Thyroxine enhancement and the role of reactive oxygen species in tadpole tail apoptosis.
Our objective is to clarify the role of reactive oxygen species (ROS) in the atrophying tail of anuran tadpoles (tail apoptosis). Changes in catalase, superoxide dismutase (SOD) and caspase activity, genomic DNA, and nitric oxide (NO) generation were investigated biochemically using Rana japonica tadpole tails undergoing regression during thyroid hormone enhancement. DNA fragmentation and ladder formation with concomitant shortening of tadpole tail were induced by DL-thyroxine (T4) in culture medium. Catalase activity was also decreased by T4 treatment. T4 was also found to increase NO synthase (NOS) activity in cultured tadpole tail with concomitant increase in the concentration of NO2- plus NO3- (NOx) in the culture medium. Additional treatment with N-monomethyl-L-arginine (NMMA), a potent inhibitor of NOS, suppressed the enhancing effects of T4 on tail shortening and catalase activity reduction. It was also found that treatment with isosorbide dinitrate (ISDN), a NO generating drug, alone also had an enhancing effect on tail shortening and catalase activity reduction similar to that seen with T4. Both NO and an NO donor (ISDN) strongly suppressed catalase activity. Kinetic analysis revealed that catalase activity decreased and caspase-3-like activity increased during normal tadpole tail atrophy (apoptosis). These results suggested that T4 enhances NO generation, thereby strongly inhibiting catalase activity, resulting in an increase in hydrogen peroxide, and that the oxidative stress elicited by excess hydrogen peroxide might activate cysteine-dependent aspartate-directed protease-3 (caspase-3-like protease), which is thought to cause DNA fragmentation, leading to apoptosis.
Kashiwagi
A
A
Laboratory for Amphibian Biology, Faculty of Science, Hiroshima University, Higashihiroshima, Japan.
Hanada
H
H
Yabuki
M
M
Kanno
T
T
Ishisaka
R
R
Sasaki
J
J
Inoue
M
M
Utsumi
K
K
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D020169:Caspases / Q000378:metabolism; D002374:Catalase / Q000378:metabolism*; D053938:DNA Fragmentation; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D007814:Larva; D009569:Nitric Oxide / Q000378:metabolism; D009924:Organ Culture Techniques; D011898:Ranidae; D017382:Reactive Oxygen Species / Q000502:physiology*; D013482:Superoxide Dismutase / Q000378:metabolism*; D013623:Tail; D013974:Thyroxine / Q000494:pharmacology*; D013997:Time Factors; D019323:omega-N-Methylarginine / Q000494:pharmacology
D017382:Reactive Oxygen Species; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D020169:Caspases; D013974:Thyroxine