The relation of nucleolus diameter to cell body diameter in mammalian dorsal root ganglion cells

The present study correlates cell body and nucleolar sizes for dorsal root ganglion cells in the rat. To do this, we measured cell body areas and calculated their diameters and measured nucleolar diameters for 719 cells. These data indicate that there is a correlation in that increasing cell size is associated with increasing nucleolar size. However, there is considerable variability of cell body size for each nucleolar diameter and vice versa. Nevertheless, when nucleolar diameters are grouped, the function 1n D = 1.687 + 0.334 dr, where D is the diameter of the cell body and dr the rounded diameter of the nucleolus, produced an almost straight line. Thus this formula provides a good estimate of the relation of nucleolar and cell body sizes for dorsal root ganglion cells of the rat. In addition, estimates of the variances of cell body size at each nucleolar diameter are provided.     

TNF-α在CCD模型大鼠背根神经节中的表达及机制

目的:探讨TNF-α和NF-κB在背根神经节慢性压迫(CCD)模型大鼠背根神经节(DRG)中的表达变化及其对疼痛学行为的影响。方法:建立CCD大鼠模型,采用von Frey纤维丝监测机械痛阈的改变;通过Western blotting检测TNF-α和NF-κB在DRG中的表达变化趋势,分析其与疼痛行为之间的相关性;并采用免疫荧光双染技术研究TNF-α在DRG中的表达位置。结果:CCD组的50%机械缩足阈值在术后1 d即开始明显下降(P0.01),7~14 d达到高峰,其后逐渐上升,直至术后35 d仍明显低于术前及sham组(P0.01)。而DRG上的TNF-α及NF-κB于造模后各时点均显著增多(P0.01),且TNF-α的表达趋势与50%机械缩足阈值显著相关(P0.05)。结论:DRG慢性压迫可促进其上的TNF-α和NF-κB的合成和分泌,进而诱发机械痛觉过敏。因此,TNF-α/NF-κB信号通路可能是CCD模型疼痛形成的重要通路之一。     

Chronic compression of mouse dorsal root ganglion alters voltage-gated sodium and potassium currents in medium-sized dorsal root ganglion neurons

Chronic compression (CCD) of the dorsal root ganglion (DRG) is a model of human radicular pain produced by intraforaminal stenosis and other disorders affecting the DRG, spinal nerve, or root. Previously, we examined electrophysiological changes in small-diameter lumbar level 3 (L3) and L4 DRG neurons treated with CCD; the present study extends these observations to medium-sized DRG neurons, which mediate additional sensory modalities, both nociceptive and non-nociceptive. Whole-cell patch-clamp recordings were obtained from medium-sized somata in the intact DRG in vitro. Compared with neurons from unoperated control animals, CCD neurons exhibited a decrease in the current threshold for action potential generation. In the CCD group, current densities of TTX-resistant and TTX-sensitive Na(+) current were increased, whereas the density of delayed rectifier voltage-dependent K(+) current was decreased. No change was observed in the transient or "A" current after CCD. We conclude that CCD in the mouse produces hyperexcitability in medium-sized DRG neurons, and the hyperexcitability is associated with an increased density of Na(+) current and a decreased density of delayed rectifier voltage-dependent K(+) current.     

Purinergic activation of dorsal root ganglion neurones in vivo

Functional relevance of non-synaptic purinergic receptors on dorsal root ganglion cells was tested in vivo by the influence of ATP using 2P-LSM and Ca imaging. Within a few seconds after local application of ATP, neurones in dorsal root ganglion were activated indicated by an increase of their calcium signal. The signal reached its maximum within a few seconds and declined to control values after about 30 s. Purinergic action seems to include non-synaptic cell-to-cell communication within dorsal root ganglia.     

Morphological changes of dorsal root ganglion cells in the process-forming period

Rat DRG cells at early stages of development were observed to clarify the forming process from bipolar to pseudounipolar cells.

Dissociated DRG cells in vivo and in vitro were observed by light and scanning electron microscopy. The percentage of cell types in the 13th prenatal to the 1-day postnatal DRG in vivo were investigated. It is noteworthy that about 5% of ganglion cells were still bipolar in the 1-day postnatal rat. When dissociated DRG cells were seen in long-term culture, completely unipolarized DRG cells were revealed by silverimpregnation. But when fibroblast growth was suppressed in long-term culture, no pseudounipolar cells were observed.

Dorsal root ganglia on day 14 of gestation were organ cultured. The incidence of pseudounipolar cell appearance was estimated time-sequentially by silver-impregnations (Table 2). Unipolar cells increased to about 80% at the 22nd day of culture.

Cultured DRG cells were examined by transmission electron microscopy and the morphological change from bipolar cells to pseudounipolar cells was investigated. No evidence was obtained which showed that the processes of bipolar cells fuse to make a stem process.

In conclusion, observations of developing DRG cells by light and electron microscopy revealed that the stem process of the pseudounipolar DRG cells is derived from the elongated cytoplasm of the cell body and is not derived from the fusion of the opposite two processes.     

Dendrotoxin-sensitive K+ channels in dorsal root ganglion cells

In primary sensory cells, a K current active at resting potential is selectively blocked by the convulsant snake toxin, dendrotoxin. Using the patch-clamp technique, we have examined the characteristics of this K current at the unitary level. The voltage-activated K+ channels were found to have a maximum conductance of 5-10 pS in a 'physiological' K+ gradient. They show negligible sensitivity to calcium at the inner membrane aspect. Blockade by dendrotoxin seems likely to be due to direct action on the K+ channel.     

Isolation of specific peptides that home to dorsal root ganglion neurons in mice

We isolated peptides that home to mouse dorsal root ganglion (DRG) from a phage library expressing random 7-mer peptides fused to a minor coat protein (pIII) of the M13 phage. An in vitro biopanning procedure yielded 113 phage plaques after five cycles of enrichment by incubation with isolated DRG neurons and two cycles of subtraction by exposure to irrelevant cell lines. Analyses of the sequences of this collection identified three peptide clones that occurred repeatedly during the biopanning procedure. Phage-antibody staining revealed that the three peptides bound to DRG neurons of different sizes. To determine if the peptides would recognize neuronal cells in vivo, we injected individual GST-peptide-fusion proteins into the subarachnoid space of mice and observed the appearance of immunoreactive GST in the cytosol of DRG neurons with a similar size distribution as that observed in vitro, indicating that the GST-peptide-fusion proteins were recognized and taken up by different DRG neurons in vivo. The identification of homing peptide sequences provides a powerful tool for future studies on DRG neuronal function in vitro and in vivo, and opens up the possibility of neuron-specific drug and gene delivery in the treatment of diseases affecting DRG neurons.     

外周神经系统中背根神经节的研究进展

背根神经节是机体内、外环境与脊髓联结的纽带,也是痛觉传入的第一级神经元,在感觉传递途径中起着重要的作用。外周神经损伤可导致背根神经节神经元减少、神经元之间的连接出现紊乱,造成感觉传导异常,引起神经病理性疼痛;NGF、BDNF、NT-3等因子在与痛觉相关的背根节小型神经元上表达增加。电压门控钠离子通道在疼痛产生的机制中也起到了举足轻重的作用。本文对背根神经节的形态结构特点、体外培养以及与神经病理性疼痛发生的关系予以综述。     

Expression of interleukin-6 in human dorsal root ganglion cells

The expression of Interleukin-6 (IL-6) was studied in normal dorsal root ganglia (DRG) of juvenile and foetal humans, using immunohistochemistry and in situ hybridization techniques. There was an expression of IL-6-like immunoreactivity in more than 75% out of neuronal cells in the juvenile ganglia with a peripheral localization, and also an expression in the foetal ganglion cells. There was a co-localization of IL-6 with substance P (SP) and calcitonin gene-related peptide (CGRP) in more than 60% of the DRG cells, respectively. By in situ hybridization 0.9% of the cells in the juvenile ganglia and 1.1% of the cells in the foetal ganglia showed a positive signal for IL-6. In addition, expression of IL-6 was found in juvenile medulla spinalis, preferentially in the white matter.     

Effects of electro-acupuncture on NT-4 expression in spinal dorsal root ganglion and associated segments of the spinal dorsal horn in cats subjected to adjacent dorsal root ganglionectomy

It is well known that neuroplasticity occurs in the central nervous system in response to injury. Electro-acupuncture (EA) may also promote neuroplasticity. But little is known about the underlying molecular mechanisms for the beneficial effects of EA. This study investigated the effects of EA on neurotrophin-4 (NT-4) expression in L(6) spinal dorsal root ganglion (DRG) and associated segments of the spinal dorsal horn in cats subjected to unilateral removal of L(1)-L(5) and L(7)-S(2) DRG. NT-4 protein was normally present in the cytoplasm of the L(6) DRG neurons and L(3) and L(6) spinal dorsal horn neurons and glia. Adjacent ganglionectomy leads to a significant decrease in NT-4 expression in the L(6) DRG, but no change in the spinal dorsal horn. Following EA treatment a significant increase occurred in the L(6) DRG at 14 days post-operation (dpo) as well as the L(6) cord segment at 7 and 14 dpo. These findings pointed to a possible association between NT-4 expression and EA promoted spinal cord plasticity in adult cats subjected to partial ganglionectomy.     

Serotonin receptor mRNA expression in rat dorsal root ganglion neurons

In the present study, we have used in situ hybridization to examine the distribution of serotonin (5-HT) receptors in rat dorsal root ganglion (DRG) neurons. Within DRG neurons, mRNAs for 5-HT1B, 5-HT1D, 5-HT2A, 5-HT2B, 5-HT3B and 5-HT4 receptors were readily detected in small (<25 microm), medium (25-45 microm) and large (>45 microm) diameter neurons. In contrast mRNAs for 5-HT1A, 5-HT1E, 5-HT2C, 5-HT5A, 5-HT5B, 5-HT6 and 5-HT7 receptors were undetectable in these neurons. The present study provides an insight into the molecular profile of 5-HT receptor subtypes in neurons responsible for modulating sensory information.     

Proton-induced sodium current in frog isolated dorsal root ganglion cells

1. The proton-induced current was examined in isolated frog dorsal root ganglion (DRG) cells by the use of the "concentration-clamp" technique, which allows intracellular perfusion and rapid change of external solution with various pH (pHo) within 2 ms under single-electrode voltage-clamp condition. 2. Over one-half of the examined neurons showed no response for a "step" reduction of pHo even in a Ca2(+)-free external solution. In smaller neurons having a diameter less than 20 microns, the persistent and reliable proton-induced responses were obtained, though the current amplitude and the activation and inactivation varied considerably for each cell. 3. The decrease of external Na+ concentration ([Na+]o) reduced the proton response. The proton response reversed the direction and the Na+ equilibrium potential (ENa). 4. With decreasing pHo from 7.4, proton response increased in a sigmoidal fashion. The threshold was around pH 7.0 and the maximum response appeared at pH 5.2, whereas pKa and Hill coefficient were 6.0 and 1.97, respectively. 5. The activation and inactivation phases of the proton-induced current behaved as a single exponential function. The time constants of activation (tau a) and inactivation (tau i) were not affected by changing either the holding membrane potential (VH) or the low external Ca2+ concentration [( Ca2+]o) between 10(-6) and 5 X 10(-3) M. But the decrease of pHo up to 5.2 decreased both tau a and tau i in a saturable manner. 6. In the inactivation curve of proton-induced current obtained by decreasing pHo from various conditioning pHo to 5.5, half inactivation occurred at pHo 7.45.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sensory fiber regeneration after dorsal root ganglion section in rats

Department of Histology, S. M. Kirov Gor'kii Medical Institute. Laboratory of Neuronal Structure of the Brain, Brain Research Institute, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR D. S. Sarkisov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 110, No. 10, pp. 429–430, October, 1990.     

小鼠背根神经节中痒觉神经元的分布特性

目的观察小鼠背根神经节(DRG)中痒觉特异性Mrgpr A3+神经元的分布特征。方法采用遗传学方法将Mrgpr A3+神经元特异性标记强化绿色荧光蛋白(EGFP)和td Tomato;选取3只纯合Mrgpra3EGFP-Cre;ROSA26td Tomato成年转基因小鼠,分离皮肤和背根神经节组织;采用激光扫描共焦成像技术观察Mrgpr A3+神经元的外周神经纤维在小鼠躯体皮肤的投射分布特征;采用双光子成像技术观察Mrgpr A3+神经元在整体背根神经节中的三维空间分布情况。结果脸颊、背部和脚掌皮肤的Mrgpr A3+神经纤维分布密集,粗且长,分布广泛;颈部和腹部皮肤的Mrgpr A3+神经纤维分布稀疏,短且小,呈散点状分布;Mrgpr A3+神经纤维在皮肤的有毛和无毛区域都有投射分布,且不同部位分布特点不同;几乎所有的Mrgpr A3+神经元都为小直径感觉神经元,且在颈、胸、腰、尾段背根神经节中均有分布;颈段、胸段、腰段和尾段的Z轴成像深度分别为350μm、250μm、400μm和200μm;躯体不同部位背根神经节中的Mrgpr A3+神经元的三维空间分布在不同节段存在明显差异。结论小鼠躯体不同部位皮肤的Mrgpr A3+神经纤维的分布特征和整体背根神经节中Mrgpr A3+神经元的三维空间分布特征都存在较大差异,痒觉神经元和末梢分布的差异可能是不同区域存在痒觉生理反应差异的结构基础。     

CLP36 interacts with palladin in dorsal root ganglion neurons

CLP36, a member of the α-actinin-associated LIM protein (ALP)/enigma protein family, plays a role in neurite outgrowth in the peripheral nervous system. However, the underlying molecular mechanisms are not known. In this study, we performed yeast two-hybrid screening of an E18 mouse whole-body cDNA library with CLP36 as the bait and isolated palladin as a CLP36-binding protein. Palladin is an actin-binding protein and it was shown to have a role in the extension of cortical neurons. A coimmunoprecipitation study showed that CLP36 and palladin formed a complex in the dorsal root ganglion (DRG). In addition, CLP36 and palladin were colocalized in the neurites and cell bodies of primary DRG neurons. Furthermore, sciatic nerve transection increased the expression of both CLP36 and palladin mRNAs in DRG neurons, with the increase in CLP36 mRNA being more prominent. This implies that CLP36 has a more specific role in nerve regeneration than palladin. Our results suggest that CLP36 may interact with palladin to influence neurite outgrowth during sciatic nerve regeneration.     

Endothelial cell influence on dorsal root ganglion cell formation

Although endothelial cells are known to produce neutrotrophic factors, endothelial cell influence on growth and survival of ganglion cells has not been documented. For this reason, a long-term culture technique was modified to obtain dorsal root ganglion (DRG) cells. Cells, among them neurons, were released from clusters into the medium for more than four weeks. These cells were grown together with endothelial cells either (1) in close contact as contiguous co-culture, or (2) on porous inserts for non-contiguous co-culture, and, finally, (3) without endothelial cells for ganglion cell culture. Samples from the cultures were stained for the nuclear Ki-67-antigen to detect proliferating cells, and for neurofilaments (NF) to verify the presence of DRG cells with and without mitotic figures. The contiguous co-culture contained three times as many mitotic DRG cells as other culture set ups. Nerve growth factor had no mitotic effect on the different DRG cultures, although it supported the growth of endothelial cells. It is concluded that a subpopulation of DRG cells is easily harvested from long-term DRG cultures. These DRG cells undergo mitosis when in direct contact with endothelial cells.