Ablation of central nervous system progenitor cells in transgenic rats using bacterial nitroreductase system.
Specific ablation of central nervous system (CNS) progenitor cells in the brain of live animals is a powerful method to determine the functions of these cells and to reveal novel avenues for the treatment of several CNS-related disorders. To achieve this goal, we generated a line of transgenic rats expressing a bacterial enzyme, Escherichia coli nitroreductase gene (NTR), under control of the nestin promoter. In this system, NTR(+) cells are selectively eliminated upon application of prodrug CB1954, through activation of programmed cell death machineries. At 5 days of age, which is a time when cerebellar development is occurring, transgenic rats bearing the nestin-NTR/green fluorescent protein (GFP) gene are overtly normal and express NTR/GFP in neuronal stem cells, without any toxicity in these cells. The functional consequence of progenitor cell ablation was demonstrated by administering prodrug CB1954 into the cerebellum at this 5-day time point. Stem cell ablation in these neonates resulted in sensorimotor abnormalities, cerebellar degeneration, overall reduction in cerebellar seize, and manifestation of ataxia. In adult rats, GFP expression was not seen in the hippocampal progenitor cells and seen only at very low levels in the lateral ventricles, indicating a different NTR/GFP expression pattern between neonates and adults. In addition, application of CB1954 by intraventricular delivery reduced the number of 5-bromo-2'-deoxyuridine-labeled proliferating cells in the lateral ventricle but not hippocampus of NTR/GFP rats. These findings shows that targeted expression of NTR under a specific promoter might be of significant value in addressing the function of distinct cell population in vivo.
Kwak
Seung P
SP
High Q Foundation, New York, NY, USA.
Malberg
Jessica E
JE
Howland
David S
DS
Cheng
Ke-Yi
KY
Su
Jianying
J
She
Yin
Y
Fennell
Myles
M
Ghavami
Afshin
A
D000818:Animals; D030801:Animals, Genetically Modified; D000831:Animals, Newborn; D000970:Antineoplastic Agents / Q000494:pharmacology; D001388:Aziridines / Q000494:pharmacology; D001522:Behavior, Animal / Q000187:drug effects / Q000502:physiology; D001973:Bromodeoxyuridine / Q000378:metabolism; D045744:Cell Line, Tumor; D002490:Central Nervous System / Q000166:cytology*; D004305:Dose-Response Relationship, Drug; D004597:Electroshock / Q000009:adverse effects; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007276:Injections, Intraventricular / Q000379:methods; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009601:Nitroreductases / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
D000970:Antineoplastic Agents; D001388:Aziridines; D029968:Escherichia coli Proteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D049452:Green Fluorescent Proteins; C100099:tretazicar; C116053:NfsA protein, E coli; D009601:Nitroreductases; D001973:Bromodeoxyuridine