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Paul A. Dijkhuizen Wim T. J. M. C. Hermens Marc A. T. Teunis Joost Verhaagen 《Developmental neurobiology》1997,33(2):172-184
The neurotrophins are a family of proteins that promote neuronal survival and neurite outgrowth during development and can also enhance the regeneration of injured adult neurons. The local and continuous delivery of these proteins at the site of injury is problematic, since this requires repeated intraparenchymal injections or the use of invasive canula-micropump devices. In the present study we report the generation and characterization of an adenoviral vector for a member of the neurotrophins, neurotrophin-3 (Ad-NT-3). Using Ad-NT-3, we examined the expression and biological activity of NT-3 in dorsal root ganglia (DRG) explant cultures. Gene transfer with Ad-NT-3 results in the synthesis of genuine NT-3 and in a dosage-dependent neurite outgrowth response in DRG explants. Transduction of DRG explants with a viral vector dosage of 5 × 105 to 5 × 106 plaque-forming units induced the formation of a dense halo of neurites comparable to outgrowth observed following the addition of 100 ng/mL exogenous NT-3. In addition, a single infection with Ad-NT-3 produced biologically active NT-3 for at least 20 days in culture, as evidenced by continued neurite extension. This indicates that adenoviral vector-mediated expression of NT-3 results in high-level production of biologically active NT-3 and could therefore be used as a strategy to promote the regeneration of injured peripheral and central nerve projections. © 1997 John Wiley & Sons, Inc. J Neurobiol 33: 172–184, 1997 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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催产素对大鼠背根神经节分离细胞GABA激活电流的调制作用 总被引:8,自引:0,他引:8
在急性分离的大鼠背根神经节(dorsal root ganglion,DRG)细胞上,应用全细胞膜片箝技术观察了预知催产素(oxytocin,OT)对GABA激活电流的调制作用。结果如下:(1)大多数细胞(48/52,90.5%)对GABA敏感。(2)OT可引起51.3%(20/39)的受检细胞出现外向膜电流;43.6%(17/39)无明显膜反应;5.1%(2/39)出现内向膜电流。(3)预加OT 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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Masato Kano Hideo Tashiro Tadashi Kawakami Toshifumi Takenaka Hideki Gotoh 《Cellular and molecular neurobiology》1995,15(3):297-306
Summary 1. Growth cones of cultured dorsal root ganglion neurons from mice were irradiated using a mercury lamp.2. The flux of particles of fast retrograde axoplasmic transport decreased promptly after light irradiation without a change in velocity.3. That of anterograde transport decreased as well, but with a significant latency. The decrease in the anterograde flux was attributed to decreased velocity of particles.4. Video-enhanced contrast microscopy of growth cones revealed transient swelling of growth cones and transient stagnation of particles in growth cones.5. The longer the neurite, the larger the latency of the change of the anterograde transport; peripheral information was calculated to be conveyed to the cell body at a speed of 6 µm/min.6. The mechanism of this information conveyance and the export of materials from the cell body are discussed. 相似文献
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对GPI-1046是否具有神经营养作用目前有两种不同的认识。Steiner等认为GPI-1046能促进体外培养的感觉神经节神经元突起生长。但Harper等却没能证明GPI-1046有这样的作用。由于GPI-1046在临床上具有重要应用价值和前景,我们重新评价了GPI-1046对体外培养鸡胚神经节的神经营养作用,发现在低浓度神经生长因子(nerve growth factor,NGF)存在下,GPI-1046能明显促进鸡背根神经节神经突起的生长。 相似文献
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In this study, we have examined the properties of synaptic transmission between dorsal root ganglion (DRG) and dorsal horn (DH) neurons, placed in co-culture. We also examined the effect of the anti-hyperalgesic gabapentinoid drug pregabalin (PGB) at this pharmacologically relevant synapse. The main method used was electrophysiological recording of excitatory post synaptic currents (EPSCs) in DH neurons. Synaptic transmission between DRG and DH neurons was stimulated by capsaicin, which activates transient receptor potential vanilloid-1 (TRPV1) receptors on small diameter DRG neurons. Capsaicin (1 μM) application increased the frequency of EPSCs recorded in DH neurons in DRG-DH co-cultures, by about 3-fold, but had no effect on other measured properties of the EPSCs. There was also no effect of capsaicin in the absence of co-cultured DRGs. Application of PGB (100 μM) for 40-48 h caused a reduction in the capsaicin-induced increase in EPSC frequency by 57%. In contrast, brief preincubation of PGB had no significant effect on the capsaicin-induced increase in EPSC frequency. In conclusion, this study shows that PGB applied for 40-48 h, but not acute application inhibits excitatory synaptic transmission at DRG-DH synapses, in response to nociceptive stimulation, most likely by a presynaptic effect on neurotransmitter release from DRG presynaptic terminals. 相似文献
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Laurel M. Donahue 《Neurochemical research》1995,20(6):713-717
Tetrodotoxin-insensitive (TTX-I) sodium currents have been recorded from newborn and adult rat sensory neurons, but the sodium channel gene(s) responsible for the TTX-I current are unknown. Because SkM2, one of six voltage-sensitive sodium channel genes cloned from rat, encodes the only cloned channel that is relatively resistant to tetrodotoxin, we sought to test whether the TTX-I current in rat sensory neurons is due to the SkM2 channel. We hypothesized that the TTX-I current might be generated from (1) an RNA splicing variant of SkM2, (2) post-translational modification of the SkM2 protein, or (3) interaction with altenate additional channel subunits. SkM2 mRNA expression was examined in newborn rat dorsal root ganglia (DRG) by RNase arotection assay. No SkM2 expression was detected. Therefore, we conclude that the TTX-I sodium current in DRG is unlikely to result from the expression of the SkM2 gene. 相似文献
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Postsynaptic currents and action potentials recorded from neurons in a mixed culture of rat dorsal root ganglion and spinal
cord cells are described. The existence of mutual synaptic connections between the above two types of neurons is demonstrated.
Neirofiziologiya/Neurophysiology, Vol. 38, No. 4, pp. 358–360, July–August, 2006. 相似文献
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Qian Bai Jinping Shao Jing Cao Xiuhua Ren Weihua Cai Songxue Su Sanjeeth George Zhiyong Tan Weidong Zang Tieli Dong 《Journal of cellular biochemistry》2020,121(1):768-778
Previous studies have found that increased expression of Nav1.9 and protein kinase C (PKC) contributes to pain hypersensitivity in a couple of inflammatory pain models. Here we want to observe if PKC can regulate the expression of Nav1.9 in dorsal root ganglion (DRG) in rheumatoid arthritis (RA) pain model. A chronic knee joint inflammation model was produced by intra-articular injection of the complete Freund's adjuvant (CFA) in rats. Nociceptive behaviors including mechanical, cold, and heat hyperalgesia were examined. The expression of Nav1.9 and PKCα in DRG was detected by a quantitative polymerase chain reaction, Western blot, and immunofluorescence. The in vitro and in vivo effects of a PKC activator (phorbol 12-myristate 13-acetate [PMA]) and a PKC inhibitor (GF-109203X) on the expression of Nav1.9 were examined. Moreover, the effects of PKC modulators on nociceptive behaviors were studied. Increased mechanical, heat, and cold sensitivity was observed 3 to 14 days after CFA injection. Parallel increases in messenger RNA and protein expression of Nav1.9 and PKCα were found. Immunofluorescence experiments found that Nav1.9 was preferentially colocalized with IB4+DRG neurons in RA rats. In cultured DRG neurons, PMA increased Nav1.9 expression while GF-109203X prevented the effect of PMA. PMA increased Nav1.9 expression in naïve rats while GF-109203X decreased Nav1.9 expression in RA rats. In naïve rats, PMA caused mechanical and cold hyperalgesia. On the other hand, GF-109203X attenuated mechanical and cold hyperalgesia in RA-pain model. Nav1.9 might be upregulated by PKCα in DRG, which contributes to pain hypersensitivity in CFA-induced chronic knee joint inflammation model of RA pain. 相似文献
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Sasaki H Hozumi Y Hasegawa H Ito T Takagi M Ogino T Watanabe M Goto K 《Cell and tissue research》2006,326(1):35-42
The dorsal root ganglion (DRG) and dorsal horn of the spinal cord are areas through which primary afferent information passes enroute to the brain. Previous studies have reported that, during normal neuronal activity, the regional distribution of a second messenger, diacylglycerol (DG), which is derived from phosphoinositide turnover, is diverse in these areas. However, the way that DG is regulated in these organs remains unknown. The present study was performed to investigate mRNA expression and protein localization of DG kinase (DGK) isozymes, which play a central role in DG metabolism. Gene expression for DGK isozymes was detected with variable regional distributions and intensities in the spinal cord. Among the isozymes, most intense signals were found for DGKζ and DGKι in the DRG. By immunohistochemical analysis, DGKζ immunoreactivity was detected heterogeneously in the nucleus and cytoplasm of small DRG neurons with variable levels of distribution, whereas it was detected exclusively in the cytoplasm of large neurons. On the other hand, DGKι immunoreactivity was distributed solely in the cytoplasm of most of the DRG neurons. Double-immunofluorescent imaging of these isozymes showed that they coexisted in a large population of DRG neurons at distinct subcellular sites, i.e., DGKζ in the nucleus and DGKι in the cytoplasm. Thus, DGK isozymes may have different functional roles at distinct subcellular sites. Furthermore, the heterogeneous subcellular localization of DGKζ between the nucleus and cytoplasm implies the possible translocation of this isozyme in small DRG neurons under various conditions.The work was supported by grants-in-aid from the Ministry of Education, Science, Culture, and Sports of Japan (M.T., K.G.) and from the Ono Medical Research Foundation, Kato Memorial Bioscience Foundation, and Janssen Pharmaceutical (K.G.) and by the 21st Century Center of Excellence Program of the Ministry of Education, Culture, Sports, Science and Technology of Japan. 相似文献
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To find a promising alternative to neurons or schwann cells (SCs) for peripheral nerve repair applications, this study sought
to isolate stem cells from fetal rat dorsal root ganglion (DRG) explants. Molecular expression analysis confirmed neural stem
cell characteristics of DRG-derived neurospheres in terms of expressing neural stem cell-specific genes and a set of well-defined
genes related to stem cell niches and glial fate decision. Under the influence of neurotrophic factors, bFGF and NGF, the
neurospheres gave rise to neurofilament-expressing neurons and S100-expressing Schwann cell-like cells by different pathways.
This study suggests that a subpopulation of stem cells that reside in DRGs is the progenitor of neurons and glia, which could
directly induce the differentiation toward neurons, or SCs. 相似文献
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Masuda T Tsuji H Taniguchi M Yagi T Tessier-Lavigne M Fujisawa H Okado N Shiga T 《Developmental biology》2003,254(2):289-302
Initial trajectories of dorsal root ganglion (DRG) axons are shaped by chemorepulsive signals from surrounding tissues. Although we have previously shown that axonin-1/SC2 expression on DRG axons is required to mediate a notochord-derived chemorepulsive signal, Dev. Biol. 224, 112-121), other molecules involved in the non-target-derived repulsive signals are largely unknown. Using coculture assays composed of tissues derived from the chick embryo or mutant mice treated with function-blocking antibodies and phosphatidylinositol-specific phospholipase C, we report here that the chemorepellent semaphorin 3A (Sema3A) and its receptor neuropilin-1 are required for mediating the dermamyotome- and notochord-derived, but not the ventral spinal cord-derived, chemorepulsive signal for DRG axons. The dermamyotome-derived chemorepulsion is exclusively dependent on Sema3A/neuropilin-1, whereas other molecules are also involved in the notochord-derived chemorepulsion. Chemorepulsion from the ventral spinal cord does not depend on Sema3A/neuropilin-1 but requires axonin-1/SC2 to repel DRG axons. Thus, differential chemorepulsive signals help shape the initial trajectories of DRG axons and are critical for the proper wiring of the nervous system. 相似文献
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电生理学特性得到确定的大鼠脊髓背根神经节细胞内谷氨酸和P物质的共存 总被引:2,自引:0,他引:2
在离体灌流带脊髓和坐骨神经经的标本上,对脊髓背根神经节(DRG)细胞的电生理学特性、对P物质(SP)受体激动剂的反应及谷氨酸(Glu)/SP共存的特点进行了研究。(1)对135个细胞进行了细胞内记录,并依纤维传导速度将其分为Aα/β(〉12m/s)和C(〈1.3m/s)两大类,它们的动作电位的快速后超极化(fAHP)有明显区别,C类的fAHP幅度小、时程长,Aα/β类的fAHP幅度大、时程短;(2 相似文献
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Enhanced neuronal regeneration by retinoic acid of murine dorsal root ganglia and of fetal murine and human spinal cord in vitro 总被引:2,自引:0,他引:2
Summary This study demonstrates that retinoic acid (RA), an active metabolite of vitamin A, can act to enhance regeneration of neurites,
at physiologic concentrations, in vitro. Explanted fragments of mouse dorsal root ganglia (DRG) and mouse and human spinal
cord (SC) were maintained, in vitro, for periods up to 11 d. Murine DRG neurons were exposed to RA concentrations ranging
from 100 μM to 1 nM, whereas neurons within murine and human SC explants were exposed to 10 μM to 10 nM RA. Results show that RA significantly (P<0.001) increases mean neurite length but not neurite number. Specifically, murine DRG neurons showed increases in mean neurite
length of 30.7% with individual explants showing increases of up to 133.5%. Murine and human SC showed mean enhancements of
43.4 and 58.1%, respectively, but did so at lower concentrations of RA. The results indicate that RA may play a potentially
critical role in neuronal regeneration. 相似文献