Laminin, a major component of all basement membranes (BM) is a heterotrimer of a , b and g subunits which trimerize along their long arms to form a cruciform with three short arms. There are pleiotropic effects on survival, reproduction and behavior when any laminin subunit is missing. Here we address the issue of BM assembly and function. Our understanding of the roles of the subunits during development, allows us to address a long standing issue of whether polymerized laminin is important for a functional BM. Biochemical studies have shown that trimers can polymerize via short arm interactions to form a 2-dimensional network. This can be blocked by adding trimers lacking short arms or by adding short arm fragments to compete for polymerization sites. It is not clear to what extent polymerization takes place in vivo and how important it is for a functional BM. If important, the biochemical studies predict that transgenes expressing short arm deletions in any subunit or over expressing a short arm ought to act in a dominant negative fashion. We tested this hypothesis by using the
hsp16-2 promoter to drive expression of truncated versions of
lam-1( b subunit) that encode only the short arm or have deletions in the short arm to varying extents. Transgenic embryos exhibit embryonic and L1 lethality with phenotypes similar to those caused by strong
epi-1 ( a subunit) alleles. Heat shocked L1 animals survive and grow, but show defects in gonad migration and germline development. We see gonad arms without sperm or oocytes, with reduced germ cell proliferation, and with sperm but no oocytes. Gonadal sheath cells provide cues for germline development. In laminin mutants they often peel away from the arms due to defects in the BM. It is likely that perturbation of the BMs by these transgenes has the same effect on the sheath and hence on germline development. We are now examining the integrity of the BM in the heat shocked animals by electron microscopy and by a functional b :GFP fusion which labels all the BMs. These results provide the first evidence that laminin polymerizes in vivo and that polymerized laminin is required for BM functions b .