High level Purkinje cell specific expression of green fluorescent protein in transgenic mice

Zhang, Xulun; Baader, Stephan L.; Bian, Feng; Müller, Wolfgang; Oberdick, John

doi:10.1007/s004180100283

Cited by 25 publications

(14 citation statements)

References 61 publications

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“…Apart from the cerebellum, expression of hTBP was also identified in the cortex, brainstem, and hippocampus of transgenic mice. The widespread expression pattern indicates that the shorter L7/ pcp2 promoter fragment used here is less specific than the fragments used in other SCA models (Burright et al 1995;Tomomura et al 2001;Zhang et al 2001). On the other hand, the near-ubiquitous expression of L7-hTBP in the mouse brain mimics the mutant TBP expression pattern in SCA17 patients.…”

Section: Discussionmentioning

confidence: 83%

“…Of the several cell types in cerebellum, Purkinje cells are the target cells in most SCA patients. We thus tried to establish a SCA17 mouse model with hTBP gene over-expression driven by L7/pcp2 promoter for selective expression in Purkinje cells (Burright et al 1995;Zhang et al 2001). Apart from the cerebellum, expression of hTBP was also identified in the cortex, brainstem, and hippocampus of transgenic mice.…”

Section: Discussionmentioning

confidence: 99%

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Neuroprotective effects of granulocyte‐colony stimulating factor in a novel transgenic mouse model of SCA17

Chang¹,

Cao²,

Hsu³

et al. 2011

Journal of Neurochemistry

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J. Neurochem. (2011) 118, 288–303. Abstract Spinocerebellar ataxia type 17 (SCA17) is an autosomal dominant inherited disorder characterized by degeneration of spinocerebellar tracts and selected brainstem neurons owing to the expansion of a CAG repeat of the human TATA‐binding protein (hTBP) gene. To gain insight into the pathogenesis of this hTBP mutation, we generated transgenic mice with the mutant hTBP gene driven by the Purkinje specific protein (Pcp2/L7) gene promoter. Mice with the expanded hTBP allele developed ataxia within 2–5 months. Behavioral analysis of L7‐hTBP transgenic mice showed reduced fall latency in a rotarod assay. Purkinje cell degeneration was identified by immunostaining of calbindin and IP3R1. Reactive gliosis and neuroinflammation occurred in the transgenic cerebellum, accompanied by up‐regulation of GFAP and Iba1. The L7‐hTBP transgenic mice were thus confirmed to recapitulate the SCA17 phenotype and were used as a disease model to explore the potential of granulocyte‐colony stimulating factor in SCA17 treatment. Our results suggest that granulocyte‐colony stimulating factor has a neuroprotective effect in these transgenic mice, ameliorating their neurological and behavioral deficits. These data indicate that the expression of the mutant hTBP in Purkinje cells is sufficient to produce cell degeneration and an ataxia phenotype, and constitutes a good model for better analysis of the neurodegeneration in SCA17.

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Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Neuroprotective effects of granulocyte‐colony stimulating factor in a novel transgenic mouse model of SCA17

Chang¹,

Cao²,

Hsu³

et al. 2011

Journal of Neurochemistry

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“…In a transgenic mouse line that expressed high levels of humanized GFP under the control of the original L7-1 promoter, aggressive behavior reflected by bitten females was observed. 39 Therefore, we intravenously injected high-titer AAV-PHP.B expressing GFP under the control of the L7-6 to adult male (n = 4) and female (n = 4) mice, and observed their behavior over a period of 2 months. Based on the results, none of the treated mice displayed aberrant behavioral phenotypes during the observation period, suggesting little influence of intravenously administered AAV-PHP.B and L7-6-driven GFP expression on the behavior of adult mice.…”

Section: Resultsmentioning

confidence: 99%

Minimal Purkinje Cell-Specific PCP2/L7 Promoter Virally Available for Rodents and Non-human Primates

Nitta

Matsuzaki

Konno

et al. 2017

Molecular Therapy - Methods & Clinical Development

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Cell-type-specific promoters in combination with viral vectors and gene-editing technology permit efficient gene manipulation in specific cell populations. Cerebellar Purkinje cells play a pivotal role in cerebellar functions. Although the Purkinje cell-specific L7 promoter is widely used for the generation of transgenic mice, it remains unsuitable for viral vectors because of its large size (3 kb) and exceedingly weak promoter activity. Here, we found that the 0.8-kb region (named here as L7-6) upstream of the transcription initiation codon in the first exon was alone sufficient as a Purkinje cell-specific promoter, presenting a far stronger promoter activity over the original 3-kb L7 promoter with a sustained significant specificity to Purkinje cells. Intravenous injection of adeno-associated virus vectors that are highly permeable to the blood-brain barrier confirmed the Purkinje cell specificity of the L7-6 in the CNS. The features of the L7-6 were also preserved in the marmoset, a non-human primate. The high sequence homology of the L7-6 among mouse, marmoset, and human suggests the preservation of the promoter strength and Purkinje cell specificity features also in humans. These findings suggest that L7-6 will facilitate the cerebellar research targeting the pathophysiology and gene therapy of cerebellar disorders.

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“…Notably, we have found that neurons from other transgenic mouse types [(e.g. L7‐GFP); Zhang et al. , 2001] were fluorescent in slices but not in culture (J. E. Slemmer, unpublished observations).…”

Section: Discussionmentioning

confidence: 99%

Glutamate‐induced elevations in intracellular chloride concentration in hippocampal cell cultures derived from EYFP‐expressing mice

Slemmer

Matsushita

Zeeuw

et al. 2004

Eur J of Neuroscience

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The homeostasis of intracellular Cl(-) concentration ([Cl(-)](i)) is critical for neuronal function, including gamma-aminobutyric acid (GABA)ergic synaptic transmission. Here, we investigated activity-dependent changes in [Cl(-)](i) using a transgenetically expressed Cl(-)-sensitive enhanced yellow-fluorescent protein (EYFP) in cultures of mouse hippocampal neurons. Application of glutamate (100 microm for 3 min) in a bath perfusion to cell cultures of various days in vitro (DIV) revealed a decrease in EYFP fluorescence. The EYFP signal increased in amplitude with increasing DIV, reaching a maximal response after 7 DIV. Glutamate application resulted in a slight neuronal acidification. Although EYFP fluorescence is sensitive to pH, EYFP signals were virtually abolished in Cl(-)-free solution, demonstrating that the EYFP signal represented an increase in [Cl(-)](i). Similar to glutamate, a rise in [Cl(-)](i) was also induced by specific ionotropic glutamate receptor agonists and by increasing extracellular [K(+)], indicating that an increase in driving force for Cl(-) suffices to increase [Cl(-)](i). To elucidate the membrane mechanisms mediating the Cl(-) influx, a series of blockers of ion channels and transporters were tested. The glutamate-induced increase in [Cl(-)](i) was resistant to furosemide, bumetanide and 4,4'-diisothiocyanato-stilbene-2,2'-disulphonic acid (DIDS), was reduced by bicuculline to about 80% of control responses, and was antagonized by niflumic acid (NFA) and 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB). We conclude that membrane depolarization increases [Cl(-)](i) via several pathways involving NFA- and NPPB-sensitive anion channels and GABA(A) receptors, but not through furosemide-, bumetanide- or DIDS-sensitive Cl(-) transporters. The present study highlights the vulnerability of [Cl(-)](i) homeostasis after membrane depolarization in neurons.

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High level Purkinje cell specific expression of green fluorescent protein in transgenic mice

Cited by 25 publications

References 61 publications

Neuroprotective effects of granulocyte‐colony stimulating factor in a novel transgenic mouse model of SCA17

Neuroprotective effects of granulocyte‐colony stimulating factor in a novel transgenic mouse model of SCA17

Minimal Purkinje Cell-Specific PCP2/L7 Promoter Virally Available for Rodents and Non-human Primates

Glutamate‐induced elevations in intracellular chloride concentration in hippocampal cell cultures derived from EYFP‐expressing mice

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