Estradiol modulation of pineal gland activity in the wild bandicoot rat, Bandicota bengalensis

In the present study, the effect of estradiol dipropionate on the cytology and mitotic activity of the pineal gland was evaluated in adult ovariectomized and juvenile bandicoot rats, Bandicota bengalensis. Estradiol treatment for 5 days inhibited the ovariectomy-induced hypertrophy of the pineal gland, and increased the nuclear diameters of pinealocytes in juvenile males while having no effect on the pineal cytology of juvenile females. Estradiol injection induced mitosis in the pinealocytes of adult ovariectomized and juvenile bandicoot rats. Thus, estradiol exhibited a dual action (stimulatory and inhibitory) on the pineal gland of this wild rat.     

Effect of clomiphene citrate on the rabbit ovary

Summary The effect of clomiphene citrate on the rabbit ovary was studied in mature nulliparous rabbits pretreated with three consecutive doses ranging from 0.01–10.0 mg/kg per day. With increasing doses a trend of decrease in mean ovarian weight (mg/kg body weight) is observed 2 days after termination of treatment. Five days later a significant increase occurs, which then subsides again to control values on day 12 after termination of treatment. During this period, no matings or injections of luteinizing hormone were performed to trigger ovulation; consequently no ovulations are observed. Folliculogenesis appears as normally; number and morphology of follicles are within normal ranges. No endogenous, spontaneous gonadotropin surges are detected in blood serum up to the 7th day after termination of treatment (2 and 10 mg doses). The surface epithelium of the ovary resembles normal germinal epithelium; however, after treatment with high doses a secretory-like activity is observed, accompanied with ultrastructural changes.Supported by the Deutsche Forschungsgemeinschaft, Special Program on Biology and Clinics of Reproduction. (Grant Be 524/7-7).Visiting Scientist from the Department of Obstetrics and Gynecology, Hadassah University Hospital, Jerusalem, Israel.     

Conditioned taste aversion in lesser bandicoot rat, Bandicota bengalensis

The lesser bandicoot rat after ingesting a sublethal dose of 0.025% zinc phosphide, in preferred food millet (Pennisetum typhoides) grains, for 4 days, showed aversion for 5-6 days towards plain millet offered in choice with the less preferred sorghum (Sorghum vulgare) grains. The aversion response to nontoxic bait was stronger (aversion index greater than 0.7) for first 3-4 days in individual and for 1-2 days in paired rats. 100% or more shift in aversion index from pre-treatment to post-treatment periods indicated that the aversive and naive partners of the heterosexual and unisexual female pairs mutually influence the feeding preferences of each other as a result of which they showed aversion for first 2-3 days to the plain food in which poison was given to one of the partner earlier.     

Mitotic activity and its 24-hour rhythm in the ovarian follicles during the estrous cycle of a wild rat, Bandicota bengalensis

Mitotic activity in ovarian follicles was studied in relation to the size of the follicles during a 24-hour period (10.00, 16.00, 22.00 and 04.00 h) throughout the estrous cycle of the wild bandicoot rat (Bandicota bengalensis) to ascertain the cell proliferation rate and its 24-hour rhythm in the follicular tissue. In the bandicoot ovary, mitotic activity in the granulosa and thecal cells was highest in the follicles ranging from 201 to 400 micron in diameter. During the estrous cycle, mitotic activity of the granulosa cells was highest at estrus in follicles less than 601 micron, and at diestrus in follicles greater than 600 micron; while the mitotic trough was at proestrus in all the follicles. Thecal mitosis was significantly lower than mitosis of the granulosa cells. In most of the follicles, mitotic activity in the thecal cells was highest at diestrus and lowest at metestrus. In both the granulosa and thecal cells, the timing of mitotic peaks and troughs varied according to the size of the follicles and the stages of the estrous cycle. In the granulosa cells mitotic peaks were maximal in the daytime (10.00 h, 16.00 h) and in some cases at night (04.00 h); and mitotic troughs were primarily during the night (22.00 h, 04.00 h) and in some cases in the day (10.00 h). In the thecal cells, however, mitotic activity in most of the follicles was distinctly higher in the daytime (16.00 h) than at night (22.00 h, i.e., evening). Thus, a prominent 24-hour mitotic rhythm was noticed in the ovarian follicles of the bandicoot rat.     

Toxicity of Nerium indicum Miller seed extract on bandicoot rat, Bandicota bengalensis Gray

Crude extract (12.5 ml/kg) of N. indicum seed gave 100% mortality of B. bengalensis. Humanness assessment study revealed that this plant orgin chemical caused low pain and sufferings to the target pests.     

Laboratory evaluation of a toxicant-sterilant, alpha-chlorohydrin, for the control of Indian mole rat Bandicota bengalensis

A toxicant-sterilant, alpha-chlorohydrin, was evaluated against the Indian mole rat Bandicota bengalensis and the ship or house rat Rattus rattus. It caused 100% mortality of B. bengalensis at 100 mg/kg but no mortality was observed in R. rattus even at 300 mg/kg. The acute oral LD₅₀ for B. bengalensis was found to be 82 mg/kg. The survivors of B. bengalensis, which received 60 - 90 mg/kg of alpha-chlorohydrin by oral intubation, showed dose-dependent decreases in testicular weight, and cauda epididymal sperm concentration, live sperm count and sperm motility. The values of these parameters indicated that mole rats receiving the above doses would be sterile. In contrast, these changes were observed in R. rattus only at doses of 200 and 300 mg/kg. The toxic and antifertility effects of alpha-chlorohydrin observed on B. bengalensis suggest that it should be evaluated for the management of this species under held conditions.     

Variation of constitutive heterochromatin in the sex chromosomes of the rodent Bandicota bengalensis bengalensis (Gray)

Bandicota bengalensis bengalensis (Gray) trapped from different localities of India and Nepal exhibited a marked variation in the size and morphology of sex chromosomes. Three types of X's were found; A) simple acrocentric, B) composite subtelocentric and C) composite submetacentric X with their relative sizes 5.9%, 7.5% and 9.6% of the genome respectively. The autosomes remained unaltered. It was shown that this variation in the size of sex chromosomes was caused by deletion of constitutive heterochromatin. The Y chromosome was also found to be variable. Usually a large X was combined with a large Y. The preponderance of homozygotes for each type of X chromosome in populations, suggested the probable role of sex chromosomes heterochromatin in speciation.     

Effects of clomiphene citrate on the pituitary gland in chronically estrogenized rat

Effects of clomiphene citrate (clomiphene) on the pituitary gland of chronically estrogenized ovariectomized rats were investigated. Estradiol-17 beta (E2) pellet implanted subcutaneously in castrated rats for 7 days caused significant increases in pituitary weight and serum prolactin (PRL) level but suppressed serum luteinizing hormone (LH) level. In the estrogenized rats about 40% of estrogen receptor (ER) found in whole pituitary cells (65 +/- 7 fmol/10 mg tissue) was observed in the nucleus, while 60% of ER was present in the cytosol fraction. A single injection of 5 micrograms E2 translocated cytosol ER immediately to nuclear compartment; amounts of ER found in cytosol and nuclear fractions were 16 +/- 1 and 37 +/- 4 fmol/10 mg tissue, respectively, at 1 h. However, the distribution of ER returned to the pre-injection level within 4 h. In the non-estrogenized castrated rats, the nuclear retention of ER was significantly longer than that in the estrogenized rats. A single administration of 200 micrograms clomiphene in the estrogenized rats, on the other hand, increased nuclear ER gradually. Nuclear ER reached the peak level at 4 h (62 +/- 5 fmol/10 mg tissue) and the level remained almost unchanged for 24 h. Cytosol ER decreased and reached a nadir at 4 h (4.3 +/- 0.3 fmol), and the replenishment of cytosol ER could not be detected for 24 h. Similar patterns of cytosol and nuclear ER following the clomiphene injection were also found in the castrated rats. The clomiphene administration in the estrogenized rats resulted in a significant reduction of the pituitary weight 48 h after the administration. The present results seem to show the antiestrogenic action of clomiphene in the pituitary gland.     

Follicular growth and kinetics during the estrous cycle, pregnancy and postpartum in the Indian mole rat (Bandicota bengalensis)

Follicular growth and kinetics were studied in detail in the ovaries of the Indian mole rat (Bandicota bengalensis) during various stages of the estrous cycle; days 7, 12, 15, 19, and 21 of pregnancy; and day 2 postpartum. The sizes of follicles, oocytes, nuclei, and nucleoli were measured. In all rats, regression coefficients, a, and intercepts, b, were calculated in oocyte/follicle, oocyte nucleus/follicle and oocyte nucleus/oocyte regressions. The oocyte reached its maximum size when the average follicle diameter was 117 microns in nonpregnant rats and 131 microns in pregnant rats. The oocyte nucleus attained maximum size when the follicle diameter was 110 microns during the estrous cycle and 111 microns during pregnancy and postpartum. Maximum values of the diameter of the largest antral follicle and average diameter of the four largest antral follicles were observed during proestrus (473 and 442 microns, respectively) and on day 21 of pregnancy (611 and 538 microns, respectively). Chi 2 analysis showed that distribution of various types of follicles was not independent of the stage of the estrous cycle and pregnancy. In estrus and metestrus most of the follicles were between stages I and V. However, by diestrus and proestrus, follicles of all size groups developed. The numbers of stage I and II follicles did not differ as pregnancy advanced. More stage V follicles were present on day 12 than on day 7 of pregnancy; however, their numbers decreased by day 15. Afterwards, progressive increase of stage V and (VI + VII) follicles was observed until day 21. This was accompanied by the shift of follicles from stage (III + IV) on days 19 and 21 of pregnancy and even of stage II on day 2 postpartum. Wherever possible, the results have been compared with previous observations in various rodent species.     

Effect of clomiphene citrate upon periovulatory endometrial development in the baboon

Endometrial histology in baboons (Papio cynocephalus anubis) was evaluated after five days of clomiphene citrate (CC) treatment initiated on either cycle day six (n = 8) or ten (n = 4). Biopsies performed before and after a human chorionic gonadotropin (hCG) induced ovulation were compared to non-CC treated controls. Although CC had a definite antiestrogenic effect on perineal skin, no significant effect of endometrial proliferation was demonstrable.     

The effect of clomiphene citrate on LH release by dispersed rat anterior pituitary cells

Preincubation of pituitary cells for 48 hours with estrogen or clomiphene citrate did not enhance LH release of dispersed pituitary cells after incubation for 4 hours without the secretagogues. In the presence of GnRH, estrogen increased release, but neither the racemic mixture of clomiphene nor its two trans and cis isomers had any appreciable effect on LH release. It is concluded that contrary to published reports, clomiphene in the rat pituitary may behave differently from estrogen in its effects on gonadotrphin release by dispersed cells.     

Effect of estradiol and clomiphene citrate on Erk activation in breast cancer cells

Abstract

The mitogen-activated protein kinase (MAPK) signaling pathway plays key roles in the transmission of proliferative signals in normal and dysregulated cells. Nevertheless, some studies have shown that activation of the extracellular regulated kinases 1/2 (Erk1/2) is involved in apoptosis. In this study, we evaluate the effect of two fertilizing drugs, clomiphene citrate and estradiol, on the activation of Erk1/2 and the viability of two breast cancer cell lines, MCF-7 (hormone dependent) and BT20 (hormone independent).We show that both drugs induce Erk1/2 phosphorylation in MCF-7 and BT20 cells despite their opposite effect on cell viability. In fact, clomiphene citrate is significantly proapoptotic while estradiol promotes cell proliferation. The fact that phospho-Erk1/2 is a common element to both mechanisms suggests that specific factors deciding between proliferation and apoptosis must be operative downstream of this signaling pathway.     

Effects of clomiphene citrate on early pregnancy in guinea-pigs

Clomiphene citrate (2 mg/kg body wt) given on the day of mating can block or interrupt pregnancy in guinea-pigs. Corpus luteum function, uterine histology, implantation and embryo development were studied in clomiphene-treated and control animals on Days 5, 9 and 20 of pregnancy. Following treatment, only 25% of the females were regularly pregnant, presenting large and healthy foetuses. The other females examined showed either pregnancy with embryos undergoing resorption or no sign of pregnancy. In these females, corpus luteum size was reduced, progesterone concentrations were very low and the endometrial glands and the epithelium were often altered. It is concluded that clomiphene causes a reduction in fertility by altering the uterus and, by directly or indirectly inducing luteolysis, causes later pregnancy loss.     

Effects of clomiphene citrate on ovarian function in hypophysectomized rats

On Days 28-30 of age, hypophysectomized rats were treated with oestradiol-17 beta (0.1 mg/day) and/or clomiphene citrate (0.1 mg/day). Subsequent treatment with PMSG (10 i.u., on Day 31) and hCG (10 i.u., on Day 33) was identical for all animals. Rats were killed on Day 34. Treatment with oestradiol-17 beta alone resulted in ovulations of 45.1 +/- 5.5 oocytes/rat (mean +/- s.e.m.). There were no ovulations among animals treated with clomiphene citrate alone but treatment with oestradiol-17 beta and clomiphene citrate resulted in a significant (P less than 0.05) reduction (23.1 +/- 7.6 oocytes/rat) in ovulatory response. Similarly, ovarian weights and serum progesterone concentrations were highest in the oestradiol-17 beta-treated rats, intermediate in those given oestradiol plus clomiphene citrate and the lowest in rats receiving clomiphene citrate alone. We suggest that clomiphene citrate exerts direct ovarian antiovulatory and oestrogen-antagonist actions.     

Effects of clomiphene citrate, a nonsteroidal antiestrogen, on brain monoaminergic mechanisms in female goldfish, Carassius auratus

The antiestrogen, clomiphene citrate ( Merrell -National Laboratories) was administered to female goldfish in order to test the hypothesis that this drug may act on brain monoaminergic mechanisms. Brain monoamine oxidase (MAO) activity and hypothalamic serotonin (5-HT) content were measured fluorometrically after i.p. administration of 0, 5 or 25 micrograms clomiphene citrate/g body wt. Brain MAO activity was significantly inhibited by the high dose of clomiphene citrate, whereas hypothalamic 5-HT content was significantly elevated by both low and high doses of the antiestrogen. These data support the idea that brain monoamines in teleost fish are influenced by estrogen feedback mechanisms which can be blocked by clomiphene citrate.     

Effect of clomiphene citrate on pituitary responsiveness to gonadotropin releasing hormone in rams and wethers

The objective of this study was to determine the effect of clomiphene citrate (clomid) on pituitary responsiveness to gonadotropin releasing hormone (GnRH) in rams and wethers. Doses of 200 mg clomid per ram and 1 mug GnRH per 50 kg body weight were used in studies on 12 rams and 4 wethers. The experimental design involved bleeding each animal at 15-minute intervals for 6.5 hours. At the end of the first hour, GnRH was injected IV. The second GnRH challenge was administered 0.5 hours after an injection of clomid or vehicle (4.5% sorbitol solution) which was given on the third hour. The relative response to clomid or vehicle was calculated as the mean increase in concentration of LH during the two-hour period after the second GnRH injection. Each treatment (clomid and vehicle) was given to all animals with a 14-day recovery period between treatment days. The relative response for the rams receiving vehicle (1.80 +/- 0.65) was greater (P < 0.05) than the response during clomid treatment (0.34 +/- 0.22). This suppression of LH response by clomid was observed in 10 of the 12 rams. In contrast to the rams, the concentrations of LH in wethers after the second GnRH injection were lower than those observed after the first GnRH injection. Similar to the rams, the relative response following clomid treatment of wethers (0.04 +/- 0.04) was less than the relative response (P > 0.05) following vehicle (0.40 +/- 0.16). The results suggest that clomid at this dosage inhibits GnRH-induced release of LH from the pituitary of rams but not of wethers.