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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: 63, 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: 955, 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: 504, 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: 671, 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: 687, 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: 1321, 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: 1556, 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 ...

Registration and Alignment Between in vivo Functional and Cytoarchitectonic Maps of Mouse Visual Cortex

Authors: Jun Zhuang, Quanxin Wang, Marc Takeno and Jack Waters, date: 02/20/2018, view: 1265, Q&A: 0

[Abstract] This protocol describes a method for registration of in vivo cortical retinotopic map with cytochrome c oxidase (CO) labeled architectonic maps of the same mouse brain through the alignment of vascular fiducials. By recording surface blood vessel pattern and sequential alignment at each step, this method overcomes the challenge imposed by ...

Determination of DNA Damage in the Retina Photoreceptors of Drosophila

Authors: Damulewicz Milena and Pyza Elzbieta, date: 02/05/2018, view: 1641, Q&A: 0

[Abstract] The retina is sensitive for light damages, because of direct light exposure, especially intense blue and UV light, which increase level of ROS and other toxic phototransduction products in photoreceptor cells. In our previous work (Damulewicz et al., 2017a and 2017b), we used 8-oxo-deoxyguanosine (8-OHdG) as a marker for oxidative stress ...

A Novel Protocol to Quantitatively Measure the Endocytic Trafficking of Amyloid Precursor Protein (APP) in Polarized Primary Neurons with Sub-cellular Resolution

Authors: Florent Ubelmann, Tatiana Burrinha and Cláudia Guimas Almeida, date: 12/05/2017, view: 2157, Q&A: 0

[Abstract] Alzheimer’s disease’s established primary trigger is β-amyloid (Aβ) (Mucke and Selkoe, 2012). The amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at endosomes depends on its sorting for degradation in lysosomes (Haass et al., 1992; Morel et al. ...

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