DNA fragmentation in leukocytes following repeated low dose sarin exposure in guinea pigs.
The objective of this study was to determine levels of DNA fragmentation in blood leukocytes and parietal cortex from guinea pigs following repeated low-level exposure to the chemical warfare nerve agent (CWNA) sarin. Guinea pigs were injected (s.c.) once a day for 10 days with saline, or 0.1, 0.2, or 0.4 LD50 (50% mean lethal dose) sarin dissolved in sterile physiological saline. Blood and parietal cortex was collected after injection at 0, 3, and 17 days recovery and evaluated for DNA fragmentation using single-cell gel electrophoresis (Comet assay). Cells were imaged using comet analysis software and three parameters of DNA fragmentation measured: tail length, percent DNA in the tail, and tail moment arm. Repeated low-dose exposure to sarin produced a dose-dependent response in leukocytes at 0 and 3 days post-exposure. There was a significant increase in all measures of DNA fragmentation at 0.2 and 0.4 LD50, but not at 0.1 LD50. There was no significant increase in DNA fragmentation in any of the groups at 17 days post-exposure. Sarin did not produce a systematic dose-dependent response in parietal cortex at any of the time points. However, significant increases in DNA fragmentation at 0.1 and 0.4 LD50 were observed at 0 and 3 days post-exposure. All measures of DNA fragmentation in both leukocytes and neurons returned to control levels by 17 days post-exposure, indicating a small and non-persistent increase in DNA fragmentation following repeated low-level exposure to sarin.
Dave
J R
JR
Division of Psychiatry and Neuroscience, Walter Reed Army Institute of Research, 503 Robert Grant Ave., Silver Spring, Maryland 20910-7500, USA. jit.dave@na.amedd.army.mil
Connors
R A
RA
Genovese
R F
RF
Whipple
R A
RA
Chen
R W
RW
DeFord
S M
SM
Moran
A V
AV
Tortella
E C
EC
D000818:Animals; D002619:Chemical Warfare Agents / Q000633:toxicity*; D020552:Comet Assay; D053938:DNA Fragmentation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D006168:Guinea Pigs; D007928:Lethal Dose 50; D007962:Leukocytes / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D012524:Sarin / Q000008:administration & dosage / Q000633:toxicity*
D002619:Chemical Warfare Agents; D012524:Sarin