Joe E. Springer

Nerve growth factor receptors in the central nervous system.

Nerve growth factor (NGF) is well known to be involved in the development, survival, and maintenance of sympathetic and neural crest-derived sensory neurons in the peripheral nervous system. Over the last 10-15 years, however, the role of NGF as a necessary trophic substrate for magnocellular cholinergic neurons in the central nervous system (CNS) has emerged. Because the trophic effects of NGF are initiated by its interaction with membrane-bound receptors, the characterization, localization, and function of these specific NGF receptors are essential to understanding the many actions of NGF. The first part of this review will summarize briefly the presence and possible role of NGF in the CNS, with the remainder of the review focusing on what is known about the receptor to NGF.

cDNA sequence and differential mRNA regulation of two forms of glial cell line-derived neurotrophic factor in Schwann cells and rat skeletal muscle

Total RNA from rat Schwann cells grown in culture and adult rat skeletal muscle was reverse transcribed, amplified for glial cell line-derived neurotrophic factor (GDNF) messenger RNA (mRNA) using the polymerase chain reaction (PCR), and the PCR products sequenced. Two forms of GDNF were detected in the PCR step, one of a predicted size (GDNF633) and a second smaller form missing a 78-base pair sequence (GDNF555). Sequence analysis demonstrated that GDNF633 is similar to the published sequence of GDNF differing only at three nucleotides. Southern and Northern blot analyses reveal that the two forms are probably derived from a single RNA species that is alternatively spliced. Interestingly, GDNF633 mRNA was found to be selectively upregulated in denervated rat skeletal muscle at 1-2 weeks following axotomy, providing evidence that the innervation status of the muscle may determine the expression profile of the two alternatively spliced forms. Given these findings, we suggest that GDNF may function as a target-derived trophic factor for neuronal populations innervating skeletal muscle, including sensory neurons and spinal cord motoneurons.

Spinal cord motoneurons express p75NGFR and p145trkB mRNA in amyotrophic lateral sclerosis

In the present study, in situ hybridization was used to examine the expression of nerve growth factor (NGF) receptor (p75NGFR), trk (p140trk) and trkB (p145trkB) mRNA in spinal cord sections from patients with amyotrophic lateral sclerosis (ALS). We report that the expression of p75NGFR and p145trkB mRNA is elevated in alpha motoneurons in ALS sections. However, p140trk mRNA was not expressed in either ALS or control sections.

Neurotrophic factor mRNA expression in dentate gyrus is increased following in vivo stimulation of the angular bundle.

Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are two structurally-related neurotrophins synthesized in dentate gyrus granule cells and pyramidal neurons of the hippocampal formation. These neurons receive excitatory glutamatergic afferents from the entorhinal cortex via the angular bundle/perforant path. In the present study, we tested whether electrophysiological stimulation of this glutamatergic pathway modifies NGF or BDNF messenger RNA (mRNA) expression in vivo. Within hours following brief trains of high frequency angular bundle stimulation, the levels of mRNA encoding both neurotrophins were increased exclusively in granule cells of the ipsilateral dentate gyrus. The increase in neurotrophic factor mRNA expression was found to be mediated through the N-methyl-D-aspartate (NMDA) glutamate receptor subtype, and occurred in the absence of seizure. These findings provide evidence that neurotrophic factor mRNA levels in the hippocampal formation are increased by direct activation of excitatory afferents originating in the entorhinal cortex. We suggest that the function of some neurotrophin-responsive neuronal populations may depend upon the integrity and activity of neurons in the entorhinal cortex, a population of neurons reported to be compromised in patients with Alzheimers disease.

Regulation of nerve growth factor mRNA in the hippocampal formation : effects of N-methyl-D-aspartate receptor activation

In the hippocampal formation, nerve growth factor (NGF) is produced in granule cells of the dentate gyrus and a few pyramidal cells of Ammons horn. Both neuronal populations express N-methyl-D-aspartate (NMDA) receptors and receive putative glutamatergic afferents originating in the entorhinal cortex and projecting via the perforant path. We report in this study that intra-hippocampal or intraventricular injections of NMDA increase NGF mRNA levels in dentate gyrus granule cells as determined using in situ hybridization histochemistry and a solution hybridization assay. NGF mRNA induction is detected within 2 h following NMDA treatment and returns to control levels within 24 h. This NMDA effect is dose-dependent and blocked by pretreatment with 2-amino-5-phosphonopentanoic acid, a competitive NMDA antagonist. Finally, the induction of NGF mRNA is observed in the absence of detectable neurotoxicity or seizure activity. We postulate that normal physiological events associated with the activation of hippocampal NMDA receptors may regulate mRNA expression of this neurotrophic factor.

Experimental rationale for the therapeutic use of neurotrophins in amyotrophic lateral sclerosis

Current therapeutic efforts to treat chronic and progressive neurodegenerative disease include, for the first time, attempts to regenerate affected nervous tissue using neurotrophic factors. The rationale for using trophic factors includes the understanding that they support neuronal survival and regrowth processes. The potential benefits of trophic factor therapy will be no more realized in the near future than in the treatment of amyotrophic lateral sclerosis (ALS). ALS is pathologically characterized by the selective degeneration of specific populations of cranial and spinal motoneurons. Evidence for the existence of factors that support motoneurons has come from studies demonstrating that motoneurons receive trophic influences from various tissues, both central and peripheral, within their local environment. Although the identity of these putative tissue-derived factors has remained enigmatic, recent studies have demonstrated that several previously characterized trophic factors exhibit trophic influences on motoneurons. Among these are several members of the neurotrophin family, most notably brain-derived neurotrophic factor. These neurotrophins meet most of the criteria to be considered motoneuron trophic factors: they are locally available to motoneurons in vivo; motoneurons express specific receptors for these factors; and exogenous application of these factors mimicks the effects of the uncharacterized endogenous agents. The clinical use of these factors for the treatment of ALS, therefore, appears to be scientifically justified.

Neurotrophic factor mRNA expression in dentate gyrus is increased following angular bundle transection

In the central nervous system, the highest levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNA are found in the hippocampal formation. In the present study, we report that a unilateral transection of the angular bundle, which relays cortical information via the entorhinal cortex to the hippocampal formation, increases NGF and BDNF mRNA in the ipsilateral dentate gyrus. Within 4 hours following transection, the hybridization signal for NGF and BDNF mRNA increases in stratum granulosum 3- and 5-fold, respectively, compared to control levels. This lesion-induced increase of both mRNA returns to control levels within 24 hours and is maintained for at least 5 days. The induction is not prevented by pretreatment with AP-5, CNQX, or cholinergic denervation due to transection of the fimbria-fornix. Finally, the induction of neurotrophin mRNA is preceded by an increase in c-fos mRNA. These results provide evidence that transection of the cortical input to the hippocampal formation upregulates NGF and BDNF mRNA selectively in stratum granulosum. We suggest that the increased expression of NGF and BDNF mRNA may be an early step in the synaptic rearrangement of neurotrophin responsive cholinergic afferents observed following damage to the entorhinal cortex.

Co-grafts of embryonic dopamine neurons and adult sciatic nerve into the denervated striatum enhance behavioral and morphological recovery in rats.

We have recently demonstrated that a diffusible factor(s) derived from explanted adult rat sciatic nerve can increase the number and neurite outgrowth of embryonic rat dopamine (DA) neurons in culture. The present study extends this finding to compare DA neuron-sciatic nerve co-grafts to grafts of DA-rich neural tissue alone for behavioral and morphological effects in rats with unilateral nigrostriatal lesions of the DA pathway. Our results indicate that the presence of a co-grafted segment of sciatic nerve increased the likelihood of rapid behavioral recovery and promoted complete recovery mediated by small grafts that yielded only modest behavioral changes in the absence of co-grafted nerve. These behavioral effects were accompanied by a modest increase in survival of grafted tyrosine hydroxylase-positive neurons in the striatum and a more pronounced increase in the area and density of striatal reinnervation provided by grafted DA neurons in co-grafted animals. This evidence supports the view that a diffusible product of explanted peripheral nerve acts as a growth-promoting factor for embryonic DA neurons and that the presence of this factor augments the behavioral efficacy of grafted DA neurons.