MOLECULAR ANAYLSIS OF NEURAL CELL DEVELOPMENT

Project: Research project

Project Details

Description

The long term goal of this project is to understand at the molecular level
the genetic and epigenetic factors responsible for cell determination and
differentiation in the mammalian Central Nervous System. This knowledge is
essential for understanding and treating many diseases caused by mutation
or injury that result in abnormal brain development and for manipulating
cell phenotypes in neural regeneration and replacement therapies.

This project builds on past findings and has four specific aims. First, a
continued analysis using transgenic mice will test the hypothesis that a
specific DNA element found in many photoreceptor specific genes, RET 1, is
both sufficient and necessary to direct photoreceptor expression of genes.
Using specific DNA sequences, consisting of either RET 1 alone or the
opsin promoter in which RET 1 has been mutated, linked to a lac Z marker
gene, the spatial and temporal expression of the marker will be examined
using X-gal histochemistry. If expression is detected in any cells other
than photoreceptors, these will be examined for the RET 1 binding protein
and for negative regulators that normally block expression of opsin.

Second, the protein binding to the RET 1 element has been purified and it
is now proposed to clone it and study its structure and activity in more
detail. The cDNA sequence will categorize this molecule as a member of a
new or an existing family of transcriptional regulators, the genomic
sequence will identify promoter elements controlling its expression and
antibodies will allow a precise description of the cellular extent of its
expression and will provide tools to isolate associated proteins. These
experiments will provide both new information about neuronal
transcriptional regulators and probes to study molecular events closer to
the time of cell birth and commitment.

Third, retinoic acid and basic fibroblast growth factor have been shown to
affect the binding activity of nuclear proteins interacting with the RET
1 sequence, and it is now proposed to study the ways in which these
factors influence binding and thus provide insights into the molecular
mechanisms by which they can influence neuronal development.

Fourth, the hypothesis that transcription factors expressed during earlier
phases of forebrain and retinal development are essential for terminal
differentiation of neurons such as photoreceptors, and expression of cell
type -specific molecules such as opsin, will be tested using a culture
system in which a retina is formed by transdifferentiation of the adjacent
retinal pigment epithelium.

Together these specific aims represent a focussed effort to define
specific and general rules governing the formation of neural cell types
and the transcription of cell-type specific genes of known function in the
mammalian Central Nervous System.
StatusFinished
Effective start/end date12/31/892/28/99

Funding

  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke
  • National Institute of Neurological Disorders and Stroke

Fingerprint

Explore the research topics touched on by this project. These labels are generated based on the underlying awards/grants. Together they form a unique fingerprint.