Karl Deisseroth

Prefrontal cortical (PFC) circuits provide top-down control of threat reactivity. This includes ventromedial PFC (vmPFC) circuitry, which plays a role in suppressing fear-related behavioral states. Dynorphin (Dyn) has been implicated in mediating negative affect and mal-adaptive behaviors induced by severe threats and is expressed in limbic circuits, including the vmPFC. However, there is a critical knowledge gap in our understanding of how vmPFC Dyn-expressing neurons and Dyn transmission detect threats and regulate expression of defensive behaviors. Here, we demonstrate that Dyn cells are broadly activated by threats and release Dyn locally in the vmPFC to limit passive defensive behaviors. We further demonstrate that vmPFC Dyn-mediated signaling promotes a switch of vmPFC networks to a fear-related state. In conclusion, we reveal a previously unknown role of vmPFC Dyn neurons and Dyn …

The anterior cingulate cortex (ACC) has emerged as a brain region of focus toward understanding the complex interplay between pain and opioid use disorders. However, little is known about the anatomical location and molecular identity of ACC nociceptive cell-types expressing the mu-opioid receptor (MOR)—the molecular target of opioid analgesics—and if this cortical architecture is conserved across species. Using single nuclei RNA sequencing of murine ACC, we found that three cell-subtypes of Slc17a7 glutamatergic neurons were the most transcriptionally active to noxious stimuli (identified by weighted expression of immediate early genes (IEGs) and differential gene expression analysis between nuclei from uninjured and chronic neuropathic pain mice). These cell-subtypes all co-expressed Oprm1 and single genetic identifiers, which we spatially resolved to label layer-specific, distinct neurons in L2, L5a …

The anterior cingulate cortex plays a pivotal role in the cognitive and affective aspects of pain perception. Both endogenous and exogenous opioid signaling within the cingulate mitigate cortical nociception, reducing pain unpleasantness. However, the specific functional and molecular identities of cells mediating opioid analgesia in the cingulate remain elusive. Given the complexity of pain as a sensory and emotional experience, and the richness of ethological pain-related behaviors, we developed a standardized, deep-learning platform for deconstructing the behavior dynamics associated with the affective component of pain in mice, LUPE (Light aUtomated Pain Evaluator). LUPE removes human bias in behavior quantification and accelerated analysis from weeks to hours, which we leveraged to discover that morphine altered attentional and motivational pain behaviors akin to affective analgesia in humans. Through activity-dependent genetics and single-nuclei RNA sequencing, we identified specific ensembles of nociceptive cingulate neuron-types ex-pressing mu-opioid receptors. Tuning receptor expression in these cells bidirectionally modulated morphine analgesia. Moreover, we employed a synthetic opioid receptor promoter-driven approach for cell-type specific optical and chemical genetic viral therapies to mimic morphine's pain-relieving effects in the cingulate, without reinforcement. This approach offers a novel strategy for precision pain management by targeting a key nociceptive cortical circuit with on-demand, non-addictive, and effective analgesia.

Understanding the nature and role of communication between distinct cellular populations in the brain will require simultaneous measurement and control of activity within those populations during behavior. Current optogenetic tools, present limitations such as spectral incompatibility with sensors and modest efficacy. To address these issues, we engineered novel red-shifted high potent cation-selective channelrhodopsin, rsChRmine, and K+-selective channelrhodopsins (KCRs), KALIs, with enhanced K+ selectivity based on our cryo-EM structures. By integrating these new opsins with multiple genetically-encoded Ca2+ indicators, we can selectively control specific neural circuits while simultaneously observing the responses of other elements within the same network. We applied this method to the mPFC of freely-behaving mice, quantifying dynamic information transmission between excitatory and inhibitory …

Cell type-specific interfaces within living animals will be invaluable for achieving communication with identifiable cells over the long term, enabling applications across many scientific and medical fields. However, biological tissues exhibit complex and dynamic organization properties that pose serious challenges for chronic cell-specific interfacing. A new technology, combining chemistry and molecular biology, has emerged to address this challenge: genetically targeted chemical assembly (GTCA), in which specific cells are genetically programmed (even in wild-type or non-transgenic animals, including mammals) to chemically construct non-biological structures. Here, we discuss recent progress in genetically targeted construction of materials and outline opportunities that may expand the GTCA toolbox, including specific chemical processes involving novel monomers, catalysts and reaction regimes both de …

Provided herein is a macroscope comprising an objective apparatus comprising a multifocal widefield optics comprising a plurality of optical components configured to focus on a plurality of planes. Also provided herein are methods for analyzing a three-dimensional specimen, the method comprising obtaining, via a macroscope, synchronous multifocal optical images of a plurality of planes of the three-dimensional specimen, wherein the macroscope comprises an objective apparatus comprising a multifocal widefield optics comprising a plurality of optical components configured to focus on a plurality of planes. The three-dimensional specimen can be a biological specimen, such as brain.

Psychiatric diseases are strongly influenced by genetics, but genetically guided treatments have been slow to develop, and precise molecular mechanisms remain mysterious. Although individual locations in the genome tend to not contribute powerfully to psychiatric disease incidence, genome-wide association studies (GWAS) have now successfully linked hundreds of specific genetic loci to psychiatric disorders [, –]. Here, building upon results from well-powered GWAS of four phenotypes relevant to psychiatry, we motivate an exploratory workflow leading from GWAS screening, through causal testing in animal models using methods such as optogenetics, to new therapies in human beings. We focus on schizophrenia and the dopamine D2 receptor (DRD2), hot flashes and the neurokinin B receptor (TACR3), cigarette smoking and receptors bound by nicotine (CHRNA5, CHRNA3, CHRNB4), and alcohol use and …

Endocannabinoid (eCB)–mediated suppression of inhibitory synapses has been hypothesized, but this has not yet been demonstrated to occur in vivo because of the difficulty in tracking eCB dynamics and synaptic plasticity during behavior. In mice navigating a linear track, we observed location-specific eCB signaling in hippocampal CA1 place cells, and this was detected both in the postsynaptic membrane and the presynaptic inhibitory axons. All-optical in vivo investigation of synaptic responses revealed that postsynaptic depolarization was followed by a suppression of inhibitory synaptic potentials. Furthermore, interneuron-specific cannabinoid receptor deletion altered place cell tuning. Therefore, rapid, postsynaptic, activity-dependent eCB signaling modulates inhibitory synapses on a timescale of seconds during behavior.