Prenatal Stress and Adaptive Behavior of Offspring: The Role of Placental Serotonin.

The effect of mild prenatal stress in mice, leading to an increase in the placental serotonin level, on the formation of adaptive behavior in male offspring at the age of 35 days was studied. It was shown that, in BalbC mice, daily immobilization for 1 h during the period from 11 to 14 days of pregnancy led to an increase in placental and fetal serotonin levels on the 15th day of prenatal development. According to "resident-intruder" behavioral test, the prenatally stressed mice showed more reactive behavior in adulthood and low tendency to defend their territory. Thus, placental serotonin, formed under the stress condition, may act as a mediator between the environment and the fetuses and determine the adaptive behavior of offspring.

Disruptions in the reproductive system of female rats after prenatal lipopolysaccharide-induced immunological stress: role of sex steroids.

Stress signals during fetal or early postnatal periods may disorganize reproductive axis development at different levels. This study was aimed to test the hypothesis that prenatal immunological stress induced by bacterial endotoxin, lipopolysaccharide (LPS), has impact on structure and function of the reproductive system in female offspring. Adult female Wistar rats were divided into two groups, a control group (n = 5) and a LPS group (n = 12). Rats were injected with LPS 50 µg/kg body or 0.9% saline intraperitoneally on the 12th day of pregnancy. After birth the female pups (n = 20 in each group) were divided into four groups: (group 1) 0.9% saline prenatally, sesame oil (vehicle) postnatally; (group 2) LPS prenatally, sesame oil postnatally; (group 3) LPS prenatally, fulvestrant postnatally; (group 4) LPS prenatally, flutamide postnatally. Pups were injected subcutaneously into the neck with fulvestrant (estrogen receptor antagonist), 1.5 mg/kg in sesame oil, from postnatal day (PND) 5 to PND14; or flutamide (androgen receptor antagonist), 20 mg/kg in sesame oil, from PND14 to PND30. Rats of the control group were injected with sesame oil during the same time period. Parameters were evaluated by ELISA (serum estradiol and testosterone) and ovarian histology. The main findings were: (1) prenatal stress during the critical period resulted in delayed vaginal opening, decreased body weight and serum concentrations of sex steroids, and significant disorders in ovarian development; (2) postnatal estradiol and testosterone antagonist treatments decreased follicular atresia through increasing the number of healthy follicles and restored endogenous steroid production. Lay summaryImmunological stress, caused by simulating infection through exposure to a bacterial toxin (LPS), during a critical period of fetal development in laboratory rats results in delayed reproductive maturity, decreased body weight and decreased secretion of sex steroids in female offspring, and abnormalities in the ovaries like those in polycystic ovarian syndrome. These prenatally toxin-induced sexual disorders in females could be corrected by estradiol/testosterone antagonists during the postnatal period.

[The experimental evaluation of the protective effect of the extract of the cyclodextrin-containing phytotherapeutic composition on the development of osteoporosis].

OBJECTIVE: The relevance of the problem considered in the present communication arises to-day from the widespread prevalence of osteoporosis (OP), the rather low effectiveness of the methods currently available for the treatment of this condition, and iatrogenic effects of its medication therapy. The great diversity of phytotherapeutic modalities of traditional medicine are is only poorly supported by the results of the scientific studies results; moreover, most of them are coming from the foreign literature publications. AIM:  The objective of the present study was to evaluate the influence of the phytotherapeutic preparation (Phytocost) having cyclodextrin in its composition on the restoration of the cartilaginous and bony tissues based on the experimental research with the use of the OP experimental model. MATERIAL AND METHODS:  The experimental studies were performed on 5 month-old Balb/c mice used as the prednisolone-induced osteoporosis model. The animals were divided into four study groups according to the daily doses administered to them: 0.005; 0.05; 0.5 and 5.0 mg/mouse during 30 days and 3 control groups: intact, 14 and 30 days after prednisolone administration, naïve mice. RESULTS:  The study has demonstrated the dose-dependent protective effect of cyclodextrin-containing Phytocost. The most pronounced effect in the form of reduction of osteoblast number and the increase in the number of osteocytes was obtained at a Phytocost dose of 0.5 mg/mouse. No undesirable adverse reactions were documented during the study. CONCLUSION:  The domestically produced Phytocost composition differs from its foreign analogues in that it contains a significantly greater number of constituent components because the ultimate goal of the study was to create a medication acting on all currently known mechanisms of OP pathogenesis, whereas our Chinese colleagues proceed from the ancient knowledge gained by traditional medicine that it is sometimes difficult to understand and explain in the light of the modern concepts. All the plants used as raw materials for Phytocost production grow at the territory of the Russian Federation which provides the possibilities for the efficient import substitution of the components necessary for the manufacture of the preparation in question.

The inhibition of dopamine synthesis in fetuses changes the pattern of T-lymphocyte maturation in the thymus of adult rats.

The mRNA for dopamine receptors of type D1, D3, D5, but not type D2, was detected in the thymus of rats starting from day 16 of embryonic development (E16). Dopamine at concentrations of 10-8-10‒6 M inhibited fetus thymocyte response to mitogen, confirming the functionality of the receptors and the possibility of a direct effect of dopamine on the developing thymus. Pharmacological inhibition of catecholamine synthesis in the crucial period of thymus development leads to long-term changes in the T-system immunity due to increased production of natural regulatory T-lymphocytes. The presence and functional activity of dopamine receptors in the fetal thymus indicates its ability to influence the development of the immune system of rats during ontogeny.

[New phytotherapeutic composition for restoring bone and cartilage. Experimental study].

Presents a new import-substituting the composition of the powder mixture 60 medicinal plants and dead bees. In prednisolonbuy models of osteoporosis have shown dose-dependent regenerative effects bone and cartilage of the hip joints of mice. Unwanted side effects when taking composition was observed. It is assumed the possibility of effective application of the composition as a complementary treatment for osteoporosis.

[The role of serotonin in the immune system development and functioning during ontogenesis].

In this study, we investigated the influence of serotonin on the development and functioning of T- and B-cell-mediated immunity during ontogenesis using the pharmacological model of serotonin depletion in rat fetuses. It has been demonstrated that prenatal serotonin deficiency resulted in a decrease in thymus and spleen weights, changes in their cellular composition, and long-lasting disturbances in cell-mediated and humoral immunity in postnatal ontogenesis. The data obtained suggest that serotonin may be considered a morphogenic factor in development of the immune system.

[The comparative assessment of the wound-healing effects of the treatment with the use of Bioptron, Minitag, Orion+ apparatuses and hollow cathode lamps (experimental study)].

The objective of the present experimental study was the comparative assessment of the wound-healing effects of radiation emitted from Bioptron, Minitag, Orion+ apparatuses and hollow cathode lamps (HCL). The emitters of any type were shown to be equally efficacious in that they accelerated wound epithelization by 30% on the average compared with control. Based on the difference between spectral and power characteristics of different sources of radiation and dynamics of their wound-healing efficacy (including that of two types of HCL), the authors arrived at the conclusion that the further development of the proposed approach to wound healing is a promising line of research in the field of spectral phototherapy.

[Effect of bacterial endotoxin on migration of gonadotropin-releasing, hormone producing neurons in rat embryogenesis].

The effect of bacterial lipopolysaccharide endotoxin (LPS), immune system activator, on differentiation and migration of gonadotropin-releasing, hormone producing neurons in rat embryogenesis has been studied. Intraperitoneal introduction of LPS (18 jg/kg) to pregnant rats on the 12th day of pregnancy led to 50% decrease in total number of GRH-neurons in the forebrain of 17-day-old embryos and 17% decrease in 19-day-old embryos. At the same time, the number of GRH-neurons in the nasal area of the head of 17- and 19-day-old embryos increased by 40 and 50%, respectively, whereas it increased by 20% in olfactory bulbs of 17-day-old embryos and did not changed in olfactory bulbs of 19-day-old embryos. Neither the total number of neurons nor their distribution patterns were affected by the introduction of LPS into pregnant rats on the 15th day of pregnancy. Singular localization of GRH-neurons in embryo forebrain was observed after LPS administration, whereas the neurons were located by groups of 3-4 cells in rostral areas. Therefore, at the early stages of pregnancy, LPS was shown to suppress initial stages of differentiation and migration of GRH producing neurons. The effects observed in our study may be mediated by LPS-induced, proinflammatory cytokines.

[Reparative regeneration of rat skin under influence of hollow cathode lamp (HCL) with manganese and copper line spectrum emission].

Influence of local light exposure by hollow cathode lamp with typical manganese and copper (HCL-Mn, Cu) line emission spectrum on posttraumatic regeneration rate of rat skin has been investigated. We performed the comparative analysis of the morphology and the differentiation ability of rat skin on the 15th and 24th days after full-thickness skin wound had been inflicted on rat dorsums. On the 15th day after injury, the experimental group (daily 30 s exposure for two weeks) showed scab loss, re-epithelialization, and hair regrowth, in contrast to the control rats, where scabs were still observed on the 24th day. Histological analysis revealed that in contrast to the control group the treatment with HCL-Mn, Cu resulted in the increased number of hair follicles and sebaceous glands, the decreased number of blood vessels and horizontal orientation of collagen fibers. The immunohistochemistry for OX-62 revealed that the number of dermal dendritic cells in the experimental groups was maximal on the 15th day, and then decreased to the 24th day after injury. The number of dermal dendritic cells was significantly lower in the control group. The immunohistochemistry for pan-keratins in the control animals revealed a high number of cells expressing different types of keratins, distributed in the main part of the epidermis on the 15th day after surgery, whereas in the experimental group the number of such cells was significantly lower and the cells were concentrated more close to the external part of the epidermis. The number of cells stained for keratin 19 was higher in the experimental group on the 15th day after surgery, whereas this number decreased in this group on the 24th day after surgery as compared to the control group. Thus, typical manganese and copper line spectrum emission emitted by hollow cathode lamp stimulates innate immunity, accelerates restoration of derma, skin epithelium and other skin derivates, and stimulates wound healing in general.

Transcutaneus photophoresis of metal ions using emitters of band spectrum of chemical elements.

Local exposure to light with hollow cathode lamp radiating band spectrum typical of manganese, copper, potassium, sodium, calcium, and magnesium enhances migration of these elements from the solution applied to the skin to the blood in rats. This effect is most pronounced at low initial blood level of manganese. Its serum concentration increased 17-fold after application of manganese salts and exposure to hollow cathode lamp radiating manganese spectrum.

The brain is one of the most important sources of dopamine in the systemic circulation in the perinatal period of ontogenesis in rats.

This study was designed to test the authors' hypothesis that dopamine passes from dopamine-synthesizing cells in the brain to the systemic circulation prior to the formation of the blood-brain barrier during ontogenesis. High-performance liquid chromatography studies demonstrated that peripheral blood dopamine levels before formation of the blood-brain barrier-in rat fetuses and neonates-are significantly higher than after formation of the barrier in adult rats, providing indirect evidence in support of the hypothesis. Furthermore, formation of the blood-brain barrier is accompanied by a significant increase in dopamine levels in the rat brain. Direct evidence for the hypothesis was obtained in the form of a sharp decrease in blood dopamine levels in fetuses after lesioning of dopamine-synthesizing neurons in the brain by encephalectomy.

[Migration and differentiation of neurons producing gonadotropin-releasing hormone under conditions of serotonin excess in the brain of mouse embryos].

The work has been carried out on mice of the Tg8 line with knockout of gene of monoamineoxidase A with an increase of serotonin and noradrenaline content in the brain, and on mice of the C3H line with unchanged genome and normal concentration of monoamines. An immunocytochemical study has been performed of development of neurons producing gonadotropin-releasing hormone (GnRH) under conditions of excess of serotonin and noradrenaline in the mice in embryogenesis. The GnRH-neurons were revealed at the 18th day of embryonic development in telencephalon along trajectory of their migration from olfactory bulbs to the retrochiasmatic area. In telencephalon of mouse embryos of the Tg8 line, a redistribution of the GnRH-neurons along their migration trajectory was observed as compared with embryos of the C3H line mice. The percent of the GnRH-neurons in the Tg8 mouse embryos in caudal parts of the migration trajectory was lower than in rostral parts, the opposite distribution of the neurons being observed in the C3H line mouse embryos; at the excess of serotonin and noradrenaline in the Tg8 line mouse embryos, the total amount of GnRH-neurons in the brain was lower than in the C3H mice. In males of the Tg8 line mice under conditions of excess of serotonin and noradrenaline the optical density of neurons, which correlated with the GnRH concentration in the cell, was higher than in control mice. Thus, in the Tg8 mice under conditions of the serotonin and noradrenaline excess, migration of the GnRH-neurons to their final anlage in hypothalamus is accelerated as well as the total number of the GnRH-neurons decreases, which indicates a decrease of proliferation of cells-precursors and the earlier differentiation of neurons.

[The brain is one of the most important sources of dopamine in general circulation during perinatal ontogenesis in rats].

This study was aimed to test our hypothesis that dopamine synthesized in the neurons of the brain is delivered to the general circulation in rats during prenatal and early postnatal periods, i.e. before the establishment of the blood-brain barrier. Using the high performance liquid chromatography, it was demonstrated that the dopamine concentration and content in the peripheral blood in fetuses and neonatal rats (i.e. before the establishment of the blood-brain barrier) greatly exceeded those in adult rats. Moreover, the establishment of the blood-brain barrier was accompanied by the significant increase of the dopamine concentration in the brain. A drop of the dopamine concentration in fetal plasma after the microsurgical lesion of the forebrain and mesencephalon (encephalectomy) are considered as direct evidence in favour of our hypothesis.

[Catecholamines in regulation of development of GnRH neurons of rat fetuses].

The contents of dopamine, serotonin, and noradrenaline in rat fetuses developing under the conditions of their deficiency induced by administration of alpha-methyl-para-tyrosine to females during 11th to 16th or 20th day of pregnancy and in fetuses, whose mothers were given saline at the same time, were determined using HPLC with subsequent electrochemical detection. Administration of alpha-methyl-para-tyrosine led to decreased levels of dopamine and noradrenaline in the areas of migration of GnRH-neurons in fetuses on days 17 and 21 of prenatal development. The concentration of serotonin remained unchanged, except in the head nasal area in males on day 21. The areas of interaction between the brain catecholaminergic systems and migrating and differentiating GnRH-neurons were determined by double immunohistochemical labeling. Close topographical location of GnRH-immunoreactive neurons and tyrosine hydroxylase-immunoreactive in the area of nucleus accumbens on days 17 and 20, as well as in the median eminence on day 20. The GnRH concentration in the caudal areas of migration of GnRH-neurons under the normal conditions and in the case of catecholamine deficiency was determined using radioimmunoassay. After administration of alpha-methyl-para-tyrosine the GnRH concentration in the anterior hypothalamus decreased in females. The data obtained suggest the involvement of catecholamines in the regulation of development of GnRH-Neurons during prenatal development. In addition, the adequacy and efficiency of the used model of catecholamine deficiency for studying the development of such neurons was confirmed.

[Effect of catecholamines on migration and differentiation of gonadotropin releasing hormone-neurons in rats]. / Vliianie kateholaminov na migratsiiu i differentsirovku gonadotropin-relizing gormon-neironov u krys.

The GnRH producing neurons are the key link of neuroendocrine regulation of the adult reproductive system. Synthesis and secretion of GnRH are, in turn, under the afferent catecholaminergic control. Taking into account that catecholamines exert morphogenetic effects on target cells during ontogenesis, this study was aimed at investigation of the effects of catecholamines on development of GnRH neurons in rats during ontogenesis. We carried out comparative quantitative and semiquantitative analyses of differentiation and migration of GnRH neurons in fetuses of both sexes under the conditions of normal metabolism of catecholamines (administration of saline) or their pharmacologically induced deficiency (administration of alpha-methylparatyrosine). The inhibition of catecholamine synthesis from day 11 of embryogenesis led to an increasing number of GnRH neurons in rostral regions of the trajectory of their migration over the brain: in the area of olfactory tubercles on day 17 and in the area of olfactory bulb on days 18 and 21. In addition, the optical density of GnRH neurons located in the rostral regions of migration was higher in the fetuses after administration of alpha-methylparatyrosine during embryogenesis, as compared to the control. It has been concluded that catecholamines stimulate the migration of GnRH neurons and affect their differentiation.