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1.
Immunoreactivity to calbindin D-28k, a vitamin D-dependent calcium-binding protein, is expressed by neuronal subpopulations of dorsal root ganglia (DRG) in the chick embryo. To determine whether the expression of this phenotypic characteristic is maintained in vitro and controlled by environmental factors, dissociated DRG cell cultures were performed under various conditions. Subpopulations of DRG cells cultured at embryonic day 10 displayed calbindin-immunoreactive cell bodies and neurites in both neuron-enriched or mixed DRG cell cultures. The number of calbindin-immunoreactive ganglion cells increased up to 7-10 days of culture independently of the changes occurring in the whole neuronal population. The presence of non-neuronal cells, which promotes the maturation of the sensory neurons, tended to reduce the percentage of calbindin-immunoreactive cell bodies. Addition of horse serum enhanced both the number of calbindin-positive neurons and the intensity of the immunostaining, but does not prevent the decline of the subpopulation of calbindin-immunoreactive neurons during the second week of culture; on the contrary, the addition of muscular extract to cultures at 10 days maintained the number of calbindin-expressing neurons. While calbindin-immunoreactive cell bodies grown in culture were small- or medium-sized, no correlation was found between cell size and immunostaining density. At the ultrastructural level, the calbindin immunoreaction was distributed throughout the neuroplasm. These results indicate that the expression of calbindin by sensory neurons grown in vitro may be modulated by horse serum-contained factors or interaction with non-neuronal cells. As distinct from horse serum, muscular extract is able to maintain the expression of calbindin by a subpopulation of DRG cells. 相似文献
2.
Dorsal root ganglion (DRG) neurons may give origin to ascending branches that terminate in the dorsal column nuclei (DCN); uncertainties still exist with regard to the proportion of these neurons in different DRGs and to the type of these neurons. The percentage and size of neurons that project to the DCN were determined in a large number of DRGs by means of the retrograde transport of colloidal gold-labeled wheat germ agglutinin conjugated to enzymatically inactive horseradish peroxidase (WGAapoHRP-AU). A total of 16,239 neurons was tallied in 80 DRGs from nine rats; 3,240 (20%) of these were retrogradely labeled by the tracer injected in the DCN. Percentages of DCN projecting neurons vary considerably at different segmental levels: they are higher in cervical (up to 63%) than in thoracic (up to 31% for T1, up to 12% for thoracic DRGs below T1) or lumbar DRGs (up to 15%). At cervical levels highest percentages were encountered in C6, C7, and C8 and lowest percentages in C2-C4. At lumbar levels highest percentages were encountered in L4 and lowest in L1 and L6. When considering the soma size of DRG neurons it appears that: 1) there are more large cells, labeled and unlabeled, at cervical (38%) than at lumbar levels (30%) and more at lumbar than at thoracic levels (23%); 2) at every level, most labeled, i.e., projecting, neurons are large; and 3) DRGs with the highest proportions of large vs. small cells contain the highest percentages of DCN projecting neurons. These results represent the first attempt at establishing the percentages and soma size of DCN projecting neurons from a large number of DRGs and at comparing the contribution to these nuclei from cervical, thoracic, and lumbar DRGs. Some of the differences in the ratio of projecting neurons at different levels may be explained on the basis of well-known anatomical features, e.g., the projections to the Clarke's column of many DRG neurons in lumbar ganglia. The contribution of virtually exclusively large DRG neurons to the DCN, suggested by indirect or incomplete evidence, is demonstrated by the present retrograde labeling and soma size measurements. The results relate to the functional component of peripheral receptors that relay their input via the dorsal columns and do not seem to support a recent suggestion that a sizeable fraction of unmyelinated primary afferents ascend in the dorsal columns to terminate in the DCN. 相似文献
3.
We have studied the subcellular localization of peroxidase-labeled organelles after anterograde axonal transport by chick retinal ganglion cells that had been exposed 23-25 h earlier to wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). After intravitreal injection of WGA-HRP, we found in the optic tectum that 82% of labeled organelles were located within axons and axon terminals. The organelles included: tubules and cisternae of the smooth endoplasmic reticulum, hypolemmal cisternae, vesicles, dense bodies and multivesticular bodies. We also measured the distances between the centers of the labeled organelles and the plasma membrane of these profiles. The density of organelles (number of organelles/micron 2) was plotted as a function of distance from the plasma membrane. Irrespective of the dose of lectin-peroxidase injected, labeled organelles were most densely concentrated in a 30 nm wide annular zone centered 75 nm in from the plasma membrane. In axon terminals the labeled organelles were most concentrated 75-90 nm in from the plasma membrane. Assuming that the peroxidase label indicates the presence of WGA-HRP, we conclude that after anterograde axonal transport the lectin accumulates in lysosomal organelles and elements of the smooth endoplasmic reticulum. Therefore, in contrast to the more restricted localization of [125I]WGA as inferred from electron microscopic autoradiography after uptake and transport by the same cell type, WGA-HRP-labeled organelles are found more diffusely within the axoplasm, particularly in axon terminals. Furthermore, peroxidase-labeled organelles in dendritic, glial or neuronal cell bodies in the tectum were seen less frequently than expected based on evidence of frequent transfer to second cells after intravitreal injections of [125I]WGA. Thus, we infer that at these concentrations WGA labeled with HRP may not be transferred intercellularly as efficiently as even lower concentrations of iodinated WGA are apparently transferred. 相似文献
4.
Increase of preprotachykinin mRNA and substance P immunoreactivity in spared dorsal root ganglion neurons following partial sciatic nerve injury 总被引:5,自引:0,他引:5
Complete sciatic nerve injury reduces substance P (SP) expression in primary sensory neurons of the L4 and L5 dorsal root ganglia (DRG), due to loss of target-derived nerve growth factor (NGF). Partial nerve injury spares a proportion of DRG neurons, whose axons lie in the partially degenerating nerve, and are exposed to elevated NGF levels from Schwann and other endoneurial cells involved in Wallerian degeneration. To test the hypothesis that SP is elevated in spared DRG neurons following partial nerve injury, we compared the effects of complete sciatic nerve transection (CSNT) with those of two types of partial injury, partial sciatic nerve transection (PSNT) and chronic constriction injury (CCI). As expected, a CSNT profoundly decreased SP expression at 4 and 14 days postinjury, but after PSNT and CCI the levels of preprotachykinin (PPT) mRNA, assessed by in situ hybridization, and the SP immunoreactivity (SP-IR) of the L4 and L5 DRGs did not decrease, nor did dorsal horn SP-IR decrease. Using retrograde labelling with fluorogold to identify spared DRG neurons, we found that the proportion of these neurons expressing SP-IR 14 days after injury was much higher than in neurons of normal DRGs. Further, the highest levels of SP-IR in individual neurons were detected in ipsilateral L4 and L5 DRG neurons after PSNT and CCI. We conclude that partial sciatic nerve injury elevates SP levels in spared DRG neurons. This phenomenon might be involved in the development of neuropathic pain, which commonly follows partial nerve injury. 相似文献
5.
Following intravitreal injection, affinity purified, iodinated wheat germ agglutinin ([125I]WGA) is taken up by chick retinal ganglion cells and transported in an anterograde direction to nerve terminals in the optic tectum. The accumulation of axonally transported label in the tectum may be measured quantitatively. Using such an approach, we find that co-injection of [125I]WGA with an excess of unlabeled WGA reduces the amount of axonally transported labeled lectin. Since co-injection of comparable levels of soybean agglutinin or Ulex Europeanus-I fails to reduce tectal labeling to a similar extent, and since native WGA at the same concentration does not appear to be toxic to retinal ganglion cells, these results support the hypothesis that the uptake and subsequent anterograde axonal transport of WGA by these cells is a selective process, dependent on a limited number of extra- or intracellular binding sites. 相似文献
6.
Horseradish peroxidase conjugates of either the lectin wheat germ agglutinin (WGA-HRP) or choleragenoid (B-HRP) have been shown to be sensitive neuroanatomical tracers. In the present study a comparison was made between these two conjugates as transganglionic tracers in trigeminal primary sensory neurons following injection into the rat mystacial vibrissae skin. Differences between the two tracers were observed in the labeling of cell bodies in the trigeminal ganglion. Injection of WGA-HRP resulted in labeling of predominantly small cell bodies, whereas B-HRP gave rise to labeling of somewhat larger cell bodies. By increasing the concentration of the injected WGA-HRP solution the number of labeled cells increased substantially, while a corresponding increase in the concentration of B-HRP resulted in a relatively small increase in the number of labeled cells. WGA-HRP injection resulted in labeling of primary afferents mainly in the substantia gelatinosa of the trigeminal subnucleus caudalis. When the concentration of the injected WGA-HRP solution was increased, labeling was also observed in the marginal and magnocellular zones. Following B-HRP injection, labeling was only observed in the magnocellular zone and innermost part of the substantia gelatinosa. This general pattern of labeling was the same when the concentration of the B-HRP solution was increased. 相似文献
7.
Following intravitreal injection, affinity purified, iodinated wheat germ agglutinin ([125I]WGA) is taken up by chick retinal ganglion cells and transported in an anterograde direction to nerve terminals in the optic tectum. The accumulation of axonally transported label in the tectum may be measured quantitatively. Using such an approach, we find that co-injection of [125I]WGA with an excess of unlabeled WGA reduces the amount of axonally transported labeled lectin. Since co-injection of comparable levels of soybean agglutinin or Ulex Europeanus-I fails to reduce tectal labeling to a similar extent, and since native WGA at the same concentration does not appear to be toxic to retinal ganglion cells, these results support the hypothesis that the uptake and subsequent anterograde axonal transport of WGA by these cells is a selective process, dependent on a limited number of extra- or intracellular binding sites. 相似文献
8.
Afferents to the zona incerta in the rat: a combined retrograde and anterograde study 总被引:1,自引:0,他引:1
In a first set of experiments, the retrograde transport of horseradish peroxidase (HRP) was utilized to investigate the afferent projections to the zona incerta (ZI) in the hooded rat. HRP was introduced in its crystalline form into various sectors of the ZI of seven subjects. The largest contingent of afferents arises from the following centers: the cingulate and somatosensory cortices, central amygdaloid nucleus, ventromedial hypothalamic nucleus, posterior thalamic nucleus, anterior pretectal nucleus, peripeduncular area, deep and intermediate layers of the superior colliculus, dorsal and ventral parabrachial nuclei, principal and interpolar trigeminal subnuclei, and cuneate nucleus. Other centers less systematically or more sparsely labeled were the lateral hypothalamic area, ventrobasal complex, lateral geniculate nucleus pars ventralis, medial geniculate nucleus, interstitial nucleus of Cajal, Darkschewitsch nucleus, perirubral fields, cuneiform, tegmental pedunculopontine, and deep mesencephalic reticular nuclei, pontine reticular nucleus pars oralis, lateral and interpositus cerebellar nuclei, and gracile nucleus. In a second set of experiments, an anterograde tracer (WGA-HRP) was injected in several centers projecting to the ZI in order to localize their terminal fields within this structure. It has been thus possible to distinguish a ventral zone (ventral sector of pars caudalis and pars ventralis) in which the somesthetic (somatosensory cortex, trigeminal complex, and dorsal column nuclei (DCN), collicular, and cerebellar projections terminate, from a dorsal zone (pars dorsalis) to which a limbic input (cingulate cortex and ventromedial hypothalamic nucleus) is directed. In most cases, the labeled terminal fields consisted of well-delimited, narrow bands disposed obliquely, parallel to the cerebral peduncle or the internal capsule. The contingent of somatosensory afferents is relatively large and there is a high degree of overlapping between the different somatosensory terminal fields within the ventral ZI. This suggests a participation of this structure in the treatment of somesthetic information and/or in the transmission of noxious stimuli. 相似文献
9.
The anterograde and retrograde rates of axonal transport of the lectin wheat germ agglutinin (WGA) were investigated using native and derivatized lectins in embryonic (stage 39) and posthatch chickens. Anterograde transport rates in the retinotectal projection of posthatch animals ranged from 168 mm/day for native WGA to 345 mm/day for horseradish peroxidase conjugated WGA. Anterograde transport rates in embryos were at least 258 mm/day based on experiments employing tritium and horseradish peroxidase conjugates. Retrograde rates measured by appearance of label in the isthmo-optic nucleus in both embryonic and posthatch chickens were in the range of 150-180 mm/day. A fluorescein isothiocyanate conjugate of WGA was transported retrogradely but not anterogradely. When the extraocular eye muscles were labeled accidentally during injection, cells in the oculomotor or trochlear nuclei were more intensely labeled than neurons in the isthmo-optic nucleus. It was concluded that at least some conjugates of WGA, and possible the native lectin as well, travel in the fastest component of axonal transport. In view of the known intercellular movement of WGA from labeled presynaptic processes, it is recommended that survival times be kept short in experiments using WGA as a neuronal tracing agent (less than 24 h) to minimize the possibility of uptake and redistribution of the lectin by nearby cells. 相似文献
10.
Projection neurons in the rat dorsolateral septal nucleus (DLSN) were retrogradely labeled following intraseptal injection of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP). Injections of WGA-HRP centered in the medial septum (MS) and parts of the intermediate and ventrolateral subdivisions of the lateral septum retrogradely labeled only a few centrally scattered multipolar-shaped neurons. In contrast, injections placed in the nucleus of the diagonal band of Broca (DBB) consistently resulted in labeling of DLSN neurons of all sizes and shapes. Large injections in rostral DBB appeared to retrogradely label every DLSN neuron, while similar injections in caudal DBB only labeled neurons in restricted regions of the nucleus. A collection of small cells forming the ventricular border of caudal DLSN and a group of larger cells situated in the dorsolateral tip of rostral DLSN were consistently labeled following each DBB injection. The pattern of retrogradely labeled neurons in the DLSN appeared in a complementary fashion to that seen in the other lateral septal nuclei. Our findings support the conclusion that the DLSN is a morphologically heterogeneous nucleus consisting almost entirely of projection neurons. The pattern of retrograde labeling in the lateral septum suggests that these projection neurons may be topographically organized since distinct subpopulations of cells were labeled following different injections in the MS/DBB complex. © 1996 Wiley-Liss, Inc. 相似文献
11.
The effect of des-Tyr1-gamma-endorphin (beta-LPH62-77, DT gamma E) and des-Tyr1-alpha-endorphin (beta-LPH62-76, DT alpha E), administered intracerebroventricularly (icv) in doses of 0.01, 0.1, 1 and 10 micrograms, was studied on the alpha-methyl-p-tyrosine-(alpha-MPT) induced disappearance of catecholamines in a number of microdissected rat brain regions, which were selected on the basis of the neuroanatomy of the dopamine systems in the brain and of previous observations. A dose-dependent increase in the disappearance of dopamine in alpha-MPT-pretreated rats was observed following icv administration of DT gamma E in the nucleus interstitialis striae terminalis, the paraventricular nucleus, the anterior hypothalamic nucleus and the zona incerta. In these same brain regions a decrease in the alpha-MPT-induced disappearance of dopamine was found following the administration of DT alpha E, but only after doses of 0.01, 0.1 and 1 microgram. In none of these regions were effects observed after 10 micrograms of DT alpha E. No effects were seen on dopamine utilization in the nucleus accumbens, caudate nucleus and median eminence after any of the doses of DT gamma E or DT alpha E. The alpha-MPT-induced disappearance of noradrenaline was significantly enhanced in the anterior hypothalamic nucleus of rats treated with DT gamma E. It is concluded that DT gamma E and DT alpha E induce opposite changes in the utilization of dopamine selectively in brain regions which are predominantly innervated by neurons belonging to the intradiencephalic dopamine systems. 相似文献
12.
Calretinin immunostaining was performed on chick lumbosacral dorsal root ganglia during embryonic development. Calretinin-immunopositive neurons were first observed at around the 9th day of incubation. Quantitative evaluation revealed a close correlation between the number of immunopositive cells and the duration of incubation. Morphometric measurements disclosed that calretinin-immunoreactive cells belong in the large or intermediate categories of dorsal root ganglion neurons. It was concluded that the appearance of calretinin immunopositivity in spinal ganglion cells during development may be associated with both the morphological and functional maturation of this particular population of primary sensory neurons. 相似文献
13.
We have investigated the rate of anterograde axonal transport of affinity-purified [125I]wheat germ agglutinin (WGA) in chick retinal ganglion cells. By 2 lines of evidence, we found the rate to be 22–44 mm/day. This rate is slower than that reported for many other endogenously synthesized proteins or for horseradish peroxidase, the only other exogenously supplied protein studied as an anterograde transported marker. This difference in rate of transport of [125I]WGA may reflect a novel pathway important in membrane recycling. 相似文献
14.
G G Chen A E Cole A B MacDermott G D Lange J L Barker 《The Journal of neuroscience》1987,7(8):2412-2422
Dorsal root ganglion (DRG) neurons from embryonic mice grown in coculture with dissociated skeletal muscle or in skeletal muscle conditioned medium (CM) showed an increased incidence of repetitive firing of action potentials when injected with sustained (60-100 msec) depolarizing current. This is in contrast to DRG neurons grown in monoculture and normal medium, which exhibit such behavior far less frequently. The first action potential showed less sensitivity to block with TTX and more sensitivity to Ca2+ channel blockers than the subsequent action potentials. The increased incidence of repetitive firing occurred when CM was added after as few as 2 or as many as 22 d in culture and with as little as 1-7 hr exposure to CM. This effect of CM cannot be mimicked by NGF or by coculture with cells from embryonic spinal cord (Peacock et al., 1973), can be eliminated by heating the CM at 56 degrees C for 30 min, and partially reversed following short exposure to CM. These results indicate that skeletal muscle releases some heat-labile factor(s) that can cause repetitive firing and, in addition, significant decrease in input resistance in the CM-treated neurons and a depression of the anomalous rectification, neither of which could account for the increase in repetitive firing. 相似文献
15.
Lee-Yuan Liu-Chen Steven A. Gillespie Thorkild V. Norregaard Michael A. Moskowitz 《The Journal of comparative neurology》1984,225(2):187-192
Neurons containing both wheat germ agglutinin (WGA) and substance P (SP) immunoreactivities were found in the ophthalamic division of the ipsilateral trigeminal ganglia following application of the axonally transported lectin WGA to cat middle cerebral artery. Immunohistochemistry was accomplished by staining for WGA and SP on adjacent sections by using the chromogen diaminobenzidine, or by staining on a single section with sequential application of WGA and SP antisera using the chromogens diaminobenzidine and 4-chloro-1-naphthol, respectively. These observations confirm the results of trigeminal lesion studies indicating that trigemino-vascular projections to the middle cerebral artery in the cat contain the neurotransmitter SP. In addition, other neurotransmitters may be present in this pathway since less than 50% of WGA-labeled cells contained SP. 相似文献
16.
Small amounts of wheat germ agglutinin-horseradish peroxidase conjugate were injected into the L5 spinal ganglion of adult rats. The conjugate was transported peripherally by the sensory neurons to their cutaneous receptors in the hind paw. Nerve plexa in the skin were heavily labeled, and different types of nerve endings could be clearly distinguished. This method gives new possibilities to study peripheral fibers and nerve endings originating in single sensory ganglia. 相似文献
17.
Focal cortical seizures prevent HRP and HRP-WGA labeling only in neurons bidirectionally connected to the cortex 总被引:1,自引:0,他引:1
Intracortical implants of polyacrylamide gel containing horseradish peroxidase labeled cortical efferents and perikarya in some cortical areas and a number of subcortical formations. When epileptogenic penicillin was added to the gel, no labeling was seen in the efferents and cell bodies of the cortex, thalamus, or claustrum, whereas the magnocellular nuclei of the basal forebrain, raphe nuclei and locus coeruleus did contain the label. 相似文献
18.
M. de Len A.A. Welcher R.H. Nahin Y. Liu M.A. Ruda E.M. Shooter C.A. Molina 《Journal of neuroscience research》1996,44(3):283-292
Peripheral nerve trauma induces the expression of genes presumed to be involved in the process of nerve degeneration and repair. In the present study, an in vivo paradigm was employed to identify molecules which may have important roles in these processes. A cDNA library was constructed with RNA extracted from rat dorsal root ganglia (DRG) 3 days after a sciatic nerve crush. After differential hybridization to this library, several cDNAs were identified that encoded mRNAs that were upregulated in the DRG ipsilateral to the crush injury, as opposed to the contralateral or naive DRG. Approximately 0.15% of all the clones screened were found to be induced. This report presents the types of induced sequences identified and characterizes one of them, DA11. The 0.7 kb DA11 full length cDNA clone contains a 405 nucleotide open reading frame that encodes a putative protein of 15.2 kDa (135 amino acid residues) and is a member of the family of fatty acid binding proteins (FABP). The DA11 protein differs by one amino acid residue from the sequence of the C-FAPB protein and by eight residues from the sequence of mal1, proteins found in rat and mouse skin, respectively. Northern and Western blot analyses showed that the DA11 mRNA and protein were induced in the injured DRG. Furthermore, studies using antibodies generated against DA11 found that the DA11-like immunoreactivity was more pronounced in the nuclei of neurons located in the DRG ipsilateral to the sciatic cut than those located in the contralateral DRG. The induction of DA11 mRNA and protein in DRG neurons suggests, for the first time, the involvement of a neuronal FABP in the process of degeneration and repair in the nervous system. © 1996 Wiley-Liss, Inc. 相似文献
19.
The calcium-binding proteins calbindin D-28k (CalB) and calretinin (CalR) have been shown to be useful markers of neuronal subpopulations located mainly in layers II–III of the neocortex of a variety of species, including human. Double labeling immunocytochemical studies of CalB, CalR, and GABA in experimental animals have shown that CalB and CalR are present in separate subpopulations of neurons. However, there are no studies of colocalization of these calcium-binding proteins and GABA in the human neocortex. The principal goal of the present work was to investigate the degree of colocalization of these substances in layers II–III of the human temporal neocortex, using a postembedding immunocytochemical method. The patterns of staining for CalB, CalR, and GABA in the human cortex were similar to those found in monkey neocortex. However, the degree of colocalization for certain combinations was different from that reported in the monkey and other experimental animals. A relatively large proportion of CalB- and CalR-immunoreactive cells (approximately 71% and 74%, respectively) were found to be immunoreactive for GABA. However, the degree of colocalization of CalB with CalR was low (between 4% and 6%). Thus, our quantitative and qualitative data suggest that these calcium-binding proteins are present in similar cortical circuits in all primates, but that in the human neocortex, there might be additional GABAergic and perhaps also non-GABAergic interneurons with unique chemical characteristics. © 1996 Wiley-Liss, Inc. 相似文献
20.
Spinal motoneuron neuroprotection by deprenyl was previously reported; the present study was carried out to evaluate neuroprotectivity in the dorsal root ganglion sensory neuron. The total neuron counts were calculated, and the axotomized sensory neurons of the dorsal root ganglion were significantly lower than those of the unaxotomized sides. Three secondary and three tertiary parameters were used. The secondary parameters were: the percentages of sensory neuron increase at the axotomized side (PNIA) and at the unaxotomized side (PNIU), and the percentage of neuronal response (PNR). The tertiary parameters were: the percentages of maximal response at the axotomized side (PMRA) and at the unaxotomized side (PMRU), and the percentage of maximal relative response (PMRR). Nonlinear statistical analysis using Gaussian, quadratic and logistic models of the tertiary parameters suggested that the data were bell-shape, which indicated that the data were biphasic. The data were divided into ascending and descending sets, and linear regression. They were analyzed according to Bent-hyperbola model and the ascending set was considered as a neurotrophic phase, while the descending one as a neurotoxic phase. The slops of PMRA were higher than that of PMRU, which indicates that the axotomized neurons were more sensitive than the unaxotomized neurons to the protective and neurotoxic effect of deprenyl. Moreover, the results showed that deprenyl had a proliferative effect on the dorsal root ganglion sensory neuron. 相似文献