Chronic excess of corticosterone increases serotonergic fibre degeneration in aged rats.

Evidence is presented for the potentiating role of corticosterone on axonal degeneration of serotonergic neurones during ageing. Aged rats, 24 months old, were implanted subcutaneously with 2 x 100 mg pellets of corticosterone. Serotonergic and cholinergic (ChAT- and NADPHd-positive) fibre degenerations in the anteroventral thalamic nucleus (AVT) were measured 2 months after corticosterone implantation. Numbers of immunoreactive serotonergic raphe and mesolimbic cholinergic neurones were also quantified. Basal plasma corticosterone and adrenocorticotropin (ACTH) concentrations were assayed at 2, 4, 6, and 8 weeks after implantation in the plasma and at 1, 2, 4 and 6 weeks in urine. The degree of serotonergic fibre aberrations in the AVT increased significantly after corticosterone exposure, while that of ChAT-positive and NADPHd-stained axon aberrations showed a modest but nonsignificant increase. A positive correlation between the magnitudes of serotonergic and cholinergic fibre aberrations appeared in the AVT, but only in the corticosterone-treated rats. The number of serotonin immunopositive neurones in the raphe nuclei after corticosterone decreased marginally, while that of mesopontine ChAT-positive neurones was not influenced. Measurements of basal plasma corticosterone and ACTH, as well as urine corticosterone, revealed that the steroid implantation increased the plasma corticosterone level for at least 4 weeks and decreased ACTH level for at least 6 weeks. By the week 8, the pituitary-adrenal function was apparently restored. However, at sacrifice, both the weight of adrenal glands and that of thymus remained reduced, indicating the long-lasting effects of corticosterone on target tissues. It is concluded that the raphe serotonergic neurones and their projecting fibres are sensitive to corticosterone excess in aged rats and become more vulnerable to degeneration processes than under normal ageing conditions. Cholinergic neurones of brainstem origin, which also express massive NADPHd activity, are more resistant against corticosterone, but their axon degeneration correlates to serotonergic fibre degeneration.

Changes in serotonin-immunoreactivity in the dorsal and median raphe nuclei of rats exposed to 2,4-dichlorophenoxyacetic acid through lactation.

Comparison of serotonin-immunoreactive (SER-IR) neurons in nucleus raphe dorsalis (NRD) and median raphe nucleus (MRN) of 25-d-old rat pups exposed to 70 mg/kg/d 2,4-dichloro-phenoxyacetic acid through mothers milk and control pups was made using an immunohistochemical analysis. Significant 2,4-D-treatment-related increase in size and density of SER-IR neuronal somata as well as in fiber length were observed. We postulate that exposure to 2,4-dichlorophenoxyacetic acid on the first day of life would modify the synthesis of 5-HT or the maturation of the brain serotonergic system.

Pattern and time course of immediate early gene expression in rat brain following acute stress.

The pattern and time course of brain activation in response to acute swim and restraint stress were examined in the rat by in situ hybridization using complementary RNA probes specific for transcripts encoding the products of the immediate early genes c-fos, c-jun and zif/268. A widespread pattern of c-fos messenger RNA expression was detected in response to these stressors; surprisingly, the expression patterns were substantially similar following both swim and restraint stress. A dramatic induction of c-fos messenger RNA was observed in numerous neo- and allocortical regions, the lateral septal nucleus, the hypothalamic paraventricular and dorsomedial nuclei, the anterior hypothalamic area, the lateral portion of the retrochiasmatic area, the medial and cortical amygdaloid nuclei, the periaqueductal gray, and the locus coeruleus; however, a prominent induction of c-fos was also seen in numerous additional subcortical and brainstem regions. Although not as widely expressed in response to stress as c-fos, induction of zif/268 messenger RNA was also detected throughout many brain areas; these regions were largely similar to those in which c-fos was induced, although in a number of regions zif/268 was expressed in regions devoid of c-fos messenger RNA. Few brain areas showed increased expression of c-jun following stress; these regions also showed induction of c-fos and/or zif/268. The time courses of expression of all three immediate early genes were similar, with peak levels observed at the 30 or 60 min time point, and a markedly reduced signal evident at 120 min post-stress. However, in a number of cases a delayed and/or prolonged induction was noted that may be indicative of secondary neuronal activation. A number of recent studies have attempted to define neural pathways which convey stress-related information to the hypothalamic-pituitary-adrenal axis. The present results reveal a widespread pattern of neuronal activation in response to acute swim or restraint stress. These findings may aid in the identification of stress-specific neural circuits and are thus likely to have important implications for our understanding of neuronal regulation of the stress response.

Central nervous system innervation of the penis as revealed by the transneuronal transport of pseudorabies virus.

Transneuronal tracing techniques were used in order to identify putative spinal interneurons and brainstem sites involved in the control of penile function. Pseudorabies virus was injected into the corpus cavernosus tissue of the penis in rats. After a four day survival period, rats were perfused with fixative and virus-labelled neurons were identified by immunohistochemistry. Postganglionic neurons were retrogradely labelled in the major pelvic ganglia. In the spinal cord, sympathetic and parasympathetic preganglionic neurons were labelled transneuronally. Presumptive interneurons were also labelled in the lower thoracic and lumbosacral spinal cord in locations consistent with what is currently known about such interneurons. In the brainstem, transneuronally labelled neurons were found in the medulla, pons and hypothalamus. Regions consistently labelled included the nucleus paragigantocellularis, parapyramidal reticular formation of the medulla, raphe pallidus, raphe magnus, A5 noradrenergic cell group, Barrington's nucleus and the paraventricular nucleus of the hypothalamus. This study confirmed previous studies from our lab and others concerning the preganglionic and postganglionic neurons innervating the penis. The number, morphology and location of these neurons were consistent with labelling seen following injection of conventional tracers into the penis. The brainstem nuclei labelled in this study were also consistent with what is currently known about the brainstem control of penile function. The labelling appeared to be highly specific, in that descending systems involved in other functions were not labelled. These results provide further evidence that the pseudorabies virus transneuronal tracing technique is a valuable method for identifying neural circuits mediating specific functions.

[Relationship between acupuncture analgesia and neurotransmitters in nucleus raphe magnus].

Nucleus Raphe Magnus (NRM) is a complex cell group. 5-HT, SP and ENK neurons in the NRM were identified by immunocytochemistry method. The afferent fibers containing 5-HT, SP, M-ENK, L-ENK, B-EP and SRIF were observed in NRM, the efferent fibers containing 5-HT, SP, ENK and TRH from NRM to spinal cord were studied. Two neurotransmitters (such as 5-HT with SP, ENK or TRH) were found in same neuron, fiber or vesicle. The neurons and axo-dendritic synapses of the NRM were analysed during Electroacupuncture (EA). The NRM increased their synaptic releases and the neurons were in active functional state during EA "Zusnanli". Studies show that NRM is one of important positions in EA analgesia.

[Ultrastructure identification of raphe-spinal serotonergic pain modulating system in rats].

5,6-dihydroxytryptamine was microinjected into the nucleus raphe magus (NRM) of rats. The sections of upper cervical segments were processed for immunoreactive substance P (SP). The degenerated axon terminals and immunoreactive fibers were identified by electron microscopy. The results show that the degenerated axon terminals and immunoreactive positive dendrites and axons were found in the laminae I and II of the dorsal horn. The electron-dense axon terminals were in contact with unlabelled dendrites, some also in contract with immunoreactive dendrites. In lamina II, one of degenerated axon terminals as a center was contacted by several unlabelled axonal boutons. SP labelled terminals were synapsed with unlabelled dendrites and dendritic spine; in addition, unlabelled axo-axonal synapse was found in lamina II. These results suggest that 5-HT axon terminals from NRM directly innervate SP and non-SP neurons in the laminae I and II.

[Electron microscopic observation on the effect of electroacupuncture (EA) on the ultrastructure of nucleus raphe dorsalis (NRD) in rats].

Nucleus raphe dorsalis (NRD) plays an important role in acupuncture analgesia. The aim of this investigation is to determine the effects of electroacupuncture (EA) analgesia on the ultrastructure of NRD. 12 Wistar rats (220-250g) were divided into control and experiment groups. Pain threshold was determined by potassium iontophoretic colorimetry. EA was applied at bilateral "Zusanli" points. The effective analgesia animals and control animals were sacrificed and the NRD were taken out for electron microscopic observation. In EA analgesia groups, NRD was observed the number of the clear round vesicle-containing terminals and clear round vesicles with the granular vesicle-containing terminals showed a significant decrease and sometimes vesicles were emptied. The area of presynaptic terminals were expanded. Some synaptic gaps appeared narrow profiles. Part of the mitochondria and endoplasmic reticulum in neuron and neurogliocytes appeared the expanded profiles. These ultrastructural change of the NRD in EA analgesia might indicate the neurons and neurogliocytes were in active functional state.

[Electron microscopic observation on the effect of electroacupuncture (EA) on the ultrastructure of periaqueductal gray (PAG) and nucleus raphe magnus (NRM) in rats].

12 Wistar rats (220-250g) were divided into control and experimental groups. Pain threshold was determined by WQ-9E Pain Threshold Measuring Apparatus. EA was applied at bilateral "Zusanli" points. The animals showing EA analgesia and the control animals were sacrificed and the ventrolateral part of PAG and the NRM were taken out for electron microscopic observation. The peripheral region of ventrolateral PAG (PAG-PR) and the NRM contained a wide variety of cell types and synaptic relationships. The neurons and axo-dendritic synapses of the PAG-PR and NRM were analysed in EA and control groups. In EA analgesia group, the clear round vesicle-containing presynaptic boutons and the presynaptic bouton with clear round vesicles and granular vesicles showed a significant decrease of their vesicle content. Part of the mitochondria and endoplasmic reticulum in the neurons appeared the expanded profiles. These ultrastructural changes of the PAG and the NRM during EA analgesia may indicate an increased release of acetylcholine, biogenic amines and/or peptide neurotransmitters and that the neurons were in active functional state.