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Development in Drosophila Ruth Lehmann New York University/Howard Hughes Medical Institute lehmann@saturn.med.nyu.edu Model Organisms and Innovative Approaches in Developmental Biology Juquehy, Brazil-2005 Practical session: Ruth Lehmann Stained Chromatin Showing Superficial Cleavage in a Drosophila Embryo Gastrulation Overview of the Stages of Development 5 11 6 12 7 14 8 15 9 10 16 17 From V. Hartenstein Atlas of Drosophila CSH press Yellow: germ cells; green mesoderm,VF ventral furrow, red midgut (PMG and AMG anterior and posterior midgut), light blue foregut and hindgut, dark blue neuroblast, purple nervous system, pv proventriculus, go, gonad, vnc: ventral nerve cord, sg salivary gland, br brain Maternal effect genes: • Bicoid transcription factor localized to anterior • Nanos translational repressor localized to posterior • Torso receptor tyrosine kinase activated at anterior and posterior termini • Dorsal transcription factor localized to ventral nuclei Three maternal gene systems control AP patterning Head/acron Anterior thorax telson abdomen abdomen Posterior head thorax Head/acron thorax telson telson Terminal abdomen telson Head/acron thorax abdomen telson bicoid nanos torso Maternal effect genes: •Observe bicoid and nanos RNA localization Method: RNA in situ hybridization, DIC optics •Analyze gastrulation patterns of embryos from: •wildtype mutant mothers •bicoid mutant mothers •nanos mutant mothers •torso mutant mothers •“dorsal” mutant mothers •Method: Methanol/Formaldehyde fixed embryos (2 + 02hrs AEL) DIC optics Germ cell formation 1) Reduction in germ cell number in embryos from tudor mutant mothers germ cells are visualized by Vasa antibody staining 2) Transcriptional derpression of tll and slam in embryos from pgc mutant mothers tll and slam RNA in situ hybridization Method: RNA in situ hybridization, DIC optics Oskar organizes germ plasm Osk-/- No germplasm/ No germ cells Wild type Osk-bcd3’UTR 4X 0sk+ More germ plasm/ More germ cells Drosophila germ plasm pathway Germ cell migration in Drosophila Attachment to gonadal mesoderm Formation St 5 Gastrulation St 8 Migration through posterior midgut St 10 Migration on posterior midgut St 10/11 St 11 Germband retraction St 12 Line formation St 13 Gonad coalescence St 14 aVasa Reviewed in Starz-Gaiano and Lehmann (2001) Mech Dev. 105, 5-18 germ cells posterior midgut somatic gonadal precursors
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