The vesicular glutamate transporter-1 upstream promoter and first intron each support glutamatergic-specific expression in rat postrhinal cortex

Zhang, Guorong; Xu, Li; Zhao, Hua; Geller, Alfred I.

doi:10.1016/j.brainres.2010.12.040

Cited by 18 publications

(16 citation statements)

References 69 publications

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“…In particular, expression levels from the VGLUT1 promoter might be improved. An initial analysis of the VGLUT1 promoter identified a basal promoter that is not glutamatergic-specific, and glutamatergic-specific elements, likely enhancers, in both the upstream promoter and first intron (Zhang et al, 2011). Thus, substitution of the VGLUT1 basal promoter with a strong basal promoter, and/or use of cancatamers of the glutamatergic-specific upstream elements might increase expression levels.…”

Section: Discussionmentioning

confidence: 99%

Neurons can be labeled with unique hues by helper virus-free HSV-1 vectors expressing Brainbow

Zhang

Zhao

Abdul-Muneer

et al. 2015

Journal of Neuroscience Methods

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Background-A central problem in neuroscience is elucidating synaptic connections, the connectome. Because mammalian forebrains contain many neurons, labeling specific neurons with unique tags is desirable. A novel technology, Brainbow, creates hundreds of hues by combinatorial expression of multiple fluorescent proteins (FPs).New method-We labeled small numbers of neurons, and their axons, with unique hues, by expressing Brainbow from a helper virus-free Herpes Simplex Virus (HSV-1) vector.Results-The vector expresses a Brainbow cassette containing four FPs from a glutamatergicspecific promoter. Packaging HSV-brainbow produced arrays of seven to eight Brainbow cassettes, and using Cre, each FP gene was in a position to be expressed, in different cassettes. Delivery into rat postrhinal (POR) cortex or hippocampus labeled small numbers of neurons with different, often unique, hues. An area innervated by POR cortex, perirhinal (PER) cortex, contained axons with different hues. Specific axons in PER cortex were matched to specific cell bodies in POR cortex, using hue.Comparison with existing methods-HSV-Brainbow is the only technology for labeling small numbers of neurons with unique hues. In Brainbow mice, many neurons contain the same hue. Brainbow-adeno-associated virus vectors require transduction of the same neuron with multiple vector particles, confounding neuroanatomical studies. Replication-competent Brainbowpseudorabies virus vectors label multiple neurons with the same hue.Conclusions-Attractive properties of HSV-Brainbow include each vector particle contains multiple cassettes, representing numerous hues, recombination products are stabile, and experimental control of the number of labeled neurons. Labeling neurons with unique hues will benefit mapping forebrain circuits.

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Section: Discussionmentioning

confidence: 99%

Neurons can be labeled with unique hues by helper virus-free HSV-1 vectors expressing Brainbow

Zhang

Zhao

Abdul-Muneer

et al. 2015

Journal of Neuroscience Methods

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“…Because the INS-TH-NFH promoter supports long-term expression in multiple types of neurons in multiple brain areas, additional transcription factors, besides LSD1 and CLOCK, may modulate expression from the INS-TH-NFH promoter, particularly in neuron types that lack LSD1 and/or CLOCK. Moreover, a number of neuron type-specific promoters support significant levels of long-term expression from HSV-1 plasmid vectors, including the TH, ENK, GAD, phosphate-activated glutaminase, and vesicular glutamate transporter-1 promoters (Jin et al, 1996; Kaplitt et al, 1994; Rasmussen et al, 2007; Song et al, 1997; Wang et al, 1999; Zhang and Geller, 2010; Zhang et al, 2011). It remains to be determined if either LSD1 or CLOCK can improve long-term expression from any of these neuron type-specific promoters.…”

Section: Discussionmentioning

confidence: 99%

“…Neuron type-specific promoters that support significant levels of long-term expression from HSV-1 plasmid vectors include the tyrosine hydroxylase (TH) promoter, preproenkephalin (ENK) promoter, glutamic acid decarboxylase (GAD) promoter, phosphate-activated glutaminase (PAG) promoter, vesicular glutamate transporter-1 (VGLUT1) promoter, and specific fragments of the VGLUT1 promoter (Jin et al, 1996; Kaplitt et al, 1994; Rasmussen et al, 2007; Song et al, 1997; Wang et al, 1999; Zhang and Geller, 2010; Zhang et al, 2011). Interestingly, large fragments of the rat TH promoter (6.8 or 9 kb) support long-term, catecholaminergic-specific expression (Jin et al, 1996; Song et al, 1997; Wang et al, 1999), but a small fragment of this promoter (766 bp) supports only short-term expression (Wang et al, 1999), indicating that upstream sequences in the TH promoter support long-term expression.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of either lysine-specific demethylase-1 or CLOCK, but not Co-Rest, improves long-term expression from a modified neurofilament promoter, in a helper virus-free HSV-1 vector system

2012

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Long-term expression from helper virus-free Herpes Simplex Virus (HSV-1) vectors is required for many specific neural gene therapies and studies on neuronal physiology. We previously developed a promoter that supports long-term, neuron-specific expression by fusing the chicken ß-globin insulator (INS), followed by an upstream enhancer from the rat tyrosine hydroxylase (TH) promoter, to a neurofilament heavy gene (NFH) promoter. Here, we examined the capability of specific transcription factors to further improve long-term expression from this promoter. Following a HSV-1 virus infection, the virus genome is localized to promyelocytic leukemia protein (PML) nuclear bodies (NB). At these sites, specific cellular transcription factors interact with HSV-1 encoded transcription factors, and together regulate HSV-1 gene expression. Importantly, lysine-specific demethylase-1 (LSD1), CLOCK, and Co-Rest each activate HSV-1 gene expression. However, gene expression from HSV-1 vectors differs in a number of important aspects from the virus, including no HSV-1 genes are expressed. Nonetheless, these observations raise the possibility that specific transcription factors may improve long-term expression from specific promoters in HSV-1 vectors. Here, we show that overexpression of either LSD1 or CLOCK improves long-term expression from the INS-TH-NFH promoter, but overexpression of Co-Rest supports levels of long-term expression similar to those supported by a control vector. Further, overexpression of LSD1 is compatible with neuron-specific expression. Thus, overexpressing specific transcription factors can improve long-term expression from specific cellular promoters in HSV-1 vectors, and the chromatin structure of the vector has an important role in enabling expression.

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“…HSV-1 vectors containing specific promoters restrict expression to all neurons, or catecholaminergic, enkephalinergic, GABAergic, glutamatergic, or subtypes of glutamatergic neurons (Jin et al, 1996; Kaplitt et al, 1994; Rasmussen et al, 2007; Song et al, 1997; Zhang et al, 2000; Zhang and Geller, 2010; Zhang et al, 2011). Novel specificities of expression may be obtained by performing targeted gene transfer with vectors that contain specific promoters.…”

Section: Discussionmentioning

confidence: 99%

Antibody-mediated targeted gene transfer of helper virus-free HSV-1 vectors to rat neocortical neurons that contain either NMDA receptor 2B or 2A subunits

Zhang

Geller

2011

Brain Research

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Because of the numerous types of neurons in the brain, and particularly the forebrain, neuron type-specific expression will benefit many potential applications of direct gene transfer. The two most promising approaches for achieving neuron type-specific expression are targeted gene transfer to a specific type of neuron and using a neuron type-specific promoter. We previously developed antibody-mediated targeted gene transfer with Herpes Simplex Virus (HSV-1) vectors by modifying glycoprotein C (gC) to replace the heparin binding domain, which mediates the initial binding of HSV-1 particles to many cell types, with the Staphylococcus A protein ZZ domain, which binds immunoglobulin (Ig) G. We showed that a chimeric gC--ZZ protein is incorporated into vector particles and binds IgG. As a proof-of-principle for antibody-mediated targeted gene transfer, we isolated complexes of these vector particles and an anti-NMDA NR1 subunit antibody, and demonstrated targeted gene transfer to neocortical cells that contain NR1 subunits. However, because most forebrain neurons contain NR1, we obtained only a modest increase in the specificity of gene transfer, and this targeting specificity is of limited utility for physiological experiments. Here, we report efficient antibody-mediated targeted gene transfer to NMDA NR2B- or NR2A-containing cells in rat postrhinal cortex, and a neuron-specific promoter further restricted recombinant expression to neurons. Of note, because NR2A-containing neurons are relatively rare, these results show that antibody-mediated targeted gene transfer with HSV-1 vectors containing neuron type-specific promoters can restrict recombinant expression to specific types of forebrain neurons of physiological significance.

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The vesicular glutamate transporter-1 upstream promoter and first intron each support glutamatergic-specific expression in rat postrhinal cortex

Cited by 18 publications

References 69 publications

Neurons can be labeled with unique hues by helper virus-free HSV-1 vectors expressing Brainbow

Neurons can be labeled with unique hues by helper virus-free HSV-1 vectors expressing Brainbow

Overexpression of either lysine-specific demethylase-1 or CLOCK, but not Co-Rest, improves long-term expression from a modified neurofilament promoter, in a helper virus-free HSV-1 vector system

Antibody-mediated targeted gene transfer of helper virus-free HSV-1 vectors to rat neocortical neurons that contain either NMDA receptor 2B or 2A subunits

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