acts to inhibit estradiol and androstenedione production.     

Effects of neonatal treatment with phytoestrogens, genistein and daidzein, on sex difference in female rat brain function: estrous cycle and lordosis

It is well known that neonatal exposure to estrogen induces masculinization or defeminization of the brain. In this study, the effects of neonatal treatment with two kinds of soybean isoflavone aglycone, genistein (GS) and daidzein (DZ), on the estrous cycle and lordosis behavior were investigated. Female rats were injected subcutaneously with 1 mg GS, 1 mg DZ, 100 microg estradiol (E2), or oil daily for 5 days from birth. As a result, vaginal opening was advanced in GS- or E2-treated females. A vaginal smear check indicated that oil- or DZ-treated females showed a constant 4- or 5-day estrous cycle, whereas GS- or E2-treated rats showed a persistent or prolonged estrus. Ovariectomy was performed in all females at 60 days of age. The ovaries in the GS- or E2-treated groups were smaller than those in the oil- and DZ-treated groups and contained no corpora lutea. In the DZ group, although corpora lutea were seen, ovaries were smaller than that of control females. Behavioral tests were carried out after implantation of E2-tubes. All of the oil- or DZ-treated females showed lordosis with a high lordosis quotient (LQ). On the other hand, as male rats, LQs were extremely low in the E2-treated group, when compared to the oil-treated group. In the GS-treated group, the mean LQ was lower than that in the oil-treated group, but higher than those in the E2-treated female or male groups. These results suggest that genistein acts as an estrogen in the sexual differentiation of the brain and causes defeminization of the brain in regulating lordosis and the estrous cycle in rats. In addition, neonatal daidzein also has some influence on ovarian function.     

Studies on the effect of manual massage of the ovaries on the reproductive activity of the mare

The aim of this study was to determine the influence of hand massage of mares' ovaries on breeding activity and hormonal changes in the winter anestrous period and during the luteal phase of the oestrus cycle. The experiment was conducted on 5 experimental and 5 control mares. In winter, (January) the experimental mares underwent 30-sec daily massage of both ovaries, for 30 d, and in summer (August) from the 6th day of the cycle to the occurrence of estrus. The sexual behavior of all mares was determined each day by individual teasing by a vigorous stallion, and the ovaries were checked by palpation per rectum and with an USG. Every second day blood samples were drawn from each mare to determine progesterone and estradiol in the plasma. Ovarian massage during deep winter anestrus had no significant effect on acceleration of the mares' active breeding season. Nevertheless, a higher concentration of estradiol was observed in the experimental group. These differences occurring on the 11th, 17th and 20th days were found to be significant (P ≤ 0.05). It was shown that during the summer period, in the luteal phase of the cycle, ovarian massage shortened the length of the estrous cycle, and ovulation was brought on somewhat earlier.     

Luteal regression in the normally cycling rat: apoptosis, monocyte chemoattractant protein-1, and inflammatory cell involvement

In hypophysectomized rats, prolactin induces regression of the corpora lutea. Luteal regression is accompanied by infiltration of monocytes/macrophages, declines in luteal mass and plasma progestins, and increased staining for monocyte chemoattractant protein-1 (MCP-1). We investigated whether similar events are induced during the estrous cycle, after the proestrous prolactin surge. Rats were killed on proestrus or on estrus, and one ovary was frozen for immunohistochemical detection of MCP-1, monocytes/macrophages (ED1-positive), and differentiated macrophages (ED2-positive) and for in situ detection of apoptotic nuclei. Corpora lutea of the current (proestrus) or preceding (estrus) cycle were dissected from the ovaries of additional rats and frozen for the same analyses and for determination of total protein content. In sections of whole ovaries, intensity and distribution of MCP-1 staining were increased in corpora lutea of multiple ages on estrus as compared to proestrus, as were numbers of differentiated macrophages and apoptotic nuclei per high-power field. Sections of isolated corpora lutea showed these increases on estrus, and the number of monocytes/macrophages per high-power field was also significantly increased. Accompanying these inflammatory/immune events, the corpora lutea on estrus showed decreased weight and total protein per corpus luteum, as compared to corpora lutea on proestrus. These changes are consistent with a proposed role for prolactin in the initiation of luteal apoptosis and of a sequence of inflammatory/immune events that accompany regression of the rat corpus luteum during the normal estrous cycle.     

Effect of CRESTAR on estrus synchronization and the relationship between fecal and plasma concentrations of progestagens in buffalo cows

In buffaloes estrus synchronization provides an opportunity for enhanced use of AI; however, changes in hormone secretion during synchronization are poorly understood. The aim of this investigation was to determine if the concentration of progesterone metabolites in feces would correlate with the concentration of progesterone in blood and thus, could be used for noninvasive monitoring of the reproductive status in buffalo cows. Additionally, the influence of a norgestomet-estradiol treatment (CRESTAR-ear implant) was investigated. According to the clinical examination and the progesterone profile in blood samples during the three wks before the treatment, the 17 animals were allotted to 3 groups: 1) CL = presence of corpus luteum throughout the period of 3 wks before the treatment (n = 8); 2) CY = cyclic, corpus luteum present for less than 3 wks (n = 6); and 3) AE = anestrous, with inactive ovaries (n = 3). In the first group, 4 animals started an estrous cycle after implant withdrawal and conceived after natural mating. In the second group one of the cyclic cows showed estrus two d after implant withdrawal, the other 3 had a delayed estrus (12 to 16 d). The two cows which had had inactive ovaries at the beginning but were cyclic before the treatment started, remained cyclic after implant withdrawal but did not become pregnant. The 3 anestrous cows of the third group remained anestrous after the treatment. The progesterone concentration in blood clearly correlated with the concentration of the metabolites in feces. Therefore, this noninvasive method is a valuable tool for determining the luteal status, and such information may be useful for developing estrus synchronization regimens in buffalo cows.     

Regulation of estrous cycle in hysterectomized albino rats by activation of follicular development.

Wistar strain albino rats were hysterectomized and the estrous cycle was compared with sham operated controls. Duration of estrous cycle in hysterectomized rats increased markedly with significant delay in the luteal phase and this was correlated to the inhibited follicular development of ovary. When these rats were treated with PGF2 alpha and PMSG and subjected to physical exercises, the estrous cycle was synchronised and the ovaries of such animals had active follicular development. Thus the deranged operation of sexual cycle in hysterectomized rats was regulated through physical exercises.     

Impact of single neonatal allylestrenol treatment on the estrus cycle of rats treated with FSH+LH or TSH.

Neonatal allylestrenol treatment administered to female rats significantly increases the duration of estrus phase in the sexual cycle. Treatment with follicle stimulating hormone (FSH) + luteinizing hormone (LH) in adulthood prolongs the duration of estrus even on its own; the effect, however, is more pronounced in those animals who were treated (imprinted) with allylestrenol neonatally. When administered to the control animals, the chemically related thyreotrop hormone (TSH) is either indifferent or it even decreases the estrus index. In animals having received neonatal allylestrenol treatment, however, TSH administration increases significantly the duration of the estrus phase. Either with or without FSH+LH treatment, the ratio of estrogenic to gestagenic phase increases following neonatal allylestrenol treatment. The experiments call attention to the potential functional risks inherent in neonatal allylestrenol treatment. The actual risks, however, seem to be smaller than the effects seen at the receptor level.     

Gonadotropin-releasing hormone binding sites in ovaries of normal cycling and persistent-estrus rats

The gonadotropin-releasing hormone (GnRH) binding capacity in ovaries and pituitaries of normal cycling rats at different stages of the estrous cycle and in ovaries of persistent-estrus rats was measured using radioligand-receptor assay (RRA). Persistent estrus was induced either by neonatal administration of testosterone propionate (1.25 mg s.c.) on the second day of life or by a hypothalamic suprachiasmatic frontal cut made with Halász' knife. All animals were killed during the critical period (1400-1600 h), and GnRH receptor was assayed. GnRH receptor levels in both ovaries and pituitaries changed during the estrous cycle. The total number of ovarian GnRH binding sites was significantly higher in proestrus than in diestrus 1, the stage in which the lowest level was found. When binding sites were expressed in fmol/mg ovary, the highest level was observed in diestrus 2; however, no changes were observed during the estrous cycle when GnRH binding sites were expressed as fmol/mg protein. Changes noted were very similar to those demonstrated in pituitary GnRH receptors in our present and previous experiments. Higher levels of pituitary binding sites were found in diestrus 2 and proestrus than in estrus and diestrus 1. The changes in the GnRH receptor levels were more striking in the pituitary than in the ovaries. It appears that the total number of ovarian GnRH binding sites was not altered in either of the two persistent-estrus groups, but that their concentration was significantly higher (expressed in fmol/mg ovary or fmol/mg protein) than on any day during the estrous cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Biological sexual maturation in rats: Interaction among genetic line,sex, age,and body weight

Interaction among genetic line, sex, age and body weight was studied for biological sexual maturation in rats. In female rats vaginal opening, first estrus, and duration of estrus cycle were determined. In male rats first presence of sperm was determined. Age and body weight at vaginal opening, first estrus or first detection of sperm were determined. Differences among five genetic lines were found on all measures of sexual maturation. The random bred control rats differed from the selectively bred lines of rats on majority of the measures, thus suggesting the influence of selective breeding on sexual maturation. The Maudsley lines selected for open-field defecation and Roman lines selected for two-way active avoidance learning showed different patterns of sexual maturation. Age was found to be crucial for the sexual maturation of female rats, whereas body weight was more important for the sexual maturation of male rats. This study is the first to investigate systematically the biological sexual maturation in live male rats and related sex differences. Findings suggest an important genetic component, and effect of selective breeding on the biological sexual maturation of rats.     

Characterization of the estrous cycle in Octodon degus

We characterized the reproductive cycle of Octodon degus to determine whether reproductive maturation is spontaneous in juveniles and if ovarian cyclicity and luteal function are spontaneous in adults. Laboratory-reared prepubertal and adult females were monitored for vaginal patency and increased wheel-running. Sexual receptivity was assessed by pairing adult females with a male 1) continuously, 2) at the time of vaginal patency, or 3) following estradiol treatment. Blood samples were assayed for estradiol and progesterone concentrations on Days 1, 4, 8, and 16 relative to vaginal opening. Ovarian tissues were collected 6 and 16 days after behavioral estrus and 6 days after copulation for histology. In juveniles, the onset of cyclic vaginal patency and increased wheel-running activity was spontaneous, occurred in the absence of proximal male cues, and appeared at regular intervals (17.5 ± 1.4 days). In adults, vaginal patency and increased wheel-running occurred cyclically (21.2 ± 0.6 days) in the absence of proximal male cues, and these traits predicted the time of sexual receptivity. Corpora lutea develop spontaneously and are maintained for 12-14 days. The ovaries had well-developed corpora lutea 6 days after mating and 6 days after estrus without mating. Progesterone concentrations were highest in the second half of the cycle when corpora lutea were present and estradiol concentrations peaked on the day of estrus. Thus, female degus appear to exhibit a spontaneous reproductive cycle consistent with other Hystricognathi rodents. Octodon degus is a novel model with which to examine the mechanisms underlying different reproductive cycles.     

Correlation between sexual maturation and induction of persistent estrus by continuous illumination.

The rats brought up under continuous illumination from the neonatal day persist in the rhythmic estrous cycle for long time and do not easily reveal so-called continuous vaginal estrus. On the contrary, immature rats at about 30 days of age show the persistent estrus soon after exposure to continuous illumination as well as adult rats. The purpose of this paper was to examine the relation of the age placed under continuous illumination and the time required to reach the persistent estrus. As the results, it was found that the younger rats need proportionally the longer time to reach the persistent estrus; the rats which were transferred into continuously illuminated environment at 0 (group A), 5 (group B), 10 (group C), 15-16 (group D) and 30 (group E) days of age required a mean time of 132.1 +/- 8.4, 126.8 +/- 8.9, 89.7 +/- 6.2, 91.5 +/- 3.8 and 70.3 +/- 3.0 days after birth to reach the persistent estrus, respectively. In the present data, a significant difference was seen between group B and C. Though all rats were killed after the same length of persistently estrous periods, actual endocrine level appeared to differ among the groups; the rats required longer time to reach the persistent estrus had smaller ovaries and adrenals.     

Biochemical status of ovaries after induction of superovulation on different days of estrus cycle in mice

To investigate the role of ovarian status and to find out a suitable hormonal dose for induction of superovulation and its effect on biochemical status of the ovaries, the mice were injected with PMSG in doses of 5, 7.5, and 10 IU on different days of the estrous cycle i.e. proestrus, estrus, metestrus and diestrus followed by hCG injection 48 hr later. All these treatments increased the mean ovarian weight and ovulation rate when compared with that of control animals. Maximum response was observed by treatment with 7.5 IU PMSG on the day of estrus. This treatment resulted in a non-significant decrease in total proteins but a significant increase in the lipid concentrations while no change in cholesterol content of the ovaries of superovulated mice. The activity of acid phosphatase and lactate dehydrogenase significantly increased and alanine aminotranseferase significantly decreased in the ovaries of mice after superovulatory treatment when compared with that of control animals. This reveals that treatment with PMSG and hCG results in metabolic alterations in the ovaries which may perhaps be inducing biosynthetic deficiencies in oocytes as indicated by increased prenatal mortality in superovulated pregnant mice when compared with that of controls in the present studies.     

Ovarian and behavioral cyclicity of the laboratory maintained cat

This study examined the sexual behavior-ovarian activity relationship of the laboratory cat maintained under consistent environmental conditions. Mean (±SEM) lengths of estrus for nonovulating, mated ovulating, and human chorionic gonadotrophin (HCG)-induced ovulating queens were 5.8 ± 0.17, 6.5 ± 0.75, and 10.0 ± 0.65 days, respectively. The duration of estrus was significantly (P < 0.01) longer in HCG-treated queens in comparison to the nonovulating or mated ovulating groups. No differences were observed in the number of behavioral estrous periods detected monthly. Duration of estrus was affected by season with significantly shorter periods of estrus observed in June (P < 0.05), September (P < 0.05), October (P < 0.05), and November (P < 0.01) compared to the March, April, May, August, or December averages. Length of estrous cycle (from Day 1 of estrus to Day 1 of next estrus) was variable (range: 6–120 days); however, 49.8% of the estrous cycles were 12 to 21 days in length. Although the ovaries of queens displaying sexual receptivity always contained developing or mature foilicles, ovaries of cats showing no estrous behavior also showed patterns of follicular development and regression at 14- to 19-day intervals. These results suggest that the estrous cycle of the laboratory maintained cat varies from 2 to 3 weeks in duration and that reproductive behavioral patterns alone are not always an accurate indication of ovarian activity or duration of the reproductive cycle.     

The role of the uterus in regulation of heat duration in cycling rats

These experiments examined the effects of hysterectomy on heat duration and on the reinduction of estrous behavior by progesterone (P) following the termination of spontaneous heat in 4-day cycling rats. Hysterectomy did not affect the onset of estrus but prolonged heat duration. The average duration of sexual receptivity for hysterectomized (H) and sham-hysterectomized (SH) rats was 18.2 and 13.0 hr, respectively. Furthermore, H animals injected with either 0.5 mg P within 2 hr, or 4.0 mg P 24 hr following the termination of natural estrus showed significantly higher lordosis and solicitation responses than SH rats similarly treated. These behavioral findings were correlated with the level of hypothalamic progestin receptors. That is, H animals had a significantly higher concentration of progestin receptors than SH rats immediately following the termination of spontaneous heat and also 24 hr later. Both in estrous-cycling rats and in gonadectomized animals treated with estradiol benzoate (EB), hysterectomy resulted in higher serum estradiol (E2) levels. The results of these experiments suggest that prolongation of the period of sexual receptivity and the facilitated behavioral responses to P following the cessation of estrus in hysterectomized animals may be due to a lowered clearance rate of circulating estradiol which presumably enhances the estrogen conditioning of the neural substrates.