Update on deficiency screens Joohong Ahnn and Andrew Fire Carnegie Institution of Washington, Department of Embryology, Baltimore, MD 21210 We have been screening available chromosomal deficiencies in order to identify genetic loci whose zygotic expression is required for early events in embryogenesis including formation of body wall muscle cellsl. A surprising result of deficiency screens (oursl and others2 3) has been the extensive tissue differentiation that occurs in many of the deficiency homozygote embryos. In our initial analysis we found only one region (on the left tip of LGV) which was required for accumulation of body-wall myosin. This gazette article reports updates on our deficiency analysis. Analysis of additional characterized deficiencies: Embryos homozygous for nDf3 (II)4 and nDf4 (Jl)4 arrest elongation before comma stage (although a small fraction develop to the 1-112 fold stage). These embryos form pharyngeal structures, have gut granules and stain with antibodies to
unc-54 and
myo-3 produsts. Deficiency sDf23 (IV)s homozygous embryos arrest before comma stage, have gut granules but no pharyngeal structures; these embryos do not show any movement but stain with both anti-myosin antibodies. nDf42 (V)6 homozygous embryos show an early arrest with no movement, no gut granules, and no pharyngeal structures. Homozygous embryos from nDfl 7 (m)4 and nDfl 9 (X)7 both arrest at 2-fold stage, show twitching movement, have gut granules, form pharynx and stain with myosin antibodies (n.b. mapping data on ACEDB suggests nDfl 7 and nDfl 9 could be compound deficiencies). Producing a bank of Dfs of VL: In order to isolate small deficiencies in the region of LG VL which appears to be essential for myogenesis, gamma-ray induced mutagenesis has been conducted. Mutagenized N2 males were crossed into
unc-34 dpy-l l animals, and rare Unc Non- dpy F1 progeny were cloned. Seven deficiencies have been isolated among 17,000 haploid chromosomes screened so far. Further mapping of these deficiencies by physical and genetic means should allow us to narrow down the myogenic locus (or loci) in the region. A very early zygotic requirement ? In a more general screen for early arrest mutants, several very early lethals linked on LG m have been isolated. These segregate a high fraction of dead embryos, suggesting a chromosomal break or rearrangement. About 1/4 of the embryos from these strains show a characteristic very early terminal phenotypes. These embryos have 200-2S0 nuclei, with no overt tissue differentiation. The embryos fail to stain with myosin antibodies and do not have gut granules. Embryos from two of these strains were observed with 4-D microscopy to look for defects during early embryogenesis. The early cleavage pattern of early-arrest embryos look normal, but these embryos fail to complete last 1-2 cell divisions. In some cases, nuclear divisions appear to complete without subsequent cytokinesis. It is interesting to compare these terminal phenotypes with those of
emb-29 (V).
emb-29 produces embryos with comparable numbers of nuclei, but these embryos exhibit extensive differentiation8. This suggests that the putative chromosome m defect is not simply due to blockage or failure of cell divisions. WBG 12: 2, 106 Genetics (137:483-498) 2. F. Storfer-Glazer & W. Wood Genetics (137: 499-508) '93 C. elegans Meeting (Abt. 497) 4. I. Greenwald & R. Horvitz unpublished 6. M. Hengartner & R. Horvitz unpublished 7. V. Ambros & R. Horvitz unpublished 8. R. Hecht et. al J. of Cell Science (87:305- 314)