...improve the study of specific neuronal cell types involved in neurodegenerative diseases, the Cole Lab selected...linked to disease, including motor neurons (-3mnx1/Hb9), pan neuronal (elavl3/HuC), microglial/macrophage...specific neuronal subtypes was not available until now. In order to target specific neuronal subtypes ...infecting GABA-ergic interneurons here. Dimidschstein J, et al. Nature Neuroscience. 2016. PubMed PMID:...generate zebrafish transgenic lines to study neurobiology and neuorodgenerative diseases. The Tol2 Gateway-Compatible...), and astrocytic (gfap).This toolbox adds new neuronal tools to the expanding gateway compatible vectors... PMID: 25772473 Targeting and manipulating interneurons using a viral approach Genetically modified...

...great in a GABA neuron, for instance, may not transduce the adjacent norepinephrine neuron at all. There... for Defense Behaviors. Neuron, 93(1), 33–47. https://doi.org/10.1016/j.neuron.2016.11.045 More resources...expression can still occur (Fig. 3) - for example, some neurons can transiently express TH - so you’ll need to ...expression in both oligodendrocytes (Olig2) and neurons (NeuN). Image from Griffin et al., 2019, Figure...central nervous system. Frontiers in Molecular Neuroscience, 7. https://doi.org/10.3389/fnmol.2014.00076...cFork anterograde tracing system. Experimental Neurobiology, 29(3), 219–229. https://doi.org/10.5607/en20006...

...named NeuroStack was used (Orguc et al., 2020). Using a programmable microcontroller, NeuroStack coordinates...and stimulate or manipulate responses through neurological means like electric stimulation, drug administration...optogenetic tools, calcium sensors, and other common neuroscience tools into the brains of model organisms, including...tissue. They performed optical stimulation using NeuroStack and the fibers as previously described, then ...trended with expression of the opsin in the target neurons, validating the functionality of these inputs and...Success! They also observed reliable optically evoked neuron activity for at least several months after implantation...enteroendocrine cells within the intestines known as neuropods. They observed increased vagus nerve firing in...

...biologists, while neuroscientists would like to monitor calcium specifically in their favorite neurons. This specificity... V., Fluorescent Biosensors for Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers... or animal system. This ‘thing’ could be a neurotransmitter, such as norepinephrine, or an electrolyte...contraction, and calcium channels are activated for neuronal action potentials; it’s an ion of interest for...—keep reading to find out how!   Neurotransmitters Many canonical fluorescent as well as FRET-based...been engineered to detect a wide array of neurotransmitters ranging from dopamine to glutamate, all the... Frontiers in Cellular Neuroscience. 2019 Oct Vol. 13; 1662-5102. DOI: 10.3389/fncel.2019.00474 Resources...

...cell types (astrocytes, microglia, neurons, etc.) and even neuronal phenotype in a mixed population (Burger...Many neuroscience experiments that require gene expression in a specific cell type rely on transgenic...tyrosine hydroxylase (TH) promoter for catecholamine neurons or glial fibrillary acidic protein (GFAP) for astrocytes...vectors for off-target expression.  Some classes of neurons transiently express TH during development or have...there is promoter activity. Additionally, some neurons express low levels of GFAP, which can lead to off-target...off-target expression of the packaged transgene in neurons and glia.   Check the recombination rate The first...on Helper Virus Concentrations. Front Synaptic Neurosci 12. https://doi.org/10.3389/fnsyn.2020.00006 O...

...oligodendrocytes, microglia, astrocytes, neurons, and inhibitory neurons. Your choice of AAV will depend upon... as a new tool for neuroanatomical tracing (11). With the ability to produce neuron specific transfection...contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University of Manitoba. ... These AAVs may move toward the cell bodies of neurons (retrograde transport) or towards the synapse (...Cre-lox, this new method could allow for labeling of neurons showing a very specific phenotype. Previous neural... Katrina Armstrong. Katrina Armstrong is a Neurophysiology Msc Student at the University of Manitoba. ... Floris G., et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring...

...coffee does it take to fuel more than 30,000 neuroscientists over a 5 day conference? I feel like here at... at the annual Society for Neuroscience meeting in Washington, D.C., we must be making a dent in the global...conference, I have been impressed by how the neuroscience community pulls off such an inspiring, thought-provoking...made in reprogramming astrocytes to functional neurons in vivo. And I’ve seen evidence of the infiltration...infiltration of CRISPR-Cas tools into neuroscience experiments. Speaking of CRISPRs, we were excited to have... the importance of scientific sharing to the neuroscience community. The scientists we've spoken with ... for a free plasmid! Also check out the PLOS Neuroscience Community blog to read reports from bloggers...

... directed by a gRNA. 2A-Puro In this plasmid, SpCas9 is fused to the puromycin resistance gene through...pSpCas9(BB)-2A-Puro (PX459) V2.0 from it's predecessor, which had a point mutation in the puromycin resistance...most request plasmid in 2016 was... pSpCas9(BB)-2A-Puro (PX459) V2.0       Similar to last year, the top...through a 2A peptide. The puromycin resistance gene enables selection of cells with the plasmid post transfection...causes the ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This...mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro (PX459) V2.0: What's not in the name? Not included...

...., et al. (2023) Neuron, 111(10), 1547–1563.e9. doi: https://doi.org/10.1016/j.neuron.2023.03.009. PMID...microglia The REVeRT Dual AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation... vectors for CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression of BFP2.   ...fidelity during action potentials of fast-spiking neurons. The same A-to-D mutation in the related voltage...vectors and are suitable for extended imaging of neurons in vivo. Because they are negative-going sensors... the vector to silence transgene expression in neurons and other non-microglia cells.      Figure...42386-0. PMID: 37852949. New and notable Neurodegeneration Collection updates Last but not least, check...

...continues to supply neuroscience with ever more efficient tools for light mediated neuronal control and we ...pAAV-Ef1a-fDIO-GCaMP6s Neurotransmitter sensors: Dopamine and norepinephrine Neuroscience research relies on...photostimulation of axons and dendrites on neighboring neurons. We have also packaged hChR2(H134R) plasmids from...calcium dynamics that govern such activities as neurotransmitter release is vital in studying neural networks... on accurate monitoring of neurotransmitters and we are excited to offer AAV for the newest GRAB sensors...tool from Loren Looger’s lab at Janelia for neuroscientists to measure glutamate levels. iGluSnFR.A184S...

...Oliver Griesbeck of the Max Planck Institute for Neurobiology has been working on genetically encoded indicators...monitoring the activity and biochemistry of living neurons in an effort to understand the connection between...considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors Griesbeck’s...available beam splitters. In addition to imaging neurons, Griesbeck adds that this color change has a particular...refined by a secondary screen in rat hippocampal neuron cultures. The optimized Twitch sensors allowed ...researchers to see tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes...them a versatile tool for application in both neuroscience and immunology. Don’t be shy: Ratiometric imaging...

...be used for sparse labeling of neurons for tracing experiments.  Neuronal tracing experiments: While synthetic...broad number of cell types, such as inhibitory neurons, rods, cones, or retinal ganglion. Rather than ...that produce sparse labeling would be useful for neuronal tracing experiments where lower levels of labeling...labeling makes it easier to trace a neuron’s projections. Generating animal models: Transgenic animals allow...for retinal gene therapy Find more about precise neuronal targeting using AAV technologies Browse all of... H.P., Krol, J., & Roska, B. (2019). Targeting neuronal and glial cell types with synthetic promoter AAVs... mice, non-human primates and humans. Nature Neuroscience, 22, 1345-1356. https://doi.org/10.1038/s41593...

...pushed the boundaries in neuroscience by granting targeted control over neuronal activity. While they serve...scale of milliseconds, and can precisely link a neuron's activation to behaviors. Scientists continue to... mutated to have faster off kinetics and allow neurons to repolarize faster.  Chemogenetics does not offer...Administering the drug once can result in several hours of neuronal excitation or inhibition. This sustained activation...tissues, cell types or even subcellular regions of a neuron. Optogenetic approaches excel at discrete cell ...precise spatiotemporal control of a small subset of neurons is necessary, use optogenetics. If broad control...

... reagent resource for the neuroscience community called NABOR (Neuroscience AntiBody Open Resource). We...Brain Research through Advancing Innovative Neurotechnologies® (BRAIN) Initiative, will allow Addgene to...thousands of high-quality antibodies/nanobodies for neuroscience research and is an expert at validating these...While the focus of the NABOR platform will be neuroscience reagents and data, the platform can be expanded...are among the most commonly used reagents in neuroscience research today, but there is a critical unmet... to create recombinant antibodies for use in neuroscience research. More broadly, advances in gene synthesis...

...are powerful tools to modulate the activity of neurons and other brain cells. Since the opsins or chemogenetic... we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or mice (Galvan et al., 2019)... electron microscope showed us that, in monkey neurons, the large majority of the gold particles were ...found a larger proportion on the membrane in mice neurons, most of them were still cytoplasmic. Use of ‘small...hM4Di‐mCherry at the plasma membrane in monkey neurons, we next hypothesized that a different tag fused...instance, when the fluorescence expressed in the neuropil conceals identification of opsin-expressing cells...Ultrastructural localization of DREADD s in monkeys. Eur J Neurosci 50:2801–2813 . https://doi.org/10.1111/ejn.14429...

...presynaptic region of one neuron and postsynaptic region of a different neuron, come into proximity in ...Druckmann et al., Neuron. 2014. Feb 5;81(3):629-40.  Next-Gen Brainbow toolkit for neuronal imaging Joshua...perform independent two-color optical excitation of neurons. The paper describes five previously unknown ...allowing labs to precisely and non-invasively control neuronal signaling. For more information on DREADDs, ...Addgene's Genome Engineering Guide. Fire up those neurons: mGRASP A Nature 2012 article by Jinny Kim and... use of another set of mGRASP plasmids in their Neuron 2014 paper. Kim et al., Nat Methods. 2011. Dec...high-resolution fluorescent imaging of individual neurons. The technology generates a unique spectral identity...

...norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype. ...challenge to scientists. What works well in one cell or neuron type may not work well for a neighboring cell type...it to publish!  Working with AAV vectors in neuroscience Let’s consider my first challenge when I started...express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested four serotypes containing an expression...84%) were both excellent at infecting our target neurons (Figure 1). However, this one set of experiments...in Deep Brain Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https://doi.org/10.3389...

...particularly well-suited for neuroscience investigations. For example, neuronal activity can be tracked in...available from strain distribution centers in the US, Europe, and China. In addition, the Zebrafish Model Organism...example, specific tissues such as blood cells or neurons can be fluorescently labeled, allowing researchers...high-resolution mapping of the brain by uniquely labeling neurons. Zebrafish are also amenable to manipulation of...consequences of activating or silencing specific neurons in living larvae. In addition, zebrafish have been...PubMed PMID: 18845710. Kim, Dal Hyung, et al. "Pan-neuronal calcium imaging with cellular resolution in freely...

...expressed may be found at very different levels (Mauro and Chappell, 2014). Due to this variation in tRNA...scientists with the ability to selectively silence neurons in vivo (Slimko and Lester, 2003).  Addgene has...modification sites and even affect protein function (Mauro and Chappell, 2014). Different rates of translation...experiments and in biotechnology and therapeutics (Mauro and Chappell 2014).  Ultimately, it is important...s12859-017-1793-7 Dresios J, Chappell SA, Zhou W, Mauro VP (2005) An mRNA-rRNA base-pairing mechanism for...604–615 . https://doi.org/10.1101/gr.139758.112 Mauro VP, Chappell SA (2014) A critical analysis of codon...dramatically improves expression levels. Journal of Neuroscience Methods 124:75–81 . https://doi.org/10.1016/...

...spinal muscular atrophy (SMA)." European Journal of Paediatric Neurology 19 (2015): S64-S65. Additional...results in the degeneration of spinal cord motor neurons. This disease is estimated to affect anywhere from...screens for small molecules that block the neurodegenerative effects of smn1 disruption. As part of his...for use in the clinic and thereby restore motor neuron function in spinal muscular atrophy patients. Noushin... University of British Columbia Department of Urological Sciences. Dr. Nabavi studies peritoneal mesothelioma...