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1.
The myelin-associated glycoprotein (MAG) has been postulated to play a crucial role during myelin formation. Evidence supporting this hypothesis was provided by infecting rat Schwann cells with a retrovirus expressing MAG antisense RNA; these Schwann cells showed reduced levels of MAG expression and failed to myelinate DRG neurons in vitro. However, when MAG expression was disrupted by generating MAG-deficient mice, normal myelin sheaths were formed in peripheral nerves in vivo. In the present study we investigated whether myelination is compromised in MAG-deficient Schwann cells in vitro, i.e., under similar conditions where Schwann cells expressing MAG antisense RNA failed to myelinate. We show that MAG-deficient Schwann cells do myelinate DRG neurons in vitro and express the myelin-specific glycolipid galactocerebroside (Gal-C) and the myelin proteins P0 and MBP. Furthermore, myelin sheaths appear morphologically normal with both compacted and uncompacted aspects when investigated by electron microscopy. Quantitative analysis revealed that the number of myelin sheaths was similar in cultures from MAG-deficient and wild-type mice. These findings support the view that MAG is not essential for myelin formation in the PNS. GLIA 22:213–220, 1998. © 1998 Wiley-Liss, Inc. 相似文献
2.
Robert F Guennoun R Désarnaud F Do-Thi A Benmessahel Y Baulieu EE Schumacher M 《The European journal of neuroscience》2001,13(5):916-924
In peripheral nerves, progesterone synthesized by Schwann cells has been implicated in myelination. In spite of such an important function, little is known of the regulation of progesterone biosynthesis in the nervous system. We show here that in rat Schwann cells, expression of the 3 beta-hydroxysteroid dehydrogenase and formation of progesterone are dependent on neuronal signal. Levels of 3 beta-hydroxysteroid dehydrogenase mRNA and synthesis of [3H]progesterone from [3H]pregnenolone were low in purified Schwann cells prepared from neonatal rat sciatic nerves. However, when Schwann cells were cultured in contact with sensory neurons, both expression and activity of the 3 beta-hydroxysteroid dehydrogenase were induced. Regulation of 3 beta-hydroxysteroid dehydrogenase expression by neurons was also demonstrated in vivo in the rat sciatic nerve. 3 beta-hydroxysteroid dehydrogenase mRNA was present in the intact nerve, but could no longer be detected 3 or 6 days after cryolesion, when axons had degenerated. After 15 days, when Schwann cells made new contact with the regenerating axons, the enzyme was re-expressed. After nerve transection, which does not allow axonal regeneration, 3 beta-hydroxysteroid dehydrogenase mRNA remained undetectable. The regulation of 3 beta-hydroxysteroid dehydrogenase mRNA after lesion was similar to the regulation of myelin protein zero (P0) and peripheral myelin protein 22 (PMP22) mRNAs, supporting an important role of locally formed progesterone in myelination. 相似文献
3.
A technique for isolation of adult Schwann cells (ScC) from dorsal root ganglia (DRG) is described. Decapsulated DRG explants embedded into type I collagen gels were cultured for 3 days in serum-free medium during which ScC migrated from the explant. These explants were then grown in serum-supplemented medium to allow ScC proliferation. On day 10 the number of ScC isolated from DRG explants per mouse was about 2.5 × 105, and the purity was greater than 95%. This culture system provided sufficient numbers of highly purified adult ScC in a shorter culture period (2–3 times) than other methods. We used ScC from this method to determine the age-related changes in attachment, growth, and survival of ScC cultured in serum-free medium. The attachment capacity of adult ScC on type I collagen or polylysine was similar to that of newborn ScC. However, the collagen promoted growth and survival of adult ScC but not that of neonatal ScC, indicating age-related differences of ScC properties in vitro. © 1993 Wiley-Liss, Inc. 相似文献
4.
Daniel M. Ventre Avery Cluff Christian Gagnon David Diaz Vera Ryan A. Koppes Abigail N. Koppes 《Journal of neuroscience research》2021,99(1):374-391
Satisfactory treatment of peripheral nerve injury (PNI) faces difficulties owing to the intrinsic biological barriers in larger injuries and invasive surgical interventions. Injury gaps >3 cm have low chances of full motor and sensory recovery, and the unmet need for PNI repair techniques which increase the likelihood of functional recovery while limiting invasiveness motivate this work. Building upon prior work in ultrasound stimulation (US) of dorsal root ganglion (DRG) neurons, the effects of US on DRG neuron and Schwann cell (SC) cocultures were investigated to uncover the role of SCs in mediating the neuronal response to US in vitro. Acoustic intensity‐dependent alteration in selected neuromorphometrics of DRG neurons in coculture with SCs was observed in total outgrowth, primary neurites, and length compared to previously reported DRG monoculture in a calcium‐independent manner. SC viability and proliferation were not impacted by US. Conditioned medium studies suggest secreted factors from SCs subjected to US impact DRG neuron morphology. These findings advance the current understanding of mechanisms by which these cell types respond to US, which may lead to new noninvasive US therapies for treating PNI. 相似文献
5.
Dynamic changes in glypican-1 expression in dorsal root ganglion neurons after peripheral and central axonal injury 总被引:2,自引:0,他引:2
Glypican-1, a glycosyl phosphatidyl inositol (GPI)-anchored heparan sulphate proteoglycan expressed in the developing and mature cells of the central nervous system, acts as a coreceptor for diverse ligands, including slit axonal guidance proteins, fibroblast growth factors and laminin. We have examined its expression in primary sensory dorsal root ganglion (DRG) neurons and spinal cord after axonal injury. In noninjured rats, glypican-1 mRNA and protein are constitutively expressed at low levels in lumbar DRGs. Sciatic nerve transection results in a two-fold increase in mRNA and protein expression. High glypican-1 expression persists until the injured axons reinnervate their peripheral targets, as in the case of a crushed nerve. Injury to the central axons of DRG neurons by either a dorsal column injury or a dorsal root transection also up-regulates glypican-1, a feature that differs from most DRG axonal injury-induced genes, whose regulation changes only after peripheral and not central axonal injury. After axonal injury, the cellular localization of glypican-1 changes from a nuclear pattern restricted to neurons in noninjured DRGs, to the cytoplasm and membrane of injured neurons, as well as neighbouring non-neuronal cells. Sciatic nerve transection also leads to an accumulation of glypican-1 in the proximal nerve segment of injured axons. Glypican-1 is coexpressed with robo 2 and its up-regulation after axonal injury may contribute to an altered sensitivity to axonal growth or guidance cues. 相似文献
6.
The aim of these experiments was to determine the state of maturity of dorsal root ganglia and axons in opossums (Monodelphis domestica) at birth and to assess quantitatively changes that occur in early life. Counts made of dorsal root ganglion cells at cervical levels showed that the numbers were similar in newborn and adult animals, approximately 1,600 per ganglion. In cervical dorsal root ganglia of newborn animals, division of neuronal precursors cells had ceased. The number of axons in cervical dorsal roots was similar in newborn and adult animals (about 4,500). For each ganglion cell body, approximately three axons were counted in the dorsal root. At birth, dorsal roots contained several bundles about 30 μm in diameter consisting of small axons (0.05–2 μm in diameter). A few non-neural cells were identified as Schwann cell perikarya, each enclosing a number of neurites. Later, marked changes occurred in Schwann cells and in their relationship to axons in the roots. Thus, at 12 days, an increase occurred in the number of Schwann cells and fibroblasts, and the bundles had enlarged to about 80 μm with little increase in axon diameter (0.1–2 μm). By 18 days, the bundles were larger, and myelination had already started. At 23 days, the dorsal root contained more than 500 myelinated axons that could reach 5 μm in diameter. The adult dorsal root enclosed about 900 myelinated axons. Throughout this time, the relationship between the Schwann cells and axons changed. Together, these results indicate that the number of axons and cell bodies of sensory dorsal root ganglia in opossum do not show major changes after birth. In addition, these results set the stage for quantitative studies of regeneration of dorsal column fibers in injured neonatal opossum nervous system. J. Comp. Neurol. 396:338–350, 1998. © 1998 Wiley-Liss, Inc. 相似文献
7.
目的研究Nogo—A在成年正常大鼠脊髓和背根节的分布。方法免疫组织化学方法(ABC法)和免疫荧光双标记法。结果正常成年大鼠的脊髓灰质分布有大量的Nogo—A免疫阳性的寡突胶质细胞、运动神经元和中间神经元,免疫阳性反应产物主要分布于细胞的胞体和部分突起中。Nogo—A广泛分布于穿行于脊髓白质的纤维包裹的髓鞘和轴突上。在脊髓前根、后根和坐骨神经的运动和感觉的有髓和无髓纤维也可观察到Nogo—A的表达。而背根神经节的神经元也大量表达Nogo—A,其强度由弱至强不等,广泛分布于大、中、小各类感觉神经元的胞质及突起中。结论Nogo—A在成年大鼠脊髓,背根神经节和外周神经纤维的广泛存在提示其在正常状态下的神经功能中可能起重要作用。 相似文献
8.
Calbindin-immunoreactive sensory neurons of dorsal root ganglion project to skeletal muscle in the chick 总被引:1,自引:0,他引:1
In the chicken dorsal root ganglia, two neuronal subpopulations referred to as A1 and B1 share in common an immunoreactivity to antisera raised to calbindin D-28k but are distinguished by their cytological and ultrastructural characteristics. To determine the peripheral targets innervated by calbindin-immunoreactive neurons in lumbosacral dorsal root ganglia, cryostat sections of various hindlimb tissues were treated with anticalbindin antisera. Calbindin-immunostained axons were clearly detected in skeletal muscle. Large myelinated nerve fibres and afferent axon terminals in neuromuscular spindles were calbindin-immunoreactive; thin unmyelinated nerve fibres were also immunostained in nerve bundles of the perimysium. Since motoneurons and neurons of the autonomic nervous system were devoid of calbindin immunostaining, it was suggested that the immunoreactive axons found in skeletal muscle originate from sensory neurons expressing a calbindin immunoreaction in the dorsal root ganglia. This hypothesis was corroborated after introduction of wheat germ agglutinin coupled with horseradish peroxidase or colloidal gold particles into the sartorius muscle. The retrogradely transported tracer was collected only in ganglion cell bodies which displayed the ultrastructural characteristics of A1 and B1 sensory neurons. On the basis of calbindin immunoreaction and of tracer retrograde transport, it is concluded that ganglion cells of subclasses A1 and B1 contribute to the sensory innervation of skeletal muscle in the chicken. 相似文献
9.
Experiments were undertaken to examine anatomical correlates of physiological effects of rabbit sera raised against nerve growth factor (anti-NGF) on nociceptive afferents. This antiserum has been shown to deplete the population of A-δ high threshold mechanoreceptors and to reduce neurogenic vasodilatation. Because numerous studies implicate calcitonin gene related peptide (CGRP)- containing sensory neurons in these effects, immunocytochemical and anatomical techniques were used to examine the normal development of CGRP-immunoreactive (-IR) neurons in the dorsal root ganglion (DRG) of rats from 13 days to 19 weeks of age, and to compare this to the development in rats treated neonatally (postnatal days 2-14) with anti-NGF. In controls the rate of increase in the mean diameter of CGRP-IR cells was substantially greater between 13 days and 5 weeks of age than it was between 5 weeks and 19 weeks, in contrast to CGRP-negative neurons whose rate of growth remained relatively constant. Anti-NGF had no significant effect on growth rate, but rats treated with anti-NGF exhibited a reduced proportion of CGRP-IR neurons at 5 weeks. This deficit was reversed by 19 weeks unlike the physiological changes. These results indicate independent regulation of CGRP expression and nociceptor physiology by NGF. J. Comp. Neurol. 392: 489–498, 1998. © 1998 Wiley-Liss, Inc. 相似文献
10.
The distribution of intramembranous particles within the axolemma of cultured dorsal root ganglion neurons was determined by freeze-fracture microscopy. Utilizing culture conditions which eliminate Schwann cells, the particle distribution of the P-face, 735 +/- 119 microns2, and E-face, 100 +/- 39 microns2 resembled that of pre- and non-myelinated axons in vivo and no node-like E-face particle patching was seen. These results indicate that cultured neurite development is similar to that seen in vivo and that axons maintained in a glial-free environment do not develop nodal, E-face membrane specializations. 相似文献
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13.
Warm cells were identified by Fura-PE3-based microfluorimetry of Ca2+ in cultured dorsal root ganglion (DRG) neurons. In response to a physiologically relevant stimulus temperature (43°C), a subpopulation of small DRG neurons from new born rats increased the intracellular Ca2+ concentration ([Ca2+]i). Seven percent of the cells responded to the warm stimulus. The stimulus evoked elevation in [Ca2+]i from 52.5±9.5 nM (mean±S.D., n=18) to 171.0±15.6 nM in cells between 15 and 25 μm in diameter. The depletion of extracellular Ca2+ diminished the Ca2+ elevation. The Na+-free condition also diminished the response. We concluded that the heat stimulation opens nonselective cation channels in putative warm cells from DRG neurons. 相似文献
14.
Effect of nerve crush on perikaryal number and volume of neurons in adult rat dorsal root ganglion. 总被引:5,自引:0,他引:5
Assumption-free stereological methods were applied to assess the effect of nerve crush on perikaryal number and mean volume of neuronal subpopulations in adult rat dorsal root ganglion (DRG). The L5 spinal nerve of 20 Wistar rats was crushed approximately 7 mm distal to the DRG, and the contralateral spinal nerve and DRG were left intact and used as controls. After four, 15, 45, and 120 days, the rats were killed, and the tissue was fixed and processed for subsequent preparation of 30-microm-thick sections. Estimates of neuron number were obtained with the optical fractionator technique and estimates of the mean perikaryal volume with the vertical planar rotator principle. Perikaryal loss was progressive during the early study period but stabilized 45 days after nerve injury. The mean number (n) of all neurons in intact L5 DRG was 16,400 (S.D. = 2,000). The loss of perikarya was 16% (P < 0.05) after four days, 15% (P < 0.05) after 15 days, 30% (P = 0.059) after 45 days, and 34% (P < 0. 05) after 120 days. B cells were lost at an earlier time than were A cells, and the B cell loss was more pronounced (39% vs. 22%, respectively, after 120 days). For A cells, the mean perikaryal volume was initially reduced but was normalized at the end of the study. Distributions of perikaryal volume showed that the curves of both A and B cells were uniformly displaced toward smaller values 15 and 45 days after injury. Neuronal loss caused by crush seems similar to that seen in rats exposed to permanent axotomy (Vestergaard et al. [1997] J Comp Neurol 388:307-312) at the same location, indicating that survival of perikarya is not dependent on possibility for fiber growth. 相似文献
15.
Serum from patients with the acute form of the Guillain-Barré syndrome was applied to cultures containing only rat dorsal root ganglion neurons and Schwann cells. Serum taken from 4 of 10 patients during the first 1-3 weeks of clinical onset had previously been shown to have significant demyelinating activity in this culture system when observed at the light microscopic level. More detailed assessment made at the ultrastructural level showed that: (1) wide-spread myelin-related Schwann cell lysis occurred in concert with vesicular myelin breakdown; (2) non-myelin-related Schwann cells avidly phagocytized necrotic cell debris and fragments of compact myelin; and (3) neurites and non-myelin-related Schwann cells remained structurally undamaged. Cultures treated with convalescent phase serum from patients whose acute phase serum had cytolytic activity displayed no significant ultrastructural damage to either neurites or Schwann cells. This is the first electron microscopic study to provide direct evidence that acute Guillain-Barré serum can be cytolytic for myelin-related Schwann cells and peripheral myelin in an experimental setting free of leukocytes, lymphocytes and mononuclear phagocytes. 相似文献
16.
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
17.
Studies have shown that retreatment of the distal stoma after nerve grafting can stimulate nerve regeneration. The present study attempted to verify the effects of reanastomosis of the distal stoma, after nerve grafting, on nerve regeneration by assessing brain-derived neurotrophic factor expression in 2-month-old rats. Results showed that brain-derived neurotrophic factor expression in L 2-4 dorsal root ganglia began to increase 3 days after autologous nerve grafting post sciatic nerve injury, peaked at 14 days, decreased at 28 days, and reached similar levels to the sham-surgery group at 56 days. Brain-derived neurotrophic factor expression in L 2-4 dorsal root ganglia began to increase 3 days after reanastomosis of the distal stoma, 59 days after autologous nerve grafting post sciatic nerve injury, significantly increased at 63 days, peaked at 70 days, and gradually decreased thereafter, but remained higher compared with the sham-surgery group up to 112 days. The results of this study indicate that reanastomosis of the distal stoma after orthotopic nerve grafting stimulated brain-derived neurotrophic factor expression in L 2-4 dorsal root ganglia. 相似文献
18.
Lehmann HC Köhne A Bernal F Jangouk P Meyer Zu Hörste G Dehmel T Hartung HP Previtali SC Kieseier BC 《Glia》2009,57(5):479-489
Matrix metalloproteinases (MMPs) comprise a large family of endopeptidases that are capable of degrading all extracellular matrix components. There is increasing evidence that MMPs are not only involved in tissue destruction but may also exert beneficial effects during axonal regeneration and nerve remyelination. Here, we provide evidence that MMP-2 (gelatinase A) is associated with the physiological process of myelination in the peripheral nervous system (PNS). In a myelinating co-culture model of Schwann cells and dorsal root ganglia neurons, MMP-2 expression correlated with the degree of myelination as determined by immunocytochemistry, zymography, and immunosorbent assay. Modulation of MMP-2 activity by chemical inhibitors led to incomplete and aberrant myelin formation. In vivo MMP-2 expression was detected in the cerebrospinal fluid (CSF) of patients with Guillain-Barré syndrome as well as in CSF and sural nerve biopsies of patients with chronic inflammatory demyelinating polyneuropathy. Our findings suggest an important, previously unrecognized role for MMP-2 during myelination in the PNS. Endogenous or exogenous modulation of MMP-2 activity may be a relevant target to enhance regeneration in demyelinating diseases of the PNS. 相似文献
19.
Schwann cells transiently express the transmembrane heparan sulfate proteoglycan syndecan-3 during the late embryonic and early postnatal periods of peripheral nerve development. Neonatal rat Schwann cells released soluble syndecan-3 into the culture medium by a process that was blocked by inhibition of endogenous matrix metalloproteinase activity. When Schwann cells were plated on a substratum that binds syndecan-3, the released proteoglycan bound to the substratum adjacent to the cell border. Membrane-anchored syndecan-3 was concentrated in actin-containing filopodia that projected from the lateral edges of the Schwann cell membrane. Membrane shedding was specific for syndecan-3 and was not observed for the related proteoglycan syndecan-1. Analysis of Schwann cells transfected with wild-type and chimeric syndecan-1 and syndecan-3 cDNAs revealed that membrane shedding was a property of the syndecan-3 ectodomain. Inhibition of syndecan-3 release significantly enhanced Schwann cell adhesion and process extension on dishes coated with the non-collagenous N-terminal domain of alpha4(V) collagen, which binds syndecan-3 and mediates heparan sulfate-dependent Schwann cell adhesion. Matrix metalloproteinase-dependent syndecan-3 shedding was also observed in newborn rat peripheral nerve tissue. Syndecan-3 shedding in peripheral nerve tissue was age specific, and was not observed during later stages of postnatal nerve development. These results demonstrate that Schwann cell syndecan-3 is subject to matrix metalloproteinase-dependent membrane processing, which modulates the biological function of this proteoglycan. 相似文献