Generation of a transgenic zebrafish model of Tauopathy using a novel promoter element derived from the zebrafish eno2 gene.
The aim of this study was to isolate cis-acting regulatory elements for the generation of transgenic zebrafish models of neurodegeneration. Zebrafish enolase-2 (eno2) showed neuronal expression increasing from 24 to 72 h post-fertilization (hpf) and persisting through adulthood. A 12 kb eno2 genomic fragment, extending from 8 kb upstream of exon 1 to exon 2, encompassing intron 1, was sufficient to drive neuronal reporter gene expression in vivo over a similar time course. Five independent lines of stable Tg(eno2 : GFP) zebrafish expressed GFP widely in neurons, including populations with relevance to neurodegeneration, such as cholinergic neurons, dopaminergic neurons and cerebellar Purkinje cells. We replaced the exon 2-GFP fusion gene with a cDNA encoding the 4-repeat isoform of the human microtubule-associated protein Tau. The first intron of eno2 was spliced with high fidelity and efficiency from the chimeric eno2-Tau transcript. Tau was expressed at approximately 8-fold higher levels in Tg(eno2 : Tau) zebrafish brain than normal human brain, and localized to axons, neuropil and ectopic neuronal somatic accumulations resembling neurofibrillary tangles. The 12 kb eno2 promoter drives high-level transgene expression in differentiated neurons throughout the CNS of stable transgenic zebrafish. This regulatory element will be useful for the construction of transgenic zebrafish models of neurodegeneration.
Bai
Qing
Q
Pittsburgh Institute for Neurodegenerative Diseases, Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Garver
Jessica A
JA
Hukriede
Neil A
NA
Burton
Edward A
EA
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D004195:Disease Models, Animal; D008969:Molecular Sequence Data; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020449:Repetitive Sequences, Amino Acid; D017422:Sequence Analysis, DNA; D024801:Tauopathies / Q000235:genetics*; D024363:Transcription Initiation Site; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism; D016875:tau Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D029961:Zebrafish Proteins; D016875:tau Proteins; D010751:Phosphopyruvate Hydratase; C525219:enolase 2, zebrafish