Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling.
OBJECTIVE
To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells.


METHODS
The apoptosis of duck embryo fibroblasts (DEF) induced by NGVEV was investigated by fluorescence-activated cell sorter (FACS) and fluorescence microscope after the cells were stained with Annexin V-FITC and propidium iodide (PI).


RESULTS
By staining cells with a combination of fluorescein annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze non-apoptotic cells (Annexin V-FITC negative/PI negative), early apoptotic cells (Annexin V-FITC positive/PI negative), late apoptotic/necrotic cells (Annexin V-FITC positive/PI positive) and dead cells (Annexin V-FITC negative/PI positive) through flow cytometry and fluorescence microscope. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 120 h after infection, while the percentage of non-apoptotic cells decreased.


CONCLUSIONS
NGVEV can induce the infected DEF cells to undergo apoptosis and the apoptosis occurs prior to necrosis.
Chen
Shun
S
Avian Diseases Research Centre, College of Veterinary Medicine of Sichuan Agricultural University, Yaan 625014, Sichuan Province, China.
Cheng
An-Chun
AC
Wang
Ming-Shu
MS
Peng
Xi
X
D000818:Animals; D017304:Annexin A5 / Q000494:pharmacology*; D017209:Apoptosis; D002469:Cell Separation; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D019793:Fluorescein / Q000494:pharmacology*; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology*; D009336:Necrosis; D011419:Propidium / Q000494:pharmacology; D013997:Time Factors; D014780:Viruses / Q000378:metabolism*
D017304:Annexin A5; D011419:Propidium; D016650:Fluorescein-5-isothiocyanate; D019793:Fluorescein