Levels Of p75NTR Research Articles

HoloTransferrin but Not ApoTransferrin Prevents Schwann Cell De-Differentiation in Culture

Schwann cells (SCs) in culture, without the presence of axons, become de-differentiated, reaching a condition similar to that of their precursor cells. The cytoplasmic accumulation of transferrin (Tf) in the myelinated peripheral nerve has been reported and data in the literature support a role for apoTf in myelination in the CNS. In the present report, we used SC cultures to evaluate the capacity of apoTf and holoTf to prevent cell de-differentiation promoted by fetal calf serum deprivation. SCs incubated in a serum-free medium showed a decrease in the expression of myelin basic protein (MBP) and P₀, markers of mature myelin-forming SCs, together with an increase in the levels of p75NTR and glial fibrillary acidic protein, markers of immature SCs. Treatment with holoTf prevented the decrease in expression of MBP and P₀ and the increase in p75NTR. ApoTf was unable to prevent these changes except when iron was added to the cultures. These results suggest a role for holoTf in the regulation of myelin formation by SCs.

Expression of p75 NTR in photoreceptor cells of dystrophic rat retinas

Although a gene mutation in the Royal College of Surgeons (RCS) dystrophic rat results in defective phagocytosis and in accumulation of debris in the subretinal space, the molecular mechanisms leading to photoreceptor cell death remain unclear. In this study, the expression of p75 NTR, the low-affinity neurotrophin receptor incriminated in the apoptosis of developing neurons, was investigated at various stages of retinal degeneration in dystrophic rats using immunohistochemistry, in situ reverse transcription polymerase chain reaction (RT-PCR), Western blot, and relative RT-PCR. In normal adult retinas, p75 NTR immunolabeling was observed mainly in the outer limiting membrane, with punctate labeling in the inner nuclear and ganglion cell layers. In 18- to 30-day-old dystrophic retinas, the immunostaining appeared to increase especially in the photoreceptor outer and inner segments. Dense staining was also observed in the retinal pigment epithelium (RPE) and choroid. In 60-day-old dystrophic rat retinas, the density of immunolabeling for p75 NTR increased dramatically in the remaining inner retina, especially in the inner nuclear, inner plexiform, and ganglion cell layers. Post-embedding immunogold labeling of normal retinas verified the distribution of p75 NTR in photoreceptor cells within the inner segments, cell bodies, and outer segments. The apparent increased intensity in p75 NTR immunostaining in dystrophic retinas was verified by Western blots and densitometric analyses. In situ RT-PCR and relative RT-PCR further established increased synthesis of p75 NTR in dystrophic retinas. The increased levels of p75 NTR in the RPE and photoreceptor cells, the initial sites of injury, during retinal degeneration in dystrophic rats strongly suggest that altered expression of p75 NTR may be directly involved in photoreceptor death.

Amyloid β Binds Trimers as Well as Monomers of the 75-kDa Neurotrophin Receptor and Activates Receptor Signaling

p75(NTR), a nerve growth factor co-receptor that has been implicated in apoptosis of neurons, is structurally related to Fas and the receptors for tumor necrosis factor-alpha that display ligand independent assembly into trimers. Using embryonic day 17 fetal rat cortical neurons and p75(NTR)-expressing NIH-3T3 cells, we now show that p75(NTR) exists as a trimer as well as a monomer. Furthermore, we have reported and others have confirmed that amyloid beta binds p75(NTR), and that this binding leads to apoptotic cell death. We now report that amyloid beta binds to trimers of p75(NTR) as well as to p75(NTR) monomers but not to the p140(trkA), the nerve growth factor co-receptor that mediates neuronal survival. Furthermore, amyloid beta activates p75(NTR), strongly inducing the transcription of c-Jun mRNA and stimulating the stress-activated c-Jun NH(2)-terminal kinase, as measured by phosphorylation of its substrate (glutathione S-transferase-c-Jun-(1-79)). Our data suggest that p75(NTR) may be present as a preformed trimer that binds amyloid beta to induce receptor activation, and support the hypothesis that p75(NTR) activation by amyloid beta is causally related to Alzheimer's disease.

Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts

A reduced ability to regenerate peripheral axons may be partly responsible for diabetic neuropathy. The source of the impairment has not been narrowed down to axonal or Schwann cell failure. We used nerve grafts from control or diabetic donor rats transplanted into control or diabetic hosts to pursue this differential diagnosis. An isograft between the left sciatic nerves of inbred Lewis rats was performed 8 weeks after STZ treatment and on age-matched controls. The nerve exchanges were control–control, control–diabetic, diabetic–control and diabetic–diabetic. At postsurgical day 14, nerves were excised and analysed for levels of axonal markers, total and phosphorylated neurofilament, and Schwann cell receptors, ErbB2 and p75 NTR, using immunohistochemistry and Western blotting. The aim was to measure ingress of axonal markers into the graft and judge the appropriateness of Schwann cell phenotype changes. Transfer of nerve from diabetic to control rats resulted in a doubling in neurofilament, both phosphorylated and nonphosphorylated (both P<0.05). ErbB2 was decreased in grafts from diabetic rats (53% of control, P<0.05) and p75 NTR levels were increased in both types of graft in diabetic rats (to 300–400% of controls, P<0.05). Schwann cells in diabetic nerve grafts showed receptor levels more similar to controls when placed into a normal environment and the converse also appeared to hold. TUNEL staining revealed increased apoptosis in diabetic nerve distal to the graft. The data show that alterations in Schwann cell phenotype in diabetes are reversed by transfer to control rats and develop in normal nerve after transfer to a diabetic host.

Dexamethasone Induces TrkA and p75NTR Immunoreactivity in the Cerebral Cortex and Hippocampus

Nerve growth factor (NGF) plays a crucial role in synaptic plasticity during brain development and adulthood by activating a dual receptor system composed of TrkA and p75 (p75NTR) receptors. Exogenous NGF modulates the expression of both receptors. Little is known about the ability of endogenous NGF to regulate the expression of these receptors in basal forebrain cholinergic terminals. The ability of glucocorticoids to increase NGF expression in the hippocampus prompted us to investigate whether the synthetic glucocorticoid dexamethasone (DEX) increases TrkA and p75NTR expression in NGF-target cholinergic neurons in developing rats. We first examined the effect of DEX on NGF mRNA by in situ hybridization. DEX given systemically (0.5 mg/kg, sc) for 1 week to 7-day-old rats elicited an increase in NGF mRNA levels in the dentate gyrus of the hippocampus and superficial layers II and III of the cerebral cortex. Immunohistochemical analysis of p75NTR and TrkA levels revealed a dramatic increase in p75NTR immunoreactivity (IR) in both basal forebrain and hippocampus and TrkA IR in the hippocampus. Interestingly, in DEX-treated rats more axonal terminals were immunopositive for p75NTR in the hippocampus and cortex, suggesting an increase in p75NTR IR in cell bodies as well as in terminals. Our data indicate that the endogenously produced NGF elicits biological changes similar to those of the exogenously delivered NGF. We suggest that glucocorticoids might regulate and coordinate cholinergic neuronal maturation by increasing the biosynthesis of NGF.

Expression of p75 NTR, trkB and trkC in nonmanipulated and axotomized motoneurons of aged rats

Several lines of evidence indicate that adult neurons remain dependent on neurotrophins and that changes in tissue expression of neurotrophins and/or their receptors may play a role in senile neurodegeneration. We have studied the expression of p75 NTR, trkB and trkC, respectively, in lumbar motoneurons of young adult (2–3 months) and aged (30 months) rats subjected to sciatic transection using in situ hybridization and immunohistochemistry. Nonmanipulated age-matched animals were processed in parallel. In nonmanipulated aged rats, high levels of p75 NTR could be seen in a number of motoneurons (10–15%), while in young adult animals no p75 NTR could be detected. Seven days following sciatic axotomy, a conspicuous ipsilateral upregulation p75 NTR was observed in young adult rats. Also in aged rats there was a marked ipsilateral increase in number of p75 NTR expressing neurons (≈100%). In comparison to young adult rats, aged rats showed a decreased expression of both trkB (5/6 animals) and trkC (6/6 animals). Furthermore, in response to sciatic transection, 3 out of 5 aged rats did not show an increased expression of trkB. In aged rats, axotomy did not induce any significant change in trkC expression. In the young adult rats, we recorded a side-to-side effect with lower values ipsilaterally, however, it cannot be excluded that this difference was caused by an upregulation in the contralateral motoneurons. Oligonucleotide probes against BDNF and NT3 mRNA showed only very few faintly positive neurons in both age groups. Our results indicate that the pattern of regulatory changes of NT receptors in response to axotomy is different in aged and young adult rats. The lack of covariation between p75 NTR and trkB and trkC regulation in aged rats indicates a changed role for p75 NTR in senescent motoneurons.

Level of p75 receptor expression in sensory ganglia is modulated by NGF level in the target tissue.

Neurotrophins play an essential role in sensory development by providing trophic support to neurons that innervate peripheral targets. Nerve growth factor (NGF), neurotrophin-3, neurotrophin-4, and brain-derived neurotrophin exert their survival effect by binding to two transmembrane receptor types: trk receptors, which exhibit binding specificity, and the p75NTR receptor, which binds all neurotrophins. To determine how target-derived neurotrophins affect sensory neuron development and function, we used transgenic mice that overexpress NGF in the skin to examine the impact of NGF overexpression on receptor expression. Previous studies of trk expression in trigeminal ganglia of adult NGF transgenics showed that the percentage of trkA neurons doubled and their number increased fivefold. The present study focused on the p75 receptor and shows that the percentage of neurons expressing p75NTR also increase in NGF ganglia, but only by 10%. This increase did not encompass the small, BS-IB-4 isolectin-positive cells as they remained p75 negative in transgenic ganglia. Interestingly, levels of trkA protein were not increased on a per-cell level, whereas levels of p75NTR increased nearly threefold. These results show that in sensory systems, target-derived NGF modulates the level of p75NTR receptor expression, and in so doing, may act to regulate the formation of functional receptor complexes and subsequent trophic action.

P75NTR andTrk receptors are expressed in reciprocal patterns in a wide variety of non-neural tissues during rat embryonic development, indicating independent receptor functions

The p75 kDa neurotrophin receptor (p75NTR) has been detected in a number of non-neural tissues, especially during development. Reports of Trk receptor transcripts in non-neural tissues raise the possibility that the sites of p75NTR expression during development may correlate with Trk receptor expression. Coexpression of p75NTR with the Trk receptors in developing non-neural tissues would support the hypothesis that there is a cooperative function between the two receptor subclasses. To address these questions, p75NTR was localized relative to the three known Trk receptors in adjacent sections of rat embryos at stages of development when the highest levels of p75NTR have been observed in the muscle, maxillary pad, kidney, and lung. Using in situ hybridization and immunhistochemical analyses, we show here that the Trk receptors are expressed extensively in non-neural tissues during cell differentiation and tissue morphogenesis but in patterns that are generally reciprocal to that of p75NTR. The results indicate p75NTR most likely functions independently of the Trk receptors in most developing non-neural tissues. However, the p75NTR consistently appears in non-neural cells adjacent to those expressing Trk receptors. The reciprocal patterns of expression indicate that the separate activities of the two receptors most likely complement each other in regulating cell-cell interactions important for the innervation of developing non-neural tissues.

Binding of beta-amyloid to the p75 neurotrophin receptor induces apoptosis. A possible mechanism for Alzheimer's disease.

Alzheimer's disease is a neurodegenerative disorder characterized by the extracellular deposition in the brain of aggregated beta-amyloid peptide, presumed to play a pathogenic role, and by preferential loss of neurons that express the 75-kD neurotrophin receptor (p75NTR). Using rat cortical neurons and NIH-3T3 cell line engineered to stably express p75NTR, we find that the beta-amyloid peptide specifically binds the p75NTR. Furthermore, 3T3 cells expressing p75NTR, but not wild-type control cells lacking the receptor, undergo apoptosis in the presence of aggregated beta-amyloid. Normal neural crest-derived melanocytes that express physiologic levels of p75NTR undergo apoptosis in the presence of aggregated beta-amyloid, but not in the presence of control peptide synthesized in reverse. These data imply that neuronal death in Alzheimer's disease is mediated, at least in part, by the interaction of beta-amyloid with p75NTR, and suggest new targets for therapeutic intervention.

Induction of apoptosis by p75 neurotrophin receptor in human neuroblastoma cells.

The low-affinity nerve growth factor receptor p75NTR belongs to a membrane receptor superfamily whose members, in certain cell types, are able to transduce an apoptotic signal. To investigate the effect of p75NTR expression in neuroblastoma cells, we transfected the p75NTR cDNA into SK-N-BE cells, a neuroblastoma cell line that lacks expression of both p75NTR and TrkA. Cell clones expressing elevated levels of p75NTR showed a high degree of cell death by apoptosis, even in serum-supplemented medium. Moreover, the level of apoptosis correlated directly with the expression level of the receptor, indicating that p75NTR could activate the cell death program by itself. Clones expressing p75NTR showed a dramatic increase of cell death when switched into serum-free medium; these cultures rapidly extinguished. This apoptotic effect was greatly inhibited by NGF treatment. Our results support the hypothesis that p75NTR, when it is not bound by NGF, may play a role in neuronal selection during embryonic development and suggest that neuroblastomas may arise from immature neuroblasts that escape programmed cell death. Therefore, the loss of p75NTR expression in developing neural crest cells might be a primary event in the genesis of neuroblastoma.

Schwann cells responding to primary demyelination in vivo express p75NTR and c-erbB receptors: a light and electron immunohistochemical study.

We have used quantitative and qualitative light microscope immunohistochemistry to examine the expression of p75NTR and c-erbB receptors in Schwann cells in a demyelinating lesion induced by the intraneural injection of lysophosphatidyl choline (LPC). We report that levels of p75NTR, c-erbB2 and c-erbB4, as assessed using image analysis of immuno-peroxidase labelled sections, and c-erbB3, as assessed by eye, increased within each lesion site soon after the initiation of myelinolysis, peaked between 5 and 8 days after induction of demyelination and fell to undetectable levels at the onset of remyelination. Pre-embedding immunoelectron microscopy confirmed that Schwann cells ensheathing demyelinated axons were p75NTR positive. Immunolabel decorated overlapping processes of neighbouring Schwann cells, suggesting that in this context p75NTR could play a role in juxtacrine signalling between reacting cells. We conclude that upregulation of p75NTR and c-erbB receptors is a constitutive Schwann cell response to an acute disruption of the axon-Schwann cell relationship.

Evidence that the p75 neurotrophin receptor mediates perineural spread of desmoplastic melanoma

Background: The desmoplastic variant of melanoma has a striking propensity for neurotropism. This neurotropism may reflect Schwannian differentiation. The migration of Schwann cells along embryonic nerves is reportedly regulated by the 75 kd neurotrophin receptor (p75 NTR) and one of its cognate ligands, nerve growth factor (NGF). Objective: The purpose of this study was to test the hypothesis that the mechanism of perineural spread in desmoplastic melanomas is analogous to that of neurotropism in Schwann cells, which apparently depends on expression of p75 neurotrophin receptor and its ligands. Methods: Immunolabeling of p75 NTR in histologic sections of spindle cell and desmoplastic melanomas was compared and contrasted with that of epithelioid melanomas. Results: The histologic material consisted of spindle cell melanoma specimens from 11 patients, of which seven exhibited features of desmoplastic melanoma. All spindle cell melanoma specimens expressed the p75 NTR in at least 10% of the cells, and most expressed p75 NTR in more than 50% of cells. In contrast, 10 of 11 control melanomas of conventional epithelioid phenotype expressed lower levels of p75 NTR (0% to 10% of cells). Conclusion: There is a strong correlation between expression of p75 NTR and the desmoplastic phenotype, supporting the hypothesis that the propensity of these melanomas for neurotropism involves p75 NTR and its ligands. Immunolabeling for p75 NTR may be a useful marker for the neurotropic phenotype, as well as for detecting perineural spread in histologic sections of melanomas.