In Vivo Imaging of Neuronal Activity by Targeted Expression of a Genetically Encoded Probe in the Mouse

Bozza, Thomas; McGann, John P.; Mombaerts, Peter; Wachowiak, Matt

doi:10.1016/s0896-6273(04)00144-8

Cited by 336 publications

(401 citation statements)

References 48 publications

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“…One prominent exception this is in the olfactory bulb, where thousands of odorant-specific olfactory sensory neurons fasciculate to target individual glomeruli. The olfactory marker protein (OMP) promoter has been used to drive synaptopHluorin expression across olfactory sensory neurons (OSN), providing a functional map of OSN responsiveness to odorant presentation in vivo [28]. Intriguingly, results from this imaging study were different from those obtained using optical imaging or 2-deoxyglucose analysis, possibly because these methods are lower resolution, requiring greater signal averaging within and between animals, or because these methods may be more sensitive to post-synaptic activity (rather than presynaptic release).…”

Section: Phluorins Ph-based Sensorsmentioning

confidence: 99%

Visualizing circuits and systems using transgenic reporters of neural activity

Barth

2007

Current Opinion in Neurobiology

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Genetically encoded sensors of neural activity enable visualization of circuit-level function in the central nervous system. Although our understanding of the molecular events that regulate neuronal firing, synaptic function, and plasticity has expanded rapidly over the past fifteen years, an appreciation for how cellular changes are functionally integrated at the circuit level has lagged. A new generation of tools that employ fluorescent sensors of neural activity promises unique opportunities to bridge the gap between cellular and system-levels analysis.This review will focus on genetically-encoded sensors. A primary advantage of these indicators is that they can be non-selectively introduced to large populations of cells using either transgenic or viral-mediated approaches. This ability removes the non-trivial obstacles of how to get chemical indicators into cells of interest, a problem that has dogged investigators who have been interested in mapping neural function in the intact CNS. Five different types of approaches and their relative utility will be reviewed here: 1) reporters of immediate-early gene (IEG) activation using promoters such as c-fos and arc, 2) voltage-based sensors, such as GFPcoupled Na + and K + channels, 3) Cl − based sensors, 4) Ca ++ -based sensors, such as Camgaroo and the troponin-based TN-L15, and 5) pH-based sensors, which have been particularly useful for examining synaptic activity of highly convergent afferents in sensory systems in vivo. Particular attention will be paid to reporters of IEG expression, since these tools employ the built-in threshold function that occurs with activation of gene expression, provoking new experimental questions by expanding the timescale of analysis for circuit and system-level functional mapping. Fluorescent reporters of inducible gene expressionImmediate-early gene expression (IEG) can be a reliable marker of elevated neuronal firing, reporting activity over time scales that range from minutes to hours compared to ms and seconds for voltage-or ion-based sensors. In ion-based approaches, indicators are constitutively present in the cell, and the fluorescent protein must be engineered to be a sensor with rapid onset and offset kinetics to achieve temporal fidelity with the process being measured. In contrast, monitoring IEG expression can be directly coupled to transcription of the reporter gene. However, because maturation of the fluorophore is required for detection, a temporal delay may be introduced into reporter detection. This delay in reporting activity is advantageous in that it expands the types of experimental questions that can be addressed. For example, do IEGexpressing cells remain more excitable after stimulus cessation? Are cells activated by two temporally distinct stimuli (i.e. training and recall in a memory task) overlapping or distinct?

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Section: Phluorins Ph-based Sensorsmentioning

confidence: 99%

Visualizing circuits and systems using transgenic reporters of neural activity

Barth

2007

Current Opinion in Neurobiology

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“…SynaptopHfluorin (SpH) and intrinsic signal experiments were performed on adults (8-12 weeks) B6.129P2-Omp tm2(spH)Mom/J mice (Bozza et al, 2004) available from The Jackson Laboratory (Bar Harbor, Maine). In addition, for intrinsic signal some adult C57B16J mice were also used.…”

Section: Experimental Setup Animals and Surgical Proceduresmentioning

confidence: 99%

“…1D) on a mouse model that genetically express the synaptopHfluorin (SpH) protein in the sensory neurons synaptic terminals (Fig. 1E, Bozza et al, 2004). SpH corresponds to the fusion protein between the synaptic vesicle protein VAMP-2 and a green fluorescent protein pH-sensitive (Miesenbock et al, 1998).…”

Section: In Vivo Imaging Of Odour Evoked Signalsmentioning

confidence: 99%

“…Lately, the possibility of a compromise between non-invasiveness and signal specificity has emerged. Promising results have been obtained with genetically modified animals that express reporter fluorescent proteins in targeted neuronal populations (Bozza et al, 2004;Hasan et al, 2004), which then can be observed optically with very little preparation even in vivo.…”

Section: Introductionmentioning

confidence: 99%

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Wavelet-based multi-resolution statistics for optical imaging signals: Application to automated detection of odour activated glomeruli in the mouse olfactory bulb

Bathellier¹,

Ville²,

Blu³

et al. 2007

NeuroImage

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“…It is evident that ORNs expressing the same receptor may transmit different patterns of signals to the same glomerulus under certain conditions. In vivo glomerular imaging studies show that the dynamic range of ORN input into a defined glomerulus is larger than that reported for single isolated ORNs [3,4]. Subsequent studies further demonstrate that ORN input is responsible for, or partially responsible for, the diverse spatial and temporal dynamics observed in glomeruli [5,6].…”

Section: Introductionmentioning

confidence: 99%

Gap junctions in olfactory neurons modulate olfactory sensitivity

Zhang

2010

BMC Neurosci

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BackgroundOne of the fundamental questions in olfaction is whether olfactory receptor neurons (ORNs) behave as independent entities within the olfactory epithelium. On the basis that mature ORNs express multiple connexins, I postulated that gap junctional communication modulates olfactory responses in the periphery and that disruption of gap junctions in ORNs reduces olfactory sensitivity. The data collected from characterizing connexin 43 (Cx43) dominant negative transgenic mice OlfDNCX, and from calcium imaging of wild type mice (WT) support my hypothesis.ResultsI generated OlfDNCX mice that express a dominant negative Cx43 protein, Cx43/β-gal, in mature ORNs to inactivate gap junctions and hemichannels composed of Cx43 or other structurally related connexins. Characterization of OlfDNCX revealed that Cx43/β-gal was exclusively expressed in areas where mature ORNs resided. Real time quantitative PCR indicated that cellular machineries of OlfDNCX were normal in comparison to WT. Electroolfactogram recordings showed decreased olfactory responses to octaldehyde, heptaldehyde and acetyl acetate in OlfDNCX compared to WT. Octaldehyde-elicited glomerular activity in the olfactory bulb, measured according to odor-elicited c-fos mRNA upregulation in juxtaglomerular cells, was confined to smaller areas of the glomerular layer in OlfDNCX compared to WT. In WT mice, octaldehyde sensitive neurons exhibited reduced response magnitudes after application of gap junction uncoupling reagents and the effects were specific to subsets of neurons.ConclusionsMy study has demonstrated that altered assembly of Cx43 or structurally related connexins in ORNs modulates olfactory responses and changes olfactory activation maps in the olfactory bulb. Furthermore, pharmacologically uncoupling of gap junctions reduces olfactory activity in subsets of ORNs. These data suggest that gap junctional communication or hemichannel activity plays a critical role in maintaining olfactory sensitivity and odor perception.

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In Vivo Imaging of Neuronal Activity by Targeted Expression of a Genetically Encoded Probe in the Mouse

Cited by 336 publications

References 48 publications

Visualizing circuits and systems using transgenic reporters of neural activity

Visualizing circuits and systems using transgenic reporters of neural activity

Wavelet-based multi-resolution statistics for optical imaging signals: Application to automated detection of odour activated glomeruli in the mouse olfactory bulb

Gap junctions in olfactory neurons modulate olfactory sensitivity

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