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

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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.

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----------------------
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.


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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.

   FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES

                               R&D Systems
                               1-800-343-7475


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