We thank Dan McWhorter for narrating the protocol in the video. This work is supported by NIH Grant
5R01NS062182, a grant from American Federation for Aging Research, and institutional funds awarded to HTG.

I.	Procedures
The following techniques should be performed under sterile conditions, in a laminar flow hood, using sterilized tools.
Preparation of glass bottom dishes for organotypic slices
<ol>
<li>Dishes must be prepared in sterile environment and using sterilized tools.</li>
<li>A 150&mu;L drop of slice medium (see recipes) is placed in the center of the glass-bottom part of the dish with care to avoid air bubbles in the medium.</li>
<li>With a disposable s...

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Neuronal migration in the RMS is an essential component of postnatal neurogenesis in the olfactory bulbs 1. Migration through the RMS occurs in a plane tangential to the surface of the brain. Tangentially migrating neuroblasts are distinct from radially migrating cells based on the location of their progenitor source, as well as the divergent fate of their final neuronal products 1, 2, 3. The relatively pure population of tangentially migrating cells in the postnatal RMS makes this anatomically de...

an organotypic slice assay for high-resolution time-lapse imaging of neuronal migration in the postnatal brain

Neurogenesis in the postnatal brain depends on maintenance of three biological events: proliferation of progenitor cells, migration of neuroblasts, as well as differentiation and integration of new neurons into existing neural circuits. For postnatal neurogenesis in the olfactory bulbs, these events are segregated within three anatomically distinct domains: proliferation largely occurs in the subependymal zone (SEZ) of the lateral ventricles, migrating neuroblasts traverse through the rostral migratory stream (RMS), and new neurons differentiate and integrate within the olfactory bulbs (OB). The three domains serve as ideal platforms to study the cellular, molecular, and physiological mechanisms that regulate each of the biological events distinctly. This paper describes an organotypic slice assay optimized for postnatal brain tissue, in which the extracellular conditions closely mimic the in vivo environment for migrating neuroblasts. We show that our assay provides for uniform, oriented, and speedy movement of neuroblasts within the RMS. This assay will be highly suitable for the study of cell autonomous and non-autonomous regulation of neuronal migration by utilizing cross-transplantation approaches from mice on different genetic backgrounds.

Watch this Scientific Journal Video about An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain at JoVE.com

An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain

This protocol describes an organotypic slice assay optimized for the postnatal brain and high-resolution time-lapse imaging of neuroblast migration in the rostral migratory stream.

Neurogenesis in the postnatal brain depends on maintenance of three biological events: proliferation of progenitor cells, migration of neuroblasts, as ...

an-organotypic-slice-assay-for-high-resolution-time-lapse-imaging

Research

JoVE Journal

Neuroscience

11.8K Views.  North Carolina State University. Questo protocollo descrive un saggio organotipica fetta ottimizzato per il cervello postnatale e ad alta risoluzione time-lapse imaging di migrazione neuroblasti nel flusso migratorio rostrale.