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.Voltage-Sensitive Dye Imaging of Cortical Dynamics
Voltage-sensitive dyes can be topically applied to the surface of the brain in order to label membranes in the supragranular layers of the neocortex. Fluorescence signals from voltage-sensitive dyes in vivo correlate linearly with membrane potential of excitatory layer 2/3 neurons with millisecond temporal resolution and with a spatial resolution resolving single cortical columns. Voltage-sensitive dye imaging across large cortical areas in awake behaving mice reveals interactions between somatosensory and motor cortex that depend strongly on behavioral state. The recent development of genetically encoded voltage-sensitive...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
In Vivo Population Imaging of Dendritic Integration in Neocortex
Translating the advances seen recently in recording from neurons in vivo to dendritic recordings presents special difficulties. In vivo two-photon imaging of dendrites was achieved over a decade ago and is still the method of choice for recording from small dendritic compartments in single neurons but has proven more difficult to apply to many dendrites simultaneously or to awake, freely moving preparations. An alternative that can be applied to layer 5 neocortical pyramidal neurons is the use of a fiber-optic method combined with bolus loading of fluorescent calcium indicator. This method takes advantage of the fact that ...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
In Vivo Dendritic Mapping of Sensory Inputs in Cortical Neurons
Mammalian cortical neurons integrate sensory information that arrives through numerous synaptic inputs located on their dendrites. Here we introduce an approach to identify sensory-evoked dendritic input sites in cortical neurons in vivo involving the use of two-photon calcium imaging combined with targeted whole-cell recordings. We provide basic technical descriptions as well as experimental procedures of this method. First, we discuss various scanning modes for two-photon imaging of cortical neuron dendrites in vivo. Second, we focus on practical aspects of dye-loading by means of whole-cell recordings in vivo. Third, we...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Neocortical Circuit Interrogation with Optogenetics
Optogenetic tools have gained popularity for enabling manipulation of specified populations of neurons on a precise temporal scale. These opsins, as the proteins are known, utilize retinal (Vitamin A) as a co-factor—in a form related to the same molecule used by the human retina as a light-sensing moiety; opsin function does not require (at least in vertebrates) exogenous supplementation of this molecule [1]. Taken together, these properties of optogenetic tools situate them ideally for the interrogation of neural circuits in the settings of acute slice preparations, in vivo single and multiunit recordings, and behav...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Chronic Two-Photon Imaging of Neural Activity in the Anesthetized and Awake Behaving Rodent
Chronic in vivo two-photon imaging of genetically encoded sensors has recently enabled the measurement of activity from the same individual neurons repeatedly in different imaging sessions over months, opening new possibilities to investigate the function and plasticity of neuronal activity in both anesthetized and awake animals. A successful chronic imaging experiment involves the combination of several key techniques, including expression of genetically encoded indicators, optical measurement with cellular resolution through a cranial window, and, in awake animals, behavioral paradigms adapted to the two-photon microscop...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Optogenetic Tools for Control of Neural Activity
Understanding the contribution of defined elements to the dynamics of neural circuitry requires not only control on a time scale relevant to neural activity, but also specificity such that the experimental manipulation leaves other components of the circuit unaltered [1]. Electrical stimulation, while providing precise temporal control, modulates neurons based only on location, regardless of identity. Conversely, pharmacological control manipulates the activity of neurons based on specific properties, but in the absence of precise temporal control and with limited spatial control. The adaptation of light-activated ion chan...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Fluorescent Indicators for Functional Optical Imaging
Optical imaging is developing towards the method of choice for monitoring the activities of large numbers of cells in the cortex of living rodents. During the last decade this approach was fueled by methodological progress in both optical instrumentation and fluorescent indicators of neuronal activities. In the first part of this chapter, we provide information required to reach an informed decision regarding the most suitable fluorescent indicators for the different imaging modalities and the experimental approaches for imaging cortical circuit dynamics in vivo. In particular, an overview is given of the available palette...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Optical Properties of Neural Tissue
The optical properties of neural tissues play critical roles in all types of optical imaging methods. The wavelength-dependent absorption and scattering properties of tissue influence imaging resolution, penetration depth, and often provide sources of contrast. Therefore, quantitative interpretation of imaging data requires knowledge of the optical properties of neural tissues. Light scattering in tissue arises from nanometer-scale spatial variations in refractive index and requires a thorough electromagnetic description of light propagation through this complex medium. Unfortunately, the complexity of neural tissues and t...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Principles and Fundamentals of Optical Imaging
In this chapter I will give a brief general introduction to optical imaging and then discuss in more detail some of the methods specifically used for imaging cortical dynamics today. Absorption and fluorescence microscopy can be used to form direct, diffraction-limited images but standard methods are often only applicable to superficial layers of cortical tissue. Two-photon microscopy takes an intermediate role since the illumination pathway is diffraction-limited but the detection pathway is not. Losses in the illumination path can be compensated using higher laser power. Since the detection pathway does not require image...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
Neocortex in the Spotlight: Concepts, Questions, and Methods
When mammals such as mice, cats, monkeys, or humans act in the world, they continually make behaviorally relevant decisions based on perceived sensory information and memorized experiences and they constantly adapt to outside challenges through learning. These cognitive capabilities largely arise from neural processing in the outermost thin sheet of the forebrain called the neocortex. Although the mammalian neocortex has been studied extensively, the astounding complexity of both its structure and dynamics has precluded a comprehensive understanding of its function so far. Higher cortical function emerges from the interpla...
Source: Springer protocols feed by Neuroscience - January 27, 2014 Category: Neuroscience Source Type: news
In Vivo Gene Silencing by Virally Delivered MicroRNA
Loss of function is a standard approach to elucidate the function of a specific protein. Among these multiple strategies for silencing genes in living animals, genetic knockout in mice have been so far most frequently used. However, short hairpin RNAs (shRNAs) and microRNAs (miRNAs) delivered into the brain by viruses can achieve region-specific gene knockdown in any species at much lower cost and with shorter turnaround time. Recent advances in understanding of the endogenous miRNA function enabled the design of miRNAs as well as miRNA-adapted shRNA that efficiently enter the miRNA-processing pathway and mediate the gene ...
Source: Springer protocols feed by Neuroscience - October 30, 2013 Category: Neuroscience Source Type: news
Viral Manipulation of Neural Stem/Precursor Cells
Neural stem/precursor cells (NPCs) are defined according to their ability to self-renew and differentiate into all the major cellular types of the central nervous system (CNS). They can survive and proliferate in defined culture conditions as spherical aggregates (neurospheres) for a virtually indefinite number of passages, while remaining phenotypically and karyotypically stable. Besides classical stem cell-based approaches for regenerative medicine, NPCs hold a great potential also as system for delivering secreted factors and/or therapeutic genes in vivo, after transplantation. Solid evidence exist that NPCs survive tra...
Source: Springer protocols feed by Neuroscience - October 30, 2013 Category: Neuroscience Source Type: news
Viral Vector-Based Techniques for Optogenetic Modulation In Vivo
Optogenetics is a technical methodology that allows direct light-based manipulation of genetically specified cells. Optogenetic methods have provided novel insights into the role of defined neuronal populations in brain function and animal behavior. An expanding palette of single-component optogenetic tools provides powerful interventional strategies for modulating the function of targeted neurons in awake, behaving mammals and for detailed interrogation of circuit physiology in vitro. Although several genetic methods can be utilized for delivering these genes into target cell populations, the use of viral vectors for deli...
Source: Springer protocols feed by Neuroscience - October 30, 2013 Category: Neuroscience Source Type: news
Viral Vectors for Optogenetics of Hypothalamic Neuropeptides
During the last decade, viral technologies have progressively been used in the studies of different aspects of brain functions. Relatively recently, viral techniques have been combined with optogenetics, which allows the study of modifications of the activity of different brain regions and circuits in vitro in brain slices and in behaving animals. In the vast majority of these cases, the viral/optogenetic approach has been used to manipulate the release of the classical neurotransmitters, l-glutamate, and GABA. Recently, several studies have focused on viral/optogenetic manipulation of “nonclassical” neurotrans...
Source: Springer protocols feed by Neuroscience - October 30, 2013 Category: Neuroscience Source Type: news
Viral Vectors in Primate Research: Examples from Parkinson’s Disease Research
Neurodegenerative diseases, such as Parkinson’s disease (PD), offer unique opportunities for the therapeutic application of viral vector gene transfer. In this chapter, we discuss the current state of viral vector preclinical development in nonhuman primate with special focus on vectors derived from adeno-associated virus (AAV) and lentivirus (LV), as they are the only ones that have reached clinical trials. We also outline desired features for improving the routes of delivery, tropism specificity, and other parameters that influence achievement of successful therapeutic goals. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - October 30, 2013 Category: Neuroscience Source Type: news
