Basement membranes are specialized extracellular matrices found in all metazoans and involved in many important developmental processes, including; cell adhesion, cell and axon migration, and cell differentiation. They are also involved in certain pathologic conditions, such as tumor metastasis, wound healing, blistering diseases of the skin, and forms of nephritis. The broad goal of this application is to elucidate basement membrane functions utilizing the genetic tools available in the nematode Caenorhabditis elegans. A major conserved basement membrane protein is nidogen, which, via its binding to laminin, type IV collagen and perlecan, was thought to be critical for basement membrane assembly. Using deletion mutations of the C.elegans hid-1 nidogen gene, we have shown that is not required for viability or basement membrane assembly. However, we find it has significant functions in axonal guidance, synaptic morphogenesis, and neuromuscular coordination.
The first aim of this proposal will determine which domains of NID-1 are required for these various functions, by transgenic expression of modified nid-1 constructs in nid-1 null animals. We will also examine when and where nid-1 expression is required.
The second aim will identify new alleles of nid-1 and of genes that interact with, or function in the same processes as, hid-1. We will identify genes that become critical in the absence ofnidogen, by screening of an RNAi feeding library encompassing most C. elegans genes.
The third aim i s to characterize the synthetic lethality that results when nid-1 is combined with mutations in the dystroglycan gene dgn-1 or the transcription factor unc-39. These interactions reveal potential nid-1 functions in epithelial polarity, that are not apparent in the single mutants. The cellular phenotypes underlying lethality will be characterized at the light and electron microscope levels, and mutations that can suppress lethality will be sought. These studies will elucidate features of nidogen that are critical for it specific functions and identify other molecules with which it interacts.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD027211-12
Application #
6736740
Study Section
Special Emphasis Panel (ZRG1-PC (02))
Program Officer
Moody, Sally Ann
Project Start
1990-09-01
Project End
2008-11-30
Budget Start
2004-02-03
Budget End
2004-11-30
Support Year
12
Fiscal Year
2004
Total Cost
$290,571
Indirect Cost
Name
Northwestern University at Chicago
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
Ihara, Shinji; Hagedorn, Elliott J; Morrissey, Meghan A et al. (2011) Basement membrane sliding and targeted adhesion remodels tissue boundaries during uterine-vulval attachment in Caenorhabditis elegans. Nat Cell Biol 13:641-51
Muriel, Joaquin M; Xu, Xuehong; Kramer, James M et al. (2006) Selective assembly of fibulin-1 splice variants reveals distinct extracellular matrix networks and novel functions for perlecan/UNC-52 splice variants. Dev Dyn 235:2632-40
Johnson, Robert P; Kang, Seong Hoon; Kramer, James M (2006) C. elegans dystroglycan DGN-1 functions in epithelia and neurons, but not muscle, and independently of dystrophin. Development 133:1911-21
Sherwood, David R; Butler, James A; Kramer, James M et al. (2005) FOS-1 promotes basement-membrane removal during anchor-cell invasion in C. elegans. Cell 121:951-62
Inoue, Takao; Sherwood, David R; Aspock, Gudrun et al. (2002) Gene expression markers for Caenorhabditis elegans vulval cells. Mech Dev 119 Suppl 1:S203-9
Inoue, Takao; Sherwood, David R; Aspock, Gudrun et al. (2002) Gene expression markers for Caenorhabditis elegans vulval cells. Gene Expr Patterns 2:235-41
Ackley, B D; Crew, J R; Elamaa, H et al. (2001) The NC1/endostatin domain of Caenorhabditis elegans type XVIII collagen affects cell migration and axon guidance. J Cell Biol 152:1219-32
Kang, S H; Kramer, J M (2000) Nidogen is nonessential and not required for normal type IV collagen localization in Caenorhabditis elegans. Mol Biol Cell 11:3911-23
Kramerova, I A; Kawaguchi, N; Fessler, L I et al. (2000) Papilin in development; a pericellular protein with a homology to the ADAMTS metalloproteinases. Development 127:5475-85
Gupta, M C; Graham, P L; Kramer, J M (1997) Characterization of alpha1(IV) collagen mutations in Caenorhabditis elegans and the effects of alpha1 and alpha2(IV) mutations on type IV collagen distribution. J Cell Biol 137:1185-96

Showing the most recent 10 out of 13 publications