Role of water in chromosome spreading and swelling induced by acetic acid treatment: a FTIR spectroscopy study

  • D. Ami | diletta.ami@unimib.it Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milano, 20126, Italy, Italy.
  • M. Di Segni Laboratory of Medical Genetics - Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico, Milan, Italy., Italy.
  • M. Forcella Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milano, 20126, Italy, Italy.
  • V. Meraviglia Centro Cardiologico Monzino, Italy.
  • M. Baccarin Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Italy.
  • S.M. Doglia University of Milano-Bicocca, Italy.
  • G. Terzoli Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Italy.

Abstract

The so called chromosome preparation is a procedure consisting of three strictly connected stages that enables to obtain chromosomes of quality suitable for cytogenetic analysis. Interestingly, experimental evidence strongly suggested that chromosome spreading and swelling (key processes that allow their counting and detailed structural analysis) are induced in the last fixative-evaporation stage by the interaction, mediated by acetic acid, between water from the environmental humidity, and the cytoplasmic matrix and the chromatin. However, since a considerable variation in the quality of chromosome preparations is observed, strongly depending on the environmental conditions in which the procedure takes place, a better comprehension of the mechanisms underlying chromosome preparation is required. To this aim, here we analysed intact lymphocytes before and at each stage of the chromosome preparation protocol by Fourier transform infrared (FTIR) spectroscopy, a technique widely used for the study not only of isolated biomolecules, but also of complex biological systems, such as whole cells. Interestingly, we found that the chromosome preparation protocol induces significant structural changes of cell proteins and DNA, in particular due to the interaction with acetic acid. Moreover, noteworthy, through the monitoring of changes in the water combination band between 2300 and 1800 cm–1, we provided evidence at molecular level of the crucial role of the bound water to the cytoplasmic matrix and to the chromatin in determining the chromosome spreading and swelling. Our FTIR results, therefore, underline the need to perform the last fixative-evaporation stage in standardized and optimized temperature and relative humidity conditions, thus providing chromosomes of high quality for the cytogenetic analysis that would lead in this way to more reliable results.

Dimensions

Altmetric

PlumX Metrics

Downloads

Download data is not yet available.

Author Biographies

V. Meraviglia, Centro Cardiologico Monzino
Vascular Biology and Regenerative Medicine Lab
M. Baccarin, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico
Laboratory of Medical Genetics
G. Terzoli, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico
Laboratory of Medical Genetics
Published
2014-02-11
Info
Issue
Section
Original Papers
Keywords:
Chromosome preparation, chromosome spreading and swelling, DNA conformational transition, FTIR (micro)spectroscopy, protein and DNA hydration.
Statistics
  • Abstract views: 2367

  • PDF: 369
  • Supplementary: 107
  • HTML: 693
How to Cite
Ami, D., Di Segni, M., Forcella, M., Meraviglia, V., Baccarin, M., Doglia, S., & Terzoli, G. (2014). Role of water in chromosome spreading and swelling induced by acetic acid treatment: a FTIR spectroscopy study. European Journal of Histochemistry, 58(1). https://doi.org/10.4081/ejh.2014.2330