New applications of laser microbeam irradiation to cell and developmental biology include a new instrument with a tunable wavelength (217- to 800-nanometer) laser microbeam and a wide range of energies and exposure durations (down to 25 × 10 -12 second). Laser microbeams can be used for microirradiation of selected nucleolar genetic regions and for laser microdissection of mitotic and cytoplasmic organelles. They are also used to disrupt the developing neurosensory appendages of the cricket and the imaginal discs of Drosophila .

Cells of a V79 subline of the Chinese hamster were microirradiated at wavelength 365 nm in the presence of the psoralen derivative, trioxsalen. Microirradiation was accomplished by a pulsed argon laser microbeam either in anaphase or in interphase 3 hr after mitosis. Inhibition of clonal growth and formation of micronuclei at the first postirradiation mitosis were observed after microirradiation of anaphase chromosomes and of small parts of the interphase nucleus. Microirradiation of the cytoplasm beside the interphase nucleus or between the sets of chromosomes moving apart from each other in anaphase did not produce these effects. Anaphase experiments showed that only the daughter cell which received microirradiated chromatin exhibited an abnormal growth pattern. Most interestingly, shattering of the whole chromosome complement could be induced by microirradiation of small parts of the interphase nucleus and post-treatment with caffeine. Since microirradiation of chromatin in the absence of psoralen was not effective, we consider formation of psoralen photoadducts to nucleic acids in microirradiated chromatin to be the specific cause of the effects. We suggest that DNA photolesions in chromosome segments present in the microirradiated part of the nucleus can induce shattering of all the chromosomes in the microirradiated nucleus. Several possibilities are discussed to explain this unexpected finding.

No abstract

No abstract

Abstract. 4′‐Aminoethyl‐4,5′,8‐trimethylpsoralen, a DNA photoreagent, was used in conjunction with near UV light to modify cell growth and morphology. Since near UV radiation is needed to trigger the photoreactions, various doses of classical and laser near UV light were exposed to cells growing in media containing psoralen. It was found that near UV light caused a reduction in cell viability as indicated by complete inhibition of growth. The specificity of psoralens for nucleic acids was also investigated by using a tritium labeled psoralen derivative and tracing its appearance in different fractions of DNA from treated and untreated cells.

No abstract

The light-activated, nucleic acid-binding drugs, psoralens, were used in conjunction with a 365-nm laser microbeam to selectively bind to any nucleic acids in the centriolar region. 4′-aminomethyl-4,5′,8—trimethyl-psoralen (AMT) has a high affinity for both RNA and DNA and can be shown to cause mitotic abortion when centriolar regions of prophase PTK2 cells and reacted with AMT and 365-nm laser light. Other psoralen derivatives which have a high affinity for DNA and a low affinity for RNA are not effective in blocking mitosis in dividing PTK2 cells. Examination of psoralen-bound centriolar regions by single-cell electron microscopy shows that at various times after treatment, the number of microtubules associated with the irradiated poles is much lower than in normal, dividing cells. Light-activated psoralen binding of the centriolar regions does not seem to affect the condensation or structure of mitotic chromosomes. It is concluded that there is an RNA in the centriolar region that is responsible for the formation of the spindle in dividing cells.

No abstract