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1.
Vlachová V Lyfenko A Vyklický L Orkand RK 《Physiological research / Academia Scientiarum Bohemoslovaca》2002,51(2):193-198
The effect of suramin, an inhibitor of G protein regulated signalling, was studied on the membrane currents induced by noxious heat and by capsaicin in cultured dorsal root ganglia neurones isolated from neonatal rats. Whole-cell responses induced by a heat ramp (24-52 degrees C) were little affected by suramin. The noxious heat-activated currents were synergistically facilitated in the presence of 0.3 microM capsaicin 13.2-fold and 6.3-fold at 40 degrees C and 50 degrees C, respectively. In 65% of neurones, the capsaicin-induced facilitation was inhibited by 10 microM suramin to 35 +/- 6% and 53 +/- 6% of control at 40 degrees C and 50 degrees C (S.E.M., n = 15). Suramin 30 microM caused a significant increase in the membrane current produced by a nearly maximal dose (1 microM) of capsaicin over the whole recorded temperature range (2.4-fold at 25 degrees C and 1.2-fold at 48 degrees C). The results demonstrate that suramin differentially affects the interaction between capsaicin and noxious heat in DRG neurones and thus suggest that distinct transduction pathways may participate in vanilloid receptor activation mechanisms. 相似文献
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Summary This study describes three-dimensional aspects of the development and pseudo-unipolarization of neuroblasts and the maturation of satellite cells in prenatal rat dorsal root ganglia, using scanning electron microscopy, after removal of extracellular connective tissue components by trypsin digestion and HC1 hydrolysis.At 14 days of gestation, the vast majority of neurons are spindle-shaped or bipolar and only 3% are unipolar, while at 16 and 18 days this percentage has increased to 30% and 91%, respectively. The initial portions of the central and peripheral neuronal processes gradually approach each other and form a common initial portion. Finally, the cytoplasm of this common initial portion becomes thinner and elongates to form the stem process of the mature cell.Satellite cells are present from the beginning of the period studied, but intricate networks of branching satellite cell processes only develop after about day 17. 相似文献
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Summary Putative aspartergic and glutamatergic sensory neurons in the rat were identified by autoradiography and immunocytochemistry respectively. Approximately 3% of large L4 dorsal root ganglion neurons (diameter 18–52 m) accumulated radiolabelled aspartate, whereas all satellite glia had high affinity for the amino acid. Glutamate-immunofluorescent (Glu-FITC) dorsal root ganglia neurons comprised 38.3% at S1, 35.6% at L2 33.9% at C5 and 28.8% at T6. Numbers of immunoreactive neurons were higher with the more sensitive peroxidase-anti-peroxidase (Glu-PAP) method; and the cell counts totalled 42% (S1), 41.2% (L4), 35% (C5) and 34.6% (T6). The trigeminal ganglion (TG) contained 24% Glu-FITC and 32.3% Glu-PAP positive cells. The majority of glutamate-immunoreactive sensory neurons were small, ranging from 10–35 m with median diameters of 17.5m (C5), 21m (S1), 24.2m (TG) and 28.5 m (L2). It is evident therefore, that a subgroup of class B cells are glutamatergic. Glutamate immunoreactivity in the spinal cord was similar in all segments and was localized in the superficial lamina and substantia gelatinosa of the dorsal horn. Stained interneurons were located among the immunoreactive fibres. The dorsolateral funiculus contained dense plexus of immunoreactive fibres which increased in prominence after intraperitoneal injection of L-glutamate, but penetration of exogenous glutamate into the grey matter was limited. Instead, the meninges and basal layers of the spinal blood vessels were intensely immunoreactive. The studies describe the subtypes of acidic amino acidergic neurons and relates the immunohistochemistry to a functional subclass. 相似文献
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Summary Whole-cell patch-clamp experiments were performed with neurons cultured from rat dorsal root ganglia (DRG). Two types of Na+ currents were identified on the basis of sensitivity to tetrodotoxin. One type was blocked by 0.1 nm tetrodotoxin, while the other type was insensitive to 10 m tetrodotoxin. The peak amplitude of the tetrodotoxin-insensitive Na+ current gradually decreased after depolarization of the membrane. The steady-state value of the peak amplitude was attained several minutes after the change of holding potential. Such a slow inactivation was not observed in tetrodotoxin-sensitive Na+ current. The slow inactivation of the tetrodotoxin-insensitive Na+ current was kinetically distinct from the ordinary short-time steady-state inactivation. The voltage dependence of the slow inactivation could be described by a sigmoidal function, and its time course had a double-exponential process. A decrease of external pH partially antagonized the slow inactivation, probably through an increased diffusion potential across the membrane. However, the slow inactivation was not due to change in surface negative charges, since a shift of the kinetic parameters along the voltage axis was not observed during the slow inactivation. Due to the slow inactivation, the inactivation curves for the tetrodotoxininsensitive Na+ current were shifted in the negative direction as the prepulse duration was increased. Consequently, the window current activated at potentials close to the resting membrane potential was markedly reduced. Thus, the slow inactivation may be involved in the long-term regulation of the excitability of sensory neurons.We thank Prof. Hirosi Kuriyama for his support and advice and Dr. M. Yoshii for helpful discussions. This study was supported by the Japanese Ministry of Education (Scientific Research 02670090). 相似文献
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Previous studies have shown that a subpopulation of large dorsal root ganglion neurons contains neurotrophin-3 (NT3)-like
immunoreactivity. It is not known, however, whether these NT3 immunoreactive neurons also express the high affinity receptor
for NT3, trkC. In the present study, the distribution and morphology of trkC immunoreactive neurons have been correlated with
those of NT3 immunoreactive neurons in the dorsal root ganglia. Size and segmental distributions of both antigens indicate
that they are present in the same group of large sensory neurons. Almost twice the number of these neurons are present in
the cervical and lumbar spinal ganglia than in the thoracic. Co-localization study indicates that 94% of NT3 immunoreactive
neurons express trkC. Our findings support the proposal that NT3 in these neurons is derived from their peripheral targets
rather than synthesized in situ.
Special issue dedicated to Dr. Hans Thoenen. 相似文献
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The neurotoxic effect of capsaicin has been shown to be selective on a subpopulation of small dorsal root ganglion neurons in newborn animals. The aim of this study was to provide evidence of the long lasting effect of capsaicin and its ultrapotent analog resiniferatoxin (RTX) on sensory peptidergic neurons maintained in organotypic cultures. The effects of the two irritants were examined on neurons that contained substance P (SP) and calcitonin gene-related peptide (CGRP). Exposure of the cultures to 10 microM capsaicin and 100 nM RTX for periods of 2 days or longer resulted in almost complete elimination of SP-immunoreactive (IR) neurites and reduction, but not elimination, of CGRP-IR neurites. In addition, both 10 microM capsaicin and 100 nM RTX significantly reduced the number of SP- and CGRP-IR cell bodies within DRG explants. Capsaicin in 100 microM concentration produced complete elimination of SP-IR fibers and a greater decrease in the number of CGRP-IR fibers, but failed to completely eliminate IR cell bodies. Exposure of the cultures to the irritants in the same concentrations for 90 min did not produce a measurable effect on SP- or CGRP-IR in neurites or cell bodies. It is important to establish that the effect of capsaicin and RTX on cultured neurons was of long duration (longer than 4 days) and is therefore different from depletion of peptides. These findings demonstrate that processes of cultured sensory neurons are much more sensitive to capsaicin and RTX than cell bodies. Furthermore, our results show that SP-IR neuronal elements are more sensitive to capsaicin than CGRP-IR elements. These data suggest that cultured sensory neurons express the functional properties of differentiated sensory neurons in vivo. 相似文献
10.
Role of neurotrophin signalling in the differentiation of neurons from dorsal root ganglia and sympathetic ganglia 总被引:2,自引:0,他引:2
Uwe Ernsberger 《Cell and tissue research》2009,336(3):349-384
Manipulation of neurotrophin (NT) signalling by administration or depletion of NTs, by transgenic overexpression or by deletion
of genes coding for NTs and their receptors has demonstrated the importance of NT signalling for the survival and differentiation
of neurons in sympathetic and dorsal root ganglia (DRG). Combination with mutation of the proapoptotic Bax gene allows the
separation of survival and differentiation effects. These studies together with cell culture analysis suggest that NT signalling
directly regulates the differentiation of neuron subpopulations and their integration into neural networks. The high-affinity
NT receptors trkA, trkB and trkC are restricted to subpopulations of mature neurons, whereas their expression at early developmental
stages largely overlaps. trkC is expressed throughout sympathetic ganglia and DRG early after ganglion formation but becomes
restricted to small neuron subpopulations during embryogenesis when trkA is turned on. The temporal relationship between trkA
and trkC expression is conserved between sympathetic ganglia and DRG. In DRG, NGF signalling is required not only for survival,
but also for the differentiation of nociceptors. Expression of neuropeptides calcitonin gene-related peptide and substance
P, which specify peptidergic nociceptors, depends on nerve growth factor (NGF) signalling. ret expression indicative of non-peptidergic
nociceptors is also promoted by the NGF-signalling pathway. Regulation of TRP channels by NGF signalling might specify the
temperature sensitivity of afferent neurons embryonically. The manipulation of NGF levels “tunes” heat sensitivity in nociceptors
at postnatal and adult stages. Brain-derived neurotrophic factor signalling is required for subpopulations of DRG neurons
that are not fully characterized; it affects mechanical sensitivity in slowly adapting, low-threshold mechanoreceptors and
might involve the regulation of DEG/ENaC ion channels. NT3 signalling is required for the generation and survival of various
DRG neuron classes, in particular proprioceptors. Its importance for peripheral projections and central connectivity of proprioceptors
demonstrates the significance of NT signalling for integrating responsive neurons in neural networks. The molecular targets
of NT3 signalling in proprioceptor differentiation remain to be characterized. In sympathetic ganglia, NGF signalling regulates
dendritic development and axonal projections. Its role in the specification of other neuronal properties is less well analysed.
In vitro analysis suggests the involvement of NT signalling in the choice between the noradrenergic and cholinergic transmitter
phenotype, in the expression of various classes of ion channels and for target connectivity. In vivo analysis is required
to show the degree to which NT signalling regulates these sympathetic neuron properties in developing embryos and postnatally.
U.E. is supported by the DFG (Er145-4) and the Gemeinnützige Hertie-Stiftung. 相似文献
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Previously, we demonstrated that sphingosine 1-phosphate (S1P) increased the excitability of small-diameter sensory neurons, in part, through activation of S1P receptor 1 (S1PR(1)), suggesting that other S1PRs can modulate neuronal excitability. Therefore, studies were undertaken to establish the expression profiles of S1PRs in the intact dorsal root ganglion (DRG) and in defined single isolated sensory neurons. To determine mRNA expression of S1PRs in the DRG, SYBR green quantitative PCR (qPCR) was used. To determine the expression of S1PR mRNAs in single neurons of defined diameters, a preamplification protocol utilizing Taqman primer and probes was used to enhance the sensitivity of detection. The preamplification protocol also permitted detection of mRNA for two hallmark neuronal receptor/ion channels, TRPV1 and P(2)X(3). Expression profiles of S1PR mRNA isolated from lung and brain were used as positive control tissues. In the intact DRG, the order of expression of S1PRs was S1PR(3)>R(1)≈R(2)>R(5)≈R(4). In the single neurons, the expression of S1PRs was quite variable with some neurons expressing all five subtypes, whereas some expressing only one subtype. In contrast to the DRG, S1PR(1) was the highest expressing subtype in 10 of the 18 small-, medium-, and large-diameter sensory neurons. S1PR(1) was the second highest expressor in ~50% of those remaining neurons. Overall, in the single neurons, the order of expression was S1PR(1)>R(3)≈R(5)>R(4)>R(2). The results obtained from the single defined neurons are consistent with our previous findings wherein S1PR(1) plays a prominent but not exclusive role in the enhancement of neuronal excitability. 相似文献
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The size of the vanilloid receptor was examined by high-energy radiation inactivation analysis of the binding of [3H]resiniferatoxin to pig dorsal root ganglion membranes; it was found to be 270 +/- 25 kDa. This value most likely represents the size of a receptor complex rather than of an individual subunit. Other ligand-gated cation channel complexes have reported molecular weights in this range, e.g. 300 kDa for the acetylcholine receptor. 相似文献
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Franke-Radowiecka A 《Histology and histopathology》2011,26(12):1509-1517
The present study investigated the chemical coding of mammary gland-projecting dorsal root ganglia (DRG) neurons using double-labelling immunohistochemistry. Earlier investigations revealed the presence of Fast blue - positive (FB+) neurons in Th9-Th12 DRG after injection of the tracer into the second, right thoracic mamma. Neurons projecting to the last right abdominal mamma were found in L1-L3 DRG. In the present study, the cryostat sections from these ganglia were stained for calcitonin gene-related peptide (CGRP), substance P (SP), nitric oxide synthase (NOS), galanin (GAL) and pituitary adenylate cyclase activating polypeptide (PACAP). Immunohistochemistry revealed that the vast majority of FB+ mammary gland-projecting neurons contained immunoreactivity to CGRP (68.87±0.7%), SP (63.4±0.9%), NOS (32.47±0.9%), GAL (16.28±0.8%) and less numerous nerve cells stained for PACAP (5.87±0.5%). The present results largely correspond with findings dealing with immunohistochemical characterization of nerve fibres supplying porcine mammary gland structures described earlier. 相似文献
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GDNF对体外运动神经元和感觉神经元的影响 总被引:5,自引:0,他引:5
目的:探讨胶质细胞源性神经营养因子(GDNF)对正常胎鼠脊髓运动神经元(SMN)和背根神经节神经元(DRG)生长活性的作用.方法:建立大鼠胚胎SMN和DRG单细胞培养体系,观察1 μg/L、10 μg/L、50 μg/L和100 μg/L GDNF对SMN和DRG存活及突起生长的影响.结果: GDNF组培养的SMN和DRG存活数目明显增加,神经元突起长度比对照组明显增长,且具有剂量依赖趋势.结论: GDNF对正常大鼠胚胎发育期运动神经元和感觉神经元具有神经营养作用. 相似文献
15.
Backström E Chambers BJ Kristensson K Ljunggren HG 《Journal of immunology (Baltimore, Md. : 1950)》2000,165(9):4895-4900
In contrast to extensive studies on the role of T and B lymphocytes in the pathogenesis of autoimmune diseases of the nervous system, little is known about NK cells and their potential role in the destruction of neural tissue. NK cells have been implicated in the selective death of sympathetic neurons resident in the superior cervical ganglia of rats after exposure to the drug guanethidine. This observation suggests that NK cells may function as principle effectors in immunological diseases of the nervous system. However, the direct mechanism of action of NK cells in this model is not known. In particular, it is not known whether NK cells can kill autologous neurons directly. The aim of the present study was to examine whether NK cells can kill directly dorsal root ganglia neurons cultured in vitro. We demonstrate that C57BL/6 (B6)-derived dorsal root ganglia neurons can be killed directly by syngenic IL-2-activated NK cells, and that this nerve cell lysis is dependent on the expression of perforin in the NK cells. NK cells were less effective in destroying neurons grown in the presence of glial cells. These observations indicate a potential role for NK cells in nerve cell degeneration in inflammatory diseases of the nervous system. 相似文献
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Lennart Enerbck Krister Kristensson Lennart A. Persson 《Experimental cell research》1979,119(2):399-403
A technique is described by which neurons from mouse dorsal root ganglia can be dispersed in single-cell suspensions suitable for quantitative cytochemical analyses. The neurons were intact as controlled by trypan blue exclusion test, and the cell size distribution of the dispersed neurons corresponded to that of untreated, intact ganglia. Horseradish peroxidase and Evans blue applied to cut sciatic nerve, were transferred by somatopetal intra-axonal transport and accumulated in corresponding dorsal root ganglia neurons. The tracers were retained during the preparation of cell suspensions. The accumulation of the fluorescent tracer Evans blue was quantitated by cytofluorometric measurements on individual neurons. 相似文献
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Growth pattern and NGF-dependent survival of dorsal root ganglia neurons of distinct glyco-phenotype
Astrid Zimmermann 《Developmental neurobiology》1998,34(3):193-207
Cell-surface glyco-phenotypes of dorsal root ganglia (DRG) neurons were specified with monoclonal antibodies (mABs) D1 and E1. D1 demarcated sensory afferents in skin but not muscle target. More than 90% of the drg neurons supported by nerve growth factor (NGF) in vitro were D1 positive (D1+). A fraction of these D1+ neurons, those of small to intermediate soma size, coexpressed a PNGase-sensitive glycoepitope E1, defined by mAB E1. In situ and in vitro, E1+/D1+ and E1−/D1+ neurons and nerve fibers were affiliated. After separation of the two glyco-phenotypes, NGF-dependent survival of E1−/D1+ neurons was no longer observed. Two interrelated concepts emerge from these findings: (a) NGFs survival functions for cutaneous sensory neurons are in part indirect and appear to be based on interneuronal cooperation for survival; and (b) interneuronal survival dependencies are likely to be a decisive factor governing nerve fiber assemblages. © 1998 John Wiley & Sons, Inc. J Neurobiol 34: 193–207, 1998 相似文献
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《Journal of receptor and signal transduction research》2013,33(2):115-120
Bupivacaine and levobupivacaine have been shown to be effective in the treatment of pain as local anesthetics, although the mechanisms mediating their antinociceptive actions are still not well understood. The aim of this study was to investigate the effects of bupivacaine and levobupivacaine on intracellular calcium ([Ca2+]i) signaling in cultured rat dorsal root ganglion (DRG) neurons. DRG neuronal cultures loaded with 5?μM Fura-2/AM and [Ca2+]i transients for stimulation with 30?mM KCl (Hi K+) were assessed by using fluorescent ratiometry. DRGs were excited at 340 and 380?nm, emission was recorded at 510?nm, and responses were determined from the change in the 340/380 ratio (basal-peak) for individual DRG neurons. Data were analyzed by using Student’s t-test. Levobupivacaine and bupivacaine attenuated the KCl-evoked [Ca2+]i transients in a reversible manner. [Ca2+]i increase evoked by Hi K+ was significantly reduced to 99.9?±?5.1% (n?=?18) and 62.5?±?4.2% (n?=?15, P?<?0.05) after the application of 5 and 50?µM levobupivacaine, respectively. Bupivacaine also inhibited Hi K+-induced [Ca2+]i responses, reduced to 98.7?±?4.8% (n?=?10) and 69.5?±?4.5% (n?=?9, P?<?0.05) inhibition of fluorescence ratio values of Hi K+-induced responses at 5 and 50?μM, respectively. Our results indicate that bupivacaine and levobupivacaine, with no significant differences between both agents, attenuated KCl-evoked calcium transients in a reversible manner. The inhibition of calcium signals in DRG neurons by levobupivacaine and bupivacaine might contribute to the antinociceptive effects of these local anesthetics. 相似文献
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The differential distribution of small and large neurons in the sacrococcygeal dorsal root ganglia of the cat was disclosed by measuring the short diameter of the perikarya. The measured values were systematically charted along the rostro-caudal axis of the ganglion. This approach permitted to delineate small and large neurons; the short diameter of the former being less, that of the latter more, than 22 micrometers. Small neurons (69% of the population) are distributed along the entire length of the ganglions, while large neurons are clustered in the distal half. The same histological specimens were appropriate to show that if only every 8th section was used for counting the number of perikarya (the number of the nucleoli was interpreted as that of the perikarya) the result was not significantly different (less than +/- 5%) from that gained by a total count. The significance of the inhomogeneous distribution of ganglion cells was discussed within the emerging concept of the topological arrangement of the perikarya in the ganglion, on the one hand, and the termination of the central branch of the neurons in the spinal cord, on the other. 相似文献