Using intra-brain drug infusion to investigate neural mechanisms underlying reward-seeking behavior in mice

Chen, Xu; Peng, Beibei; Liu, Shuai

doi:10.1016/j.xpro.2022.101221

Cited by 3 publications

(3 citation statements)

References 14 publications

(27 reference statements)

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“…We envision two paths for optimizing the use of oROS-HT 635 in live animals. First, introducing the dye into brain tissue can be aided with engineered solutions such as injection cannulas or drug delivery systems 77 , 78 . Alternatively, optimization of the oROS-HT 635 with highly bioavailable dyes (e.g.…”

Section: Discussionmentioning

confidence: 99%

Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

Berndt,

Lee,

Nguyen

et al. 2024

Preprint

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H2O2 is a key oxidant in mammalian biology and a pleiotropic signaling molecule at the physiological level, and its excessive accumulation in conjunction with decreased cellular reduction capacity is often found to be a common pathological marker. Here, we present a red fluorescent Genetically Encoded H2O2 Indicator (GEHI) allowing versatile optogenetic dissection of redox biology. Our new GEHI, oROS-HT, is a chemigenetic sensor utilizing a HaloTag and Janelia Fluor (JF) rhodamine dye as fluorescent reporters. We developed oROS-HT through a structure-guided approach aided by classic protein structures and recent protein structure prediction tools. Optimized with JF635, oROS-HT is a sensor with 635 nm excitation and 650 nm emission peaks, allowing it to retain its brightness while monitoring intracellular H2O2 dynamics. Furthermore, it enables multi-color imaging in combination with blue-green fluorescent sensors for orthogonal analytes and low auto-fluorescence interference in biological tissues. Other advantages of oROS-HT over alternative GEHIs are its fast kinetics, oxygen-independent maturation, low pH sensitivity, lack of photo-artifact, and lack of intracellular aggregation. Here, we demonstrated efficient subcellular targeting and how oROS-HT can map inter and intracellular H2O2 diffusion at subcellular resolution. Lastly, we used oROS-HT with other green fluorescence reporters to investigate the transient effect of the anti-inflammatory agent auranofin on cellular redox physiology and calcium levels via multi-parametric, dual-color imaging.

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Section: Discussionmentioning

confidence: 99%

Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

Berndt,

Lee,

Nguyen

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…We envision two paths for optimizing the use of oROS-HT 635 in live animals. First, introducing the dye into brain tissue can be aided with engineered solutions such as injection cannulas or drug delivery systems 70,71 . Alternatively, optimization of the oROS-HT 635 with highly bioavailable dyes (e.g.…”

Section: Discussionmentioning

confidence: 99%

Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

Lee,

Nguyen,

Jin

et al. 2024

Preprint

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H2O2is a key oxidant in mammalian biology and a pleiotropic signaling molecule at the physiological level, and its excessive accumulation in conjunction with decreased cellular reduction capacity is often found to be a common pathological marker. Here, we present a red fluorescent Genetically Encoded H2O2Indicator (GEHI) allowing versatile optogenetic dissection of redox biology. Our new GEHI, oROS-HT, is a chemigenetic sensor utilizing a HaloTag and Janelia Fluor (JF) rhodamine dye as fluorescent reporters. We developed oROS-HT through a structure-guided approach aided by classic protein structures and recent protein structure prediction tools. Optimized with JF635, oROS-HT is a sensor with 635 nm excitation and 650 nm emission peaks, allowing it to retain its brightness while monitoring intracellular H2O2dynamics. Furthermore, it enables multi-color imaging in combination with blue-green fluorescent sensors for orthogonal analytes and low auto-fluorescence interference in biological tissues. Other advantages of oROS-HT over alternative GEHIs are its fast kinetics, oxygen-independent maturation, low pH sensitivity, lack of photo-artifact, and lack of intracellular aggregation. Here, we demonstrated efficient subcellular targeting and how oROS-HT can map inter and intracellular H2O2diffusion at subcellular resolution. Lastly, we used oROS-HT with the green fluorescent calcium indicator Fluo-4 to investigate the transient effect of the anti-inflammatory agent auranofin on cellular redox physiology and calcium levels via multi-parametric, dual-color imaging.

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“…Standard procedures were used for drug microinfusions via chronically implanted bilateral cannulas [29]. Mice received either artificial cerebrospinal fluid (ACSF) or a long-acting competitive antagonist of NMDARs (CPP, 50 ng in 300 nL per side).…”

Section: Drug Microinfusionsmentioning

confidence: 99%

The prelimbic prefrontal cortex mediates the development of lasting social phobia as a consequence of social threat conditioning

Lozano-Ortiz,

Felix-Ortiz,

Terrell

et al. 2024

Preprint

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Social phobia is highly detrimental for social behavior, mental health, and productivity. Despite much previous research, the behavioral and neurobiological mechanisms associated with the development of social phobia remain elusive. To investigate these issues, the present study implemented a mouse model of social threat conditioning in which mice received electric shock punishment upon interactions with unfamiliar conspecifics. This resulted in immediate reductions in social behavior and robust increases in defensive mechanisms such as avoidance, freezing, darting, and ambivalent stretched posture. Furthermore, social deficits lasted for prolonged periods and were independent of contextual settings, sex variables, or particular identity of the social stimuli. Shedding new light into the neurobiological factors contributing to this phenomenon, we found that optogenetic silencing of the prelimbic (PL), but not the infralimbic (IL), subregion of the medial prefrontal cortex (mPFC) during training led to subsequent forgetting and development of lasting social phobia. Similarly, pharmacological inhibition of NMDARs in PL also impaired the development of social phobia. These findings are consistent with the notion that social-related trauma is a prominent risk factor for the development of social phobia, and that this phenomenon engages learning-related mechanisms within the prelimbic prefrontal cortex to promote prolonged representations of social threat.Abstract Figure

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Using intra-brain drug infusion to investigate neural mechanisms underlying reward-seeking behavior in mice

Cited by 3 publications

References 14 publications

Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications

The prelimbic prefrontal cortex mediates the development of lasting social phobia as a consequence of social threat conditioning

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Using intra-brain drug infusion to investigate neural mechanisms underlying reward-seeking behavior in mice

Cited by 3 publications

References 14 publications

Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

Far-red and sensitive sensor for monitoring real time H2O2 dynamics with subcellular resolution and in multi-parametric imaging applications.

Far-red and sensitive sensor for monitoring real time H2O2dynamics with subcellular resolution and in multi-parametric imaging applications

The prelimbic prefrontal cortex mediates the development of lasting social phobia as a consequence of social threat conditioning

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Far-red and sensitive sensor for monitoring real time H₂O₂dynamics with subcellular resolution and in multi-parametric imaging applications