Individual typological characteristics of hormonal responses in dogs during psychoemotional stress

Effects of psycho-emotional stress on contents of steroid hormones in the blood were studied in dogs with different typological properties of the higher nervous activity. Under stress condition, a significant correlation between the cortisol and testosterone levels and the dogs' typological characteristics was found. The more obvious fluctuations in the hormones level occurred in animals with strong and excitable type of the nervous system as compared with dogs belonging to weak and inert type.     

Changes in reproductive function during chronic isolation and increases in the population density of rats with different typologic behavioral characteristics

It has been shown on white outbred female rats that animals with a strong balanced mobile nervous system and active-search behaviour had the greatest stress resistence against the action of chronic isolation and increase of population density. Weak non-balanced excitable animals with non-directed motor restlessness had a minimal resistence against stress.     

Involvement of GABA-A receptor chloride channel complex in isolation stress-induced free choice ethanol consumption in rats

The present study revealed the effect of diazepam, a benzodiazepine, and progesterone, a pregnane precursor of neurosteroids, which act via modulating GABA-A chloride channel complex on the isolation stress-induced free choice ethanol consumption in adult rats. Isolation stress for 24 hr over a period of 6 days produced a significant increase in ethanol consumption, which persisted during the 6-day recovery period. Pretreating the animals with diazepam (5 mg/kg, i.p.), or progesterone (5 mg/kg, i.p.), blocked the isolation stress-induced increase in ethanol consumption. Bicuculline (2 mg/kg, i.p.), a GABA-A receptor antagonist significantly attenuated the effect of both diazepam and progesterone on stress-induced modulation of ethanol consumption. Isolation stress also caused an increase in total fluid consumption, which was antagonised by both diazepam and progesterone. Like ethanol consumption, this effect of diazepam and progesterone on isolation stress-induced increase in total fluid consumption was attenuated by bicuculline. Neither diazepam nor progesterone produced an increase in ethanol consumption in non-stressed rats. However, unlike diazepam, progesterone administration to non-stressed rats caused a significant increase in total fluid consumption. Results of the present study thus show that GABAergic mechanisms may be playing an important role in isolation stress-induced increase in ethanol consumption.     

The adrenal gland and progesterone stimulates testicular steroidogenesis in the rat in vivo

Administration of pharmacological doses of glucocorticoid to male rats in vivo suppresses adrenal steroidogenesis and inhibits testicular steroidogenesis by inhibiting the anterior pituitary secretion of LH. In contrast, administration of ACTH to these pharmacologically-suppressed rats stimulates the adrenal secretion of progesterone and testicular steroidogenesis. The mechanism by which ACTH increases testicular steroidogenesis is dependent on the presence of the adrenal gland and is reproduced by the administration of progesterone. The conclusion from these data is that the adrenal gland has an important role in generating external signals that modulate the hypothalamic-pituitary-gonadal axis in male rats. The adrenal secretion of glucocorticoid acts as a negative signal to testicular steroidogenesis whereas progesterone acts as a positive signal. The adrenal secretion of progesterone and its conversion to testosterone by steroidogenic enzymes in the cytoplasm of the Leydig cell may provide an alternative pathway for testosterone biosynthesis and may account for the increased plasma testosterone levels during the acute phase of stress and mating.     

Progesterone prevents corticosterone mediated inhibition of estrous behaviour in rats

Stress induced by application of electric foot shocks (300 microA/shock, five shocks per episode, 4 episodes at 1800, 1830, 1900 and 1930 hrs on the proestrus day) to rats at the time of pre-ovulatory progesterone secretion, abolished lordosis and resulted in maximum rejection co-efficient, whereas treatment with a CRF receptor antagonist (alpha-helical CRF9-41) or metapirone, an inhibitor of corticosterone synthesis, prior to application of the electric foot shocks, resulted in normal lordosis and a significant reduction in rejection coefficient. Further, administration of a single dose of corticosterone (40 microg) at 1800 hrs of proestrus caused inhibition of lordosis and resulted in maximum rejection co-efficient. On the other hand, corticosterone + progesterone treatment at 1800 hrs of proestrus resulted in normal lordosis and a significant reduction in rejection coefficient. The facts that stress induced inhibition of lordosis is prevented by CRF receptor antagonist or metapirone and that corticosterone inhibits lordosis indicate that stress induced inhibition of lordosis is mediated by corticosterone. Further, normal display of lordosis by rats treated with corticosterone + progesterone in contrast to its absence in corticosterone alone treated rats suggests that impaired progesterone secretion due to action of corticosterone leads to inhibition of lordosis.     

Progesterone restores stress induced inhibition of estrous behavior in rat

Exposure to a stressor (mild electrical shocks to foot, five times per episode, at 1800, 1830, 1900 and 1930 hrs of proestrus) coinciding with period of pre-ovulatory progesterone secretion in rats abolished estrous behavior as shown by the absence of lordosis response and a significant increase in rejection quotient compared to controls. These rats did not show spermatozoa in the vaginal smear next day morning in contrast to their presence in controls. On the other hand, rats treated with progesterone (a single injection, 500 microg in 0.1 ml olive oil at 1800 hr of proestrus) prior to exposure to stressor showed normal estrous behavior, as shown by significantly lower rejection quotient than rats exposed to stress alone, lordosis quotient similar to controls and presence of spermatozoa in the vaginal smear next day. The results, albeit indirectly, to the best of our knowledge, first time indicate that stress induced impaired steroidogenesis leads to suppression of estrous behavior.     

Mechanisms responsible for progesterone's protection against lordosis-inhibiting effects of restraint I. Role of progesterone receptors

Progestins and antiprogestins are widely used therapeutic agents in humans. In many cases, these are indicated for the treatment of reproductive activities. However, progesterone has widespread physiological effects including a reduction of the response to stress. We have reported that 5 min of restraint reduced lordosis behavior of ovariectomized rats hormonally primed with estradiol benzoate. When ovariectomized rats received both estradiol benzoate and progesterone priming, restraint had minimal effects on lordosis. Progesterone influences behavior through classical intracellular progesterone receptor-mediated nuclear events as well as extranuclear events. How these multiple events contribute to the response to stress is unclear. The current project was designed to initiate examination of the mechanisms responsible for progesterone's ability to protect against the effects of the restraint. In the first experiment, ovariectomized rats, primed with 10 μg estradiol benzoate, received 500 μg progesterone 4 h, 1 h, or 30 min before restraint. When progesterone was injected 4 h before restraint, progesterone eliminated the effects of restraint. In contrast, progesterone 30 min before restraint offered no protection. Effects of progesterone 1 h before restraint were equivocal allowing the suggestion that less than 4 h of progesterone priming might be sufficient. In the second experiment, the synthetic progestin, medroxyprogesterone, was shown to mimic effects of progesterone in preventing effects of restraint. Finally, the progesterone receptor antagonist, RU486, attenuated progesterone's protection against restraint. These findings offer evidence that ligand-activated progesterone receptor mechanisms contribute to the maintenance of lordosis behavior in the presence of mild stress.     

Ovarian responses to perphenazine-induced prolactin secretion in the rats: Effect of ovulation, stress, and steroids.

A single injection of 2.5 mg perphenazine (PH)/kg body wt to rats on the day of estrus (day 0) did not result in increased serum progesterone 24 hr later. Continued daily injections, however, resulted in a 2.5-fold increase in serum progesterone between days 1 and 3 and a 1.6-fold increase between days 3 and 5 to a final concentration of 58 plus or minus 4 ng/ml on day 5 in serially anesthetized and bled rats. Neither daily administration of 5.0 nor 10.0 mg PH/kg body wt to rats subjected to the stressful conditions of this regimen resulted in further increases in serum progesterone, but the 5.0 mg dose of PH in unstressed rats bled only on day 5 resulted in a highly significant increase in serum progesterone to 110 plus or minus 7 ng/ml. In unstressed rats the increase in serum progesterone over control values after five daily injections of 2.5 mg PH/kg body wt could be attributed to decreased 20alpha-reduction of progesterone, but when the dose of PH was increased to 5.0 mg/kg, a highly significant increase in both progesterone and total progestins occurred indicating that prolactin can increase steroidogenesis as well as reduce 20alpha-hydroxysteroid dehydrogenase activity. After inhibition of ovulation, the 5.0 mg daily dose of PH resulted in serum progesterone of only 25 plus or minus 8 ng/ml on day 5 in unstressed rats. Thus, serum progesterone in ovulating rats treated with PH originated primarily in the corpora lutea. Perphenazine, 5.0 mg/kg, administered only on estrus and the first day of diestrus was sufficient to induce pseudopregnancy of 14.5 plus or minus 1.6 days. No evidence for gonadotropin stimulation of the ovaries of any rats was observed. The effect of stress on the progesterone response was not mimicked by administration of cortisol acetate and is assumed to be medicated by suppression of prolactin secretion.     

Therapeutic administration of progesterone antagonist in a model of Charcot-Marie-Tooth disease (CMT-1A)

Charcot-Marie-Tooth disease (CMT) is the most common inherited neuropathy. The predominant subtype, CMT-1A, accounts for more than 50% of all cases and is associated with an interstitial chromosomal duplication of 17p12 (refs. 2,3). We have generated a model of CMT-1A by introducing extra copies of the responsible disease gene, Pmp22 (encoding the peripheral myelin protein of 22 kDa), into transgenic rats. Here, we used this model to test whether progesterone, a regulator of the myelin genes Pmp22 and myelin protein zero (Mpz) in cultured Schwann cells, can modulate the progressive neuropathy caused by moderate overexpression of Pmp22. Male transgenic rats (n = 84) were randomly assigned into three treatment groups: progesterone, progesterone antagonist (onapristone) and placebo control. Daily administration of progesterone elevated the steady-state levels of Pmp22 and Mpz mRNA in the sciatic nerve, resulting in enhanced Schwann cell pathology and a more progressive clinical neuropathy. In contrast, administration of the selective progesterone receptor antagonist reduced overexpression of Pmp22 and improved the CMT phenotype, without obvious side effects, in wild-type or transgenic rats. Taken together, these data provide proof of principle that the progesterone receptor of myelin-forming Schwann cells is a promising pharmacological target for therapy of CMT-1A.     

Psycho-emotional manifestations in rats with hippocampal lesions

The study aimed at revealing psycho-emotional manifestations of Wistar rats in a problem situation when they had to solve a food-getting task in a multi-alternative maze. Bilateral lesion of dorsal hippocampus did not affect the learning process while the pattern of psycho-emotional manifestations changed in all the animals irrespective of their individual properties. The pverall effect was manifested in flatness of psycho-emotional responses as a result of reduction of extreme forms of both passive and active stress responses. Individual effects depended on the behavioural phenotype. Hippocampus lesion in initially excitable rats (10%) resulted in reduction of active stress responses (act/pass = 0.35/0.5 instead of 1.0/0.4 in control). Hippocampus lesion in initially inhibited rats (30%) did not affect the basic pattern of psycho-emotional responses, while in the rats with initially depressed cognitive activity (60%) it resulted in decreasing of passive and increasing of active stress responses (act/pass = 0.45/1/4 instead of 0.3/1.9). The findings suggest that hippocampus takes part in the psycho-emotional responses while the individual psycho-emotional pattern is determined by the morpho-functional features responsible for organisation of a psycho-emotional condition.     

Effects of swim stress on mRNA and protein levels of steroid 5alpha-reductase isozymes in prefrontal cortex of adult male rats

The metabolite of progesterone, allopregnanolone, is among the most potent known ligands of the gamma-aminobutyric acid receptor complex (GABA(A)-R) in the central nervous system. This neuroactive steroid is markedly increased in an animal model of acute stress. Allopregnanolone is synthesized from progesterone by steroidogenic enzymes 5alpha-reductase (5alpha-R) and 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD), with the former being the rate-limiting enzyme in this reaction sequence. In this paper, a quantitative RT-PCR method coupled to laser-induced fluorescence capillary electrophoresis (LIF-CE) and Western blot were used to measure both mRNA and protein levels of 5alpha-R type 1 (5alpha-R1) and 5alpha-R type 2 (5alpha-R2) isozymes in prefrontal cortex of male rats after acute swim stress situations. Our results demonstrate that both 5alpha-R isozymes are significantly higher in prefrontal cortex of male rats after acute swim stress in comparison with control rats. These data may open up a new research line that could improve our understanding of the role of 5alpha-R isozymes in processes that accompany stress situations.     

Melatonin and adrenal cortex steroid production: in vivo and in vitro studies.

The present study was designed to clarify the interaction between the pineal melatonin and adrenal cortex steroid production. Experiments with male rats under chronic stress conditions (sleep deprivation) revealed that melatonin circadian pattern was fully destroyed and daytime plasma concentration were significantly elevated. Constant illumination (2500 lux) during the nighttime was not able to suppress melatonin production in the stressed animals. Plasma concentration of corticosterone were increased in the stressed rats as well. The modulatory effect of melatonin on corticosterone and progesterone production by rat adrenals was studied in a superfusion system. During melatonin challenge progesterone secretion was two-three fold elevated with no effect on corticosterone content in the plasma samples. Pineal cytoplasmic glucocorticoid and progesterone receptors were investigated as well. A specific binding was not observed in that case. Presented data support the existence of direct communication between the pineal and adrenal glands.     

Steroid hormone control of myometrial contractility and parturition

The precise temporal control of uterine contractility is essential for the success of pregnancy. For most of pregnancy, progesterone acting through genomic and non-genomic mechanisms promotes myometrial relaxation. At parturition the relaxatory actions of progesterone are nullified and the combined stimulatory actions of estrogens and other factors such as myometrial distention and immune/inflammatory cytokines, transform the myometrium to a highly contractile and excitable state leading to labor and delivery. This review addresses current understanding of how progesterone and estrogens affect the contractility of the pregnancy myometrium and how their actions are coordinated and controlled as part of the parturition cascade.     

The effect of prenatal stress on activity of the neurosteroids synthesis enzyme in the "critical period" of brain sexual differentiation in male rats

The effect of daily immobillisation stress in female rats on the 15th to 18th days of pregnancy upon synthesis enzyme for neurosteroids of alpha-reductase in their male offspring brain, was studied. A decrease in the enzyme activity in the cortex and hypothalamus of male foetuses occurred within 24 hr following the latest stress, whereas it was increased in the cortex of newborn offspring. An enhancement of the 5 alpha-reductase activity in the cortex, hippocampus and hypothalamus was also found in prenatally stressed males on the 5th day of life. A decrease in the testosterone and progesterone contents in the blood plasma of the animals under study was revealed on the 19th day of their embryonic life as well as in newborn rats, the blood level of progesterone, at that, remained decreased even at the age of 5 days. A possible part ofneurosteroids in action of prenatal stress upon sexual differentiation of the brain is discussed.     

Pubertal maturation and time of day differentially affect behavioral and neuroendocrine responses following an acute stressor

Puberty markedly influences stress responsiveness such that prepubertal animals show a more protracted corticosterone (CORT) and progesterone response following acute stress compared to adults. In both adult and juvenile rats, circadian time modulates adrenocortical steroids with basal CORT and progesterone levels rising prior to the onset of the dark phase of the light-dark cycle (i.e., active period). How time of day affects the pubertal difference in stress responsiveness and if the behaviors of prepubertal and adult animals are differentially affected by stress and time of testing remain unknown. Thus, we exposed group housed (3 per cage) prepubertal (28d) and adult (77d) male rats to 30 min of restraint in either the early portion of the behaviorally inactive, light (circadian nadir of CORT and progesterone) or behaviorally active, dark (circadian peak) phase of their light-dark cycle and measured ACTH, CORT, progesterone, and home cage behavior before and after the stressor. We found that the extended hormonal stress response demonstrated by prepubertal males occurred at both times of day. However, differences in post-stress behavior were dependent on time of testing. Specifically, although pre- and post-stress behaviors were similarly affected by the stressor in the light phase in prepubertal and adult males, during the dark phase, stress suppressed play behavior in the prepubertal males, and increased their time spent resting together (huddling), while these behaviors were unaffected by stress in the adults. These data indicate that pubertal development and time of day interact to modulate post-stress behavior and demonstrate a dissociation between post-stress hormonal and behavioral responses.     

Gravity sensing in the central nervous system

For human based space research it is of high importance to understand the influence of gravity on the properties of the central nervous system (CNS). Untill now it is not much known about how neuronal tissue can sense gravity. The aim of this study was to find out weather and how the CNS, as a complex system, can percept and react to changes in gravity. Neuronal tissue and especially the CNS fulfils all the requirements for excitable media. Consequently, self-organisation, pattern formation and propagating excitation waves as typical events of excitable media have been observed in such tissue. The Spreading Depression (SD), an excitation depression wave is the most obvious and best described of these phenomena in the CNS. In our experiments we showed that the properties of the SD and therefore the CNS in its properties as an excitable medium reacts very sensitive to changes in gravity.     

Effects of progesterone, epipregnanolone and RU 38486 on potassium uptake in cultured cortical neurons

It was previously reported that progesterone and its metabolites influence electrical properties of the CNS in many different ways. In the present study we elicited the effects of progesterone, its 5 beta reduced metabolite epipregnanolone and the anti-progestin compound RU 38486 on potassium uptake in cultured cortical neurons. K+ was substituted by the tracer substance 86Rb. When hormone treatment (10(-9)-10(-7) M/l) was performed for 3 days, addition of progesterone and epipregnanolone led to a significant decrease of 86Rb uptake whereas treatment with RU 38486 markedly increased 86Rb uptake. The effect of the anti-progestin could be reversed by the addition of increasing amounts of progesterone. Hormone actions were dose-dependent and most distinct when performed from the very first day of culture. Short-term (15 min) hormone treatment of neurons did not significantly alter 86Rb uptake. These findings suggest a specific receptor mediated progestin action which, in a long-term course, controls potassium uptake across excitable membranes.     

Effect of blastocysts on rat ovarian steroidogenesis in early pregnancy.

The biosynthesis of ovarian progesterone and 20alpha-hydroxypregn-4-en-3-one (20alpha-OHP) was studied in rats with pituitaries autotransplanted beneath the kidney capsule (APTr rats) 1 1/2 days after ovulation. Some animals underwent tubal ligation 1-2 weeks prior to mating. Incubated ovarian tissue slices showed an accumulation of progesterone of 64 plus or minus 11 mcg/g tissue, and a progesterone/20alpha-OHP ratio of .32 plus or minus .16. Ovarian tissue from APTr rats with viable blastocysts synthesized considerably more progesterone than tissue from tubal ligated APTr rats. The increased concentrations of progesterone were associated with decreased concentrations of 20alpha-OHP. The accumulation of progesterone in tissue from unmated APTr rats, APTr rats cervically stimulated at proestrus, and similar cervically stimulated rats with traumatized uteri was 42 plus or minus 5, 16 plus or minus 4 and 18 plus or minus 9 net mcg/g tissue, respectively. The progesterone accumulation in tissue from cervically stimulated rats was thus similar to that in tissue from tubal ligated animals. This inhibition by cervical stimulation was overcome by stimuli from the blastocysts. The results suggest that the blastocyst secretes a substance that is more active in promoting progesterone formation than pituitary prolactin.     

Effects of progesterone on concentrations of monoamines in hypothalamic areas and plasma prolactin levels in rats.

The role of progesterone to increase prolactin (PRL) secretion on the first estrous day in pubertal rats was compared with its role in adult cyclic rats. The first estrus was induced by the administration of pregnant mare serum gonadotrophin (5 IU) at 28 days of age. A subcutaneous administration of 2.5 or 7.5 mg of progesterone/100 g body wt significantly increased the concentration of plasma PRL in pubertal rats within 4 hr. The PRL level obtained after progesterone administration was greater than that in similarly treated adult rats. The concentration of dopamine in the arcuate nucleus-median eminence (ARC-ME) in pubertal rats significantly decreased after a lower dosage of progesterone was administered, but no change was found in the preoptic area concentration. In adult estrous rats, the concentration of dopamine in the ARC-ME showed a tendency to decrease after the administration of a larger dose of progesterone (7.5 mg/100 g body wt). No change was observed in the concentrations of indoleamines in the preoptic area and ARC-ME after the administration of progesterone in both pubertal and adult rats. The concentrations of dopamine in the preoptic area and ARC-ME were lower in pubertal rats than in adults. The concentration of 5-hydroxyindole acetic acid and the ratio of 5-hydroxyindole acetic acid to 5-hydroxytryptamine in the ARC-ME were higher in pubertal rats than in adults. These results indicate that progesterone causes a greater increase in tonic PRL secretion in pubertal rats than in adult rats and that a lower hypothalamic dopamine activity and a higher serotonin activity in pubertal rats may account for these differences.     

Effect of progesterone and estrogens on the state of the hypothalamo-hypophyseal neurosecretory system in pregnancy and labor

Morphofunctional state of the hypothalamo-hypophyseal neurosecretory system was studied in rats at late stages of pregnancy. No special differences both in function of the supraoptical and praventricular nuclei, and in the hypophysis of intact and pregnant rats at administration of progesterone were noted. When estrogenes were administered to pregnant animals, a sharp increase in functioning of the supraoptic and especially of the paraventricular nuclei was noted. The greatest activity of the supraoptic nucleus was observed at delivery that could be resulted from a stress reaction to pain.