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Differentiation of Mouse Embryonic Stem Cells to Neuronal Cells Using Hanging Droplets and Retinoic Acid

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Antisense Oligodeoxynucleotide Perfusion Blocks Gene Expression of Synaptic Plasticity-related Proteins without Inducing Compensation in Hippocampal Slices

Authors: Panayiotis Tsokas, Bruno Rivard, Changchi Hsieh, James E. Cottrell, André Antonio Fenton and Todd Charlton Sacktor, date: 10/05/2019, view: 280, Q&A: 0

[Abstract] The elucidation of the molecular mechanisms of long-term synaptic plasticity has been hindered by both the compensation that can occur after chronic loss of the core plasticity molecules and by ex vivo conditions that may not reproduce in vivo plasticity. Here we describe a novel method to rapidly suppress gene expression by ...

Estimation of the Readily Releasable Synaptic Vesicle Pool at the Drosophila Larval Neuromuscular Junction

Authors: Pragya Goel, Xiling Li and Dion Dickman, date: 01/05/2019, view: 2239, Q&A: 0

[Abstract] Presynaptic boutons at nerve terminals are densely packed with synaptic vesicles, specialized organelles for rapid and regulated neurotransmitter secretion. Upon depolarization of the nerve terminal, synaptic vesicles fuse at specializations called active zones that are localized at discrete compartments in the plasma membrane to initiate synaptic ...

Measurement of Dopamine Using Fast Scan Cyclic Voltammetry in Rodent Brain Slices

Authors: Madelyn I. Mauterer, Paige M. Estave, Katherine M. Holleran and Sara R. Jones, date: 10/05/2018, view: 3249, Q&A: 0

[Abstract] Fast scan cyclic voltammetry (FSCV) is an electrochemical technique that allows sub-second detection of oxidizable chemical species, including monoamine neurotransmitters such as dopamine, norepinephrine, and serotonin. This technique has been used to record the physiological dynamics of these neurotransmitters in brain tissue, including their ...

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: 1939, 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 ...

Obtaining Acute Brain Slices

Authors: Thomas Papouin and Philip G. Haydon, date: 01/20/2018, view: 7461, Q&A: 1

[Abstract] Obtaining acute brain slices for electrophysiology or amperometric recordings has become a routine procedure in most labs in the field of neuroscience. Yet, protocols describing the step by step process are scarce, in particular for routine acute preparations such as from the mouse hippocampus. Here we provide a detailed protocol for the ...

D-serine Measurements in Brain Slices or Other Tissue Explants

Authors: Thomas Papouin and Philip G. Haydon, date: 01/20/2018, view: 3184, Q&A: 0

[Abstract] D-serine is an atypical amino acid present in the mammalian body (most amino acids in the mammalian body are L-isomers) that is mostly known in neuroscience for its role as a co-agonist controlling the N-methyl D-aspartate receptor (NMDAR). D-serine levels are decreased in patients with schizophrenia and this is thought to mediate, at least in ...

Fluorescent Measurement of Synaptic Activity Using FM Dyes in Dissociated Hippocampal Cultured Neurons

Authors: Roman M. Lazarenko, Claire E. DelBove and Qi Zhang, date: 01/20/2018, view: 4064, Q&A: 0

[Abstract] Release and recycling of synaptic vesicles are essential for neurotransmission and synaptic plasticity. To gain mechanistic understanding of these processes, direct measurements of vesicle release and retrieval is indispensable. Styryl dyes like FM1-43 and FM4-64 have been widely used for this purpose and their loading and unloading are reliable ...

Preparation, Stimulation and Other Uses of Adult Rat Brain Synaptosomes

Authors: Jigar Modi, Howard Prentice and Jang-Yen Wu, date: 12/20/2017, view: 2635, Q&A: 0

[Abstract] In this paper, our protocol for preparation of brain synaptosomes is described. Synaptosomes are a valuable model system for analysis of structural components of the synapse as well as for investigation of synaptic function. Synaptosomal preparations are necessary for understanding molecular changes at synapses where critical post-translational ...

A Streamlined Method for the Preparation of Gelatin Embedded Brains and Simplified Organization of Sections for Serial Reconstructions

Authors: Andrew W. Liu, Sho Aoki and Jeffery R. Wickens, date: 11/20/2017, view: 6376, Q&A: 0

[Abstract] Gelatin embedding of whole brains for sectioning is a critical procedure used in neuroscience to ensure all morphological and spatial details are preserved intact. Here, we describe an inexpensive, reproducible and efficient means to embed post-fixed brains ready for sectioning in gelatin within a week’s time. The sections obtained are ...

Preparation of Crude Synaptosomal Fractions from Mouse Brains and Spinal Cords

Author: Oliver Wirths, date: 08/05/2017, view: 7089, Q&A: 0

[Abstract] The current protocol describes the preparation of crude synaptosomal fractions from mouse brain or spinal cord samples. In detail, a sequential protocol yielding crude synaptosomal and light membrane fractions is provided. This fast and easy method might be sufficient to assess the amount of synaptic proteins in down-steam applications like ...

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