Authors

Bingru Sun

Type

Text

Type

Thesis

Advisor

Ge, Shaoyu. | Shelly, Maya

Date

2015-12-01

Keywords

Biology

Department

Department of Biological Sciences.

Language

en_US

Source

This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.

Identifier

http://hdl.handle.net/11401/76962

Publisher

The Graduate School, Stony Brook University: Stony Brook, NY.

Format

application/pdf

Abstract

During neuronal development, the neuronal cell polarizes to specify the structures of axons and dendrites. This polarized architecture regulates the input/output functions of the neuronal cell. In my thesis, I studied the molecular mechanisms of dendrite formation. My main hypothesis was that dendrite specification results from local elevation of the cGMP signal. Soluble guanylyl cyclase (sGC) is the major enzyme for the synthesis of cGMP in the embryonic brain. I study the molecular mechanism that activates sGC and leads to dendrite specification during neuronal polarization.Scribble is a scaffold protein, part of a polarity complex that establish apical-basal polarity in epithelial cells. In neurons, Scribble has been shown to associate with neuronal nitric oxide synthase (nNOS) that produces the molecule NO. NO activates sGC and can induce cGMP elevation. I hypothesize that dendrite specification is regulated by a polarity protein complex consisting of Scribble, sGC, and nNOS. In my thesis, I found the following: First, I show that Scribble and sGC associate biochemically in the rat embryonic brain. Second, I further characterized the interaction domain between Scribble and sGC and found a protein domain named IMR to be significant. I found that this single domain mutant of Scribble is a dominant negative inhibitor of the function of wild-type Scribble. Last, using shRNA-mediated knockdown of Scribble and sGC in dissociated hippocampal neuron culture, I found severe effects on dendrite development compared to control. Thus, based on my biochemistry studies and functional analysis in cultured neurons, I conclude that the protein complex involving Scribble and sGC regulates dendrite development through the cGMP pathway,31 pages

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