CRASH syndrome in humans, which includes X-linked hydrocephalus, is caused by mutations in the gene encoding the Ll cell adhesion molecule. There are no good experimental models in which brain syndromes, caused by various mutations in Ll, can be studied easily. The long-term goal of this exploratory project is to create a new experimental model of anomalous brain development that can be used to study in vivo the cellular effects of different mutations in L1. The overlying hypothesis to be tested is that cell behavior can be altered in the developing chicken embryo brain by ectopic expression of specific mutated Ll cDNAs, or by attenuation of endogenous Ll using direct gene conversion or antisense sequences. Human Ll mutations can be grouped according to the type of mutation. Most, but not all, severe hydrocephalus, grave mental retardation, and early deaths are caused by mutations that produce truncations in the extracellular domain (Class 3 mutations), which probably are secreted and interfere with other adhesion and signaling systems. It is theorized that similar mutations in the chicken homologue of L1, called NgCAM, will interfere with neuronal behavior (e.g., neuronal migration) when ectopically expressed in the embryonic chick brain. We also theorize that a malformed brain will result when such mutations are expressed throughout the tissue, but that localized expression will inhibit neuronal migration and axon extension only in the immediate area.
Specific Aim 1 : Define the developmental effects of widespread NgCAM misexpression. Hypothesis 1: A generally malformed brain will be generated in the developing chick by widespread introduction of antisense-NgCAM cDNAs, Class 3 mutant NgCAM cDNAs, or of gene conversion DNA oligonucleotides by in ovo electroporation.
Specific Aim 2 : Define the developmental effects of isolated NgCAM misexpression. Hypothesis2: Introduction of mutated and antisense NgCAM cDNAs in discrete, marked cell clones by retroviral vectors will inhibit neuronal migration and axon extension in some, but not all, neurons expressing such sequences.
Specific Aim 3 : Define the effects and distribution of mammalian Class 3 Ll misexpression. Hypothesis 3: Class 3 Ll mutations can operate beyond the cells that express them.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21NS040317-01A2
Application #
6435494
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Program Officer
Spinella, Giovanna M
Project Start
2001-12-15
Project End
2004-11-30
Budget Start
2001-12-15
Budget End
2002-11-30
Support Year
1
Fiscal Year
2002
Total Cost
$176,793
Indirect Cost
Name
University of Delaware
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
059007500
City
Newark
State
DE
Country
United States
Zip Code
19716
Farach, Andrew M; Galileo, Deni S (2008) O-GlcNAc modification of radial glial vimentin filaments in the developing chick brain. Brain Cell Biol 36:191-202
Masiello, Lisa M; Fotos, Joseph S; Galileo, Deni S et al. (2006) Lysophosphatidic acid induces chemotaxis in MC3T3-E1 osteoblastic cells. Bone 39:72-82
Morgan, John C; Majors, John E; Galileo, Deni S (2005) Distinct and opposite roles for SH2 and SH3 domains of v-src in embryo survival and hemangiosarcoma formation. Clin Exp Metastasis 22:167-75
Cretu, Alexandra; Fotos, Joseph S; Little, Brian W et al. (2005) Human and rat glioma growth, invasion, and vascularization in a novel chick embryo brain tumor model. Clin Exp Metastasis 22:225-36
Castellini, Meryl; Wolf, Louise V; Chauhan, Bharesh K et al. (2005) Palm is expressed in both developing and adult mouse lens and retina. BMC Ophthalmol 5:14
Stettler, Erin M; Galileo, Deni S (2004) Radial glia produce and align the ligand fibronectin during neuronal migration in the developing chick brain. J Comp Neurol 468:441-51
Galileo, Deni S (2003) Spatiotemporal gradient of oligodendrocyte differentiation in chick optic tectum requires brain integrity and cell-cell interactions. Glia 41:25-37