Mori A, Hatano T, Inoshita T, Shiba-Fukushima K, Koinuma T, Meng H, Kubo SI, Spratt S, Cui C, Yamashita C, Miki Y, Yamamoto K, Hirabayashi T, Murakami M, Takahashi Y, Shindou H, Nonaka T, Hasegawa M, Okuzumi A, Imai Y, Hattori N. Proc Natl Acad Sci U S A. Neuroimaging 181, 71–76. Figure 10. Conversion into cDNA was performed by a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, Waltham, MA, United States) using a T100 Thermal Cycler (Bio-Rad, Hercules, CA, United States). However, little is known about the effect of thyroid hormones on mitochondrial function in adult brain cells. Brain. Biochim Biophys Acta. The samples were sonicated, activated with Aconitase Activation Solution containing equal amounts of cysteine-HCl and (NH4)2Fe(SO4)2, and subsequently transferred to 96-well plates along with standards (0, 4, 8, 12, 16, and 20 nmol/well). The results are expressed as the mean ± SEM. The intensification of the leak state in animals with hypothyroidism could indicate the uncoupling of oxidative phosphorylation from ATP synthesis in these animals. *p < 0.05 vs. the control group (Wistar rats); #p < 0.05 vs. the WKY group. 2020 Feb;61(2):143-158. doi: 10.1194/jlr.RA119000281. Natl. The total T4 hormone levels were slightly but not significantly lower in WKY rats than in Wistar rats in the Redei study (Redei et al., 2001), while we found that the level of the free form of this hormone was significantly lower in the WKY rats than in the Wistar rats. Epub 2011 Jun 15. (2018). WKY rats were also characterized as a strain with increased reactivity to stress, prolonged ACTH and corticosterone secretion, and weakened negative feedback mechanisms (Aleksandrova et al., 2019). Astrocyte-neuron lactate transport is required for long-term memory formation. 27, 120–131. Brain Res. 2011 Nov;96(1-4):109-13. doi: 10.1016/j.prostaglandins.2011.06.003. Furthermore, no effect of strain or PTU was observed in the E: ETS (CI + II) capacity after the addition of succinate in either the frontal cortex or the hippocampus (Figures 10C, 11C). Thus, the increase in oxygen consumption in the leak state in animals receiving PTU suggested that in the rats with hypothyroidism, the inner mitochondrial membrane was more permeable to protons, and consequently, oxidative phosphorylation may have become uncoupled from ATP synthesis. After completing this journal-based SA-CME activity, participants will be able to: 1. â Describe the process of neuronal uptake and cerebral biodistribution of FDG and identify normal cortical and subcortical structures at FDG PET. He noted that persons who had experienced stressful life events, depression, or âlong sorrow,â often suffered from diabetes. A. The actual test in animals was preceded by a pretest carried out on the previous day during which the animals were placed in the water-filled cylinder for 15 min. n = 9–10. Arecoline increased both parameters in brain regions of iPLA(2)β(+/+) mice but quantitatively less so in iPLA(2)β(-/-) and iPLA(2)β(+/-) mice. Clipboard, Search History, and several other advanced features are temporarily unavailable. Years later, in 1897, British psychiatrist Henrâ¦ Imaging brain signal transduction and metabolism via arachidonic and docosahexaenoic acid in animals and humans. Our study has some limitations. Human neuronal uncoupling proteins 4 and 5 (UCP4 and UCP5): structural properties, regulation, and physiological role in protection against oxidative stress and mitochondrial dysfunction. The FST was conducted in accordance with the procedure described by Porsolt et al. J. Mol. The NOR test (Taglialatela et al., 2009) was applied with modifications. Moreover, in some cellular models, UCP4 has been shown to increase mitochondrial complex II activity and glucose uptake and shift ATP production from mitochondrial respiration to glycolysis (Ramsden et al., 2012). Alzheimers Res Ther. 2010 Aug;51(8):2334-40. doi: 10.1194/jlr.M006262. Statistical parametric mapping was used to identify regions of decreased cerebral glucose metabolism. Thyroid hormone responsive protein (THRP) mediates thyroid hormone-induced cytotoxicity in primary neuronal cultures. Horm. Decrease Foods that Slow Down Metabolism:. Cannabinoids decrease the metabolism of glucose in the brain by University of the Basque Country Cannabis creates a â¦ BMR is the number of calories your body uses to maintain vital functions, such as breathing, heart rate, and brain function. Furthermore, hypothyroidism in humans does not necessarily lead to depression but increases the risk of this disease, and the administration of a high dose of T4 to patients with drug-resistant depression produces a therapeutic effect, probably by normalizing any significant changes that have occurred in depression. The mitochondria-enriched membrane fraction and cytosolic fraction were transferred to a 96-well plate and mixed with the reaction mix. Although hypothyroidism appears to be an important factor in the pathogenesis of depression, the impact of thyroid hormones on the bioenergetics of the adult brain is still poorly known. (2014). Measurement of protein using bicinchoninic acid. 2017/25/B/NZ7/01708 from the National Science Center, Poland, and in part by the statutory funds of Laboratory of Immunoendocrinology, Department of Experimental Neuroendocrinology, Maj Institute of Pharmacology PAS, Poland. A hypothesis on the mechanism of action of high-dose thyroid in refractory mood disorders. Glial cells, which make up almost 50% of the brain volume, have a much lower metabolic rate and account for less than 10% of total cerebral metabolism (Siesjo, 1984).¨. The reason why a decrease was seen in [18 F]-FTHA uptake but not in [11 C]-palmitate could be that oxidative metabolism might be more flexible in nature, whereas the synthesis of complex lipids may be more preserved due to its important structural role in the brain. Consistent with iPLA(2)β's reported ability to selectively hydrolyze DHA from phospholipid in vitro, iPLA(2)β deficiency reduces brain DHA metabolism and signaling in vivo at baseline and following M(1,3,5) receptor activation. In a human, the cerebral cortex contains approximately 14â16 billion neurons, and â¦ The frontal cortex and hippocampus were selected for current study because they are the main structures affected in depressive disorders. | Basal metabolic rate (BMR). A region â¦ | The results are expressed as the average fold change ± SEM (A–D) or as the mean ± SEM. In the hippocampus, there was no difference in DIO2 expression between the rat strains, and PTU increased this parameter (treatment effect: F1,35 = 4.27, p = 0.046) in the Wistar rats only (Figure 5B). J. (2015). Figure 4. Association between subclinical hypothyroidism and depression: an updated systematic review and meta-analysis 11 medical and health sciences 1117 public health and health services 11 medical and health sciences 1103 clinical sciences. doi: 10.1097/FBP.0000000000000431, Yankelevitch-Yahav, R., Franko, M., Huly, A., and Doron, R. (2015). In summary, the obtained results suggested that in both the depression and hypothyroidism models, a reduction in glycolysis and in the connection between glycolysis and the Krebs cycle occurred, while the weakening of oxidative phosphorylation was mainly due to a lower level of thyroid hormones. Additionally, we found that the uncoupling of oxidative phosphorylation from ATP synthesis did not result from the overexpression of uncoupling protein 4 (UCP4). But gaining and losing fat can change the way your brain regulates your body weight. Ca(2+)-independent phospholipase A(2)β (iPLA(2)β) selectively hydrolyzes docosahexaenoic acid (DHA, 22:6n-3) in vitro from phospholipid. (2014). Moderate doses of alcohol decrease glucose metabolism in the human brain, which has been interpreted to reflect alcohol-induced decreases in brain activity. doi: 10.1042/bj3020857, Kelly, T. (2016). (1985). In our study, the levels of VDAC1 in the mitochondrial fraction in both brain structures examined did not change, and the HK1 concentration did not decrease or even increased in the hippocampus of the WKY rats treated with PTU; therefore, we excluded the participation of this mechanism in the models we examined. Insulin resistance was linked to decreased brain glucose metabolism and predicted worse memory function among late-middle-aged adults at risk for Alzheimer's disease, researchers reported. Rev. As indicated by recent studies, including our research conducted in a prenatal stress animal model of depression, metabolic changes may participate in the pathogenesis of depression (Rezin et al., 2008; Regenold et al., 2012; Detka et al., 2015; Głombik et al., 2017). The decreased expression of the TRα1 isoform in the frontal cortex of the WKY rats suggested weaker TH activity in this brain structure because this receptor isoform is widely distributed and is the predominant TH receptor in the adult brain. These changes might contribute to reported altered behavior following early SSRI â¦ It is located in the head, usually close to the sensory organs for senses such as vision.It is the most complex organ in a vertebrate's body. 116, 222–226. Res. A. The administration of PTU had no effect on immobility or climbing time but attenuated the memory impairment of the WKY rats and intensified the decrease in weight gain. For thyroid hormones, data on their effects on depression-like and anxiety-like behaviors are very ambiguous because both hypothyroidism and hyperthyroidism can induce such behaviors. The metabolic action of thyroid hormones in peripheral tissues and in the developing brain is well documented, and a few studies have indicated that these hormones increase the activity of Krebs cycle enzymes and affect mitochondrial respiration in the adult brain (Katyare et al., 1994; Bauer et al., 2016). doi: 10.1016/j.steroids.2005.02.003, Crispino, M., Trinchese, G., Penna, E., Cimmino, F., Catapano, A., Villano, I., et al. doi: 10.1111/nyas.12781, Iosifescu, D. V., Bolo, N. R., Nierenberg, A. Received: 24 July 2020; Accepted: 09 November 2020;Published: 04 December 2020. Acad. The combined supernatants were then centrifuged at 17,000 × g for 12 min at 4°C, after which the mitochondria-containing pellets and supernatants (cytosolic fraction) were stored at −80°C. However, the co-occurrence of hypothyroidism and depression led to changes in the presence or abundance of some metabolic markers, such as decreased pyruvate dehydrogenase levels in the frontal cortex, decreased lactate levels in the cytosolic fraction of the hippocampus, increased PDK4 expression in the hippocampus, and decreased ETS and OXPHOS coupling efficiency. Differential stress response in rats subjected to chronic mild stress is accompanied by changes in CRH-family gene expression at the pituitary level. HDL measurements showed the impact of PTU (treatment effect: F1,32 = 13.02, p = 0.001) and the strain × PTU interaction (interaction: F1,32 = 9.60, p = 0.004). The results are presented as the preference index, which was calculated as follows: preference index = (time with novel object/[time with novel object + time with familiar object]). We have also found that the changes observed in the studied models depend on which brain structure is evaluated. Neuroscience 213, 72–80. Biochem. 2. â Discuss the role of FDG PET in the diagnosis of primary neurodegenerative disorders and recognize the patterns of hypometabolism that typically characterize these disorders. The reduced level of growth factors in an animal model of depression is accompanied by regulated necrosis in the frontal cortex but not in the hippocampus. Additionally, changes in oxidative phosphorylation were present mainly in the frontal cortex, but these disturbances were related to a deficiency of thyroid hormones rather than depression. Corticosterone concentrations were significantly lower in animals receiving PTU than in the control Wistar rats (treatment effect: F1,36 = 17.02, p = 0.0002); however, there were no differences in the levels of this hormone between the strains or between the WKY control rats and those receiving PTU (Figure 2D). (2010). Cogn. (1978). doi: 10.1016/j.cell.2011.02.018, Taglialatela, G., Hogan, D., Zhang, W.-R., and Dineley, K. T. (2009). Eur. doi: 10.1016/j.neuroscience.2015.03.046. Psychiatry 63, 1127–1134. 69, 106–115. Impact Factor 3.707 | CiteScore 5.1More on impact ›, Worcester Polytechnic Institute, United States. (2014). 105, 1–23. (2012). All biochemical experiments were carried out under exactly the same conditions for every sample, regardless of the type of animal treatment. However, since no differences in the expression of these enzymes were observed between the rat strains, only slight downward trends in these levels were observed in the WKY rats, and because PTU decreased the levels of these enzymes in the WKY and Wistar rats to a similar extent, the changes in complexes II and V may be associated with hypothyroidism but not with differences between the tested rat strains. In addition, the decreased metabolism in the frontal cortex in depression may result from decreases in the levels of T3 and TRα1 and in the expression of DIO2, an enzyme that catalyzes the conversion of T4 to the active T3 hormone. Biol. The effects of strain and PTU treatment on PDK2 and PDK4 in the mitochondria-enriched fraction of the frontal cortex (A,C) and (B,D) hippocampus. All authors contributed to the article and approved the submitted version. Inhibition of mitochondrial respiratory chain in brain of rats subjected to an experimental model of depression. Note: This post delves into the science of energy balance, thermodynamics, and metabolic â¦ Mouchlis VD, Chen Y, McCammon JA, Dennis EA. PTU is an antithyroid drug which inhibits synthesis of thyroid hormones, used in the therapy of hyperthyroidism and Graves disease. The use of triiodothyronine as an augmentation agent in treatment-resistant bipolar II and bipolar disorder NOS. PTU also had no effect on the expression of this protein. Nelson AJ, Stephenson DJ, Cardona CL, Lei X, Almutairi A, White TD, Tusing YG, Park MA, Barbour SE, Chalfant CE, Ramanadham S. J Lipid Res. The obtained supernatant (SN2) was then mixed with SN1, and the supernatant mixture was centrifuged at 9,000 × g for 15 min. J. Physiol. Figure 9. ScienceDaily. Moreover, some studies suggest that depression is connected with a decrease in the level or activity of thyroid hormones only within the brain. Front. Integr. In the frontal cortex, the decreased expression of complex II and complex V in rats with hypothyroidism suggested that in this brain structure, not only glycolysis but also oxidative phosphorylation could be reduced. No significant changes in ETS coupling efficiency were observed in the frontal cortex (Figure 10E). Comp. The manuscript has been edited by the American Journal Experts for proper English language, grammar, punctuation, spelling, and overall style. Brain Res. 322, 501–518. PTU increased the level of HDL in the WKY rats compared with those of the other groups (Figure 3B). oxygen (CMRO 2), glucose (CMR glu), or generates by-products, e.g. The administration of PTU had no effect on immobility or climbing time in either strain. Neuropsychopharmacology 24, 632–639. 30, 239–250. 281, 786–794. The association between disruptions in glucose metabolism and psychiatric disorders was first documented more than 3 centuries ago by the English doctor Thomas Willis. In the frontal cortex, there were no differences in Thrsp expression between the strains or between the PTU administration and control groups (Figure 5E), but in the hippocampus, the mRNA level of Thrsp was significantly higher in the WKY rats than in the Wistar rats (strain effect: F1,34 = 7.11, p = 0.011), and PTU decreased this parameter in the WKY rats (treatment effect: F1,34 = 7.07, p = 0.011) (Figure 5F). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. COVID-19 is an emerging, rapidly evolving situation. 2015, 1–7. Int. Neuronal expression of a thyroid hormone receptor α mutation alters mouse behaviour. The link between thyroid function and depression. It is difficult to explain that in contrast to our predictions, the administration of PTU did not lead to prolonged immobility time in the Porsolt test in either the Wistar or WKY rats. The brain â¦ doi: 10.1016/S0014-2999(97)01276-4, Lambert, H. P., Zenger, M., Azarias, G., Chatton, J. Y., Magistretti, P. J., and Lengacher, S. (2014). Aconitase activity was calculated from a standard curve and is expressed as mU/mg protein. The therapeutic effectiveness of adjunctive treatment of refractory depression with superphysiological doses of thyroid hormones is well established, but the mechanism of action of thyroid hormones on the function of the central nervous system in adults is not well known (Bauer et al., 2002, 2016). Ann. Total cholesterol and low-density lipoprotein cholesterol were significantly increased under hypothyroidism conditions in both rat strains; however, high-density lipoprotein cholesterol was elevated only in the WKY rats. Magnetic resonance imaging (MRI) studies in MA users have shown enlarged striatal volumes and positron emission tomography (PET) studies have shown decreased brain glucose metabolism (BGluM) in â¦ Epub 2012 Feb 10. After 1 h, the rats were placed in the same container in the presence of the familiar object and a novel object similar in size and height but different in shape and appearance. However, in the WKY rats receiving PTU, a simultaneous decrease in pyruvate dehydrogenase levels and pyruvate levels suggested the weakening of the Krebs cycle by limiting both its substrate and the enzyme linking glycolysis with the Krebs cycle. Horm. Another significant limitation of the current research is the use of only male rats, which did not allow us to determine the sex-related differences in the parameters tested. Mol. Cell 144, 810–823. 2019. PET measures emissions from radioactively labeled metabolically active chemicals that have been injected into â¦ doi: 10.1007/s00221-004-2027-6, Hage, M. P., and Azar, S. T. (2012). However, it should be taken into account that both impaired synaptic plasticity, which is considered crucial in the pathogenesis of depression, and hypothyroidism-induced disturbances in mitochondrial function can be caused by many factors, e.g., elevated glucocorticoid levels or a high-fat diet (HFD). Additionally, since the WKY rats are partially resistant to antidepressant drugs, these rats are considered to be a model of treatment-resistant depression. In the frontal cortex in the hypothyroidism models but not in the depression model, reduced levels of pyruvate may be caused by the increased conversion of pyruvate to lactate. Brain metabolic alterations in rats showing depression-like and obesity phenotypes. The effects of strain and PTU treatment on the total cholesterol (A), HDL (B), LDL (C), and triglyceride (D) levels measured in the plasma. For this reason, we compared the selected metabolic parameters in the WKY rats and the control Wistar rats under basal conditions and under conditions of decreased thyroid hormone synthesis (obtained by the administration of a thyroid peroxidase inhibitor). Moreover, thyroid hormone action also depends on the expression and sensitivity of thyroid hormone nuclear receptors, so thyroid hormone signaling in specific brain cells may be changed in depression. (2015). Hypothyroidism and depression: salient aspects of pathogenesis and management. In the frontal cortex, the level of PDK2 did not differ among any of the examined groups; however, in the hippocampus, a strain × PTU effect was observed (F1,30 = 6.04, p = 0.020). The effects of strain and PTU treatment on the level of Mitofusin 2, UCP4, VDAC1, and HK1 in the mitochondria-enriched fraction of frontal cortex and hippocampus. Epub 2018 Nov 5. Mutations in the PLA2G6 gene encoding this enzyme occur in patients with idiopathic neurodegeneration plus brain iron accumulation and dystonia-parkinsonism without iron accumulation, whereas mice lacking PLA2G6 show neurological dysfunction and neuropathology after 13 months. Aconitase activity was measured in mitochondria-enriched fractions derived from both examined brain structures using an Aconitase Activity Colorimetric Assay Kit (BioVision, Milpitas, CA, United States) according to the manufacturer’s instructions. Decreased thyroid hormone action was also shown to attenuate oxidative phosphorylation, and this change was greater in the WKY rats. 321, 18–27. (2008). The results are expressed as the mean ± SEM. Behav. How low-carbohydrate diets can improve brain metabolism If high blood sugar and insulin can jeopardize brain health through inflammation, oxidation, and insulin resistance, then we can hypothesize that reducing blood glucose and insulin levels could help improve brain health. Based on the findings, âmidlife may be a critical period for initiating treatments aimed at â¦ In addition, a mitochondrial respiration assay showed that PTU decreased ETS and OXPHOS coupling efficiency in the brain (ETS: F1,35 = 5.21 for the hippocampus, p = 0.0286; OXPHOS: treatment effect: F1,36 = 5.16 for the frontal cortex, p = 0.029; F1,35 = 5.17 for the hippocampus, p = 0.029), while the post hoc tests showed a significant decrease in ETS and OXPHOS coupling efficiency only in the hippocampus of the WKY rats after PTU administration and in OXPHOS coupling efficiency in the frontal cortex in the same group of animals (Figures 10F, 11E,F). The data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher. The predominant mechanism for the biological action of TH involves the regulation of gene transcription through nuclear TH receptors. There was no difference in the level of lactate in the cytosol in either the frontal cortex or hippocampus of the WKY and Wistar rats. There was no effect of PTU on RXRβ expression, and although ANOVA indicated differences between the strains (F1,36 = 4.79, p = 0.035), the post hoc test showed no significant differences between the Wistar and WKY rats or between those under the control conditions and those that received PTU (Figure 4H). After the age of 30, our â¦ Finally, rotenone (1 μM, E: ETS (CII) state) and antimycin A (2.5 μM) were added to inhibit complex I and complex II, respectively. In addition, these studies were conducted under â¦ Since a significant decrease in these parameters after PTU treatment was observed only in the WKY rats, it seems that although decreased coupling efficiency is mainly an effect of hypothyroidism, in the presence of hypothyroidism and depression, this decrease was more pronounced. The concentrations of the examined proteins in the samples were calculated from a standard curve and are expressed as ng/mg protein for pyruvate dehydrogenase and as pg/mg protein for PDK2 and PDK4. Neurosci., 04 December 2020
â¢Brain represents 2 to 3% of total body weight of an average adult, but utilizes 20 to 25% of the total Oxygen consumed by the whole organism; â¢In children up to 4 years of age, the brain utilizes about 50% of the total Oxygen consumed by the whole organism; Isoform-dependent actions of thyroid hormone nuclear receptors: lessons from knockin mutant mice. After a 30-min incubation, the absorbance was measured at a wavelength of λ = 570 nm using a spectrophotometer (Tecan Infinite M200 Pro, Männedorf, Switzerland). This is likely because the concentration of active thyroid hormone T3 in the brain depends not only on its synthesis in the thyroid gland but also, to a large extent, on the expression of transporters and deiodinase enzymes in particular brain structures (Flamant et al., 2015). This research was funded by Grant No. A brain is an organ that serves as the center of the nervous system in all vertebrate and most invertebrate animals. In the case of mitochondrial dysfunction, the intensification of fusion increases the production of ATP and is considered a neuroprotective mechanism (Martorell-Riera et al., 2014); therefore, the expression of the main fusion-mediating protein, mitofusin 2, was determined. The results are expressed as the average fold change ± SEM (A–D) or as the mean ± SEM. Then weâll talk about whether the metabolism can actually be damaged. 46/2018 of 01.02.2018). 37, 406–424. doi: 10.1016/j.biopsych.2007.11.020, Katyare, S. S., Bangur, C. S., and Howland, J. L. (1994). Psychiatry 21, 229–236. Endocrinol. The low efficacy of pharmacotherapy is probably because the antidepressant drugs currently in use mainly affect noradrenergic and serotonergic neurotransmission, while many studies have shown that this disease involves not only disturbances in neurotransmission but also changes in hormonal systems, immune systems, and energy processes that together impair synaptic plasticity, leading to numerous functional changes, including depressed mood (Horowitz and Zunszain, 2015). Paradoxical hormonal and behavioral responses to hypothyroid and hyperthyroid states in the Wistar-Kyoto rat. The reduction in UCP4 expression in the frontal cortex in the model of the co-occurrence of hypothyroidism and depression seems to be an adaptive mechanism caused by the weakening of oxidative phosphorylation in this brain structure. The results are expressed as the mean ± SEM. Disord. Similarly, in the rats receiving PTU, the expression of T3 receptors was increased in the hippocampus but not in the frontal cortex, probably as a mechanism to compensate for the lower concentration of this hormone. doi: 10.1002/cne.903220405, Martorell-Riera, A., Segarra-Mondejar, M., Muñoz, J. P., Ginet, V., Olloquequi, J., Pérez-Clausell, J., et al. Alterations in markers that determine the effect of thyroid hormones in the frontal cortex clearly indicated that in the studied model of depression, their function could be weakened (as shown by the decreases in T3, TRα1, and DIO2), while in the hippocampus, only a downward trend in the T3 level was observed, while the changes in TRβ1, RXRα, and DIO3 suggested the induction of compensatory mechanisms for enhancing T3 action. Existing studies have shown that WKY rats have lower levels of monoamines (5-HT, NE, and DA) in several brain structures, elevated levels of glutamate in the prefrontal cortex, and reduced concentrations of BDNF in the plasma (Aleksandrova et al., 2019). Pilhatsch, M., Schlagenhauf, F., Silverman, D., Berman, S., London, E. D., Martinez, D., et al. *p < 0.05 vs. the control group (Wistar rats); #p < 0.05 vs. the WKY group. 6:99. doi: 10.3389/fendo.2015.00099, Głombik, K., Detka, J., Góralska, J., Kurek, A., Solnica, B., and Budziszewska, B. U.S.A. 109, 5523–5528. J. Comp. *p < 0.05 vs. the control group (Wistar rats); #p < 0.05 vs. the WKY group. Proc. Psychiatry 7, 140–156. Behav. This test is based on the spontaneous tendency of rats to spend more time exploring a novel object than a familiar object. hprt1 (Thermo Fisher Scientific, Waltham, MA, United States) was used as the reference gene. A., Jensen, J. E., Fava, M., and Renshaw, P. F. (2008). Then, the uncoupling control was measured by the titration of the protonophore carbonylcyanide-4-(trifluoromethoxy)-phenylhydrazone (FCCP) (FCCP-uncoupled state, max ETS) (Pesta and Gnaiger, 2012; Sebastian et al., 2012). The covariance pattern of PSP consists of decreased metabolism in the prefrontal cortex, frontal eye fields, caudate nuclei, medial thalamus and upper brainstem (Eckert, et al.