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First published online November 4, 2005
Journal of Experimental Biology 208, 4263-4271 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01895

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The effects of L-arginine and L-NAME supplementation on redox-regulation and thermogenesis in interscapular brown adipose tissue

Vesna Petrovi¹, Aleksandra Kora², Biljana Buzadzi¹ and Bato Kora^1,*

¹ Department of Physiology, Institute for Biological Research, `Sinia Stankovi', University of Belgrade, Bulevar Despota Stefana 142, 11060 Belgrade, Serbia and Montenegro
² Institute of Zoology, Faculty of Biology, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia and Montenegro

View larger version (113K): [in a new window]   Fig. 1. Immunohistochemical detection of iNOS in IBAT (polyclonal antibody 1:200 v/v). (A) Control 22±1°C, (B) control 4±1°C, (C) L-Arg 22±1°C, (D) L-Arg 4±1°C, (E) L-NAME 22±1°C, (F) L-NAME 4±1°C. (G) Non-immune (Ni) control - primary antibody was omitted. Inserts show fields at higher magnification. Scale bars, 25 µm.

View larger version (59K): [in a new window]   Fig. 2. The effects of L-Arg and L-NAME supplementation and different temperatures (±1°C) on the iNOS protein level in IBAT. (A) Western blotting was performed using specific antibody against iNOS. The results of three observations from a representative experiment and the position of molecular mass markers (kDa) are shown. C, control. (B) Data obtained after quantification of iNOS bands by ImageQuant software. iNOS concentration is expressed in relative to the levels in controls acclimated to room temperature (taken as 100%). The values represent the means ± S.E.M. from three independent experiments. *Comparison of the same treatments at different temperatures, **P<0.025; ***P<0.005; comparison of different treatments with the control kept at the same temperature, P<0.025; P<0.005. Volume is the sum of all the pixel intensities within a band; 1 pixel=0.007744 mm2.

View larger version (32K): [in a new window]   Fig. 3. The effects of L-Arg and L-NAME supplementation and different temperatures on UCP1 protein level in IBAT. (A) Western blotting was performed using specific antibody for UCP1. The results of three observations from a representative experiment and the position of molecular mass markers (kDa) are shown. C, control. (B) Data obtained after quantification of UCP1 bands by ImageQuant software. Relative UCP1 concentration was expressed in relation to the control cold-acclimated group taken as 100%. The values represent the means ± S.E.M. from three independent experiments. Comparison of different treatments with the control kept at the same temperature, P<0.005. Volume is the sum of all pixel intensities within a band; 1 pixel=0.007744 mm2.

View larger version (29K): [in a new window]   Fig. 4. MnSOD activity in the IBAT. *Comparison of the same treatments at different temperatures, ***P<0.005; comparison of different treatments with the control maintained at the same temperature, P<0.05; P<0.025.

View larger version (33K): [in a new window]   Fig. 5. Glutathione (GSH) content in the IBAT. *Comparison of the same treatments at different temperatures, **P<0.025; ***P<0.005; comparison of different treatments with the control acclimated to the same temperature, P<0.025.

View larger version (179K): [in a new window]   Fig. 6. IBAT TUNEL staining. (A) Control, 22±1°C, (B) control, 4±1°C, (C) L-Arg, 22±1°C, (D) L-Arg, 4±1°C, (E) L-NAME, 22±1°C, (F) L-NAME, 4±1°C. Arrows indicate apoptotic nuclei (see text for details). Scale bars, 20 µm.