Stories related to GDNF. GDNF and Parkinsons http://neuro-www.mgh.harvard.edu/neurowebforum/ParkinsonsDiseaseArticl es/GDNFandParkinsons.html This response submitted by Roblin NZ on 11/3/96. Author's Email: TAZ @ Wave.co.nz It was reported in Vol. 34 winter 1996 issue of the New Zealand Neurological Foundation newsletter HEADLINERS that Quote In 1993 a new protien called Glial cell line derived neurotrophic factor (GDNF) which has potent dopominotrophic activity the ability to stimulate the growth of the Dopomine neurons was described in the research Journal SCIENCE. Studies of GDNF have supported its neuron growth promoting effects in animal models which raises the qwuestion as to how optimistic one could be with regard to the therapeutic use of GDNF in Parkinsons. Recent studies using a model of PD in Rhesus monkeys have shown a long lasting stimulatory affect of GDNF on Dopamine neurons of the substantia nigra. More importantly the Parkinson like symtoms were all markedly decreased following the administration of a single dose of GDNF These studies indicate that clinical trials of GDNF as a treatment for PD are warranted and that a cure may be on the way. END OF QUOTE Next Article Previous Article Return to Main Article Article complete. Click HERE to return to the Neurology Web-Forum Menu. ---------------- the Cytokine Bulletin, Summer 1996 http://www.rndsystems.com/cb/cbsu96/cbsu96a4.html GDNF Signals through GDNFR-alpha and Ret Tyrosine Kinase Glial-cell-line-Derived Neurotrophic Factor (GDNF) is a distant member of the TGF-beta superfamily (1, 2). It is a growth and survival factor for neuronal cells, especially dopaminergic neurons (1), but its mechanism of action has been a mystery. Several recent papers describe its receptor(s) and extend its functions. One component of the receptors is Ret (3, 4), a tyrosine kinase receptor with a previously unknown ligand. The other is a newly identified GDNFR-alpha (5, 6), an extracellular protein linked to the cell through a glycosylphosphatidyl inositol (GPI) anchor. In studies with GDNF-responsive motor neurons (3) or Xenopus embryo cells (4), Ret activation was induced by GDNF. GDNF non-responsive cells were converted to responsive cells by Ret transfection (3). Cultures of wild-type and Ret-deficient embryo cells demonstrated that normal Ret signaling was necessary for GDNF function (4). In separate experiments, a GDNF receptor was cloned (5, 6). It was termed GDNFR-alpha. It is an extracellular protein that attaches to cells through a GPI linkage, suggesting that signaling required an additional membrane-associated protein. Ret was a logical candidate as a co-receptor, and GDNF was shown to induce phosphorylation of Ret in the presence, but not the absence, of GDNFR-alpha. A model of GDNF receptor activation shows GDNF forming a heterotetramer of two GDNF and two GDNFR-alpha molecules interacting with and activating the Ret tyrosine kinase. References 1. Lin, L-F. H. et al. (1993) Science 260:1130. 2. Kingsley, D.M. (1994) Genes Dev. 8:133. 3. Trupp, M. et al. (1996) Nature 381:785. 4. Durbec, P. et al. (1996) Nature 381:789. 5. Jing, S. et al. (1996) Cell 85:1113. 6. Treanor, J.J.S. et al. (1996) Nature 382:80. | Home | Cytokine Bulletin Contents | Next Article | ---------------------- Cell, Vol. 85, 1113-1124, June 28, 1996, Copyright c 1996 by Cell Press. http://www.cell.com/cell/abstract/x0534.html Article GDNF-Induced Activation of the Ret Protein Tyrosine Kinase Is Mediated by GDNFR- , a Novel Receptor for GDNF Shuqian Jing,*=B6 Duanzhi Wen,=B6 Yanbin Yu,* Paige L. Holst,* Yi Luo, Mei Fang,* Rami Tamir, Laarni Antonio,=A7 Zheng Hu,* Rod Cupples, Jean-Claude Louis, Sylvia Hu, Bruce W. Altrock,* and Gary M. Fox* *Department of Immunology Department of Mammalian Cell Molecular Biology Department of Neurobiology =A7Department of Developmental Biology Amgen, Incorporated Thousand Oaks, California 91320 Summary We report the expression cloning and characterization of GDNFR-, a novel glycosylphosphatidylinositol-linked cell surface receptor for glial cell line-derived neurotrophic factor (GDNF). GDNFR- binds GDNF specifically and mediates activation of the Ret protein-tyrosine kinase (PTK). Treatment of Neuro-2a cells expressing GDNFR- with GDNF rapidly stimulates Ret autophosphorylation. Ret is also activated by treatment with a combination of GDNF and soluble GDNFR- in cells lacking GDNFR-, and this effect is blocked by a soluble Ret-Fc fusion protein. Ret activation by GDNF was also observed in cultured embryonic rat spinal cord motor neurons, a cell type that responds to GDNF in vivo. A model for the stepwise formation of a GDNF signal-transducing complex including GDNF, GDNFR-, and the Ret PTK is proposed. Corresponding author: Gary M. Fox tel: 805 447 3036 fax: 805 499 2751 Cell Press / 6-25-96 / [log in to unmask] -------------------- the Cytokine Bulletin, Spring 1995 http://cytokine.rndsystems.com/cb/cbsp95/cbsp95a5.html GDNF Does More than Expected Glial-cell-Derived Neurotrophic Factor (GDNF), a TGF beta family member, was known as a neurotrophic factor for substantia nigra dopaminergic neurons, but it has now been shown to have broader actions; it is neurotrophic in a variety of situations. Yan et al. (1) demonstrated that GDNF was bound, internalized by and retrogradely transported by spinal motor neurons in a way that suggested a specific receptor was involved. In a test of in vivo nerve degeneration, they transected facial motor neurons. In control transected rats, 6% of the nerves survived seven days; in transected rats treated locally with GDNF, virtually all survived seven days, showing a motor neuron neurotropic effect. Henderson et al. (2) reported that GDNF is a potent survival factor for spinal motor neurons from rat embryo, and Oppenheim et al. (3) reported that GDNF rescued developing avian motor neurons from programed cell death. It also completely blocked loss of motor neurons caused by axotomy of spinal motor neurons of avian embryos (3). There also has been evidence for an in vivo effect of GDNF on the dopaminergic system. Tomac et al. (4) induced a Parkinson-like symptom in mice by injection of MPTP (a drug that causes Parkinson's disease). Injection of GDNF over the substantia nigra before MPTP was protective; injection after MPTP was restorative. Along the same line, Beck et al. (5) used a rat model in which they transected axons within the medial forebrain bundle, causing loss of substantia nigra neurons that express tyrosine hydroxylase. Repeated injections of GDNF prevented this loss. It also has been reported that GDNF transcripts are expressed nearly ubiquitously. References 1. Yan, Q. et al. (1995) Nature 373:341. 2. Henderson, C.E. et al. (1994) Science 266:1062. 3. Oppenheim, R.W. et al. (1995) Nature 373:344. 4. Tomac, A. et al. (1995) Nature 373:335. 5. Beck, K.D. et al. (1995) Nature 373:339. 6. Suter-Crazzolara, C. and K. Unsicker (1994) NeuroReport 5:2486. | Home | Cytokine Bulletin Contents | Next Article (Spring 1995) | Cytokine Catalog | Molecular Biology Catalog | ------------------ the 1996 Cytokine Catalog | Home | Contents | Cytokines | Antibodies | Adhesion Molecules & Antibodies | Biotinylated Antigen-purified Antibodies | Immunoassays | Matched Antibody Pairs for ELISA | Intracellular Staining Reagents | Flow Cytometry Reagents | T Cell Columns | Free Radical Reagents | Probes | Recombinant Human GDNF http://cytokine.rndsystems.com/inserts/proteins/200-299/212-gd.html Catalog Number: 212-GD Pack Sizes: 10 ug, 50 ug Specifications and Use Source A DNA sequence encoding the human GDNF precursor (Lin L-F. et al 1993, Science 260:1130-1132) was expressed in a mouse myeloma cell line NSO. Molecular Mass Mature human GDNF is a disulfide-linked homodimer and is predicted to contain two 134 amino acid residue peptides with a molecular mass of approximately 15 kDa. NSO expressed mature human GDNF lacks 30 residues from the amino-terminus of the predicted sequence. This glycosylated recombinant mature hGDNF contains the seven conserved Cys residues found in all members of the TGF-beta superfamily and is biologically active. Purity > 97%, as determined by SDS-PAGE and visualized by silver stain. Endotoxin Level < 0.1 ng per 1 ug of the cytokine as determined by the LAL method. Activity Measured by its ability to support the survival and stimulate neurite outgrowth of cultured embryonic chick dorsal root ganglia. The ED50 for this effect is typically 1 - 3 ng/mL. Formulation Lyophilized from a sterile filtered solution in PBS, containing 50 ug of bovine serum albumin per 1 ug of cytokine. Reconstitution It is recommended that sterile PBS containing at least 0.1% human serum albumin or bovine serum albumin be added to the vial to prepare a stock solution of no less than 1 ug/mL. Storage Lyophilized samples are stable for greater than six months at -20=B0 C to -70=B0 C. Upon reconstitution, this cytokine can be stored under sterile conditions at 2=B0 - 4=B0 C for one month or at -20=B0 C to -70=B0 C for three months without detectable loss of activity. Avoid repeated freeze-thaw cycles. Human Glial Cell Line-derived Neurotrophic Factor Glial Cell Line-derived Neurotrophic Factor (GDNF) is a recently discoved neurotrophic factor that has been shown to promote the survival of various neuronal subpopulations in both the central as well as the peripheral nervous systems at different stages of their development. Neuronal subpopulations that have been shown to be affected by GDNF include motoneurons, midbrain dopaminergic neurons, Purkinje cells and sympathetic neurons. Native GDNF, a disulfide-linked homodimeric glycoprotein, is a novel member of the TGF-beta superfamily. Human GDNF cDNA encodes a 211 amino acid residue prepropeptide that is processed to yield a dimeric protein. Mature human GDNF was predicted to contain two 134 amino acid residue subunits. NSO expressed mature human GDNF lacks 30 residues from the amino-terminus of the predicted sequence. This glycosylated recombinant mature human GDNF still contains the seven conserved Cys residues found in all members of the TGF-beta superfamily and is biologically active. The GDNF sequence contains two potential glycosylation sites and insect cell-expressed recombinant rat GDNF proteins are glycosylated. Mature rat and human GDNF exhibit approximately 93% amino acid sequence identity and show considerable species cross-reactivity. Cells known to express GDNF include Sertoli cells, type 1 astrocytes, Schwann cells, neurons, pinealocytes and skeletal muscle cells. For more information on GDNF, see the Glial Cell Line-derived Neurotrophic Factor Mini-Review in the R&D Systems' Cytokine Catalog. 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