pEYFP-Nuc vector is a useful tool for three-dimensional and time-lapse observation of nuclear morphology of Jurkat cells during apoptosis.
Using a pEYFP-Nuc vector, which contains nucleic acid sequences of a nuclear localization signal, we established a Jurkat-YN cell line that expressed enhanced yellow fluorescent protein (EYFP) in the nucleus. We observed three-dimensional and time-lapse changes in nuclear morphology of Jurkat-YN cells during Fas-induced apoptosis using a confocal laser scanning microscope. The nuclear forms visualized by EYFP were almost equal in quality to those visualized by SYTO59, a nucleic acid stain for living cells. Three-dimensional deformities in the nuclear form were observed during apoptosis before chromatin condensation became apparent, indicating these deformities are characteristic morphological changes of the early stage of apoptosis. In conclusion, the pEYFP-Nuc vector is a useful tool in the time-lapse observation of nuclear morphology of living cells during apoptosis.
Hasegawa
Akira
A
Department of Radiology, Okayama University Medical School, Okayama 700-8558, Japan.
Kuroda
Masahiro
M
Wang
Yadi
Y
Akaki
Shiro
S
Asaumi
Jun-Ichi
J
Shibuya
Kohichi
K
Kawasaki
Shoji
S
Okumura
Yoshihiro
Y
Kanazawa
Susumu
S
Hiraki
Yoshio
Y
D000911:Antibodies, Monoclonal / Q000276:immunology; D017209:Apoptosis / Q000276:immunology / Q000502:physiology*; D002467:Cell Nucleus / Q000276:immunology / Q000502:physiology*; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D019169:Jurkat Cells / Q000166:cytology*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D013997:Time Factors; D019014:fas Receptor / Q000276:immunology
D000911:Antibodies, Monoclonal; D019014:fas Receptor