Author

Risha Sinha

Publication Date

2014

Document Type

Honors Thesis

Department

Biochemistry

Keywords

Developmental biology, Neurosciences, Biochemistry, Zebra danio-Development, Radial nerve, Prosencephalong, Anterior commissure, Neuroglia, Diencephalon, Zebrafish, Zebrafish radial fiber (ZRF), Astroglia, Forebrain, Post optic commissure (POC)

Abstract

In the rise of the central nervous system of bilateral organisms, neuronal axons are guided across the midline by contact attractants and repellants to form commissures, a cluster of nerve fibers, which serve to connect the two sides of the central nervous system. The Barresi lab uses the zebrafish model system to study the developmental process of commissure formation at the biochemical, molecular, and cellular levels. Previously, Barresi et.al. (2005) have implicated astroglial cells as a supportive substrate to promote axonal growth across the forebrain midline. Astroglial cells express glial fibrillary acidic protein (Gfap), an intermediate filament protein. Gfap is positively recognized by the antibody zebrafish radial fiber 1 (Zrf-1). There are three additional Zrf proteins (2-4), and all four Zrfs were prepared to understand the structuring of neural segments in embryonic zebrafish hindbrain. By labeling with these antibodies in the forebrain, we were able to show differential Zrf expression in astroglia, suggesting a heterogeneous population of astroglia may be present at the midline. Elucidating the antigenic identities of the Zrf antibodies could reveal important insights upon glial development. The goal of this project is multifold. First, we aimed to identify the proteins that are being recognized by the Zrf-2, 3, and 4 antibodies in the zebrafish embryo and adult using the biochemical techniques of immunoprecipitation, Western Blot, and liquid chromatography mass spectrometry (LC-MS). Secondly, we aimed to confirm the correlation between Zrf expression patterns and cell morphology using a three-dimensional visualization technique of immunolabeled, whole-mounted embryo forebrain samples using transplanted cells from either the Tg(gfap:eGfp) or Tg(olig2:eGfp) transgenic lines.

Language

English

Comments

66 pages : color illustrations. Honors Project-Smith College, 2014. Includes bibliographical references (pages 60-63)

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