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Previous studies have demonstrated that repeated submission of rats to mild hypobaric hypoxia reduces the persistent behavioral and hormonal depressive symptoms induced by exposure to footshock in the learned helplessness paradigm. The aim of this study was to determine whether hypoxic preconditioning of mice can also induce antidepressant- and anxiolyticlike effects that are detectable with the other commonly used behavioral tests, and to determine whether these effects are accompanied by an increase in neuropeptide Y (NPY) in the hippocampus, which may suggest the involvement of NPY in these mechanisms. The intermittent mild hypobaric hypoxia was generated by 2-h exposure of mice to 0.47 atm for 3 consecutive days. In the tail suspension test a significant decrease in the duration of immobility was observed 24 h, but not 48 h after the last hypobaric session. The elevated plus maze trials performed 48 h after preconditioning showed a significant increase in the frequency of open arm entries, a reduction in the duration of closed arm occupancy and substantially more time spent in the open arms in comparison to the control groups. The open field test demonstrated the absence of increases in general activity or unspecific exploratory behavior in hypoxia-preconditioned mice. The EIA test detected a statistically significant but relatively weak increase in the NPY content in the hippocampus 24 h after preconditioning. Together, our data demonstrate that preconditioning of mice with intermittent mild hypobaric hypoxia induces anxiolytic- and antidepressantlike effects. They are accompanied by up-regulation of NPY which may suggest its mechanistic role.
Treatment with pilocarpine (PILO) induces variable degrees of loss of mossy cells (MCs) and mossy fibre (MF) sprouting in rodents, the relationships of which have not been examined in individual animals. Our aim was to test whether the loss of MCs and MF sprouting are coupled processes in PILO-treated rodents. Animals which exhibited intense PILO-induced convulsions for at least 30 min were used in this study. After a 2-month survival period, the incidence of epileptic seizures was checked individually by neuropeptide-Y (NPY) immunohistochemistry, and the numbers of MCs were counted by means of immunohistochemistry, for calretinin (CR) in mice and calcitonin gene-related peptide (CGRP) in rats. MF sprouting was checked by using Timm's silver-sulphide method for zinc. In our comparative studies, NPY immunohistochemistry resulted in more positive animals than on zinc staining. The CR immunoreactivity remained unchanged even in those mice that displayed MF sprouting and greatly increased NPY immunoreactivity. CR immunoreactivity was also verified after transection of the fornix to exclude the extrahippocampal source of this peptide. However, the CGRP immunoreactivity was severely reduced in those rats that exhibited simultaneous increases in zinc content and NPY immunoreactivity in the supragranular layer and stratum lucidum. Our findings suggest that the MCs survive PILO treatment in mice, but not in rats. There is direct evidence of a close relationship between the loss of MCs and MF sprouting in rats, but not in mice. Thus, similar PILO seizures may result from different changes in the neuronal circuits of rodents.
The hypothalamic supraoptic (SON) and paraventricular (PVN) nuclei are activated by body salt-fluid variations. Stimulation of 2-adrenoceptors by an agonist-xylazine (XYL) activates oxytocinergic but not vasopressinergic magnocellular neurons. In this study, tyrosine hydroxylase (TH), corticoliberine (CRH), and neuropeptide Y (NPY) magnocellular phenotypes, were analysed in response to 2-adrenoceptor manipulations and sustained hyperosmolality in vasopressin deficient homozygous Brattleboro (di/di) rats. Saline (0.9% NaCl, 0.1 ml/100g/bw), XYL (10 mg/kg/bw), atipamezole (ATIP, 2-adrenoceptors antagonist, 1 mg/kg/bw), and ATIP 5 min later followed by XYL, were applied intraperitoneally. Presence of immunolabeled Fos peptide signalized the neuronal activity. Ninety minutes after injections, the rats were anesthesized and sacrificed by transcardial perfusion with fixative. Coronal sections of 30 µm thickness double immunolabeled with Fos/neuropeptide were evaluated under light microscope. Under basal conditions, di/di in comparison with control Long Evans rats, displayed significantly higher number of TH, CRH, and NPY immunoreactive neurons in the SON and PVN (except NPY cells in PVN) and more than 90%, 75%, and 86% of TH, NPY, and CRH neurons, respectively, displayed also Fos signal in the SON. XYL did not further increase the number of Fos in the PVN and SON and ATIP failed to reduce the stimulatory effect of hypertonic saline in all neuronal phenotypes studied. Our data indicate that hyperosmotic conditions significantly influence the activity of TH, CRH, and NPY magnocellular neuronal phenotypes, but 2-adrenoceptors do not play substantial role in their regulation during osmotic challenge induced by AVP deficiency.
The study was performed on six male chinchillas. The animals were anaesthetised with ether and the anaesthesia was deepened with nembuthal injected intraperitoneally. The chinchillas were then transcardially perfused with 0.4 L of 4% buffered paraformaldehyde, and testes, epididymides, and vasa deferentia were collected. The tunica albugínea from one testis from each chinchilla was stained as whole-mount preparation. The tissues were cut into 12 µm-thick cryostat sections, and processed for double-immunofluorescence method. In all organs studied, the most abundant nerve fibres were dopamine ß hydroxylase positive (DßH⁺). Some of them contained neuropeptide Y (NPY). Sporadically NPY-positive-only nerve fibres were found. Single DßH⁺ nerve terminals contained also galanine. Small numbers of the nerve fibres supplying studied organs were stained for substance P (SP) and calitonin gene related peptide (CGRP). Almost all SP⁺ fibres were also CGRP⁺, whereas single CGRP⁺ nerves were SP- immunonegative. Some nerve terminals were immunoreactive to vesicular acetylcholine transporter and vasoactive intestinal peptide. The organs studied were innervated unevenly. The highest density of the nerves was found in the areas of the tunica albuginea adjacent to the mesorchial border of the testis and their number gradually decreased towards the free border of the gonad. None of the vascular tissue of the testicular parenchyma was free of the nerve fibres, except sporadically encountered DßH⁺ nerves which supply seminiferous tubules. Within the head of the epididymis a moderate number of nerve terminals were found, but in the body and tail of the organ the number of nerves gradually increased. The vas deferens was supplied with very numerous nerve fibres. There were no differences in the density of the innervation between the funicular and abdominal part of the vas deferens.
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