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Modulatory effects of gonadorelin on GABA-induced depolarization and GABA-activated current in rat spinal ganglion neurons.

AIM: To explore the modulatory effects of gonadorelin on GABA-induced depolarization and GABA-activated current in membrane of rat primary sensory neurons. METHODS: Intracellular recordings and whole-cell patch clamp techniques were performed on neurons in rat spinal ganglia (SG) preparation and neurons freshly isolated from rat SG, respectively. Drugs were applied by superfusion and/or by bath application. RESULTS: In the majority of neurons GABA (10 mumol.L-1 -1 mmol.L-1) induced a depolarization, which was blocked by bicucullin (100 mumol.L-1). Pretreatment with gonadorelin (50 mumol.L-1) decreased the GABA-induced depolarization by 79 +/- 22% (n = 29), while gonadorelin elicited no effect or slight depolarization alone. In 6 of 11 cells, GABA-activated currents were also inhibited by pretreatment with gonadorelin (50 mumol.L-1), while in 5 of 11 cells, there was no change or a slight potentiation. CONCLUSION: Gonadorelin exerts an inhibitory effect on GABA-induced depolarization and GABA-activated current in the primary sensory neurons.

Animals↗

[The effect of veratridine on the release of neurotrophic factors in a nerve tissue culture].

A depolarising agent veratridine was shown to affect the level of neurotrophic substances in a combined culture of the newborn rats hippocampus and chick embryo spinal ganglia: the growth of sprouts of the ganglia's sensitive neurons was enhanced. The effect seems to be mediated by the veratridine effect on the hippocampal neurons. The role of an increased level of neurotrophic factors in an increase of neuronal activity, is discussed.

Animals↗

Observations on proliferating sheath cells in the regenerating nerves of lizard.

The proliferation of sheath cells (Schwann and endoneurium) of growing nerves has been studied by autoradiography after 3H-thymidine administration to lizards (Anolis carolinensis and Lampropholis delicata) with regenerating tails. Schwann cells of regenerating nerves derive from the multiplication of resident cells within the growing nerves, but labelled Schwann cells derived from the regenerative blastema also appear to ensheath the new axons. Endoneurium cells of growing nerves derive from the blastema around the nerves more than from dividing and migrating cells of the original endoneurium. After 6-20 days post-injection the number of labelled sheath cells in the older proximal regenerated nerves increased due to local division and migration. In the proximal spinal ganglia which innervate the regenerating tail some satellite cells, but not neurons, took up 3H-thymidine and multiplied.

Animals↗

N-cadherin protein distribution in normal embryos and in embryos carrying mutations in the homeobox gene Hoxa-4.

N-cadherin is a calcium-dependent adhesion molecule with a potential role in a variety of morphogenetic events. Although a dynamic pattern of expression in the mouse embryo has been suggested by in situ hybridization analysis, to date there has been no report of N-cadherin protein expression. In this immunohistochemical study we surveyed N-cadherin protein expression in the mid-late gestation mouse embryo utilizing a recently characterized monoclonal antibody. We found N-cadherin expression in a wide array of tissues, including the brain, the eye, various cranial ganglia, the spinal cord, spinal ganglia, somites, vertebral and limb cartilage and perichondria, the developing lung and kidney, the enteric nervous system, and germ cells. These results suggest that N-cadherin protein expression, as in the chick embryo, correlates with the segregation of cells and with organogenesis. As cadherins have been proposed as targets of vertebrate Hox genes, we also examined N-cadherin expression in two lines of Hoxa-4 mutant mice. We did not observe any alterations in N-cadherin expression in either Hoxa-4 null embryos or in transgenic embryos that overexpress Hoxa-4 in the mesenchyme of the gut. However, the partial overlap in expression between Hox genes and N-cadherin, and the likelihood of redundancy in the regulation of target genes, leaves open the possibility that cadherins are direct or indirect targets of Hox genes during mouse embryogenesis.

Animals↗

Regulation of neuronal death and calcitonin gene-related peptide by fibroblast growth factor-2 and FGFR3 after peripheral nerve injury: evidence from mouse mutants.

The presence and regulation of basic fibroblast growth factor and its high-affinity tyrosine kinase receptor FGFR3 in sensory neurons during development and after peripheral nerve injury suggest a physiological role of the fibroblast growth factor-2 system for survival and maintenance of sensory neurons. Here we investigated L5 spinal ganglia of intact and lesioned fibroblast growth factor-2 knock-out and FGFR3 knock-out mice. Quantification of sensory neurons in intact L5 spinal ganglia revealed no differences between wild-types and mutant mice. After sciatic nerve axotomy, the normally occurring neuron loss in wild-type mice was significantly reduced in both knock-out strains suggesting that fibroblast growth factor-2 is involved in neuronal death mediated via FGFR3. In addition, the number of chromatolytic and eccentric cells was significantly increased in fibroblast growth factor-2 knock-out mice indicating a transient protection of injured spinal ganglia neurons in the absence of fibroblast growth factor-2. The expression of the neuropeptide calcitonin gene-related peptide in sensory neurons of intact fibroblast growth factor-2 knock-out and FGFR3 knock-out mice was not changed in comparison to adequate wild-types. Fibroblast growth factor-2 wild-type and FGFR3 wild-type mice showed a lesion-induced decrease of calcitonin gene-related peptide-positive neurons in ipsilateral L5 spinal ganglia whereas the loss of calcitonin gene-related peptide-immunoreactive sensory neurons is reduced in the absence of fibroblast growth factor-2 or FGFR3, respectively. In addition, FGFR3 wild-type and knock-out mice displayed a contralateral reduction of the neuropeptide after axotomy. These results suggest that endogenous fibroblast growth factor-2 and FGFR3 are crucially involved in the regulation of survival and calcitonin gene-related peptide expression of lumbar sensory neurons after lesion, but not during development.

Animals↗

Contribution to histochemical distribution of non-specific cholinesterase activity in sensory ganglia of some mammals.

The activity of non-specific cholinesterase was demonstrated histochemically in satellite cells of the spinal ganglia from adult rat, cat, rabbit and baboon. The spinal ganglia of newborn rats displayed distinct intraneuronal reactivity for non-specific cholinesterase while a low reactivity was observed in satellite cells. The spinal and trigeminal ganglia of adult mice contained satellite cells with non-specific cholinesterase reactivity only sporadically. Most of reaction product for non-specific cholinesterase activity (from low to high intensity) was found in perikarya of the neurons. Spinal and trigeminal ganglia of the same mice embryo exhibited diffuse staining for non-specific cholinesterase activity remaining in the spinal ganglia of newborn mice. The trigeminal ganglia of newborn mice exhibited, however, more differentiated pattern of the positive reaction for non-specific cholinesterase like adult animals. The pattern of histochemical distribution of non-specific cholinesterase activity in trigeminal and spinal ganglia from mice of various ages corresponds with morphological differentiation and maturation undergoing in a rostrocaudal wave. Intraneuronal presence of non-specific cholinesterase activity in sensory ganglia during development and in adult animals gives a new possibilities for explanation of the functional involvement of this enzyme in the nervous system.

Animals↗

Cellular and regional expression of glutamate dehydrogenase in the rat nervous system: non-radioactive in situ hybridization and comparative immunocytochemistry.

In the central nervous system glutamate dehydrogenase appears to be strongly involved in the metabolism of transmitter glutamate and plays a role in the pathogenesis of neurodegenerative disorders. In order to identify unequivocally the neural cell types expressing this enzyme, non-radioactive in situ hybridization, using a complementary RNA probe and oligonucleotide probes, was applied to sections of the rat central nervous system and, for comparison with peripheral neural cells, to cervical spinal ganglia. The results were complemented by immunocytochemical studies using a polyclonal antibody against purified glutamate dehydrodenase. Glutamate dehydrogenase messenger RNA was detectable at varying amounts in neurons and glial cells (i.e. astrocytes, oligodendrocytes, Bergmann glia, ependymal cells, epithelial cells of the plexus choroideus) throughout the central nervous system and in neurons and satellite cells of spinal ganglia. In some neuronal populations (e.g., pyramidal cells of the hippocampus, motoneurons of the spinal cord and spinal ganglia neurons) messenger RNA-labelling was higher than in other central nervous system neurons. This is remarkable because the immunostaining of neurons in the central nervous system regions studied was at best weak, whereas a predominantly high level of immunoreactivity was detected in astrocytes (and Bergmann glia). Thus, in neurons of the central nervous system, the detected levels of glutamate dehydrogenase messenger RNA and protein seem to be at variance whereas in peripheral neurons of spinal ganglia both in situ hybridization labelling and immunostaining are intense.

Animals↗

Activation and circuitry of uterine-cervix-related neurons in the lumbosacral dorsal root ganglia and spinal cord at parturition.

Stimulation of the uterine cervix at parturition activates neural circuits involving primary sensory nerves and supraspinally projecting neurons of the lumbosacral spinal cord, resulting in output of hypothalamic neurohormones. Dorsal root ganglia (DRG) and spinal neurons of these circuits are not well-characterized. The objectives of this study were to detail the activation of DRG and spinal neurons of the L6/S1 levels that are stimulated at late pregnancy, verify hypothalamic projections of activated spinal neurons, and determine whether activated neurons express estrogen receptor-alpha (ERalpha). Expression of phosphorylated cyclic-AMP response element-binding protein (PCREB) and Fos immunohistochemistry were used to "mark" activated DRG and spinal neurons, respectively. Retrograde tracing identified uterine-cervix-related and spinohypothalamic neurons. Baseline PCREB expression in the DRG increased during pregnancy and peaked during the last trimester. Some PCREB-expressing neurons contained retrograde tracer identifying them as cervix-related neurons. Fos-expressing neurons were few in spinal cords of nonpregnant and day 22 pregnant rats but were numerous in parturient animals. Some Fos-expressing neurons located in the dorsal half of the spinal cord contained retrograde tracer identifying them as spinohypothalamic neurons. Some DRG neurons expressing PCREB also expressed ERalpha, and some spinal neurons activated at parturition projected axons to the hypothalamus and expressed ERalpha. These results indicate that DRG and spinal cord neurons are activated at parturition; that those in the spinal cord are present in areas involved in autonomic and sensory processing; that some spinal neurons project axons to the hypothalamus, ostensibly part of a neuroendocrine reflex; and that sensory and spinal neurons can respond to estrogens. Moreover, some activated sensory neurons may be involved in the animal's perception of labor pain.

Analysis of Variance↗

Distribution of arginine-vasopressin in the trigeminal, dorsal root ganglia and spinal cord of the rat; depletion by capsaicin.

We have measured arginine vasopressin in the neural lobe, the trigeminal ganglion (TG), dorsal root ganglia (DRG), spinal cord, trigeminal and sciatic nerves of the rat by radioimmunoassay. In control rats, the neural lobe contained 1600 pg/mg, the ganglia 52.5, 21.0, 8.5, 4.28, 3.85 pg/mg in the lumbar, sacral, cervical, thoracic, and trigeminal ganglion, respectively, the spinal cord contained 5.1, 4.3, 4.2 and 2.6 pg/mg in the lumbar, thoracic, sacral and cervical cord, respectively and the trigeminal and sciatic nerves contained 3.8 and 13 pg/mg. Neonatal capsaicin treatment depleted about 38-67% of AVP in the ganglia. Residual AVP amounted to 526.8, 30.55, 20.75, 12.88, 4.95, 2.74, 2.14, 7.94 and 2.53 pg/mg in the neural lobe, lumbar, thoracic, sacral, cervical DRG, lumbar, thoracic spinal cord, the sciatic and trigeminal nerves respectively. Capsaicin destroyed about 40.5% of total cells and 52% of AVP-immunoreactive neurons.

Animals↗

Distribution of the cells of primary afferent fibers to the cat auricle in relation to the innervated region.

The origin and peripheral distribution of the primary afferent fibers to the cat auricle were studied by the horseradish peroxidase (HRP) method. Following HRP injection into the whole region of the auricle, HRP-labeled cells were found ipsilaterally in the trigeminal ganglion, geniculate ganglion, superior ganglion of the vagus nerve and in the C1 to C4 spinal ganglia. In addition to the ganglia of cranial and spinal nerves, labeled cells were also observed in the facial nerve trunk and in the dorso-lateral portion of the superior cervical ganglion. In the case of HRP injection into the central region of the auricle, labeled cells were principally observed in the ganglia of cranial nerves and to a lesser degree in the spinal ganglia. On the contrary, in the case of HRP injection into the peripheral region of the auricle, labeled cells were principally observed in the spinal ganglia, although some were seen in the ganglia of cranial nerves. This study suggests that the cutaneous innervation of the auricle is supplied by both the cranial and spinal nerves, and that the central region of the auricle is strongly innervated by the cranial nerves and the peripheral region of the auricle is strongly innervated by the spinal nerves.

Afferent Pathways↗

Tyrosinase protein is expressed also in some neural crest derived cells which are not melanocytes.

Some neural crest cells give rise to pigment cells in early ontogenesis. We tested here whether tyrosinase--a key enzyme in melanogenesis--was present in some pigment neural crest derivatives in adult hamsters. Interestingly enough, inactive tyrosinase protein was detected, using indirect immunofluorescence, in the satellite cells of spinal ganglia and Schwann cells of sciatic and facial nerves in normal adult animals. The results of cell blotting from spinal ganglia were similar to the fluorescence findings. Thus, our results seem to support the hypothesis that Schwann cells, satellite cells of spinal ganglia, and melanocytes may be more intimately related developmentally than other neural-crest-derived cells. Moreover, since we detected tyrosinase protein in cells which normally do not produce melanosomes, it could be deduced that, during the melanocyte's differentiation from its cell precursor, the expression of tyrosinase protein might precede the point when melanosomes begin to differentiate from known cytoplasmic structures.

Animals↗

Distribution of sensory neurons of the pudendal nerve in the dorsal root ganglia and their projection to the spinal cord. Horseradish-peroxidase studies in the rat.

The morphology and distribution of the sensory neurons of the pudendal nerve within the spinal ganglia of rats were investigated by use of horseradish peroxidase (HRP). The labelling was visualized in diaminobenzidine (DAB) or tetramethyl-benzidine (TMB)-stained sections. Injection of HRP directly into the pudendal nerve labelled perikarya predominantly in the sixth lumbar DRG (L6). Following injection of HRP into the scrotal skin, however, additional cells were labelled in L5 and S1. Labelling was invariably unilateral. Approximately equal numbers of small (less than 30 micrometers) and large neurones (greater than 40 micrometers) were labelled following subcutaneous injections although injections into the nerve marked twice as many small cells as large cells. This suggests that, in the rat, most of the small-diameter fibers within the pudendal nerve ascend through L6. Although a cluster of neurones was observed in one experiment, the remaining 25 experiments did not reveal any somatotopic arrangement since the labelled perikarya were distributed evenly throughout the ganglion. Similar numbers of retrogradely labelled neurones (somatopetal transport of the tracer) were observed in both DAB- and TMB-stained sections, although TMB allowed the demonstration of anterograde (somatofungal) HRP transport by terminal labelling in the superficial laminae of the lumbar spinal cord, extending into laminae II--IV.

3,3'-Diaminobenzidine↗

[The segmental distribution of the afferent neurons of the "zusanli" point and the caecum in rabbits--a study with the horseradish peroxidase method].

Ten adult rabbits were used in this experiment. A solution of 10-20% HRP (sigma IX, RZ = 3.2) was injected into the "Zusanli" point and the subserosa of the caecum. The uptake and retrograde transmission of HRP in the afferent neurons of both the somatic and visceral nerves were traced to the spinal ganglia. The results showed that: 1. Labelled afferent neurons from the region of "Zusanli" point were found in the spinal ganglia T12-S2 with a higher concentration in L4-S2. 2. Labelled afferent neurons from the region of the caecum were found in the spinal ganglia T2-S2 with a higher concentration in T12-L2. 3. The ranges of distribution of labelled afferent neurons from the regions overlapped in the segments T12-S2.

Acupuncture Points↗

Effects of peripheral axotomy on neuropeptides and nitric oxide synthase in dorsal root ganglia and spinal cord of the guinea pig: an immunohistochemical study.

The effect of axotomy (3, 10 and 21 days) on the expression of some neuronal markers was analysed in dorsal root ganglia and spinal cord of guinea-pigs using immunohistochemistry. Three weeks following injury, substance P-like immunoreactivity (-LI) was slightly reduced in the DRGs of the ipsilateral side, whereas a marked increase in neuropeptide Y(NPY)-LI could be detected ipsilaterally and a smaller increase contralaterally. NPY-LI was mainly expressed in small, but also some medium-sized and large neuron profiles after axotomy. Galanin-LI showed a moderate bilateral increase. No significant changes could be observed in DRGs for calcitonin gene-related peptide (CGRP)-, vasoactive intestinal polypeptide-, peptide histidine isoleucine- or nitric oxide synthase-LIs. In the ventral horn CGRP-LI was slightly increased bilaterally in motoneurons, most pronounced on the injured side. Autotomy behaviour was seen in seven of the nine animals in the twenty-one day group. The present results demonstrate that also in guinea-pigs several peptides undergo distinct changes in their expression after peripheral nerve injury. However, in contrast to rats and monkeys, galanin-LI is only moderately increased in guinea-pigs. Neuropeptide Y showed a dramatic increase mainly in small neurons, in contrast to the upregulation in large neurons in the rat. Thus, distinct species differences exist with regard to the cellular response to nerve injury.

Animals↗

Microtubule-associated protein 2 appears in axons of cultured dorsal root ganglia and spinal cord neurons after rotavirus infection.

The immunohistochemical distribution of microtubule-associated protein 2 (MAP2), being normally restricted to nerve cell bodies and dendrites, became altered in rat dorsal root ganglia and spinal cord neurons in cultures infected with rhesus rotavirus. MAP2 appeared in axons of both sources of neurons as displayed with monoclonal antibodies to MAP2a + b and MAP2a + b + c at 48 hr post-infection (p.i.). Other cytoskeletal elements, i.e., tau, MAP1, MAP5, neurofilament, actin, and tubulin, did not reveal any alterations in the rotavirus-infected neurons. One of the rotavirus cytosolic proteins, the inner capsid protein vp6, was expressed in axons at 48 hr p.i. simultaneously with the appearance of MAP2, while two other viral proteins, vp4 and NS28, remained in the nerve cell bodies. By quantitative enzyme-linked immunosorbent assay (ELISA) a binding of single-shelled rotaviruses, which express vp6 on their surfaces, to purified MAP2 was found. There was no binding of these viral particles to tau or tubulin proteins. This study indicates that a selective interaction between certain viral and neuronal cytoskeletal proteins can occur and that a non-cytolytic viral infection can cause alterations in the polarized sorting of neuronal proteins.

Animals↗

Neuronal organization and cell interactions of the cat stellate ganglion.

The functional structure of the cat stellate ganglion (SG) and, in particular, its extra- and intraganglionic connections and neuronal organization, were investigated using histochemical, immunohistochemical, morphological and histological methods. Retrograde axonal transport of horseradish peroxidase was used to determine most of the extraganglionic interactions. Of the targets tested, the most extensive efferent connections of the SG were with the stemocleidomastoid muscle, trachea, esophagus and heart. Neurons of the SG also send a small number of postganglionic efferents to the thyroid and stomach. Furthermore, ganglion cells send axons to the spinal ganglia. Several afferent connections of the SG were determined. Sympathetic preganglionic neurons of segments C8-T10 of the spinal cord, sensory neurons of C8-T9 spinal ganglia, intramural ganglia of the thoracic viscera and the reticular formation of the medulla oblongata send their axons to the SG. Intraganglionic interactions of intemeurons with principal ganglionic cells were assumed to occur, based on the presence of interneurons immunoreactive to GABA and substance P. GABA- and substance P-immunoreactive fibers located around a small number of postganglionic neurons were also identified. Morphological study revealed asymmetry between the left and right ganglia: the right ganglion is larger than the left and contains more cells. This asymmetry was also reflected in basic structural parameters of neurons, such as average neuronal area and average diameter of cell somata. The present data has been used to develop a scheme for the basic inter- and intraneuronal connections of the cat SG. This ganglion is a true nervous center, with postganglionic neurons, some of which might be performing sensory functions, and interneurons. The ganglion is connected not only with the spinal cord and spinal ganglia, but also with neurons of the intramural ganglia and, by direct links, with efferent neurons of the medulla oblongata. Thus, the SG may play an essential role in viscera-visceral reflexes.

Afferent Pathways↗

Selective localization of alpha-fetoprotein and serum albumin within the sensory ganglia cells of developing chicken.

The presence of alpha-fetoprotein (AFP) and serum albumin (SA) has been described within developing brain cells of mammals and chicken. In order to avoid the influence of the choroid plexuses in the transport, via the cerebrospinal fluid, of these proteins to brain cells, we studied the presence of AFP and SA within chicken sensory ganglion cells during ontogeny using the indirect immunoperoxidase technique. Our results show that before day 9 of egg incubation, both AFP and SA can be detected within the trigeminal and spinal ganglia cells. Henceforth, the labeling for AFP starts decreasing in the spinal ganglia but not in the trigeminal ganglion. The labeling for SA remains constant in both structures. Differences between AFP and SA staining in the spinal ganglia are maximal on day 11 of incubation, when AFP is no longer detected. It is concluded that at this time, the uptake of AFP by spinal ganglia cells is switched-off while SA is still taken up. As the blood concentration of both proteins is similar at this time, the finding reported here gives support to the advanced hypothesis suggesting the presence of specific receptors for AFP and/or SA in embryonic neural cells.

Animals↗

Quantitative changes in mitochondria of spinal ganglion neurons in aged rabbits.

Within the context of our research on the age-related structural changes in spinal ganglia, we studied the mitochondria of the neuronal perikaryon in the spinal ganglia of 12-, 42-, and 79-month-old rabbits. Both the volume of the perikaryon and the total mitochondrial mass within the perikaryon increased significantly passing from young adult to old animals. Hence, there is no net loss of mitochondria in these neurons with age. Since, however, the volume of the perikaryon increased by more than 63% while the total mitochondrial mass within the perikaryon increased by only 18%, the mean percentage of perikaryal volume occupied by mitochondria decreased with age. This decrease is only in very minor part a consequence of lipofuscin accumulation, so that the ratio between the total mitochondrial mass and the functionally active volume of cytoplasm decreased with age. Possible causes of this decrease are discussed briefly. Moreover, while the mitochondrial structure did not change, mitochondrial size increased with age. Finally, in each of the three age groups both the mean percentage volume of mitochondria and the mean mitochondrial size were very similar in large light and in small dark neurons.

Age Factors↗