Brugia Malayi - Clark Science Center - Smith College

Identification of Specific Zrf (2-4) Antibody Protein Targets in the
Developing Zebrafish Brain
Paula Zaman
In the developing Zebrafish brain, axons are typically guided across the midline by attractant and repellent protein cues to form
commissures. During the summer, I looked at the formation of the postoptic commisure of the diencephalon in the Zebrafish
brain. During pathfinidng of these attractant/repellent cues, the POC axons closely contact a population of glial fibrillary acidic
proteins (Gfap) postive astroglia covering the midline which was labeled by Zrf1. There were also three other Zrf (2-4) antibodies
that we were interested in due to previous studies that have stated that they played a role in POC formation by interacting with
unknown proteins.
The purpose of this research was to determine, quantify, and further look into what these proteins are and how they play a
role in commissural formation. In this experiment I used two main techniques to determine what proteins these Zrf antibodies
bind to, they are: imminuprecipitation and co-localization. Immuniprecipitation is a technique that precipitates a protein antigen
out of solution using an antibody that specifically binds to that particular protein. This process is used to isolate a particular
protein from a sample containing many different proteins. We will also be looking at co-localization on an imager called ‘Volocity’
for specific cell populations labeled with the different Zrf antibody labeling. Co-localization allows us to view protein interaction
between known and unknown proteins. In this experiment, we will use Velocity which is a highly detailed imager that could take
3-D images of antibody localization. With this research, we hope to better the current model of axonal signaling in the POC and
learn more about the key players that make neurological pathfinding. (Supported by the Howard Hughes Medical Institute)
Advisor: Michael Barresi
2012
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