Duox expression and related H2O2 measurement in mouse thyroid: onset in embryonic development and regulation by TSH in adult.
In the thyroid, H(2)O(2) is produced at the apical pole of thyrocytes by one or two NADPH oxidases (NOX), Duox1/2 proteins. The onset of Duox expression was analysed by immunohistochemistry in the developing mouse thyroid in parallel with thyroglobulin (Tg) iodination and the expression of other thyroid differentiation markers. Duox proteins were found at embryonic day (E) 15.5 and were mainly localised at the apical pole of thyrocytes. Tg was detected 1 day before (E14.5) and Tg iodination was concomitant with the expression of both Duox and Na(+)/I(-) symporter (NIS; E15.5). The role of TSH in regulating Duox expression and H(2)O(2) accumulation was evaluated in thyroids of adult mice with reduced (Tshr(hyt/hyt) or mice treated with thyroxine) or increased (methimazole or perchlorate treatment) TSH/Tshr activity. In mice with suppressed TSH/Tshr activity, Duox expression was only partially decreased when compared with wild-type, as observed by western blot. In Tshr(hyt/hyt) strain, Duox was still expressed at the apical pole and H(2)O(2) measurements were normal. On the other hand, chronic TSH stimulation of the gland led to a decrease of H(2)O(2) measurements without affecting Duox expression. The onset of Duox protein expression is compatible with their proposed function in thyroid hormone synthesis and it can be considered as a functional marker of the developing thyroid. However, Duox expression in adult is much less regulated by TSH than NIS and thyroperoxidase. It is not always correlated with the overall thyroid H(2)O(2) accumulation, highlighting the importance of additional regulatory mechanisms which control either the production or H(2)O(2) degradation.
Milenkovic
Milutin
M
IRIBHM, Erasme University Hospital, Free University of Brussels, Route de Lennik, 808, 1070 Brussels, Belgium. mmilenko@ulb.ac.be
De Deken
Xavier
X
Jin
Ling
L
De Felice
Mario
M
Di Lauro
Roberto
R
Dumont
Jacques E
JE
Corvilain
Bernard
B
Miot
Francoise
F
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D003409:Congenital Hypothyroidism / Q000196:embryology / Q000378:metabolism; D000074623:Dual Oxidases; D005260:Female; D005420:Flavoproteins / Q000032:analysis*; D018507:Gene Expression Regulation, Developmental; D006861:Hydrogen Peroxide / Q000032:analysis*; D007150:Immunohistochemistry; D007453:Iodide Peroxidase / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008817:Mice, Mutant Strains; D023421:Models, Animal; D019255:NADPH Oxidases / Q000032:analysis*; D011247:Pregnancy; D011989:Receptors, Thyrotropin / Q000378:metabolism; D013954:Thyroglobulin / Q000378:metabolism; D013961:Thyroid Gland / Q000737:chemistry / Q000196:embryology* / Q000378:metabolism; D013972:Thyrotropin / Q000097:blood / Q000494:pharmacology
D005420:Flavoproteins; D011989:Receptors, Thyrotropin; D013972:Thyrotropin; D013954:Thyroglobulin; D006861:Hydrogen Peroxide; D000074623:Dual Oxidases; D007453:Iodide Peroxidase; D019255:NADPH Oxidases; C519130:Duox1 protein, mouse