Targeted expression of a toxin gene to D1 dopamine receptor neurons by cre-mediated site-specific recombination.
Idiopathic Parkinson's disease involves the loss of midbrain dopaminergic neurons, resulting in the presynaptic breakdown of dopaminergic transmission in the striatum. Huntington's disease and some neurodegenerative diseases with Parkinsonian features have postsynaptic defects caused by striatal cell death. Mice were generated in which an attenuated form of the diphtheria toxin gene (tox-176) was expressed exclusively in D1 dopamine receptor (D1R)-positive cells with the aim of determining the effect of this mutation on development of the basal ganglia and on the locomotor phenotype. Transgenic mice expressing Cre, a site-specific DNA recombinase, were crossed with a second line in which a transcriptionally silenced tox-176 gene was inserted into the D1R gene locus by homologous recombination. Young doubly transgenic mutant mice expressing the tox-176 gene displayed bradykinesia, dystonia, and had falls caused by myoclonic jerks. The mutant brain had evidence of apoptosis and reactive gliosis and, consistent with the D1R expression pattern, the striatum was reduced in volume, and the Islands of Calleja were absent. In contrast, the cortex was of normal thickness. D1Rs were not detectable in mutants by in situ hybridization or ligand autoradiography, whereas D2 dopamine receptor (D2R) mRNA and protein was present in the striatum. In addition, substance P and dynorphin, neuropeptides known to be expressed in D1R-positive striatonigral projection neurons were not detectable. Enkephalin, a marker found in D2-positive striatopallidal projection neurons was expressed in the mutant brain. The mutant represents a novel neurodegenerative disease model with a dramatic extrapyramidal phenotype.
Drago
J
J
Neurosciences Group, Department of Anatomy, Institute of Reproduction and Development, Monash University, Clayton, Victoria, 3168, Australia.
Padungchaichot
P
P
Wong
J Y
JY
Lawrence
A J
AJ
McManus
J F
JF
Sumarsono
S H
SH
Natoli
A L
AL
Lakso
M
M
Wreford
N
N
Westphal
H
H
Kola
I
I
Finkelstein
D I
DI
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001479:Basal Ganglia / Q000737:chemistry* / Q000166:cytology / Q000201:enzymology*; D004167:Diphtheria Toxin / Q000235:genetics; D004745:Enkephalins / Q000032:analysis; D015971:Gene Expression Regulation, Enzymologic; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005911:Gliosis / Q000503:physiopathology; D017403:In Situ Hybridization; D020287:In Situ Nick-End Labeling; D019426:Integrases / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D009069:Movement Disorders / Q000378:metabolism; D009207:Myoclonus / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D010302:Parkinson Disease, Secondary / Q000378:metabolism; D010641:Phenotype; D012333:RNA, Messenger / Q000032:analysis; D011869:Radioligand Assay; D017447:Receptors, Dopamine D1 / Q000502:physiology*; D013373:Substance P / Q000032:analysis; D014764:Viral Proteins
D004167:Diphtheria Toxin; D004745:Enkephalins; D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; D017447:Receptors, Dopamine D1; D014764:Viral Proteins; D013373:Substance P; C045073:Cre recombinase; D019426:Integrases