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Herpes Simplex Virus Type 1 Propagation, Titration and Single-step Growth Curves

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Construction of Viral Vectors for Cell Type-specific CRISPR Gene Editing in the Adult Mouse Brain

Authors: Hiroshi Yamaguchi and Luis de Lecea, date: 08/20/2019, view: 783, Q&A: 0

[Abstract] Recently developed gene editing technologies based on engineered CRISPR/Cas9 systems enables researchers to disrupt genes in a cell type-specific manner in the adult mouse brain. Using these technologies, we recently showed that the dopamine beta-hydroxylase gene in Locus Coeruleus (LC) norepinephrine neurons plays a vital role in the maintenance ...

Looking through Brains with Fast Passive CLARITY: Zebrafish, Rodents, Non-human Primates and Humans

Authors: Farzad Mortazavi, Alexander J. Stankiewicz and Irina V. Zhdanova, date: 08/05/2019, view: 1005, Q&A: 0

[Abstract] Recently developed CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ-hybridization-compatible Tis-sue-hYdrogel) technique renders the tissue transparent by removing lipids in the tissue, while preserving and stabilizing the cellular and subcellular structures. This provides effective penetration of diverse ...

Accessing Olfactory Habituation in Drosophila melanogaster with a T-maze Paradigm

Authors: Ourania Semelidou, Summer F. Acevedo and Efthimios M.C. Skoulakis, date: 06/05/2019, view: 1045, Q&A: 0

[Abstract] Habituation is the process whereby perceptual changes alter the value of environmental stimuli, enabling salience filtering. This behavioral response decrement is a form of non-associative learning, where the subject learns about the stimulus and does not involve sensory adaptation, sensory or motor fatigue. The range of behavioral responses in D. ...

High-resolution Immunoelectron Microscopy Techniques for Revealing Distinct Subcellular Type 1 Cannabinoid Receptor Domains in Brain

Authors: Nagore Puente, Itziar Bonilla-Del Río, Svein Achicallende, Patrick C. Nahirney and Pedro Grandes, date: 01/20/2019, view: 2103, Q&A: 0

[Abstract] Activation of type 1 cannabinoid (CB₁) receptors by endogenous, exogenous (cannabis derivatives) or synthetic cannabinoids (i.e., CP 55.940, Win-2) has a wide variety of behavioral effects due to the presence of CB₁ receptors in the brain. In situ hybridization and immunohistochemical techniques have been ...

Optical Clearing and Index Matching of Tissue Samples for High-resolution Fluorescence Imaging Using SeeDB2

Authors: Meng-Tsen Ke and Takeshi Imai, date: 10/20/2018, view: 3911, Q&A: 1

[Abstract] Tissue clearing techniques are useful for large-scale three-dimensional fluorescence imaging of thick tissues. However, high-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we present a water-based optical clearing and mounting media, SeeDB2, which is ...

Artificial Inhalation Protocol in Adult Mice

Authors: Thomas P. Eiting and Matt Wachowiak, date: 09/20/2018, view: 1766, Q&A: 0

[Abstract] Research in the area of in vivo olfactory physiology benefits from having direct access to the nasal airways through which odorants can be presented. Ordinarily, the passage of odorants through the airways is controlled by respiratory rhythm. This fact makes it difficult to control the timing and strength of an olfactory stimulus, since ...

Quantitative Electron Microscopic Assay Using Random Sampling from Single Sections to Test Plastic Synaptic Changes in Hippocampus

Authors: G. Mark Marcello, Lilla E. Szabó, Peter Sotonyi and Bence Rácz, date: 08/05/2018, view: 2032, Q&A: 0

[Abstract] Studies over several decades on the organization of the CA1 hippocampus–a particularly favorable model for learning, memory and certain forms of cognition–have shown that the synaptic network in this brain region is plastic (Fortin et al., 2012). Recent evidence suggests that a number of environmental and endogenous stimuli may have a ...

Embryonic Intravitreous Injection in Mouse

Authors: Liyuan Cui, Yupu Diao and Jiayi Zhang, date: 07/20/2018, view: 1897, Q&A: 0

[Abstract] Axons of retinal ganglion cells (RGCs) relay visual information from the retina to lateral geniculate nucleus (LGN) and superior colliculus (SC), which are two major image-forming visual nuclei. Wiring of these retinal projections completes before vision begins. However, there are few studies on retinal axons at embryonic stage due to technical ...

In vivo Use of Dextran-based Anterograde Cortical Tracers to Assess the Integrity of the Cortical Spinal Tract

Authors: Anna M. Wiersma and Ian R. Winship, date: 05/20/2018, view: 2776, Q&A: 0

[Abstract] When injected into the motor cortex of rats, anterograde tracers label fibers of the associated descending corticospinal tract (CST) that originate from pyramidal neurons in the tracer-injected cortex. These fibers can be assessed at the level of the spinal cord to determine the integrity of the descending CST and the spatial distribution of axons ...

Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia

Authors: Yusuke Tomina and Daniel A. Wagenaar, date: 03/05/2018, view: 3255, Q&A: 0

[Abstract] In this protocol, we introduce an effective method for voltage-sensitive dye (VSD) loading and imaging of leech ganglia as used in Tomina and Wagenaar (2017). Dissection and dye loading procedures are the most critical steps toward successful whole-ganglion VSD imaging. The former entails the removal of the sheath that covers neurons in the ...

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