[Age- and sex-related differences of neuroendocrine center reactions in the hypothalamus on preparation thymalin and alpha-tocopherol acetate].

In experiment the action of Thymalin, alpha-tocopherol and stress on the centers of hypothalamus related to the regulation of the gonadotropic function has been investigated and the age-specific features of such influence as well. The results of experiments indicate the presence of neuron's reaction of the rostra preoptical area and arcuate nucleus for experimental influences. As a stress exposure and alpha-tocopherol injection to the young animals a reduction of neuron's kernels of the arcuate nucleus was observed, a sexual distinction wasn't revealed in the process. The response age-specific feature for alpha-tocopherol injection of the nucleus neuron's rostra preoptical and arcuate area of the hypothalamus of white rats is a reduction in a degree of response. Thymalin reduces a threshold of sensitivity of the neuron's arcuate center to action of stress.

Dendritic spines in the posterodorsal medial amygdala after restraint stress and ageing in rats.

Several evidences suggest that the posterodorsal medial amygdala (MePD) can be a relevant part of the rat neural circuitry for the regulation of hypothalamic neuroendocrine secretion and for ontogenetically different behavioral displays. The dendritic spine density of Golgi-impregnated neurons from the MePD was evaluated in young rats following acute or chronic restraint stress and in aged animals (24 months old). Compared to the control group, a single 1 h restraint stress session promoted a decreased spine density (p<0.01) whereas a single 6 h restraint stress session or daily 6-h restraint sessions for 28 consecutive days did not lead to the same effect (p>0.05). Aged rats showed no difference in this dendritic spine parameter when compared to young adults (p>0.05). These results indicate that short-term stress (1 h) can affect MePD dendritic spines and that neural plasticity is involved with adaptive responses onwards in restrained rats. On the other hand, brain structural modifications related with ageing appear not to influence the number of certain postsynaptic sites in the MePD of rats.

Conserved sensory-neurosecretory cell types in annelid and fish forebrain: insights into hypothalamus evolution.

Neurosecretory control centers form part of the forebrain in many animal phyla, including vertebrates, insects, and annelids. The evolutionary origin of these centers is largely unknown. To identify conserved, and thus phylogenetically ancient, components of neurosecretory brain centers, we characterize and compare neurons that express the prohormone vasotocin (vasopressin/oxytocin)-neurophysin in the developing forebrain of the annelid Platynereis dumerilii and of the zebrafish. These neurons express the same tissue-restricted microRNA, miR-7, and conserved, cell-type-specific combinations of transcription factors (nk2.1, rx, and otp) that specify their identity, as evidenced by the specific requirement of zebrafish rx3 for vasotocin-neurophysin expression. MiR-7 also labels another shared population of neurons containing RFamides. Since the vasotocinergic and RFamidergic neurons appear to be directly sensory in annelid and fish, we propose that cell types with dual sensory-neurosecretory properties were the starting point for the evolution of neurosecretory brain centers in Bilateria.

New insights on Saccus vasculosus evolution: a developmental and immunohistochemical study in elasmobranchs.

The saccus vasculosus (SV) is a circumventricular organ of the hypothalamus of many jawed fishes whose functions have not yet been clarified. It is a vascularized neuroepithelium that consists of coronet cells, cerebrospinal fluid-contacting (CSF-c) neurons and supporting cells. To assess the organization, development and evolution of the SV, the expression of glial fibrillary acidic protein (GFAP) and the neuronal markers gamma-aminobutyric acid (GABA), glutamic acid decarboxylase (GAD; the GABA synthesizing enzyme), neuropeptide Y (NPY), neurophysin II (NPH), tyrosine hydroxylase (TH; the rate-limiting catecholamine-synthesizing enzyme) and serotonin (5-HT), were investigated by immunohistochemistry in developing and adult sharks. Coronet cells showed GFAP immunoreactivity from embryos at stage 31 to adults, indicating a glial nature. GABAergic CSF-c neurons were evidenced just when the primordium of the SV becomes detectable (at stage 29). Double immunolabeling revealed colocalization of NPY and GAD in these cells. Some CSF-c cells showed TH immunoreactivity in postembryonic stages. Saccofugal GABAergic fibers formed a defined SV tract from the stage 30 and scattered neurosecretory (NPH-immunoreactive) and monoaminergic (5-HT- and TH-immunoreactive) saccopetal fibers were first detected at stages 31 and 32, respectively. The early differentiation of GABAergic neurons and the presence of a conspicuous GABAergic saccofugal system are shared by elasmobranch and teleosts (trout), suggesting that GABA plays a key function in the SV circuitry. Monoaminergic structures have not been reported in the SV of bony fishes, and were probably acquired secondarily in sharks. The existence of saccopetal monoaminergic and neurosecretory fibers reveals reciprocal connections between the SV and hypothalamic structures which have not been previously detected in teleosts.

Glutamatergic innervation of the hypothalamic median eminence and posterior pituitary of the rat.

Recent studies have localized the glutamatergic cell marker type-2 vesicular glutamate transporter (VGLUT2) to distinct peptidergic neurosecretory systems that regulate hypophysial functions in rats. The present studies were aimed to map the neuronal sources of VGLUT2 in the median eminence and the posterior pituitary, the main terminal fields of hypothalamic neurosecretory neurons. Neurons innervating these regions were identified by the uptake of the retrograde tract-tracer Fluoro-Gold (FG) from the systemic circulation, whereas glutamatergic perikarya of the hypothalamus were visualized via the radioisotopic in situ hybridization detection of VGLUT2 mRNA. The results of dual-labeling studies established that the majority of neurons accumulating FG and also expressing VGLUT2 mRNA were located within the paraventricular, periventricular and supraoptic nuclei and around the organum vasculosum of the lamina terminalis and the preoptic area. In contrast, only few FG-accumulating cells exhibited VGLUT2 mRNA signal in the arcuate nucleus. Dual-label immunofluorescent studies of the median eminence and posterior pituitary to determine the subcellular location of VGLUT2, revealed the association of VGLUT2 immunoreactivity with SV2 protein, a marker for small clear vesicles in neurosecretory endings. Electron microscopic studies using pre-embedding colloidal gold labeling confirmed the localization of VGLUT2 in small clear synaptic vesicles. These data suggest that neurosecretory neurons located mainly within the paraventricular, anterior periventricular and supraoptic nuclei and around the organum vasculosum of the lamina terminalis and the preoptic area secrete glutamate into the fenestrated vessels of the median eminence and posterior pituitary. The functional aspects of the putative neuropeptide/glutamate co-release from neuroendocrine terminals remain to be elucidated.

[Plasticity of neuroendocrine transducers under the combined influence of the radiation and light exposure].

The structural changes of neurons of the rat hypothalamic supraoptic (SON) and of paraventricular (PVN) nucleus after 48 h of bright light exposure, of 5 Gy whole-body X-irradiation and of their combination subjected to the analysis by means of light-optic and of electron microscopy for the estimation of radimodificated effect of light exposure lasted 24 h a day and plasticity of neuroendocrine transducers interacted with the optic sensory system. The structural changes of neurons of the SON after combined action are less considerable and more prolonged in comparison with the PVN that loas defermined by their direct connection with the optic sensory system via the retinohypothalamic tract.

[Interrelations of morphological parameters in the system of neuroendocrine regulation following longterm morphine administration]. / Vzaimootnosheniia morfologicheskikh parametrov v sisteme neiroéndokrinnoi reguliatsii pri dlitel'nom vozdeistvii morfina.

To study the morpho-physiological changes of the nervous and endocrine regulatory systems following long-term influence of morphine hydrochloride, morphometric and correlation analysis of the structural components of the cerebral cortex, hypothalamus, adenohypophysis and thyroid gland was performed. The interrelation between the values of some parameters and the dose of morphine hydrochloride, that caused the tolerance to the drug, was shown. Dose-dependent effect was found in the degree of morphological remodeling of organ components that was demonstrated by the changes in cell and nuclear dimensions, staining properties of cytoplasm, ultrastructural components of thyrocytes, neurons and their processes, hormonal background, the number of the functioning blood capillaries. Interrelations of the morphological parameters are demonstrated by the correlations of different strength and direction.

Interleukin-1beta immunoreactivity in identified neurons of the rat magnocellular neurosecretory system: evidence for activity-dependent release.

Interleukin-1beta has been demonstrated in neurons of the rat hypothalamus, including cells of the magnocellular neurosecretory system and tuberoinfundibular system (Lechan et al., [1990] Brain Res. 514:135-140). Despite its potential importance to regulation of neuroendocrine function, however, neither the specific cell types that express interleukin-1beta or the conditions that may result in its release have yet been described. Therefore, we utilized a combination of immunocytochemical and immunoelectron microscopic localization, in conjunction with Western blot analysis, on normonatremic, hypernatremic, and lactating rats to assess the site of synthesis and potential secretion characteristics of interleukin-1beta in the rat magnocellular neurosecretory system. Interleukin-1beta immunoreactivity was localized within both oxytocin and vasopressin neurons in the paraventricular, supraoptic, accessory and periventricular hypothalamic nuclei. Additionally, interleukin-1beta immunoreactive fibers were localized in the zona interna and zona externa of the median eminence and in the neurohypophysis. Immunoelectron microscopic analysis revealed that interleukin-1beta immunoreactivity is associated with small spherical structures, distinct from neurosecretory granules, in neurosecretory axons within the neurohypophysis. Furthermore, stimulation of heightened neurosecretory activity via chronic osmotic challenge and lactation resulted in a marked diminution in levels of interleukin-1beta immunoreactivity in the neurohypophysis with a subsequent return to normal levels after cessation of the stimuli. Western blot analysis confirmed the existence of interleukin-1beta protein in the neurohypophysis and provided further evidence for reduction in levels of IL-1beta immunoreactivity after stimulation of secretory activity. These results suggest an endogenous neuronal source of interleukin-1beta exists within the rat magnocellular neurosecretory system under normal physiological conditions. The potential for activity-dependent release of IL-1beta and implications for the involvement of interleukin-1beta in regulation of neurosecretory activity are discussed.

Complexity of the neuroendocrine system.

This paper summarizes briefly the numerous connections and mechanisms existing within the neuroendocrine system. In the last decades it became clear that besides the originally postulated basic mechanisms there are several other, such as autocrine and paracrine mechanisms within the glands, bidirectional neural connections between the target endocrine glands and the hypothalamus and lower brainstem, which are of functional significance. Further we learned that the organization of the neural structures involved is much more complicated than originally thought. In addition, it turned out that the neuroendocrine system and the immune system are closely and intimately linked to each other. The available informations indicate clearly that a physiological integration exists between the nervous, the endocrine and immune systems.

[Nucleolar apparatus of neurosecretory cells in the hypothalamus of rats of different age during acute immobilization stress]. / Kharakteristika iadryshkovogo apparata neirosekretornykh kletok gipotalamusa u krys raznogo vosrasta pri ostroi immobilizatsii zhivotnykh.

To determine condition and adaptive capacity of cell nucleoli at ageing, the effect of acute immobilization on the hypothalamic neurosecretory cells was investigated in young and old male Wistar rats. Using immunohistochemical methods and nucleolometry we have shown that: 1) the nuclear volume in all neurosecretory cells is increased; 2) the share of cells, containing nucleoli with marginal position or multiple nucleoli in the nuclei, displays opposite changes in young and in old rats under stress condition. We suppose that adaptive mechanisms are different in young and old animals. Although, both kinds of morphological reorganization result in the increase in functional activity.

Regeneration of neurosecretory axons into various types of intrahypothalamic graft is promoted by the absence of the blood-brain barrier: a neurophysin-immunohistochemical and horseradish peroxidase-histochemical study.

In order to test the hypothesis that neurosecretory axon regeneration occurs only in the presence of specific vascular, perivascular, and glial microenvironments, isografts of neural lobe and optic nerve and autografts of sciatic nerve were transplanted into the hypothalamo-neurohypophysial tract at the lateral retrochiasmatic area of adult male rats. The integrity of the blood-brain barrier (BBB) to intravenously administered horseradish peroxidase (HRP), the regenerative process of neurosecretory axons, and functional recovery from lesion-induced diabetes insipidus were analyzed at 18 hr, 36 hr, 10 days, 30 days, and 80 days postsurgery. Neurophysin-positive axons invaded all grafts, as well as perivascular spaces of the adjacent hypothalamus. Wherever neurosecretory axon regeneration occurred, the BBB was breached. Reestablishment of the BBB was paralleled by a decrease in both density and staining intensity of regenerated neurophysin-positive axons. These observations illustrate that neurosecretory axon regeneration is tributary of the absence of BBB. It is speculated that blood-borne factors, provided when the BBB is breached, initiate and sustain neurosecretory axon regeneration. In addition, products of glial elements may enhance or complement the above stimulatory processes.

[Changes in the oxytocinergic neurosecretory cells in the accessory magnocellular neuroendocrine nuclei of the rat hypothalamus during aging]. / Izmeneniia oksitotsinergicheskikh neirosekretornykh kletok v dobavochnykh krupnokletochnykh neiroéndokrinnykh iadrakh gipotalamusa u krys pri starenii.

Histophysiology of accessory magnocellular nuclei of hypothalamus was studied in young (3-6 months) and old (14-29 months) male Wistar rats. Using non-labeled antibodies to oxytocin and karyolometry activation of protein synthesis combined with inhibition of oxytocin transport along the fibres was shown to occur in 4 (circular, fornical, dorso- and ventrolateral) of 5 accessory nuclei in old rats. This was not observed in anterior commissural nucleus. A hypothesis on compensatory role of accessory magnocellular nuclei of hypothalamus in regulation of neuroendocrine homeostasis in old rats is discussed.

Luteinizing hormone-releasing hormone (LHRH) receptors in the neuroendocrine-immune network. Biochemical bases and implications for reproductive physiopathology.

It seems apparent that the brain-pituitary-reproductive axis and the brain-thymus-lymphoid axis are linked by an array of internal mechanisms of communication that use similar signals (neurotransmitters, peptides, growth factors, hormones) acting on similar recognition targets. Moreover, such communication networks form the basis and control of each step and every level of reproductive physiology. This work has focused on the LHRH system, a primary central and peripheral clock of both neuroendocrine and immune functions. From the initiation of a sexually organized response, the detection of sexual odors, and the induction of mating behavior, extrahypothalamic and hypothalamic LHRH orchestrates the neuroendocrine modulation of gonadotropin secretion, while its expression within the ovary directly controls specific events such as follicular atresia. The presence of LHRH receptors in oocytes clearly anticipates a potential action of the decapeptide during the process of fertilization and/or implantation. Within the thymus and other peripheral immune organs, LHRH plays a unique role of immunomodulator, contributing to the sex-dependent changes in immune responsiveness during the estrous-menstrual cycle as well as pregnancy. The reciprocity of the neuroendocrine-immune signaling systems is further supported by the ability of sex steroids to modulate thymus-dependent immune functions via direct effects on specific target genes involved in the development of sex dimorphism and sex-dimorphic immune responses, including the downregulation of immune response observed during pregnancy. Such cyclic changes in immune responsiveness could have a physiological implication, such as the decrease or suppression in cell-mediated immunity observed in the postovulatory phase of the cycle and in pregnancy, respectively, and might play a role during the implantation process and the establishment of pregnancy. In this context, the ability of corticosterone to directly inhibit both GR transcript levels as well as a cell-mediated immune response within the thymus, and the modulation of such an inhibitory effect by the sex steroid hormone milieu, may offer an explanation and a molecular mechanism whereby stress may be deleterious for reproduction, also via immunomodulation. On the other hand, hormonally mediated alterations in immunity might also have a pathological implication in sexually related immune diseases. For example, in mouse and humans, lupus erythematosus is more prevalent in females and estrogen accelerates the disease process, while menstruation is known to exacerbate idiopathic thrombocytopenia purpura. Sex steroid hormone milieu might also have a role in controlling the stress response through immunomodulation. Within the placenta, an intricate network of signaling systems controls a delicate interplay between the neuroendocrine hormones, growth factors, and cytokines that are susceptible to play a major local role in the processes of implantation and the establishment and completion of pregnancy. The neuroendocrine and immunomodulatory role of LHRH continues well after parturition because the presence of LHRH-like material within the mammary gland and milk participates in the physiological modulation of hypophyseal, gonadal, and immune functions of the pups. Such a significant role played by the hypothalamic peptide in the modulation of immune responsiveness would indicate LHRH as the signal conveying information to both neuroendocrine and immune cells, with the role of informing and then transducing the messages into appropriate biological responses.(ABSTRACT TRUNCATED)

[A morphofunctional analysis of the nuclear apparatus of the nonapeptidergic neurosecretory cells in the vertebrate hypothalamus]. / Morfofunktsional'nyi analiz iadernogo apparata nonapeptidergicheskikh neirosekretornykh kletok gipotalamusa pozvonochnykh.

A vast comparative material obtained on fishes, amphibians and mammals has been analysed using ecological and experimental histophysiological approaches. The structural organization of the nuclear apparatus of nonapeptidergic neurosecretory cells, especially in the lower vertebrates has been clearly shown to differ from that of the classic neurones. Morphometrical characteristics of nonapeptidergic neurosecretory cells of hypothalamic centers can be used for a reliable evaluation of their functional conditions. A high intensity of protein metabolism in nonapeptidergic cells of magnocellular hypothalamic centers leads to a bigger nucleolus/nucleus relation for these cells in parvocellular centers. All this proves the necessity of the morpho-functional analysis of nuclear apparatus of neurosecretory cells.

[The vasopressin- and oxytocinergic systems of rat strains selected for their capacity to acquire active avoidance]. / Vazopressin- i oksitotsinergicheskie sistemy u linii krys, selektirovannykh po sposobnosti k vyrabotke aktivnogo izbeganiia.

Vasopressin- and oxytocinergic structures of the hypothalamus were stained immunochemically. Volume of the nucleoli of the neurosecretory cells in the paraventricular, supraoptic and anterior commissural nuclei was calculated as a measure of neurohormone synthesis. The amount of the neurosecretory material in the neurohaemal organs (external median eminence and posterior pituitary) was estimated cytophotometrically. It was shown that both synthesis and secretion of the neurohormones were higher in intact KLA rats as compared to KHA rats. In three days after inescapable electroshock opposite changes in synthesis and secretion of both oxytocin and vasopressin were revealed: a rise in KHA rats and a fall down in KLA ones. It is suggested that differences between KHA and KLA rats in the reaction of vasopressin- and oxytocinergic systems to stress is due to different strategy of rats behavior at electroshock.

Combined use of immunocytochemistry and lectin histochemistry for the study of the hypothalamic neurosecretory system of the snake Natrix maura (L.).

Natrix maura snakes were processed for immunocytochemistry and lectin histochemistry at both light- and electron-microscopic levels. Antisera against bovine neurophysins, vasotocin and mesotocin were used as well as concanavalin A, wheat germ and Limax flavus agglutinin lectins. The hypothalamic supraoptic and paraventricular nuclei were studied. Vasotocin neurons should contain a glycopeptide and displayed large colloid droplets consisting of large cisternae filled with packed secretory material. Mesotocin was located in different neurons.

[The oxytocinergic neurosecretory system in genetically selected rats differing by their emotionality. Morphometric research]. / Oksitotsinergicheskaia neirosekretornaia sistema u geneticheski selektirovannykh krys, razlichaiushchikhsia po émotsional'nosti. Morfometricheskoe issledovanie.

The rats selectively bred for rapid (KHA) and slow (KLA) acquisition of the avoidance response were subjected to inescapable shock (IS). Synthesis and secretion of oxytocin (OT) were higher in intact KLA rats as compared to KHA ones. Preliminary exposure to IS resulted in opposite changes of the OT synthesis and secretion. The findings suggest a dependence of the stress-reactivity of the OT-ergic system on the copying of the behaviour strategy.

Gonadal hormone regulation of glial fibrillary acidic protein immunoreactivity and glial ultrastructure in the rat neuroendocrine hypothalamus.

The influence of gonadal steroids on the ultrastructure of glial cells and on the immunoreactivity for the specific astrocytic marker glial fibrillary acidic protein (GFAP) has been assessed in the neuroendocrine hypothalamus. The following parameters were analyzed in the arcuate nucleus of adult female rats: the number and the surface density of cells immunoreactive for GFAP, the number of glial profiles showing bundles of glial filaments, the size of the bundles of glial filaments, and the proportion of neuronal perikaryal membrane apposed by glial processes. These parameters were studied during the different phases of the estrous cycle, after ovariectomy, and after the administration of estradiol or progesterone to ovariectomized rats. No significant differences were detected in the number of GFAP-immunoreactive cells among the different experimental groups. The surface density of GFAP-immunoreactive material, the number of glial profiles in the neuropil, and the proportion of neuronal perikaryal membrane covered by glia were increased in the afternoon of proestrus and in the morning of estrus compared with other phases of the estrous cycle or to ovariectomized rats and showed a rapid (5 h) and reversible increase in ovariectomized rats injected with 17 beta estradiol, with a maximal effect by 24 h after the administration of the hormone. In contrast, the size of the bundles of glial filaments was decreased in the afternoon of proestrus, in the morning of estrus, and by the administration of estradiol to ovariectomized rats. The parameters studied were not affected by the administration of progesterone. However, progesterone (300 micrograms/rat) blocked the effects of 17 beta estradiol (1, 10, and 300 micrograms). The results suggest that glial cells may be actively involved in the modulation of neuroendocrine events by the hypothalamus.

Ultrastructural study of rabbit lung tissue subsequent to capsaicin treatment over several days.

The neurogenic basis of nonspecific airway hyperresponsiveness (NAH) is poorly understood. Under experimental conditions isocyanates can elicit bronchial hyperresponsiveness in animals. The purpose of our study was to determine whether reactions of neurosecretory granules in nonmedullated C fibers might play a role in NAH. Our experiments were based on the fact that capsaicin treatment causes depletion of neurosecretory granules in vicinity of C fibers. We gave rabbits repeated subcutaneous injections of capsaicin. The animals were then treated with toluene diisocyanate (TDI), inducing airway hyperresponsiveness upon acetylcholine (ACH) inhalation. In capsaicin-treated animals the neurosecretory granules were not evident ultrastructurally and airway hyperresponsiveness did not occur in response to TDI treatment. Controls that were not treated with capsaicin displayed both neurosecretory granules ultrastructurally as well as airway hyperresponsiveness to ACH. We conclude that in the rabbit, NAH is related to the presence of neurosecretory granules adjacent to nonmedullated C fibers.

Neuronal-glial and synaptic remodelling in the adult hypothalamus in response to physiological stimuli.

Activation of certain neurosecretory systems of the mammalian hypothalamus induces remodelling of the conformation of their neurons and glial cells. During stimulation of the hypothalamo-neurohypophysial system, astrocytic coverage of oxytocinergic somata and dendrites diminishes and their surfaces become extensively juxtaposed. In the neurohypophysis and median eminence, stimulation evokes a retraction of glial processes and an increase in the contact area between neurosecretory terminals and the perivascular space. These changes are reversible and glial coverage returns to normal upon cessation of stimulation. Neuronal-astrocytic rearrangements also occur in the arcuate nucleus in response to changes in sex steroid levels. The significance of such modifications is a matter of speculation. In the hypothalamic nuclei they may permit synaptic remodelling that takes place concurrently; in the neurohaemal structures they may facilitate neuropeptide release. We know little about the cellular mechanisms involved but glia and neurons of these systems express certain molecules implicated in cell-cell interactions in the developing central nervous system, such as the polysialylated isoform of the neural cell adhesion molecule; this may allow them to manifest their capacity for morphological plasticity in adulthood. The factors inducing the changes vary in the different structures: while oxytocin, in synergy with steroids, appears essential to the induction of the changes in the oxytocinergic system, oestrogen alone is critical in the arcuate nucleus; in the neurohypophysis noradrenaline appears important.