The BioLuminescent‐OptoGenetic <i>in vivo</i> response to coelenterazine is proportional, sensitive, and specific in neocortex

Gomez‐Ramirez, Manuel; More, Alexander I.; Friedman, Nina; Hochgeschwender, Ute; Moore, Christopher I.

doi:10.1002/jnr.24498

Cited by 28 publications

(35 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the present report we demonstrate early results detectability of bioluminescence using a modified UC miniscope optimized for bioluminescence imaging (BLm Removal of the excitation light, allowed us further to re the mass, cost, power consumption and assembly comple of miniscopes. Even with the minimal modifications prese here, we were able to capture temporal dynamics that typic can be observed only when using a high-end EMCCD cam Not only were we able to detect bioluminescence using m simpler, smaller, lower cost camera, we were able to do much higher framerate (as fast as 5 FPS compared to 0.1 used with EMCCD [10]), paving the way tow measurements of calcium dynamics via bioluminesc indicators undergoing active development. Transitionin bioluminescence miniature microscopy can enable testing broader spectrum of hypotheses.…”

Section: Discussionmentioning

confidence: 95%

See 1 more Smart Citation

Miniaturized Devices for Bioluminescence Imaging in Freely Behaving Animals

Celinskis

Friedman

Koksharov

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Fluorescence miniature microscopy in vivo has recently proven a major advance, enabling cellular imaging in freely behaving animals. However, fluorescence imaging suffers from autofluorescence, phototoxicity, photobleaching and non-homogeneous illumination artifacts. These factors limit the quality and time course of data collection. Bioluminescence provides an alternative kind of activity-dependent light indicator. Bioluminescent calcium indicators do not require light input, instead generating photons through chemiluminescence. As such, limitations inherent to the requirement for light presentation are eliminated. Further, bioluminescent indicators also do not require excitation light optics: the removal of this component should make lighter and lower cost microscope with fewer assembly parts. While there has been significant recent progress in making brighter and faster bioluminescence indicators, parallel advances in imaging hardware have not yet been realized. A hardware challenge is that despite potentially higher signal-to-noise of bioluminescence, the signal strength is lower than that of fluorescence. An open question we address in this report is whether fluorescent miniature microscopes can be rendered sensitive enough to detect bioluminescence. We demonstrate this possibility in vitro and in vivo by implementing optimizations of the UCLA fluorescent miniscope. These optimizations yielded a miniscope (BLmini) which is 22% lighter in weight, has 45% fewer components, is up to 58% less expensive, offers up to 15 times stronger signal (as dichroic filtering is not required) and is sensitive enough to capture spatiotemporal dynamics of bioluminescence in the brain with a signal-to-noise ratio of 34 dB.

show abstract

Section: Discussionmentioning

confidence: 95%

“…4. For comparison, we overlaid time courses acquired using the BLmini and by EMCCD in a different experiment ( [10] ; Fig. 4A).…”

Section: B Initial In Vivo Measurements With the Blmini Capture Tempmentioning

confidence: 99%

Miniaturized Devices for Bioluminescence Imaging in Freely Behaving Animals

Celinskis

Friedman

Koksharov

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…To stimulate genetically targeted neural circuits, we employed bioluminescent optogenetics (BL-OG) ( Berglund et al., 2013 , 2016 ; Gomez-Ramirez et al., 2020 ; Park et al., 2020 ; Song et al., 2018 ; Yu et al., 2019 ; Zenchak et al., 2020 ). This approach has the distinct dual advantage of enabling both non-invasive chemogenetic manipulation and temporally selective optogenetic control with the same molecular construct.…”

Section: Resultsmentioning

confidence: 99%

“…We induced hyperexcitation in developing mouse pups once a day during post-natal days 4–14, capturing the window of significant outgrowth and development of key cortical circuits ( Romand et al., 2011 ; Sasaki et al., 2015 ). BL-OG has been used to both activate and silence neurons in adult mice and rats, using various routes of administration of CTZ ( Berglund et al., 2016 ; Gomez-Ramirez et al., 2020 ; Park et al., 2020 ; Tung et al., 2015 ; Yu et al., 2019 ). To confirm the efficacy of BL-OG in pups, we tested for bioluminescence emission and neural activity after intraperitoneal CTZ administration.…”

Section: Resultsmentioning

confidence: 99%

Selective postnatal excitation of neocortical pyramidal neurons results in distinctive behavioral and circuit deficits in adulthood

Medendorp¹,

Björefeldt²,

Crespo³

et al. 2021

iScience

Self Cite

View full text Add to dashboard Cite

Summary In genetic and pharmacological models of neurodevelopmental disorders, and human data, neural activity is altered within the developing neocortical network. This commonality begs the question of whether early enhancement in excitation might be a common driver, across etiologies, of characteristic behaviors. We tested this concept by chemogenetically driving cortical pyramidal neurons during postnatal days 4–14. Hyperexcitation of Emx1-, but not dopamine transporter-, parvalbumin-, or Dlx5/6-expressing neurons, led to decreased social interaction and increased grooming activity in adult animals. In vivo optogenetic interrogation in adults revealed decreased baseline but increased stimulus-evoked firing rates of pyramidal neurons and impaired recruitment of inhibitory neurons. Slice recordings in adults from prefrontal cortex layer 5 pyramidal neurons revealed decreased intrinsic excitability and increased synaptic E/I ratio. Together these results support the prediction that enhanced pyramidal firing during development, in otherwise normal cortex, can selectively drive altered adult circuit function and maladaptive changes in behavior.

show abstract

“…1D), thus a programmable injector is required to consistently measure the initial strong signal. For the plate reader that is not equipped with an injector, a modified Gluc that shows more stable luminescence such as slow-burn Gluc (sbGluc) and superluminescent Gluc (slGluc) would be favored for consistent measurements across multiple samples (Welsh et al, 2009;Gomez-Ramirez et al, 2020).…”

Section: Comparison With Current Methodsmentioning

confidence: 99%

Secreted reporter assay enables quantitative and longitudinal monitoring of neuronal activity

Santos

Park

2021

Preprint

View full text Add to dashboard Cite

The ability to measure changes in neuronal activity in a quantifiable and precise manner is of fundamental importance to understand neuron development and function. Repeated monitoring of neuronal activity of the same population of neurons over several days is challenging and, typically, low-throughput. Here, we describe a new biochemical reporter assay that allows for repeated measurements of neuronal activity in a cell type-specific manner. We coupled activity-dependent elements from the Arc/Arg3.1 gene with a secreted reporter, Gaussia luciferase, to quantify neuronal activity without sacrificing the neurons. The reporter predominantly senses calcium and NMDA receptor-dependent activity. By repeatedly measuring the accumulation of the reporter in cell media, we can profile the developmental dynamics of neuronal activity in cultured neurons from male and female mice. The assay also allows for longitudinal analysis of pharmacological treatments, thus distinguishing acute from delayed responses. Moreover, conditional expression of the reporter allows for monitoring cell type-specific changes. This simple, quantitative, cost-effective, automatable, and cell type-specific activity reporter is a valuable tool to study the development of neuronal activity in normal and disease-model conditions, and to identify small molecules or protein factors that selectively modulate the activity of a specific population of neurons.SignificanceNeurological and neurodevelopmental disorders are prevalent worldwide. Despite significant advances in our understanding of synapse formation and function, developing effective therapeutics remains challenging, in part due to the lack of simple and robust high-throughput screening assays of neuronal activity. Here, we describe a simple biochemical assay that allows for repeated measurements of neuronal activity in a cell type-specific manner. Thus filling the need for assays amenable to longitudinal studies, such as those related to neural development. Other advantages include its simple and quantitative nature, logitudinal profiling, cell type-specificity, and being multiplexed with other invasive techniques.

show abstract

The BioLuminescent‐OptoGenetic in vivo response to coelenterazine is proportional, sensitive, and specific in neocortex

Cited by 28 publications

References 37 publications

Miniaturized Devices for Bioluminescence Imaging in Freely Behaving Animals

Miniaturized Devices for Bioluminescence Imaging in Freely Behaving Animals

Selective postnatal excitation of neocortical pyramidal neurons results in distinctive behavioral and circuit deficits in adulthood

Secreted reporter assay enables quantitative and longitudinal monitoring of neuronal activity

Contact Info

Product

Resources

About