Ahnn J, Fire A
Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210.
Genetics. 1994 Jun;137(2):483-98. doi: 10.1093/genetics/137.2.483.
We have used available chromosomal deficiencies to screen for genetic loci whose zygotic expression is required for formation of body-wall muscle cells during embryogenesis in Caenorhabditis elegans. To test for muscle cell differentiation we have assayed for both contractile function and the expression of muscle-specific structural proteins. Monoclonal antibodies directed against two myosin heavy chain isoforms, the products of the unc-54 and myo-3 genes, were used to detect body-wall muscle differentiation. We have screened 77 deficiencies, covering approximately 72% of the genome. Deficiency homozygotes in most cases stain with antibodies to the body-wall muscle myosins and in many cases muscle contractile function is observed. We have identified two regions showing distinct defects in myosin heavy chain gene expression. Embryos homozygous for deficiencies removing the left tip of chromosome V fail to accumulate the myo-3 and unc-54 products, but express antigens characteristic of hypodermal, pharyngeal and neural development. Embryos lacking a large region on chromosome III accumulate the unc-54 product but not the myo-3 product. We conclude that there exist only a small number of loci whose zygotic expression is uniquely required for adoption of a muscle cell fate.
我们利用现有的染色体缺失来筛选那些在秀丽隐杆线虫胚胎发育过程中,其合子表达对于体壁肌肉细胞形成所必需的基因座。为了检测肌肉细胞分化,我们对收缩功能和肌肉特异性结构蛋白的表达都进行了测定。针对两种肌球蛋白重链异构体(unc-54和myo-3基因的产物)的单克隆抗体被用于检测体壁肌肉分化。我们筛选了77个缺失,覆盖了大约72%的基因组。大多数情况下,缺失纯合子用体壁肌肉肌球蛋白抗体染色,并且在许多情况下观察到肌肉收缩功能。我们确定了两个在肌球蛋白重链基因表达上表现出明显缺陷的区域。缺失染色体V左端的纯合子胚胎无法积累myo-3和unc-54产物,但表达皮下、咽部和神经发育特征性的抗原。缺失染色体III上大片段区域的胚胎积累了unc-54产物,但没有积累myo-3产物。我们得出结论,只有少数基因座,其合子表达是采用肌肉细胞命运所唯一必需的。