Aneuploid and polyploid cellular DNA heterogeneity in insect cell material of diptera species analyzed by flow cytometry.
Flow cytometric 1-parameter DNA analysis and 2-parameter DNA/protein analysis have been performed with cell material of the diptera species Chironomus thummi, Drosophila melanogaster, Calliphora vicina and Musca domestica using an impulse cytophotometer with a new quartz objective, that was especially manufactured for cytofluorometric investigations. The occurrence of heterogenous cell populations with aneuploid and polyploid DNA content within the cell material of different developmental stages of diptera species have been determined, whereby in larvae polyploid cell populations and in imagos aneuploid cell populations predominate. Partially separation of 2 C cells from other cell populations with higher DNA content can be done by Ficoll-Hypaque centrifugation as demonstrated with cell material from Chironomus larvae. For flow cytometric DNA analysis of insect cell material a simple and rapid cell preparation and staining technique is presented by using the DNA-specific fluorochrome DAPI in combination with the protein fluorochrome sulforhodamine 101. Employment of flow cytometry in diptera genetics might be a new tool for cytological and cytogenetic investigations as shown with the classical genetic objects Chironomus and Drosophila.
Ulrich
W
W
Institut für Genetik, Universität Hohenheim, Stuttgart, Bundesrepublik Deutschland.
D000782:Aneuploidy; D000818:Animals; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D002645:Chickens; D002683:Chironomidae / Q000235:genetics; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004175:Diptera / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods; D006793:Houseflies / Q000235:genetics; D007814:Larva; D011123:Polyploidy; D011679:Pupa
D004247:DNA