Rodent model for Parkinson's Disease
View Patent ↗The present invention provides an animal model for Parkinson's Disease and a method for generating such a model. The method comprises the steps of transfecting dopaminergic neurons in the substantia nigra with a dominant negative mutant of FGFR1 with deleted tyrosine kinase domain. The animals are characterized by a reduction in the number of tyrosine hydroxylase neurons in the SNc area.
1. A rodent model for a symptom of Parkinson's Disease comprising a rodent whose substantial nigra (SN) nucleus has been injected with a composition comprising a DNA sequence encoding a tyrosine kinase deficient mutant of fibroblast growth factor receptor-1 (FGFR1) and polyethyleneimine (PEI), wherein expression of the DNA sequence produces a tyrosine kinase deficient mutant that is a competitive antagonist of the wild type FGFR1 and wherein the dopamine content of the striatum is reduced when compared to control rodents.
2. The rodent model of claim 1 , wherein a reduction in the dopamine content is observed after about 4 weeks.
3. The rodent model of claim 1 , wherein the injection comprises 1.5 to 3.0 μg of plasmid DNA comprising said DNA sequence and about 6 PEI equivalents, wherein 1 PEI equivalent is the amount of PEI required to neutralize the negative charges of DNA phosphate groups in the plasmid DNA.
4. The rodent model of claim 1 , wherein the injection is into the SN nucleus of one hemisphere.
5. The rodent model of claim 1 , wherein a reporter gene operably linked to a promoter is also injected together with said DNA sequence.
6. The rodent model of claim 5 , wherein the reporter gene encodes β-galactosidase.
7. A method for producing the rodent model of claim 1 , comprising the steps of:
a) providing a rodent;
b) sterotaxically injecting a composition comprising a DNA sequence encoding a tyrosine kinase deficient mutant of FGFR1 and PEI into the SN nucleus of the rodent; and
c) expressing said DNA sequence,
wherein the dopamine content of the striatum is reduced when compared to control rodents.
8. The method of claim 7 , wherein the injection comprises 1.5 to 3.0 μg of plasmid DNA comprising said DNA sequence and about 6 PEI equivalents, wherein 1 PEI equivalent is the amount of PEI required to neutralize the negative charges of DNA phosphate groups in the plasmid DNA.
9. The method of claim 7 , wherein the injection is into the SN nucleus of one hemisphere.
10. The method of claim 9 , wherein a reporter gene operably linked to a promoter is injected into the contra-lateral SN nucleus.
11. method of claim 7 , wherein a reduction in the dopamine content of the SN nucleus is observed after about 4 weeks.
12. A method of evaluating the potential of an agent for treatment of Parkinson's disease comprising the steps of:
a) providing a rodent whose SN nucleus has been injected with a composition comprising a DNA sequence encoding a tyrosine kinase deficient mutant of FGFR1 and PEI, wherein expression of the DNA sequence produces a tyrosine kinase deficient mutant that is a competitive antagonist of the wild type FGFR1 and wherein the dopamine content of the striatum is reduced when compared to control rodents,
b) administering the test agent to said rodent;
c) comparing the dopamine content of the striatum with a control rodent; and
d) evaluating the effectiveness of the test agent based on change in the dopamine content of the striatum.
13. The method of claim 12 , wherein the injection is into the SN nucleus of one hemisphere.
14. The method of claim 13 , wherein a reporter gene operably linked to a promoter is injected into the contra-lateral SN nucleus.
15. The method of claim 12 , wherein the injection comprises 1.5 to 3.0 μg of plasmid DNA comprising said DNA sequence and about 6 PEI equivalents, wherein 1 PEI equivalent is the amount of PEI required to neutralize the negative charges of DNA phosphate groups in the plasmid DNA.