Figure S1: Regional selectivity: Studies performed on the thalamus. A - ROS production
in response to the low glucose load. ROS production in the thalamus in response to saline
(NaCl) or to 3mg/kg (G3) glucose injection towards the brain measured in obese rats. ROS
level was assessed in hypothalamic area by oxidation of DCFDA probe one minute after
intracarotid injection. Data were expressed as the mean + SEM corresponding to the % of the
ROS fluorescence observed in obese rats injected with saline. No significant difference was
observed according to the student’s unpaired t test (n=6-7 each). DCFDA, dichlorofluorescein
diacetate. B - Identical Glutathione Peroxidase activity. Glutathione Peroxidase (GPx)
activity measured in the thalamus of lean (white bar) and obese (black bar) rats (e.u.
enzymatic units). No significant difference was present according to the student’s unpaired t
test (n=6-7 each). C - Obese rats mitochondria display no hypersensitivity to glutamate.
Pharmacological settings for oxygraphic analysis on isolated thalamic mitochondria:
glutamate titration (1, 5, 20 mM) to achieve the non-phosphorylating respiration; saturating
Adenosine diphosphate (ADP) concentration to achieve state 3 respiration; full inhibition of
ATP-synthase by Carboxy-Artractylate (CAtr) gives state 4 respiration. No significant
difference was present according to the repeated measures ANOVA analysis (n=6 each
genotypes) for the glutamate titration. No significant difference was present according to the
student’s unpaired t test concerning ADP and CAtr respiration in lean vs. obese rats. D – No
difference in Respiratory Control Ratio defined as State 3 divided by State 4 (n=6 each
genotypes). No significant difference was present according to the student’s unpaired t test. E
- Thalamic mitochondria exhibit identical maximal respiration capacity. Maximal
respiration assessed by CCCP (0.4 µM). No significant difference was present according to
the student’s unpaired t test (n=6 each genotypes). F - Thalamic mitochondria exhibit no
uncoupling respiration. Uncoupling respiration assessed by uncoupling protein activation by
palmitate treatment (Palm) (300 µM). No significant difference was present according to the
student’s unpaired t test (n=6 each genotypes).
Figure S2: No toxicity of GSH perfusion : A – Daily weight gain during the 3-days of
infusion. Data were expressed as mean weight gain/day + SEM during the 3-days of vehicleor GSH- i.c.v. infusion in obese rats. No significant difference was observed according to the
student’s unpaired t test (n=5-6 each). B – Daily food intake during the 3-days of infusion.
Data were expressed as mean food intake/day + SEM during the 3-days of vehicle- or GSHi.c.v. infusion in obese rats. No significant difference was observed according to the student’s
unpaired t test (n=5-6 each).