Flow cytometry measurement of the DNA contents of G0/G1 diploid cells from three different teleost fish species.
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.
Servicio de Citometría, Laboratorio de Hematologia (1(a) planta), Hospital Universitario de Salamanca, Paseo de San Vicente s/n, 37007 Salamanca, Spain. ciudad@gugu.usal.es
Juana
J
Ciudad
Elena
E
Cid
Almudena
A
Velasco
Juan M
JM
Lara
José
J
Aijón
Alberto
A
Orfao
D000818:Animals; D004396:Coloring Agents; D003530:Cyprinidae; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D016193:G1 Phase / Q000235:genetics*; D016678:Genome; D006054:Goldfish / Q000235:genetics*; D011419:Propidium; D016192:Resting Phase, Cell Cycle / Q000235:genetics*; D013045:Species Specificity; D015027:Zebrafish
D004396:Coloring Agents; D011419:Propidium; D004247:DNA