...the brain to visualize how neurons are connected. Viral Vectors as Neuronal Tracers Due to their ability...individual neurons they infect. However, they generally are not transmitted to adjacent neurons via synapses...Converting a transsynaptic neuronal tracer into a monosynaptic (first order) neuronal tracer For RABV to crawl...nerve terminals of adjacent neurons (i.e., it is taken up by projection neurons). Research into these phenomenon...order neurons, via entry mediated by the G protein/G-protein receptor. Once in first order neurons, the...most of the projections to these new neurons came from nearby neurons. It wasn’t until later time points...mapping of direct inputs to midbrain dopamine neurons. Neuron. 2012 Jun 7;74(5):858-73. Pubmed PMID 22681690...

...Dopamine? DA is produced by specific dopaminergic neurons in a brain region called the substantia nigra, ..., which is the main source of the body’s DA. Neurons often fire at a certain frequency, and this frequency...understood, the direct effect of DA binding to a target neuron is more mysterious. Thus, scientists have developed...dynamics of optogenetically activated and inhibited neurons in live mice Report modulation of DA signaling ...brain slices Demonstrate DA detection in a single neuron in vivo in Drosophila Detect DA dynamics in response...PMC426888. Patriarchi, Tommaso, et al. "Ultrafast neuronal imaging of dopamine dynamics with designed genetically...

...Limitations of common genetically encoded neuronal tracers Neuronal tracers such as epitope tags and fluorescent...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures GFP...label somata but not neuronal processes. In contrast, smFPs can label fine neuronal structures with high...functions and others are controlled by collections of neurons that are intricately wired into circuits through...have recently made it much easier to map these neuronal networks. For almost three decades, recombinant... in order to define those that target specific neuronal populations in the brain. The Allen Institute ...quickly find the best AAV serotypes to target the neurons they’re studying. Thanks to this work it is now...

...parts of the brain is mediated via individual neurons. Neurons are composed of a cell body, with dendrites...proximal neuronal cells. In the specific example of episodic memory, a subset of hippocampal neurons projects...axons directly to the prefrontal cortex, but also indirectly via synapses to neurons in other brain regions...using these viruses to mark neuronal connectivity a large majority of neuronal cells are labeled overtime...auditory neuronal circuitry in vivo. A variant of RABV has been developed to only label a neuronal connection...AAV vectors can be equipped with strong promoters for high and neuron-specific transgene expression. Fluorescent...cell types, for example only dopaminergic neurons, neuronal tracing has the added benefit of only labelling...

...available types of degron tags. What is a degron tag and why should I use one?   Degron tags are a class...Finally, some degron tags can do more than just degrade! Read on to find out more. Common degron tags When... comes to degrons, you have options! Below we will review the 4 most common types of degrons and some ...protease is situated between the degron and the gene of interest and the degron domain is terminal. Between...the protease and degron remain attached to the tagged protein. In this event, the degron domain shuttles...In the last decade, the use of degron tags has become increasingly popular for the modulation of endogenously...expressed proteins. Here we will review what advantages degron tags can offer over other protein control methods...

...bidirectional promoters, or the creation of bicistronic or multicistronic vectors. Unlike promoters which will...upstream cistron will have a few additional residues added to the end, while the downstream cistron will ...transcripts for each gene that is expressed, multicistronic vectors simultaneously express two or more ... so in this blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work... and how to get started with them. Why Use multicistronic vectors? Detecting cells that are expressing... selectable markers from a unique promoter, multicistronic plasmids ensure that any cells that are positive...derived from the same transcript. Of course multicistronic vectors do not have to exclusively be used ...

...backstage team. Meet the microenvironment, a tumor’s backstage team. Microenvironments are cellular regions...Tumors are surrounded by microenvironments and what encircles a disease is highly prone to affect it. While...type in the tumor microenvironment is the immune cells. The contradiction is strong in this one. After...cancer microenvironment Ever since Stephen Paget’s “seed and soil” theory in 1889, microenvironments have...Isaac P. "The tumor microenvironment: the making of a paradigm." Cancer Microenvironment 2.1 (2009): 9-17...associated fibroblasts (CAFs) in tumor microenvironment." Frontiers in bioscience: a journal and virtual...precisely that for breast cancer. What are tumor microenvironments made of?  A tumor’s genesis is divided into...

...solution for discriminating between neurons for light microscopy-based neuronal reconstruction. To achieve stochastic... et al., 2016, 2018).   How to use Tetbow for neuronal labeling To use Tetbow, you can introduce the ... is interested in developing genetic tools for neuronal circuit tracing.     Marcus Leiwe is an assistant...Meng-Tsen, et al. "Super-resolution mapping of neuronal circuitry with an index-optimized clearing agent...and Takeshi Imai. "Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical...Read about Brainbow Learn about using AAVs for neuronal tracing Visit the viral vectors blog topic page...

...that your project is an innovative solution with a strong impact, or a journalist does not understand the...

...and developed the SciNote Electronic Lab Notebook. Implementing an electronic lab notebook starts with ...should have functionalities such as: electronic signatures, electronic witnessing and audit trails. How ...write things down when doing scientific research? Electronic lab notebooks and researcher's needs The familiar...traceability and reproducibility of research. Electronic lab notebooks (ELNs) are here to make lab work...explored the barriers to adoption within academic environments to better understand the issues labs are facing...traceable. However, switching from paper to an electronic lab notebook requires time and knowledge. In ...recent article, we translated our knowledge of electronic lab notebooks into 9 questions that will help...

...thrive, in the face of some of Earth’s harshest environments: corrosive pH ranges, crushing pressures, near-melting...folding at low temperatures, since mesophilic chaperones would not function well under suboptimal conditions...engineering a plasmid which contained cold tolerant chaperones (the Cpn 10/60 isolated from Oleispira antarctica...the figure below shows, E. coli expressing the chaperones protein resulted in higher levels of growth compared...help foster a more productive and collaborative environment for research.  We’ve also involved ourselves ...

... the brain, a connectome that would detail how neurons form circuits and where they synapse. View the ...Golgi staining, injection of diffusible labels, or electron microscopy of stained sections, were limited due...inversion of the construct. Sanes and Lichtman used pronuclear injection to insert these Brainbow constructs...vast array of colors needed to mark individual neurons, the system also suffered from a number of limitations...the tendency of fluorophores to aggregate in the neurons’ somata. Second, the system did not permit analysis... disproportionately expressed by a majority of neurons, limiting the number of distinct colors that could...through at least six generations. In addition to the neuronal pThy1-Brainbow constructs, Addgene also has two...

...Nicola Patron is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like...that go on between plants and their pathogens. As Patron explains it, her work has always been focused on...Modular Cloning (MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate and...Addgene: Will the MoClo tools work in any plant? Patron: The MoClo kit is entirely for dicots, not grasses...tips for those looking to get started with MoClo? Patron: The parts will be familiar to most plant scientists...there other tools you are working on for plants? Patron: Our lab and others at the Sainsbury are working... of engaging about science in that way for you? Patron: I started my Twitter account recently inspired...

...neuroscience with many brain regions, neuronal networks, and specific neuronal populations to consider. To tease...Making use of AAV’s selectivity for neurons: targeting defined neuronal populations using viral strategies... in neurons that project to different regions of the brain. A more locally defined set of neurons can ...tease apart how different neurons impact a certain output, neuroscientists have begun to use more precise...genetic tools with the manipulation of specific neuronal subtypes and determine how these changes affect...combinations of AAV-based molecular tools for targeted neuronal manipulation. The goal of the workshop was to ...practical guide which was recently published in Frontiers in Neuroanatomy.  Here are the three main takeaways...

...Elementary and Middle Schools   Image courtesy of Ron Mader, via Flickr, under the Creative Commons license...education that is provided through a structured environment such as the classroom or a defined learning program...

...miRNA regulation have yet to be fully elucidated. Ron Weiss’s lab at MIT pushed the field further with ...halves to enable VAMP2 cleavage, and suppresses neuronal transmission within minutes. Constructs using ...split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter...

...23086203. PubMed Central PMCID: PMC3699407. Milo, Ron. "What is the total number of protein molecules per..., that strongly binds the glycoprotein, avidin or a similar protein, streptavidin. This strong interaction...are approximately 2-4 million proteins per cubic micron in bacteria, yeast, and mammalian cells (Milo, ...ascorbate peroxidase (APEX), used as a genetic tag for electron microscopy, could be used for efficient proximity...of hydrogen peroxide. This radical reacts with electron-rich amino acids such as tyrosine on neighboring...Stephanie S., et al. "Directed evolution of APEX2 for electron microscopy and proximity labeling." Nature methods...peroxidase as a genetically encoded reporter for electron microscopy." Nature biotechnology 30.11 (2012)...

...these regions, a process driven by neurons. By studying how neurons are physically connected, we can begin...these connections in the brain are (drum roll) neurons! Neurons are really cool because they can transmit ...There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information to ... functionally dissect specific neurons within their natural environment, all while reciting their favorite...defined population of neurons. For example, say I wanted to access all the neurons that signaled from the...ligands activates both neurons at the injection site and any projection neurons retrogradely transduced... efficient retrograde access to projection neurons." Neuron 92.2 (2016): 372-382. PubMed PMID: 27720486...

.... What do microbial rhodopsins and neurons have in common? Neuronal stimulations are triggered by the ...the neuron fires, sending the electrical signal down the axon (Lodish et al., 2000). Most neurons in the...Vandenberghe, 2019). How to achieve strong opsin expression in specific neuronal cell types then? This is where... 2014). How does FLEx enable strong opsin expression in specific neuronal cells? This can be achieved ...enabling its expression under a strong promoter only in specific neuronal cells. FLEx ensures, therefore...et al. "Silencing neurons: tools, applications, and experimental constraints." Neuron 95.3 (2017): 504-...What is optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford University...

...specify and assemble diverse arrays of neurons, forming the neuronal circuits that will later be refined ...individual neuronal populations.  The Yang Lab’s AAV plasmids used to express Cre under neuron-specific ...kinase II alpha (aCaMKII; excitatory neurons) and Synapsin-1 (pan-neuronal) promoters (Figure 1b), and sgRNAs...response to neuronal activity (hence, “immediate-early”) and consequently regulates neuronal physiology...between Bdnf I-overexpressing neurons and Bdnf IV-overexpressing neurons. Among the upregulated genes ...contributions of transcript isoforms to neuronal physiology. The Day Lab’s neuron-optimized lentiviral vectors can...process. However, while the neuronal marker TUJ1 is useful for visualizing neuronal differentiation by immunohistochemistry...