Flow cytometry measurement of the DNA contents of G0/G1 diploid cells from three different teleost fish species

Cytometry. 2002 May 1;48(1):20-5. doi: 10.1002/cyto.10100.

Abstract

Background: Although there is a lot information in the literature about genome size in fish, a high variability among data for the same species is reported, being mainly related to methodological aspects. Flow cytometry-based fluorescence measurements of intercalating dyes is the most attractive approach due to its precision, objectivity, high speed, and relative simplicity.

Methods: We analyze the DNA content of G0/G1 diploid nuclei of three teleost species (Carassius auratus, Tinca tinca, and Danio rerio) using flow cytometry. Forty-three animals were used and up to 50,000 retinal cells were analyzed per sample. Propidium iodide-associated fluorescence was assessed using a FACSCalibur flow cytometer. Standard human leukocytes were used as a reference.

Results: Our results show that C. auratus (3.584 +/- 0.058 pg per nucleus) and D. rerio (3.357 +/- 0.074 pg per nucleus) showed similar DNA contents per cell, whereas it was significantly lower (2.398 +/- 0.038 pg per nucleus) in T. tinca. Interestingly, a low intraspecies variability was observed, the coefficient of variation being 1.608%, 2.198%, and 1.573% for C. auratus, D. rerio, and T. tinca, respectively.

Conclusions: The methodology used in this study provides an accurate and easy measurement of the genome size of a species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Coloring Agents
  • Cyprinidae
  • DNA / analysis
  • Flow Cytometry / methods*
  • G1 Phase / genetics*
  • Genome
  • Goldfish / genetics*
  • Propidium
  • Resting Phase, Cell Cycle / genetics*
  • Species Specificity
  • Zebrafish

Substances

  • Coloring Agents
  • Propidium
  • DNA