Springer protocols feed by Neuroscience 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Culturing Mouse Cerebellar Granule Neurons
The cerebellum plays an important role in motor control, motor skill acquisition, memory and learning among other brain functions. In rodents, cerebellar development continues after birth, characterized by the maturation of granule neurons. Cerebellar granule neurons (CGNs) are the most abundant neuronal type in the central nervous system, and they provide an excellent model for investigating molecular, cellular, and physiological mechanisms underlying neuronal development as well as neural circuitry linked to behavior. Here we describe a procedure to isolate and culture CGNs from postnatal day 6 mice. These cultures ...
Source: Springer protocols feed by Neuroscience - May 27, 2013 Category: Neuroscience Source Type: news
Culture of Dissociated Hippocampal Neurons
Hippocampal neurons consist mainly of pyramidal neuron and granule cell, and dissociated hippocampal neurons are a good tool to investigate the molecular and cellular mechanism of neuronal development and neuronal degenerative disease in the central neuronal system (CNS). Here, we describe a general procedure of dissociated hippocampal neuron culture. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - May 27, 2013 Category: Neuroscience Source Type: news
Nucleoside Analog Labeling of Neural Stem Cells and Their Progeny
Nucleoside analog pulse labeling is an important technique which can assess the birthdate, cell cycle maintenance, or cycling rates of cells during development. This method has evolved over several decades of use and is now applied to a multitude of tissue subtypes and systems. The methodology in this chapter covers the classic uses for analog pulse labeling as well as their use in conjunction with the newly characterized technique of in utero electroporation (IUE). (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - May 27, 2013 Category: Neuroscience Source Type: news
Neural Induction with a Dopaminergic Phenotype from Human Pluripotent Stem Cells Through a Feeder-Free Floating Aggregation Culture
Pluripotent stem cells are promising potential sources for cell replacement therapy and are useful research tools for exploring disease mechanisms. Neural cells are one of the cell types that have been most efficiently differentiated through several established protocols. This chapter describes the feeder-free floating aggregation culture system for the induction of dopaminergic neurons. This method is simple and highly efficient for the production of dopaminergic neurons. It has several advantages for application in clinical usage in comparison to the other protocols using either feeder cells or Matrigel. (Source: Springe...
Source: Springer protocols feed by Neuroscience - May 27, 2013 Category: Neuroscience Source Type: news
Isolation and Maintenance of Cortical Neural Progenitor Cells In Vitro
Neural progenitor cells (NPCs) or neural stem cells are important tools for investigating central nervous system (CNS) development. NPCs can be used in therapeutic strategies and for characterizing differentiation mechanisms. Here, we describe methods for isolating and culturing embryonic NPCs. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - May 27, 2013 Category: Neuroscience Source Type: news
A 1H NMR Assay for Measuring the Photostationary States of Photoswitchable Ligands
Incorporation of photoisomerizable chromophores into small molecule ligands represents a general approach for reversibly controlling protein function with light. Illumination at different wavelengths produces photostationary states (PSSs) consisting of different ratios of photoisomers. Thus optimal implementation of photoswitchable ligands requires knowledge of their wavelength sensitivity. Using an azobenzene-based ion channel blocker as an example, this protocol describes a 1H NMR assay that can be used to precisely determine the isomeric content of photostationary states (PSSs) as a function of illumination wavelength. ...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Photochromic Potassium Channel Blockers: Design and Electrophysiological Characterization
Voltage-gated potassium (K
v) channels are membrane proteins that open a selective pore upon membrane depolarization, allowing K+ ions to flow down their electrochemical gradient. In neurons, K
v channels play a key role in repolarizing the membrane potential during the falling phase of the action potential, often resulting in an after hyperpolarization. Opening of K
v channels results in a decrease of cellular excitability, whereas closing (or pharmacological block) has the opposite effect, increased excitability. We have developed a series of photosensitive blockers for K
...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Characterization of One- and Two-Photon Photochemical Uncaging Efficiency
The idea of using light to unleash biologically active compounds from inert precursors (uncaging) was introduced over 30 years ago. Recent efforts prompted the development of photoremovable protecting groups that have increased photochemical efficiencies for one- and two-photon excitation to allow more sophisticated applications. This requires characterization of one- and two-photon photochemical efficiencies of the uncaging processes. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Characterizing Caged Molecules Through Flash Photolysis and Transient Absorption Spectroscopy
Caged molecules are photosensitive molecules with latent biological activity. Upon exposure to light, they are rapidly transformed into bioactive molecules such as neurotransmitters or second messengers. They are thus valuable tools for using light to manipulate biology with exceptional spatial and temporal resolution. Since the temporal performance of the caged molecule depends critically on the rate at which bioactive molecules are generated by light, it is important to characterize the kinetics of the photorelease process. This is accomplished by initiating the photoreaction with a very brief but intense pulse of light ...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
A FLIPR Assay for Evaluating Agonists and Antagonists of GPCR Heterodimers
Calcium signaling plays a major role in the function of cells. Measurement of intracellular calcium mobilization is a robust assay that can be performed in a high-throughput manner to study the effect of compounds on potential drug targets. Pharmaceutical companies frequently use calcium signaling assays to screen compound libraries on G-protein-coupled receptors (GPCRs). In this chapter we describe the application of FLIPR technology to the evaluation of GPCR-induced calcium mobilization. We also include the implications of GPCR hetero-oligomerization and the identification of heteromeric receptors as novel drug targets o...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Using Yeast to Study Potassium Channel Function and Interactions with Small Molecules
Analysis of ion channel mutants is a widely used approach for dissecting ion channel function and for characterizing the mechanisms of action of channel-directed modulators. Expression of functional potassium channels in potassium-uptake-deficient yeast together with genetic selection approaches offers an unbiased, high-throughput, activity-based readout that can rapidly identify large numbers of active ion channel mutants. Because of the assumption-free nature of the method, detailed biophysical analysis of the functional mutants from such selections can provide new and unexpected insights into both ion channel gating and...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Chemical Derivatization and Purification of Peptide-Toxins for Probing Ion Channel Complexes
Ion channels function as multi-protein complexes made up of ion-conducting α-subunits and regulatory β-subunits. To detect, identify, and quantitate the regulatory β-subunits in functioning K+ channel complexes, we have chemically derivatized peptide-toxins that specifically react with strategically placed cysteine residues in the channel complex. Two protein labeling approaches have been developed to derivatize the peptide-toxin, charybdotoxin, with hydrophilic and hydrophobic bismaleimides, and other molecular probes. Using these cysteine-reactive peptide-toxins, we have specifically targeted KCNQ1-KCNE1 ...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Engineering K+ Channels Using Semisynthesis
Potassium channels conduct K+ ions selectively and at very high rates. Central to the function of K+ channels is a structural unit called the selectivity filter. In the selectivity filter, a row of four K+ binding sites are created using mainly the backbone carbonyl oxygen atoms. Due to the involvement of the protein backbone, site-directed mutagenesis is of limited utility in investigating the selectivity filter. In order to overcome this limitation, we have developed a semisynthetic approach, which permits the use of chemical synthesis to manipulate the selectivity filter. In this chapter, we describe the protocols that ...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Molecularly Imprinted Polymers: Promising Advanced Materials for In Vivo Sensing
Molecularly imprinted polymers are now well known as synthetic polymeric receptors or robust artificial antibodies (“plastibodies”) and have attracted considerable attention from the scientific and industrial community due to their inherent simplicity, reusability, robust polymer network, and cost-effectiveness. In this chapter, the concept and principles of molecularly imprinted polymers are introduced and illustrated. Their fascinating properties are further described and summarized with a focus on the medical/biological aspects followed by a comprehensive review of published research results on the sensor ap...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
Electrochemiluminescent Biosensors: Neuroscience Applications
The convergence of photonics, electrochemistry, materials, and biomedical sciences at the nanoscale opens up significant new opportunities. For example, ElectroChemiLuminescence (ECL), in which an electronically excited state is electrochemically created that then goes on to emit light, opens up new possibilities for the sensitive and selective detection of biomolecules. However, key biomarkers are often present at low concentration demanding the development of highly sensitive, even single molecule, assays. Moreover, in traditional systems, the ECL generated often overlaps significantly with blood and tissue making in viv...
Source: Springer protocols feed by Neuroscience - April 12, 2013 Category: Neuroscience Source Type: news
