Characterization of the cdy mutant phenotype.

(A) Wild-type and cdy mutant embryos stained for hemoglobin at 30 hpf and 48 hpf. Brown/orange staining witho-dianisidine indicates the presence of hemoglobin in circulating red blood cells (arrows). Note the complete lack of staining in the cdy embryos. In addition, wild-type andcdy embryos were analyzed by in situ hybridization at 36 hpf for expression of αe3-globin mRNA. Note that wild-type and mutant embryos express similar levels of αe3-globin in circulating blood cells (arrows). Similar results were seen for other embryonic globin genes (data not shown). (B) Blood was collected from wild-type and cdy mutant embryos on day 2 and day 3 of development. Cells were cytospun and stained with Wright-Giemsa. All cells in this figure are embryonic erythroid cells. Note the delay in differentiation (larger nuclei) seen in day 2 and day 3 cdy cells. (C) Peripheral blood and kidney samples from wild-type and mutant adults were stained with Wright-Giemsa. Note the less condensed nuclei ofcdy peripheral blood cells. Also, note the increased number of erythroid precursors in both the peripheral blood (*) and kidney (cells with large nuclei and deep blue rim of cytoplasm) ofcdy mutants. (D) Analysis of adult blood. Blood from a wild-type adult and a cdy mutant adult was measured for MCV and MCH. The red blood cells of the cdy mutant express lower levels of hemoglobin (MCH) and are smaller (MCV) than cells from the wild-type sibling. Tall bars correspond to the range of MCV and MCH values in wild-type animals.