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Estrous cycle-dependent changes in the expression of aromatic hydrocarbon receptor (AHR) and AHR-nuclear translocator (ARNT) mRNAs in the rat ovary and liver.

The aromatic hydrocarbon receptor (AHR) and AHR nuclear translocator protein (ARNT) mediate the toxic effects of a wide variety of halogenated and polycyclic aromatic hydrocarbons. While it can be assumed that AHR has an endogenous function, its role in reproduction is currently undefined. The present study seeks to examine the regulation of AHR and ARNT mRNAs in liver and ovarian tissues across the rat estrous cycle. Message for hepatic AHR was increased significantly on the morning of proestrus, and decreased dramatically by the evening of proestrus; while hepatic ARNT mRNA was significantly decreased between diestrus and the morning of proestrus, and between the evening of proestrus and the morning of estrus. Ovarian AHR mRNA was unchanged from diestrus to proestrus, and was decreased on the evening of proestrus. Changes in the expression of ARNT mRNA mirrored changes in the liver. To assess interaction between the AHR- and estrogen-receptor (ER)-signaling pathways and to test the hypothesis that estrogen regulates AHR mRNA, 25-day-old female rats were injected with either 17beta-estradiol, the ER antagonist ICI 182 780, or with vehicle, and hepatic AHR mRNA was measured. Treatment with estrogen or the estrogen antagonist did not alter the abundance of AHR mRNA in the liver. These data suggest that while estrogen may not be the key regulator of AHR mRNA expression, a factor associated with the rat reproductive cycle may be important in regulating the expression of both the AHR and ARNT genes in the ovary and liver.

Animals↗

Effect of stage of anestrus on the induction of estrus by the dopamine agonist cabergoline in dogs.

Beagle bitches were administered the dopamine D2 receptor agonist cabergoline in 3 groups of 5 animals each, starting on known days of the estrous cycle. Cabergoline treatment was started in either early anestrus (Days 93 to 108), mid-anestrus (Days 123 to 156), or late anestrus (Days 161 to 192) at doses of 5 ug/kg/d, per os, and was continued until the confirmation of induced proestrus or for 40 d. Reproductive parameters were compared with those in 5 control anestrous bitches (Days 90 to 150). In control bitches, the mean (+/- SEM) interval to the next proestrus (73+/-11 d) resulted in an interestrus interval (192+/-9 d) similar to that of the previous cycles (196+/-11 d). In 14 of the 15 cabergoline-treated bitches, the next proestrus occurred within 4 to 30 d, was premature in early and mid-anestrous bitches and developed with low variability within groups. The resulting intervals to proestrus in bitches treated with cabergoline in early anestrus (20+/-2 d), mid-anestrus (14+/-3 d) and late anestrus (6+/-1 d) resulted in interestrus intervals in those groups of 131+/-5, 166+/-7 and 196+/-2 d, respectively. In response to treatment, interestrus intervals were reduced (P<0.05) and more synchronous (P<0.05) in early and mid-anestrus bitches, and were more synchronous (P<0.05) in late-anestrous bitches compared with those of control bitches or those of the previous cycle. Periovulatory estradiol and progesterone profiles of induced cycles in treated bitches were similar to those of spontaneous cycles in control bitches. Four of 5 control bitches and 12 of the 14 responding cabergoline-treated bitches became pregnant and produced normal litters. Plasma prolactin concentrations at Days 2 and 5 of treatment (0.3+/-0.1 ng/mL) and at the onset of proestrus shortly before the end of treatment (0.4+/-0.1 ng/mL) were lower (P<0.05) than those present in anestrus prior to treatment (1.7+/-0.6 ng/mL) or in control bitches. Prolactin was also low at the onset of proestrus in control bitches (0.5+/-0.2 ng/mL). The results demonstrate that prolactin-lowering doses of the dopamine agonist cabergoline can terminate the normal obligate anestrus in dogs, and that the effect occurs more slowly in early anestrus than in mid or late anestrus.

Anestrus↗

Hormonal control of nuclear estradiol receptor content and the luminal epithelium in the uterus of the golden hamster.

Uteri were removed and blood was drawn from hamsters during the estrous cycle, on the eighth day of pregnancy, and after different hormonal treatments. Serum estradiol (E2) and progesterone (P) levels were determined by RIA. Portions of the uteri were prepared for light and electron microscopy. Endogenous uterine nuclear E2 receptor was measured by exchange assay. Large increments of nuclear E2 receptor occurred when the level of serum E2 was rising and that of serum P was falling (diestrus day 2 through proestrus morning); decrements occurred when the serum E2 level was falling and the serum P level was rising (proestrus evening through diestrus day 1). Ovariectomy on proestrus morning led to a decline in the amount of nuclear E2 receptor, and this decline was prevented by administration of E2 at the time of ovariectomy. P depleted nuclear E2 receptor in the presence of high levels of serum E2. The uterine luminal epithelium passed through a phase of mitosis and hypertrophy (diestrus day 2 through proestrus morning), a brief phase suggestive of secretion (proestrus evening), a phase of degeneration (estrus), and a phase of quiescence (diestrus day I). Ovariectomy on proestrus morning led within 24 h to degenerative changes identical to those seen during estrus, and these changes were prevented by treatment with E2 at the time of ovariectomy. P induced these degenerative changes in the presence of high levels of serum E2. After 8 days of pregnancy, the epithelium resembled that seen during diestrus day 1. Elevated uterine nuclear E2 receptor levels were associated with growth and hypertrophy of the luminal epithelium, and severely depressed levels were associated with the degenerative and quiescent phases of the epithelium.

Animals↗

The self-priming effect of gonadotropin-releasing hormone on luteinizing hormone release: observations using rat anterior pituitary fragments and dispersed cells continuously perifused in parallel.

To study in vitro the self-priming effect of GnRH on LH release, rat anterior pituitaries were prepared either as fragments or dispersed cells and continuously perifused in parallel chambers. The experimental groups consisted of rats killed at 0800 h on diestrus day 1, diestrus day 2, proestrus or estrus, or at 1400 h on proestrus. To insure truly independent observations, each experimental preparation was tested on three occasions. After basal LH release had stabilized, the tissue preparations were exposed to 10 nM GnRH as two 30-min challenges separated by 1 h. LH secretory rates (nanograms per min/pituitary for fragments; nanograms per min/10(7) cells for dispersed cells) were calculated 1) for basal release (during the 20-min period immediately preceding each GnRH challenge), 2) in response to GnRH, and 3) as the sum of basal and GnRH-stimulated release. Comparison of the two preparations revealed that basal and GnRH-stimulated LH release by pituitary fragments was more variable than LH release by dispersed cells. In addition, while dispersed cells responded promptly to the addition/withdrawal of stimuli, fragments did so more gradually. With respect to GnRH self-priming, the second mean secretory rate for basal LH release by fragments (range, 28.8-46.5) was significantly (0.1 greater than P greater than 0.01) higher than the first rate (range, 14.4-22.0) on diestrus day 1, diestrus day 2, proestrus at 0800 h, and estrus. With dispersed cells, the first and second basal rates were similar to each other on diestrus day 1 and estrus, but on diestrus day 2 and on proestrus at 0800 and 1400 h, the second basal rate (range, 36.8-93) was significantly (P less than 0.001) higher than the first range (range, 17.7-31.7). When fragments received GnRH, the second mean secretory rate (range, 35.2-64.2) was significantly (0.1 greater than P greater than 0.03) higher than the first rate (range, 13.4-34.1) on diestrus day 2 and proestrus at 0800 h. With dispersed cells, the mean secretory rate in response to the second GnRH challenge was higher only on diestrus day 2 (37.0 +/- 4.1 vs. 60.3 +/- 3.8; P less than 0.05). When considered as the total of basal plus GnRH-stimulated LH release, the second secretory rate by fragments (range, 54.5 - 110.8) was significantly (0.1 greater than P greater than 0.02) higher than the first rate (range, 27.9 - 51.4) on diestrus day 1, diestrus day 2, and proestrus at 0800 h.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Adenylyl cyclase activities in ovarian tissues. III. Regulation of responsiveness to LH, FSH, and PGE1 in the prepubertal, cycling, pregnant, and pseudopregnant rat.

In the ovaries of prepubertal rats, responsiveness of adenylyl cyclase (AC) to LH, FSH, and PGE1 is acquired at day 10, coinciding with the appearance of the ability to produce steroids in response to gonadotrophins. The activity and responsiveness of AC on day 11 were similar to those at puberty on day 40 and relatively stable in between. In the follicles of the cycle, the responsiveness of AC to LH and FSH was poor (ca 2-fold stimulation) on metestrus and diestrus, and became high (ca 10-fold stimulation) between 1000 h of diestrus and 1000 h of proestrus. Thereafter, the system slowly became desensitized to LH and FSH, being unresponsive by the morning of estrus. Nembutal injected at 1230 h and again at 1500 h on proestrus blocked ovulation and prevented the decline in LH- and FSH-stimulated AC activity. In the CL of the cycle, the AC was unresponsive to LH on day 1 (estrus), became responsive by the morning of day 2 (metestrus), and maintained responsiveness throughout that day. Thereafter, the responsiveness and basal AC activity declined rapidly. In the CL of pregnancy, LH-stimulated AC was indistinguishable from that of the CL of the cycle during days 1 and 2, then increased steadily until day 9, showing a transient decrease on days 10 and 11, followed by a sharp rise to maximal activity on days 15 and 16. Thereafter, activity declined as parturition approached. In the CL of pseudopregnancy (PSP), LH-stimulated AC was very similar to that of the CL of pregnancy during the first 11 days. Thereafter, it decreased coincident with the termination of PSP. Injections of PRL (100 mug SC twice daily, from metestrus through estrus, and from proestrus through proestrus) or estradiol-17 beta (20 mug, SC at 1230 h on metestrus) resulted in "rescue" of the CL-AC system, which remained at metestrus levels when measured on the days of expected proestrus or estrus. Injections of pregnant mare serum gonadotrophin into prepubertal rats at day 26 (3 IU, iv), induced by day 28 a highly responsive AC system in follicles, with activities equivalent to those found in Graafian follicles on proestrus. By day 29, synchronous ovulation had occurred with a concomitant loss of LH-stimulated AC such as seen in the 1-day-old CL of mature rats. Our results suggest that the LH-sensitive AC may be indicative of the final development of ovulability of the follicles, and that it may correlate with the functional capacity of CL during various reproductive stages of the rat.

Adenylyl Cyclases↗

Variations in angiotensin-II release from the rat brain during the estrous cycle.

To investigate the hypothesis that the release of angiotensin-II (AII) in the rat brain increases on the day of proestrus, samples of cerebrospinal fluid (CSF) and interstitial fluid from the general region of the bed nucleus of the stria terminalis pars ventralis in the preoptic-anterior hypothalamic area were monitored for AII-immunoreactive material (AII-ir) using push-pull cannulas. Samples of CSF were obtained on the day of proestrus and diestrus day 1 at 30-min intervals from 1200-1600 h. Samples of interstitial fluid were obtained at 25-min intervals from 0930-1600 h. The rate of release of AII-ir into CSF was significantly greater on proestrus compared to diestrus day 1, and in the early afternoon of proestrus compared to the late afternoon. In five of seven rats and in the overall comparison of AII-ir release from the preoptic-anterior hypothalamic area, significantly more AII-ir was released on the day of proestrus vs. diestrus day 1. These observations are consistent with previous studies suggesting that brain AII may play a role in the regulation of LH release on the day of proestrus.

Angiotensin II↗

Differential expression of growth hormone messenger ribonucleic acid by somatotropes and gonadotropes in male and cycling female rats.

Past studies have reported the appearance of cells sharing phenotypic characteristics of gonadotropes and GH cells. During diestrus and early proestrus, a subset of somatotropes (40-60%) expressed both GH antigens and gonadotropin (LH-beta, LHbeta, or FSH-beta) messenger RNAs (mRNAs) or GnRH receptors. More recently, we reported that subsets of gonadotropes identified by LHbeta or FSHbeta antigens expressed GH- releasing hormone (GHRH) binding sites. The present studies were designed to learn if these putative multipotential cells also expressed GH mRNA. Biotinylated sense and antisense oligonucleotide probes were developed and cytochemical in situ hybridization tests were optimized for the detection of GH mRNA with GH, LHbeta, and FSHbeta antigens. RNase protection assays were developed with a complementary RNA probe that detected a 380-bp region at the 5' end of the GH mRNA. Both the in situ hybridization and RNase protection assays detected changes in expression of GH mRNA during the estrous cycle with the lowest expression occurring during metestrus and peak expression occurring on the morning of proestrus. Cell counts confirmed the results of the RNase protection assays showing that increases in mRNA levels seen from metestrus to proestrus reflected increased percentages of GH mRNA-bearing cells. In addition, densitometric analyses demonstrated that the higher GH mRNA levels assayed from diestrus to proestrus reflected increased area and density of label per cell. Both types of assays showed sex differences in expression of GH mRNA; male rat cell populations had higher values than female rats in metestrus, diestrus, or estrus. However, percentages of GH cells in male rats were equal to those from proestrous female rats and levels of GH mRNA were lower in male rats than proestrous females. Dual labeling experiments showed that, in male rats and diestrous, proestrous, or estrous females, GH mRNA was expressed in over 70% of GH cells. Expression of GH mRNA was also found in 50-57% of cells with LHbeta or FSHbeta antigens in the same groups. The lowest expression was seen in the metestrous groups (30-40% of GH cells or gonadotropes expressed GH mRNA). Expression of GH mRNA was first increased from metestrus to diestrous largely in GH cells, and slightly in cells with LHbeta antigens. Further increases were seen in GH and LH cells by the morning of proestrus. In contrast, FSH gonadotropes did not show an increased expression of GH mRNA until the morning of proestrus (reaching the same peak reached by LH cells). These data confirm the working hypothesis that a multihormonal cell type develops during diestrus to support both the somatotrope and gonadotrope populations. Collectively, our studies suggest that this multihormonal cell may function to help support the regulatory functions of the gonadotrope during the periovulatory period. In addition, the appearance of significant levels of expression of GH mRNA by male rat gonadotropes suggests that this multihormonal cell may play a role in regulation of the male reproductive system as well.

Animals↗

Growth-associated protein-43 messenger ribonucleic acid expression in gonadotropin-releasing hormone neurons during the rat estrous cycle.

We have shown previously at the ultrastructural level that morphological changes occur in the external zone of the median eminence allowing certain GnRH nerve terminals to contact the pericapillary space on the day of proestrus. The present study was designed to determine whether the intrinsic determinant of neuronal outgrowth, growth-associated protein-43 (GAP-43), was expressed in GnRH neurons of adult female rats, and whether its expression varied throughout the estrous cycle. To accomplish this, we perfusion-fixed groups of adult female rats at 0800 and 1600 h on diestrous day 2 (diestrous II), at 0800 h and 1600 h on proestrus, and at 0800 and 1600 h on estrus (n = 4 rats/group) and used double labeling in situ hybridization and quantification to compare the levels of GAP-43 messenger RNA (mRNA) in cells coexpressing GnRH mRNA. GnRH mRNA was detected with an antisense complementary RNA (cRNA) probe labeled with the hapten digoxigenin, whereas the GAP-43 cRNA probe was labeled with 35S and detected by autoradiography. In addition, GAP-43 protein was identified with immunohistochemistry in the median eminence. The results show that many GnRH neurons expressed GAP-43 mRNA and that GAP-43 protein was present in many GnRH axon terminals in the outer layer of the median eminence. The number of GnRH neurons expressing GAP-43 mRNA was significantly higher on proestrus (64 +/- 5%) than on diestrous II (40 +/- 2%; P < 0.001) or on estrus (45 +/- 8%; P < 0.05), and the GAP-43 mRNA levels in GnRH neurons also varied as a function of time of death during the estrous cycle. The GAP-43 mRNA levels in GnRH neurons were higher on proestrus and estrus than on diestrous II (P < 0.05). These data show that 1) GAP-43 is expressed in adult GnRH neurons; 2) GAP-43 mRNA expression in GnRH neurons fluctuates during the estrous cycle; and 3) GAP-43 mRNA content in GnRH neurons is highest on the day of proestrus, before and during the onset of the LH surge. These observations suggest that the increased GAP-43 mRNA expression in GnRH neurons on the day of proestrus could promote the outgrowth of GnRH axon terminals to establish direct neurovascular contacts in the external zone of the median eminence and thus facilitate GnRH release into the pituitary portal blood.

Animals↗

Prolactin secretion and its response to stress during the estrous cycle of the rats.

Serum and pituitary prolactin (PRL) concentrations were measured during the estrous cycle of the rat with particular attention to the afternoons of the days of proestrus and estrus. Homogenizing machines, a Polytron and Sonifier, were used to extract PRL from the pituitary gland. The effects of ether anesthesia and restraint were also examined on the afternoons of both proestrus and estrus. The occurrence of a surge in PRL secretion during proestrus was confirmed with a peak at 1500 h, and this was accompanied by a decline in pituitary PRL content. A relatively high level of serum PRL was observed in the afternoon of estrus, during which time pituitary PRL content increased progressively. Ether anesthesia had no effect on the proestrus PRL surge, while restraint enhanced it. On the afternoon of estrus, restraint completely suppressed the rise in serum PRL, but ether anesthesia failed to suppress it completely. From these results, the following conclusions were drawn: 1) the PRL surge on the afternoon of proestrus occurs without synthesis of the hormone in the pituitary; 2) PRL secretion on the afternoon of estrus is accompanied by its synthesis in the gland; 3) the PRL response is distinct for each type of stress applied; and 4) PRL secretion is thus regulated by different mechanisms in proestrus and estrus.

Anesthesia↗

Regulation of the natriuretic peptide system in rat uterus during the estrous cycle.

Uterine natriuretic peptides may be involved in the alterations that occur in the uterus during the estrous cycle through its role in hydromineral balance. The following studies were performed to determine whether uterine natriuretic peptides and receptors follow a cyclic pattern during the estrous cycle. The results obtained show that atrial natriuretic peptide (ANP) content in rat uterine tissue was low in proestrus (8.5 +/- 2.6 pg/g) and significantly increased (P < 0.001) in estrus (71.5 +/- 16.6 pg/g), metestrus (82.6 +/- 19.7 pg/g) and diestrus (91.0 +/- 19.4 pg/g), whereas plasma ANP was not altered during the cycle. Similarly, measurement of uterine ANP mRNA by reverse transcribed polymerase chain reaction (RT-PCR) indicated lowest levels of ANP mRNA at proestrus. Measurement of C-type natriuretic peptide (CNP) by a specific and sensitive radioimmunoassay revealed that uterine CNP also varies with the estrous cycle. Uterine CNP was low in diestrus (143.2 +/- 22.4 pg/mg protein) as compared with proestrus, estrus and metestrus (305.3 +/- 51.5, 267.5 +/- 44.9, 291 +/- 41.2 pg/mg protein respectively, P < 0.05). Autoradiography performed on uterine tissue slices localized natriuretic peptide receptors to myometrial smooth muscle layers and to endometrial uterine glands. High binding of 125I-ANP was observed in proestrus and estrus with 60-75% decreases during metestrus and diestrus. Binding of 125I-tyr0CNP to uterine slices was also high during proestrus, but declined by 35% at estrus, metestrus and diestrus. The alterations in the receptors were also observed at the level of synthesis. RT-PCR detection of guanylyl cyclase A (GC-A) receptor mRNA and guanylyl cyclase B (GC-B) mRNA showed high signals at proestrus but 4- and 2-fold reductions respectively at metestrus and diestrus. In conclusion, variations in uterine ANP and CNP and their receptors during the rat estrous cycle imply the involvement of the natriuretic peptides in uterine hydromineral balance and myometrial motor activity.

Animals↗

[Activity of the basic carboxypeptidases in female rat tissues at different stages of estrus cycle].

It is revealed, that the activity of neuropeptide metabolism enzymes (carboxypeptidase H, phenylmethylsulfonilfluorid-inhibited carboxypeptidase) in the female rat tissues depends upon the stage of estrus cycle. The carboxypeptidase H activity in the pituitary gland is the highest in proestrus; it is almost 3 times higher in comparison with diestrus; it is a little bit higher in striatum on the stage of estrus, than in diestrus and proestrus, in adrenals on the stage of proestrus and estrus it is a little bit lower, than in diestrus; in the ovaries on the stage of proestrus it is much higher, than in estrus and diestrus. The activity of PMSF-inhibited carboxypeptidase in ovaries on the stage of proestrus and diestrus is 1.7-1.8 times higher, than at the stage of diestrus. The activity of carboxypeptidase M in adrenal tissue at the stage of proestrus is 35-40% of that at the stage of diestrus and estrus. The activity of carboxypeptidase M in the ovaries at the stage of diestrus is 45-50% of that at the stage of diestrus and estrus. The role of the investigated enzymes in cyclic changes of a level of biologically active peptides and in regulation of estrus cycle is discussed.

Adrenal Glands↗

Influence of the estrous cycle during carcinogen exposure on nitrosomethylurea-induced rat mammary carcinoma.

The influence of individual stages of the rat estrous cycle during exposure to I-nitroso-I-methylurea (NMU) on mammary tumor incidence, latency, number, and cytosol receptor dynamics for estrogen and progesterone was determined. Virgin female Buffalo rats were separated into three groups on Day 53 according to their vaginal smear pattern. NMU (5 mg/100 g body weight, i.v.) was administered in three monthly doses beginning at 53 to 55 days of age on diestrus, proestrus, or estrus between 9:00 and 11:00 a.m. Groups of rats had their second and third injections of NMU on the same day of the estrous cycle as their initial injection. All animals were killed during the morning on a diestrus day. Receptors for estrogen and progesterone were determined by a modified dextran-coated charcoal method and by sucrose density gradient analysis. Mean latencies to first tumor appearance in diestrus, proestrus, and estrus groups were 104.4, 83.6, and 91.4 days, respectively (p less than 0.05, diestrus versus estrus and proestrus) following the first NMU injection. The mean number of tumors per rat was significantly higher in rats injected on proestrus (4.5) or estrus (4.3) than on diestrus (2.0). Estradiol bound to receptor sedimented at 8 and 4 s and was suppressed by diethylstilbestrol and estradiol. Progesterone receptor migrated to 7.8 and 4 s regions. Estrogen receptor incidence (100%) and content (16.7 fmol/mg cytosol protein) was highest in rats injected on diestrus. In the proestrus and estrus injected groups, estrogen receptor incidence was 95 and 63% and content was 10.2 and 11.2 fmol/mg protein, respectively. The affinity of estradiol for its receptor was not significantly altered in any group. Although there were no statistically significant difference in progesterone receptor incidence or affinity between groups, progesterone receptor content (74.6 fmol/mg cytosol protein) was significantly higher in tumors from rats injected on proestrus than on diestrus. These data suggest that the prevailing hormonal milieu of the estrous cycle during NMU exposure may be critically important to the subsequent biological behavior and steroid receptor status of carcinogen-induced rat mammary tumors.

Adenocarcinoma↗

Ovariectomy at different stages of the estrous cycle modifies the pulsatile secretory pattern of prolactin in rat.

The role of estrogens in the synchronization of pulsatile prolactin secretion throughout the estrous cycle in the adult female rat is studied. Mean values of prolactin, in sham-operated animals increased from diestrus-2 to proestrus and then decreased to the values found at diestrus-1. Ovariectomy did not modify the pattern which follows mean serum levels of the hormone throughout the estrous cycle; although increased values of prolactin were found at both proestrus and estrus, when compared to control animals. No changes in mean values of the hormone were observed at any other phase of the estrous cycle. The number of peaks was fairly constant in sham-operated rats in any phase of the estrous cycle, but a decrease in its number, after ovariectomy, was observed at proestrus and estrus. The absolute amplitude of the peaks increased numerically but not statistically significant from diestrus-2 to proestrus, then decreasing to diestrus-1 in control animals whereas an increase in the absolute amplitude of the prolactin peaks was detected in proestrus and estrous after ovariectomy. The duration of the prolactin peaks was not changed either by the phase of the estrous cycle or by ovariectomy. The relative amplitude of the prolactin peaks was only changed in rats ovariectomized at diestrus-1, as compared to sham-operated animals. All these data indicated that changes in estrogen/secretion by ovariectomy throughout the estrous cycle of the rat induced a differential effect on the pulsatile pattern of prolactin, being more marked in proestrus and estrus, associated with the highest circulating estrogen levels.

Animals↗

Effects of estrus cycle on thermoregulatory responses during exercise in rats.

In female rats, rectal temperature (Tre), tail vasomotor response, oxygen uptake (VO2), and carbon dioxide production (VCO2) were measured in proestrus and estrus stages during treadmill running at two different speeds at an ambient temperature (Ta) of 24 degrees C. Experiments were performed at 2.00-6.00 a.m., when the difference in Tre was greatest between the two stages; Tre at rest in the estrus stage was 0.54 degrees C higher than in the proestrus stage. In a mild warm environment, threshold Tre for a rise in tail skin temperature (Ttail) was also higher in the estrus stage than in the proestrus stage. In contrast, no difference was seen in the threshold Tre and steady state Tre at the end of exercise between proestrus and estrus stages. These values were higher at the higher work intensity. VO2 was also similar between the two stages, except in the second 5 min after the beginning of exercise, when VO2 was greater and Tre rose more steeply in the proestrus stage. These data indicate that deep body temperature during exercise is regulated at a certain level depending on the work intensity and is not influenced by the estrus cycle.

Animals↗

Does corpus luteum function autonomously during estrous cycle in the rat? A possible involvement of LH and prolactin.

This study examined the of LH and prolactin in the control of corpus luteum function during 4-day cycles in the rat. Bromocriptine (BRC) treatment was performed on proestrus or/and estrus morning that means before or after the preovulatory release of LH. This caused complete blood prolactin depression from the time of injection until diestrus 1 afternoon. This decrease in blood prolactin concentration was associated with a rise in the tonic level of LH secretion in those females which received BRC as soon as on proestrus. We first observed that injection on the morning of proestrus of doses of BRC capable of blunting prolactin secretion on proestrus afternoon did not significantly impair the preovulatory release of LH and did not prevent ovulation occurring during the following night. The life span of the corpora lutea edified from ovarian follicles rupturing before or under BRC administration did not exceed that of those formed under physiological circumstances since 4-day cycles culminating in ovulation constantly took place in all the treated animals whatever the time of BRC injection. To determine the pattern of luteal activity in the absence of prolactin secretion, we measured blood progesterone concentration from estrus until late diestrus in female rats injected with BRC on proestrus and/or estrus at 1100 h. The initiation of the function of corpus luteum on estrus and the achievement of its full activity on diestrus 1 did not appear to be affected by BRC. By contrast the level of blood progesterone declined more rapidly on the morning of diestrus 2 in BRC-treated than in control females. The capacity for autonomous progesterone secretion by corpus luteum of the cycle was discussed in the light of previous and present observations.

Animals↗

Variations in concentration of oxytocin and vasopressin in the paraventricular nucleus of the hypothalamus during the estrous cycle in rats.

Oxytocin (OT) and arginine-8-vasopressin (AVP) were measured by radioimmunoassay in micropunched hypothalamic neurosecretory nuclei of estrous cycling female Sprague-Dawley rats. In the paraventricular nucleus (PVN): the concentration (pg/microgram protein) of OT was significantly higher in rats in diestrus than during proestrus, estrus, or metestrus, while the concentration during metestrus was significantly greater than in proestrus and estrus; the concentration of AVP was significantly lower in animals in estrus than during the other three stages; because the paraventricular OT levels dropped before proestrus, the AVP/OT ratio was significantly greater in animals in proestrus than in diestrus, metestrus, and estrus. In the supraoptic nucleus (SON) a similar trend was noted: the concentration of OT was highest during diestrus, and AVP was lowest during estrus, though neither was significantly different from other stages. Because the OT and AVP cycles in the SON were asynchronous, the ratio of AVP to OT was significantly higher in proestrus than in metestrus or diestrus and significantly greater in estrus than during diestrus. In contrast to these two areas, peptide concentrations did not vary significantly across the estrous cycle in other sites of nonapeptide synthesis, i.e. the anterior commissural nucleus (ACN) and the suprachiasmatic nuclei (SCN).

Animals↗

Inhibition of the formation of 13,14-dihydroprostaglandin F2-alpha induced by chlorpromazine in rat ovary.

The present study was designed to investigate the effects of chlorpromazine (CPZ) on the formation of 13,14H2-PGF2 alpha in rat ovary and the possibility that the "critical period" observed in the effect of CPZ for the blockade of ovulatin may be appreciated by measuring the formation of 13,14H2-PGF2 alpha as a parametor. When CPZ (4.0 mg/kg) was given s.c. at 10:00 a.m. once a day for 3 days from the first day of diestrus to the expected day of proestrus followed by sacrifice after 24 hours of the final injection, a significant inhibition was recognized in the formation of 13,14H2-PGF2 alpha in ovary. The effects of a single injection of CPZ were also determined 24 hours after injection at 10:00 a.m. on the day of diestrus I, diestrus II or proestrus. The results showed that the drug treatment on the day of proestrus significantly decreased the formation of 13,14H2-PGF2 alpha, but not on the day of diestrus I or II. When the formation of ovarian 13,14H2-PGF2 alpha was inhibited by CPZ treatment, the blockade of ovulation was also confirmed. A single injection of CPZ at 05:00 and 07:00 after the "critical period" on the day of proestrus did not inhibit the 13,14H2-PGF2 alpha formation in the rat ovary. It is concluded that the administration of CPZ before the "critical period" on the day of proestrus is effective in suppressing the ovarian 13,14H2-PGF2 alpha formation as well as on the blockade of ovulation.

Animals↗

Attenuated proestrous luteinizing hormone surges in middle-aged rats are associated with decreased pituitary luteinizing hormone-beta messenger ribonucleic acid expression.

OBJECTIVE: This study determined whether attenuated preovulatory luteinizing hormone surges in aging rats are associated with a decrease in pituitary luteinizing hormone content or luteinizing hormone beta-messenger ribonucleic acid expression on proestrus. STUDY DESIGN: Blood samples were taken every 90 minutes from 1:30 to 10:30 PM on proestrus in young (n = 8) and middle-aged (n = 12), regularly cyclic rats for plasma luteinizing hormone determination. On the next proestrus at 12 noon, rats were killed and the pituitaries were removed for luteinizing hormone content determination by radioimmunoassay and luteinizing hormone beta-messenger ribonucleic acid expression by dot blot analysis. Results were analyzed by one-way analysis of variance. RESULTS: Seven of the middle-aged rats had attenuated luteinizing hormone surges while the remaining five females had surges similar to those of young rats. On the next proestrus, all rats had similar quantities of pituitary luteinizing hormone. However, luteinizing hormone beta-messenger ribonucleic acid expression in middle-aged rats with attenuated luteinizing hormone surges was lower (p less than 0.05) than that of middle-aged and young rats with normal surges. CONCLUSION: Decreased luteinizing hormone beta-messenger ribonucleic acid expression, but not pituitary luteinizing hormone content at 12 noon on proestrus is correlated with attenuated luteinizing hormone surges in middle-aged rats.

Aging↗