Effects of 3-iodo-L-tyrosine, a tyrosine hydroxylase inhibitor, on eye pigmentation and biogenic amines in the planarian, Dugesia dorotocephala

Planarians (Dugesia dorotocephala) were evaluated as bioassay organisms to detect inhibition of tyrosine hydroxylase, the rate-limiting Enzyme in the synthesis of catecholamines. Thirty planaria per dose were exposed to 0 (control), 0.001, 0.01, 0.1, or 1 mM 3-iodo-L-tyrosine (monoiodotyrosine or MIT) in standard test media beginning 24 hr before decapitation and continuing for 13 days. Complete regeneration of normal heads occurred over the first 6 days in all dose groups, a response reported to be partially dependent on catecholamines. Beginning on Day 7, the black eye Pigments began fading in the 0.1 and 1 mM dose groups and were completely absent macroscopically and histologically by Day 11. The 0, 0.001, and 0.01 mM dose groups did not lose visible eye Pigments. On Day 13, 3 planaria/dose were harvested for histopathology; 15 planaria/dose were decapitated a second time and remained in MIT solutions; and 12 planaria/dose were left intact, placed in fresh control media, and evaluated for eye repigmentation. Normal head regeneration (including eyes) was detected grossly in all groups, even in those Animals devoid of eye Pigments at the time of decapitation. As before, eye Pigments began fading 7 days after decapitation (Day 20 of experiment) and were completely absent in 73 and 33% of the Animals in the 0.1 and 1 mM groups, respectively, on Day 25. All Animals moved to control media reformed eye Pigments, beginning within 48 hr. Analysis of the decapitated heads by HPLC-ECD on Day 13 revealed a significant decrease in dopamine and 5-hydroxytryptamine (serotonin) concentrations in MIT-exposed Animals. Tyrosine hydroxylase activity (and possibly Tyrosinase activity) was shown to be inhibited by the highest two concentrations for whole planaria homogenates in vitro.