Nice use of DREADDs to dissect GPCR signaling underlying anxiety
Great bit of work by Kash lab and many others using Gi, Gs and Gq =DREADDs to deconstruct pathways essential for anxiety in BNST
Posts
AAV's now available from ADDGENE
These are exceptionally high quality with exceptional quality control and have been validated by Rothlab. We launched the first 2 AAV on our website: http://www.addgene.org/44361/ http://www.addgene.org/44362/ We're hoping the release the non DIO versions and control vector next week!
DREADD AAV's available in EU
the Viral Vector Facility (VVF) - an service facility of the Neuroscience Center Zurich, a joint network of more than 1000 researchers of the University of Zurich and ETH Zurich - as a source for DREADD AAV vectors (at least for European-based researchers, which makes processing probably more convenient for them)? They will also make small aliquots (50 ul) of various DREADD AAV vectors available through our Viral Vector Repository. Contact information: Jean-Charles Paterna, PhD Manager of the Viral Vector Facility Neuroscience Center Zurich University of Zurich and ETH Zurich Winterthurerstrasse 190 CH-8057 Zurich http://www.vvf.uzh.ch/en.html + 41 44 635 59 91
DREADD AAV's available via Duke Vector core
For further information contact: Boris Kantor, PhD Assistant Research Professor & Viral Vector Core Director Duke Department of Neurobiology-School of Medicine 412 Research Drive Bryan Research Bldg room 213 Durham, NC 27710 Phone: 919-6811068
Update on AAVs for DREADDs
UNC is no longer offering single-unit AAV for DREADDs although we expect that these will be available via ADDGENE shortly. To get onto the ADDGENE notification list contact: Leila Haery haery@addgene.org with serotype and genotype needs for DREADD constructs. She will be helping Addgene coordinate production and distribution of DREADD viruses. In the meantime, you can order plasmids from ADDGENE and simply ask UNC to make a custom prep for you or send the plasmids to the Penn Vector core. Alternatively it looks like the Duke vector core is also offering DREADD-based AAVs. Bryan
DREADD/Opto to identify VTA dopaminergic neurons which regulate ethologically relevant sleep–wake behaviors
Very interesting paper found here.
DREADDSs demonstrate glutamatergic class of neurons in the brainstem and spinal cord, the V2a class, drives increased respiratory activity
Using a chemogenetic approach, we demonstrate that a glutamatergic class of neurons in the brainstem and spinal cord, the V2a class, is sufficient to drive increased ARM activity at rest in healthy mice.
How to find all publications related to DREADD technology
Simply go to Google Scholar and type in the following: "designer receptors exclusively" OR DREADD OR hM3D OR hM4D
Combining opto/DREADD to deconstruct medial amygdala regulation of social dynamics
interesting and comprehensive paper here.
Chemogenetic silencing of the midline and intralaminar thalamus blocks amygdala-kindled seizures
cool paper here of potential translational impact
Impaired Recall of Positional Memory following Chemogenetic Disruption of Place Field Stability
Nice use of ivermectin-actuated channels f or silencing neurons and dissecting behavior.
Genetic Isolation of Hypothalamic Neurons that Regulate Context-Specific Male Social Behavior
Nice paper in Cell Reports: Here, we identify a population of neurons in the ventral premammillary nucleus of the hypothalamus (PM V ) that are strongly activated in male intruder mice in response to a larger resident male but that are not responsive to females.
Chemogenetics to interrogate Interactions between respiratory oscillators
Nice paper combining allostatin Gi linked receptors and DREADDS in elife
DREADDs do not apparently desensitize in vivo
As I recently suggested in an article in Neuron , because of receptor reserve when DREADDs are expressed desensitization is unlikely to occur to any significant extent. We now have some nice validation of this basic principle of GPCR molecular pharmacology in a nice paper just published in Nature Communications .
DREADDs defining ETOH actions on striatal D1 and D2 pathways
Distinct Synaptic Strengthening of the Striatal Direct and Indirect Pathways Drives Alcohol Consumption In this study, we measured both glutamatergic and GABAergic activity in D1-MSNs and D2-MSNs and found that NMDA receptor (NMDAR) activity in D1-MSNs and GABAergic activity in D2-MSNs were selectively potentiated following cycles of alcohol consumption and withdrawal. Using a chemogenetic approach employing designer receptors exclusively activated by designer drugs (DREADDs), which allowed selective manipulation of D1- or D2-MSN activity (23) , we discovered that both of these cell types were not only necessary, but also sufficient, to drive alcohol consumption. Furthermore, we observed that D2R-glycogen synthase kinase-3β (GSK3β) signaling regulated GABAergic activity and thus, alcohol consumption. The findings of this study provide detailed mechanistic information indicating how different forms of neuroplasticity in distinct neuronal populations of the striatal direct and ...
Retro-DREADD and axonal silencing
Very nice use of chemogenetic and viral technology here to obtain axonal silencing of projections via Retro-DREADD in different brain areas
NATURE: memory linking rescue by activating cells with DREADDs
Nice ultimate experiment from this paper by labs of Golshani and Silva
Cool DREADD paper deconstructs cortical desynchronization and arousal
Nucleus incertus promotes cortical desynchronization and behavioral arousal
Cre dependent DREADD mice
As we deposited these mice in JAX some time ago, we have now published a fairly complete characterization of said mice. These should be useful for a large number of applications.
NEURON: using DREADDs as GPCRs
I once was meeting with a well-known neuroscientist who said DREADDs were 'cool channels'. Of course they are GPCRs and here is a nice paper showing how they can be used to identify signaling pathways in neurons responsible for behavior.
NEURON: Bidirectional DREADD modulation of Transplanted DA Neurons
Nice proof of concept story here for bi-directional modulation of transplanted neurons....one can imagine all sorts of potential uses for this.
DREADDs to deconstruct how orbitofrontal cortex modulates reward sensitivity
The Medial Orbitofrontal Cortex Regulates Sensitivity to Outcome Value
DREADD + CRISPR + iPS cells: cool paper
An interesting paper and another proof-of-concept study combining chemogenetics, CRISPR and iPS cell technology.
CURRENT BIOLOGY: The reactivation by DREADDs reproduces sex-specific aggression to ESP1
Pretty amazing movie here. Exocrine gland-secreting peptide 1 (ESP1) released into male tear fluids is a male pheromone that stimulates sexually receptive behavior in female mice via the vomeronasal sensory system. ESP1 also induces c-Fos expression in male brain regions distinct from those in females. However, behavior in males following ESP1 exposure has not been examined. In the present study, we show that ESP1, in conjunction with unfamiliar male urine, enhances male aggression via the specific vomeronasal receptor V2Rp5. In addition, male mice that secrete ESP1 but lack V2Rp5 exhibit a lower level of aggressiveness than do mice that express V2Rp5. These results suggest that ESP1 not only acts as a male pheromone in both sexes but also serves as an auto-stimulatory factor that enhances male aggressiveness by self-exposure. Finally, re-activation of ESP1-induced c-Fos-positive neurons by using the designer receptor exclusively activated by designer drug (DREADD) approach resu...
Nature Neuro: Silencing spinal interneurons inhibits immune suppressive autonomic reflexes caused by spinal cord injury
Spinal cord injury (SCI) at high spinal levels (e.g., above thoracic level 5) causes systemic immune suppression; however, the underlying mechanisms are unknown. Here we show that profound plasticity develops within spinal autonomic circuitry below the injury, creating a sympathetic anti-inflammatory reflex, and that chemogenetic silencing of this reflex circuitry blocks post-SCI immune suppression . These data provide new insights and potential therapeutic options for limiting the devastating consequences of post-traumatic autonomic hyperreflexia and post-injury immune suppression.
DREADD-induced inactivation of the lateral orbitofrontal cortex increases drinking in ethanol-dependent but not non-dependent mice
I'm seeing a lot of work with DREADDs among researchers in the ETOH field. Here's an interesting paper
Pivotal role of a dorsolateral corticospinal pathway in mediating spontaneous recovery after SCI
Nice paper combining chemo- and optogenetics.
Promoter-Specific Effects of DREADD Modulation on Hippocampal Synaptic Plasticity and Memory Formation
Interesting and carefully-controlled study recently published in J Neuroscience.
CELL: Robust chemogenetic modulation of Systemic Insulin Sensitivity via Myostatin Expression in Brown Adipose Tissue
A quite amazing paper has appeared recently here. Of note is the replication of a technology reported previously of DREADD-assisted neuronal activity mapping.
NATURE: DREADDs to deconstruct predator odor neurons
Interesting paper from the Linda Buck lab now on-line .
YUSTE LAB: More evidence for in vivo silencing via DREADDS
Nice paper just published . Key data below:
KOR-DREADD and ETOH
Another paper showing the utility of both RETRO-DREADD and KOR-DREADD technology.
CELL: DREADDs to Control Whole-Body Glucose Metabolism via VMH
Pretty interesting paper published a couple of weeks back in Cell ; key data below:
That's "Bryan" not "Brian"
From an eLife paper where DREADDs were used extensively: " For hM4D(Gi) experiments, we infected V1 of P30-60 PV-Cre mice or wild-type littermates with an AAV9-hSyn-DIO-HA-hM4D(Gi)-IRES-mCitrine virus (UNC viral core – generated by Dr. Brian Roth’s laboratory)" There are plenty of Brian and Bryan out there and here is a photo of 3 at once (Bryan Roth, Brian Kobilka [Nobel 2012], Brian Shoichet)
NATURE COMMUNICATIONS: DREADDs outside the CNS
Nice paper here using Gq-DREADD and many other approaches to deconstruct Gq signaling in various adipose tissues.
Decreasing cortical activity rescues Shank3B−/− phenotype by hM4Di
Interesting paper in Nature Neuroscience . Data below:
How does Salvia exert it's actions in humans--Kappa Receptor!
Though we discovered this some time ago , there has been lingering controversy regarding it's mechanism of action in humans. Here is a controlled study consistent with k-opioid being essential target.
NEURON PRIMER ON DREADDS: essential and (hopefully) useful information for users
On-line here . A fair amount of time was spent on this to make sure key issues were addressed --especially those which may not be obvious to those who do not study GPCRs. BTW: DREADDs are engineered GPCRs......
TOCRIS NOW HAS ALL VALIDATED DREADD LIGANDS AVAILABLE
Link is here . My lab receives no proceeds from the sales of DREADD ligands and they are available to anyone without encumbrances (IP or otherwise).
NEW DREADD: Gq/11 without arrestin activation
This will be useful for determining how and to what extent canonical (G protein) vs non-canonical (arrestin) singaling is essential for the actions of a particular biological/physiological phenomenon.
Use of M1 -DREADD to probe learning and memory
A nice paper just published using the M1-DREADD which like M3 is Gq coupled to interrogate learning and memory. As predicted M1 behaves similarly to M3.
NATURE: Equivalency of chemo- and optogenetic silencing
Several of these papers have been appearing the past few days--my guess is reviewers who are fans of one technology or the other require that both be used. At any rate here is the key figure from a paper on-line in Nature today . Nice to see all the relevant controls....in addition to citing the original DREADD paper (which sadly few do).
PNAS: visualization of in vivo silencing via hM4Di
Frequently am asked whether anyone has actually seen neurons 'silenced in vivo' via hM4Di. Here is the relevant data from a paper just out in PNAS . It shows a great example of how chemogenetics can provide the killer experiment when linked with in vivo neuronal imaging of neuronal activity.
AAV quality control for DREADDs--request input
Occasionally we get emails from users regarding AAV serotypes/lots which they believe are problematic. We normally forward these to the UNC vector core but would greatly appreciate having a 'running total' of those lots/serotypes/constructs which are problematic. That way we can take that information back to the vector core and help them to institute rigorous (and recommended) quality control above and beyond what they typically do. So here's what you should do: Simply reply to this listing the AAV lot/serotype/construct which works as expected or which does not work as expected. Example: Lot#666/8/ DEADDREADD : works fine--all cells exploded upon contact with CNO as predicted.
NATURE NEUROSCIENCE: DREADDs and PSAMs interrogate memory
PV plasticity , which involves changes in PV and GAD67 expression and connectivity onto PV neurons, was regulated by cAMP signaling in PV neurons. ... Our results reveal general network mechanisms of long-term memory consolidation that requires plasticity of PV basket cells induced after acquisition and sustained subsequently through D1/5 receptor signaling.
DREADDs therapeutically modulate experimental Alzheimer's Disease
Interesting paper here Attenuation of β-Amyloid Deposition and Neurotoxicity by Chemogenetic Modulation of Neural Activity
DREADDs and optogenetics to deconstruct opioid modulation of inputs to PAG
Nice paper recently published. Intriguingly the inputs when activated modulated hot plate but not tail flick nor locomotor activity.
Detailed genotyping instructions for FLOXED-Gq-DREADD mouse from JAX
Attached find our recommended and detailed genotyping instructions for the FLOXED-Gq-DREADD (hM3Dq) mouse
NATURE COMMUNICATIONS: DREADDs to deconstruct Gs vs Gq signaling and feeding
Very complete and compelling paper Here we show that the activation of G s -coupled receptors expressed by AgRP neurons leads to a robust and sustained increase in food intake. We also provide detailed mechanistic data linking the stimulation of this class of receptors to the observed feeding phenotype. Moreover, we show that this pathway is clearly distinct from other GPCR signalling cascades that are operative in AgRP neurons. Our data suggest that drugs able to inhibit this signalling pathway may become useful for the treatment of obesity.
Chemo- and Optogenetic Activation of Gαs Signaling in BLA Induces Anxiety
Interesting combo chemo- and optogenetics approaches yielding convergent data