Light and electron microscopy of apoptotic DNA fragmentation


https://doi.org/10.4081/microscopie.2008.4960
Vol. 9 No. 1 (2008)
Submitted: 15 January 2015
Accepted: 15 January 2015
Published: 31 March 2008
apoptosis, DNA fragmentation, TUNEL/LM, TUNEL/TEM, U-937, Molt-4, UVB, staurosporine.
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Authors

  • L. Biagiotti Istituto di Scienze Morfologiche,Università degli Studi di Urbino “Carlo Bo”, Urbino, Italy.
  • P. Ferri Istituto di Scienze Morfologiche,Università degli Studi di Urbino “Carlo Bo”, Urbino, Italy.
  • A. D’Emilio Istituto di Scienze Morfologiche,Università degli Studi di Urbino “Carlo Bo”, Urbino, Italy.
  • M.B.L. Rocchi Istituto di Biomatematica, Università degli Studi di Urbino “Carlo Bo”, Urbino, Italy.
  • E. Falcieri
    elisabetta.falcieri@uniurb.it
    Istituto di Scienze Morfologiche,Università degli Studi di Urbino “Carlo Bo”, Urbino; Istituto di Genetica Molecolare,CNR, Istituti Ortopedici Rizzoli, Bologna, Italy.
  • S. Burattini Istituto di Scienze Morfologiche,Università degli Studi di Urbino “Carlo Bo”, Urbino, Italy.
One of the most known apoptotic markers is DNA fragmentation. The cell initially produces large 50-300kbp fragments and, successively, oligonucleosimic ones. However, apoptosis without DNA fragmentation, with the typical apoptotic features, such as chromatin condensation and micronuclei, has been reported. In this work we have investigated the relationship between apoptotic morphology and the underlying DNA behaviour in two different cell lines (U937 and Molt-4), undergoing apoptosis after UVB irradiation or staurosporine treatment. TUNEL reaction was utilized both in fluorescence (by FITC) and transmission electron microscpy (by colloidal gold). While the first only highlights apoptosis presence or absence in cells, the second reveals a precise localization of DNA break points, clearly identified by colloidal gold, in diffuse or dense chromatin. Colloidal gold particles density was evaluated in the different experimental conditions and was correlated to DNA fragmentation patterns.

Biagiotti, L., Ferri, P., D’Emilio, A., Rocchi, M., Falcieri, E., & Burattini, S. (2008). Light and electron microscopy of apoptotic DNA fragmentation. Microscopie, 9(1), 45–52. https://doi.org/10.4081/microscopie.2008.4960

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