|dc.description||v, 34 p.||en_US
|dc.description.abstract||Signaling centers in the developing vertebrate brain are responsible for
determination of dorsal or ventral characteristics of neural tube cells. These
characteristics will eventually determine the final functions of the neuronal cell. Bone
morphological proteins (Bmp) expressed in the roof plate, act as dorsalizing signals in
the neural tube, patterning cells to dorsal fate. Antagonistically, centers such as the
notochord and floor plate express sonic hedgehog (Shh), a powerful ventral patterning
protein signal that competes with Bmp activity.
A method to study these signals is to antagonize them and visualize effects in
neural tissue by in situ hybridization. Dominant-negative Bmp Receptor 1 B, a mutant
inhibitor of Bmp activity, can be transferred into neural tissue in order to see these
effects. Two methods that have found success in gene transfer are in ovo
electroporation and viral injection.
Electroporation allows genes contained in plasmid vectors to be directed to
specific points in tissue. The expression of the novel gene by the embryo is quick,
but short term. Conversely, viral injection allows a gene contained in a retroviral
vector to be spread by infection throughout the neural cavity. The expression of a
gene takes longer to begin, but is permanent. In this study, we explored the signaling
involved in dorsal-ventral patterning by utilizing these two methods.
We found that electroporation of the human Efx plasmid with a ligated
alkaline phosphatase gene, gave semi-satisfactory results. The electroporations were
inconsistent in success, giving weak and imprecise direction of protein expression.
We did not succeed in electroporating a retroviral based vector, RCAS, into the
neural tissue, most likely due to the vector's large size along with poor electrodes. A
noggin/pXex construct was made to replace the RCAS as another possible vector for
Dominant-negative BMPR-IB viral stocks concentrated 6-10x and injected
into day two neural tube gave a high concentration of infection in growing cells.
Further, dorsal tissue based choroid plexus formation was slightly deterred by the
BMPR-IB; however, not the extent expected. It was concluded that viral injections
must occur at an earlier age to see greater effects. Both methods can act as efficient
methods of forebrain study in the chick.||en_US
|dc.description.sponsorship||Department of Neurobiology. University of Chicago. Chicago, Illinois.||
|dc.relation.ispartof||Kalamazoo College Health Sciences Senior Individualized Projects Collection||
|dc.relation.ispartofseries||Senior Individualized Projects. Health Sciences;||
|dc.rights||U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder. All rights reserved.||
|dc.title||An Investigation of in Ovo Electroporation and High Titer Viral Injection Gene Transfer Methods in the Study of the Dorsal-Ventral Patterning of the Forebrain of Chick||en_US
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