Spinal cord injury causes damage to ascending and descending tracts, as well as to local circuits, but relatively little is known about the effect of such injury on sensory neurons located within ...adjoining ganglia. We have therefore used immunocytochemistry for activating transcription factor‐3 (ATF3), a sensitive marker of axonal damage, in order to examine the effects of spinal cord injury in rats on dorsal root ganglion (DRG) neurons. A 50‐g static compression injury applied to the dorsal surface of the T12 thoracic spinal cord led to an up‐regulation of ATF3 that was maximal at 1 day and affected 12–14% of DRG neurons in ganglia caudal to the injury (T13–L3). A similar response was seen after a T12 hemisection that transected the dorsal columns except that compression injury, but not hemisection, also evoked ATF3 expression in ganglia just rostral to the injury (T10, T11). ATF3 was up‐regulated exclusively in DRG neurons that were of large diameter and immunoreactive for heavy neurofilament. Small‐diameter cells, including the population that binds the lectin Grifffonia simplicifolia IB4, did not express ATF3 immunoreactivity. A similar pattern of ATF3 expression was induced by dorsal rhizotomy. The data show for the first time that ATF3 is up‐regulated after spinal cord and dorsal root injury, but that this up‐regulation is confined to the large‐diameter cell population.
Abstract Reg-2 is a secreted protein that is expressed de novo in motoneurons, sympathetic neurons, and dorsal root ganglion (DRG) neurons after nerve injury and which can act as a Schwann cell ...mitogen. We now show that Reg-2 is also upregulated by DRG neurons in inflammation with a very unusual expression pattern. In a rat model of monoarthritis, Reg-2 immunoreactivity was detected in DRG neurons at 1 day, peaked at 3 days (in 11.6% of DRG neurons), and was still present at 10 days (in 5%). Expression was almost exclusively in the population of DRG neurons that expresses the purinoceptor P2X3 and binding sites for the lectin Griffonia simplicifolia IB4, and which is known to respond to glial cell line–derived neurotrophic factor (GDNF). Immunoreactivity was present in DRG cell bodies and central terminals in the dorsal horn of the spinal cord. In contrast, very little expression was seen in the nerve growth factor (NGF) responsive and substance P expressing population. However intrathecal delivery of GDNF did not induce Reg-2 expression, but leukemia inhibitory factor (LIF) had a dramatic effect, inducing Reg-2 immunoreactivity in 39% of DRG neurons and 62% of P2X3 cells. Changes in inflammation have previously been observed predominantly in the neuropeptide expressing, NGF responsive, DRG neurons. Our results show that changes also take place in the IB4 population, possibly driven by members of the LIF family of neuropoietic cytokines. In addition, the presence of Reg-2 in central axon terminals implicates Reg-2 as a possible modulator of second order dorsal horn cells.
Neuronal calcium sensor-1 (NCS-1) is a member of the EF-hand calcium-binding protein superfamily which has been implicated in the modulation of a number of neuronal functions. In this study we have ...examined the expression of NCS-1 in adult rat dorsal root ganglion (DRG) neurons. NCS-1 immunoreactivity was present in most DRG neurons, including many calcitonin gene-related peptide (CGRP) expressing ones. NCS-1 showed some colocalization with the synaptic vesicle protein synaptophysin and underwent both anterograde and retrograde axonal transport. NCS-1 immunoreactivity was also present in the dorsal horn of the spinal cord, and in peripheral cutaneous terminals innervating blood vessels, where it was coexpressed with CGRP. In addition, NCS-1 in peripheral nerves was concentrated at nodes and adjoining paranodes. These results suggest novel roles for NCS-1, particularly in relation to channel function at nodes and to the peripheral release of vasoactive peptides.
The effects of glial cell line-derived neurotrophic factor on axonal outgrowth and apoptosis were studied
in vitro using explanted dorsal root ganglia-peripheral nerve preparations of adult mice. In ...gels of matrigel or collagen type 1, glial cell line-derived neurotrophic factor increased both the numbers and lengths of axons growing out of explanted preparations, although less effectively than nerve growth factor. Stimulation of axonal outgrowth by glial cell line-derived neurotrophic factor was unaffected by K252a, a protein kinase inhibitor which blocks the effects of nerve growth factor and other neurotrophins acting through trk receptors. To determine the phenotype of the axons responding to glial cell line-derived neurotrophic factor, preparations were stained using antibodies to trkA, calcitonin gene-related peptide, 200,000 mol. wt phosphorylated neurofilaments (monoclonal antibody RT97) and the lectin
Bandeiraea simplicifolia 1B4. RT97 recognizes large diameter neurons whilst 1B4 labels small diameter neurons which broadly do not express neurotrophin receptors. In preparations cultured with glial cell line-derived neurotrophic factor, significant increases in the numbers of outgrowing axons labelled with RT97 and 1B4 were observed but the numbers of calcitonin gene-related peptide-positive axons were not significantly increased and their staining intensity was generally faint. In separate preparations it was found that in the presence of glial cell line-derived neurotrophic factor, the majority of the 1B4 labelled axons were trkA negative, indicating that this factor can stimulate axonal growth in this population of neurons which do not respond to the neurotrophins.
Spontaneous apoptosis in neurons and satellite cells occurs in explanted preparations of the type used in the present investigations, but in cryostat sections of preparations cultured in the presence of glial cell line-derived neurotrophic factor, the incidence of apoptosis was lower than in control preparations which had been cultured in the absence of this factor. This suggests that glial cell line-derived neurotrophic factor may promote survival of some adult sensory neurons
in vitro.
Down syndrome (DS) is the most common cause of mental retardation. Many neural phenotypes are shared between DS individuals and DS mouse models; however, the common underlying molecular pathogenetic ...mechanisms remain unclear. Using a transchromosomic model of DS, we show that a 30%–60% reduced expression of Nrsf/Rest (a key regulator of pluripotency and neuronal differentiation) is an alteration that persists in trisomy 21 from undifferentiated embryonic stem (ES) cells to adult brain and is reproducible across several DS models. Using partially trisomic ES cells, we map this effect to a three-gene segment of HSA21, containing
DYRK1A. We independently identify the same locus as the most significant eQTL controlling
REST expression in the human genome. We show that specifically silencing the third copy of
DYRK1A rescues
Rest levels, and we demonstrate altered
Rest expression in response to inhibition of
DYRK1A expression or kinase activity, and in a transgenic
Dyrk1A mouse. We reveal that undifferentiated trisomy 21 ES cells show
DYRK1A-dose-sensitive reductions in levels of some pluripotency regulators, causing premature expression of transcription factors driving early endodermal and mesodermal differentiation, partially overlapping recently reported downstream effects of Rest +/−. They produce embryoid bodies with elevated levels of the primitive endoderm progenitor marker
Gata4 and a strongly reduced neuroectodermal progenitor compartment. Our results suggest that DYRK1A-mediated deregulation of
REST is a very early pathological consequence of trisomy 21 with potential to disturb the development of all embryonic lineages, warranting closer research into its contribution to DS pathology and new rationales for therapeutic approaches.
To determine whether perioperative antimicrobial prophylaxis would reduce incidence of postoperative infection among dogs undergoing elective orthopedic procedures.
Randomized, controlled, blinded, ...intention clinical trial.
Dogs of any breed, sex, or age undergoing elective orthopedic surgery at a veterinary teaching hospital.
Dogs were randomly assigned to 1 of 3 groups: treatment with saline solution, treatment with potassium penicillin G, and treatment with cefazolin. Treatments were intended to be administered within 30 minutes prior to surgery; a second dose was administered if surgery lasted > 90 minutes. Dogs were monitored for 10 to 14 days after surgery for evidence of infection.
After the first 112 dogs were enrolled in the study, it was found that infection rate for control dogs (5/32 dogs) was significantly higher than the rate for dogs treated with antimicrobials (3/80 dogs). Therefore, no more dogs were enrolled in the study. A total of 126 dogs completed the study. Monte Carlo simulations indicated that compared with dogs that received antimicrobials prophylactically, dogs that received saline solution developed infections significantly more frequently. Difference in efficacy, however, was not observed between the 2 antimicrobial drugs used.
Results indicated that perioperative antimicrobial prophylaxis decreased postoperative infection rate in dogs undergoing elective orthopedic surgery, compared with infection rate in control dogs. Cefazolin was not more efficacious than potassium penicillin G in these dogs.
The aim of this study was to determine whether axonal transport of activating transcription factor-2 (ATF2) occurs in adult sensory neurons, and whether this process is under neurotrophin control. ...Antisera to both total ATF2 and to the activated (i.e., phosphorylated) form were used for immunocytochemistry and Western blotting. ATF2 was localized to predominantly nociceptive dorsal root ganglion cells in adult rats and shown to accumulate proximal and distal to a sciatic nerve ligature as a result of axonal transport. Subcutaneous injection of nerve growth factor (NGF) decreased the levels of fast retrograde axonal transport of activated ATF2 by 97% (p < 0.05) and elevated levels of retrograde axonal transport of total ATF2 by twofold (p < 0.02). In contrast, blocking endogenous NGF using an anti-NGF antibody induced an elevation in retrograde axonal transport of activated ATF2 of 4. 5-fold (p < 0.05) and decreased retrograde axonal transport of total ATF2 by 72% (p < 0.05). NGF or anti-NGF treatment had no effect on the anterograde transport levels of total or activated ATF2. This study shows that signaling by target-derived NGF to the cell bodies of sensory neurons consists, in part, of the modulation of levels and activation status of a retrogradely transported transcription factor, ATF2.
Activating transcription factor 3 (ATF3) is a widely used marker of damaged primary sensory neurons that is induced in essentially all dorsal root ganglion (DRG) neurons by spinal nerve axotomy. ...Whether such injuries induce its expression in neurons of adjacent DRGs remains unknown. Following L5 spinal nerve ligation, experimental but not sham‐operated rats develop thermal and mechanical hypersensitivity. In the L4 DRG, 11–12% of neurons were ATF3 positive by 1 day post‐surgery, and numbers remain unchanged at 2 weeks. Importantly, sham exposure of the L5 spinal nerve produced a nearly identical number of ATF3‐positive neurons in the L4 DRG and also a substantial increase in the L5 DRG, with a similar time‐course to experimental animals. There was no correlation between behaviour and magnitude of ATF3 expression. Co‐localization studies with the DRG injury markers galanin, neuropeptide Y and nitric oxide synthase (NOS) showed that approximately 75, 50 and 25%, respectively, of L4 ATF3‐positive neurons co‐expressed these markers after L5 transection or sham surgery. Additionally, increases in galanin and NOS were seen in ATF3‐negative neurons in L4. Our results strongly suggest that the surgical exposure of spinal nerves induces ATF3 in the L4–5 DRG, irrespective of whether the L5 nerve is subsequently cut. This probably reflects minor damage to the neurons or their axons but nevertheless is sufficient to induce phenotypic plasticity. Caution is therefore warranted when interpreting the phenotypic plasticity of DRG neurons in adjacent ganglia in the absence of positive evidence that they are not damaged.
Several lines of evidence suggest that neurotrophin administration may be of some therapeutic benefit in the treatment of peripheral neuropathy. However, a third of sensory neurons do not express ...receptors for the neurotrophins. These neurons are of small diameter and can be identified by the binding of the lectin IB4 and the expression of the enzyme thiamine monophosphatase (TMP). Here we show that these neurons express the receptor components for glial-derived neurotrophic factor (GDNF) signaling (RET, GFRalpha-1, and GFRalpha-2). In lumbar dorsal root ganglia, virtually all IB4-labeled cells express RET mRNA, and the majority of these cells (79%) also express GFRalpha-1, GFRalpha-2, or GFRalpha-1 plus GFRalpha-2. GDNF, but not nerve growth factor (NGF), can prevent several axotomy-induced changes in these neurons, including the downregulation of IB4 binding, TMP activity, and somatostatin expression. GDNF also prevents the slowing of conduction velocity that normally occurs after axotomy in a population of small diameter DRG cells and the A-fiber sprouting into lamina II of the dorsal horn. GDNF therefore may be useful in the treatment of peripheral neuropathies and may protect peripheral neurons that are refractory to neurotrophin treatment.
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