Spindle disappearance and chromosome behavior after partial-embryo irradiation in Cecidomyiidae (Diptera)

Summary

• 1. Currently available data on chromosome elimination in Cecidomyiidae are still inadequate to support any definite conclusions regarding its immediate causes. It seems that some evidence on the mechanism of elimination could be obtained from observation of the behavior of E chromosomes in complete absence of spindle. To bring about such absence of spindle, ultraviolet microbeams would appear promising, since they have already been used to destroy spindles in cells of other organisms.

• 2. For irradiation the embryos of Rhabdophaga batatas Walsh were chosen. It was established that in this species 2 S+E=40 and 2 S (for female)=8. Elimination of 30-odd chromosomes regularly occurs in the fifth cleavage. Embryos were irradiated with 8μ and 16μ heterochromatic ultraviolet microbeams not only in this fifth cleavage but also in the fourth, in which elimination was never observed. The irradiated regions were small parts of somatic cytoplasm or small parts of germ cells.

• 3. The effects of irradiation were multifold: complete degeneration of nuclei in the vicinity of irradiated cytoplasm; disappearance of spindle and formation of quasi-rosettes; multipolar mitosis. The polar substance present in germ cells was often severely damaged and fragmented. All these effects are evidently indirect.

• 4. In mitoses thus deprived of spindles, the separation of chromosomes into daughter chromatids was suppressed but some of their active movements persisted. All chromosomes were gathered into a quasirosette, which afterwards divided into two quasi-rosettes. The daughter quasi-rosettes moved apart and formed telophase nuclei. In both irradiated divisions (fourth and fifth) E chromosomes behaved in exactly the same manner as S chromosomes; they showed the same kind of peculiar movement and were incorporated into telophase nuclei. Suppression of elimination of E chromosomes in the fifth division can be regarded as one more indirect effect of irradiation to be added to the list above.

• 5. These results show that elimination of E chromosomes does not result from failures in chromosomal spindle fibers or from intrinsic morphological defects in the centromeres. The fact that elimination can be suppressed by indirect action of ultraviolet light strongly supports the assumption that elimination occurs because of chemical changes in adjacent cytoplasm that are injurious to the centromeres of E chromosomes. Most probably the indirect action of ultraviolet interferes with the production of some substance that is normally injurious to the centromeres in the fifth division.

• 6. Since in Cecidomyiidae the general features of the metabolism are known to be stable up to the mid-blastoderm stage, it would appear that the chemical changes involved in elimination cleavage are probably quite specific.