Abstract
Clonidine is an anti-hypertensive drug that inhibits the release of norepinephrine from pre-synaptic terminals binding to pre-synaptic α2-adrenoreceptors. Some studies suggest that this drug decreases brain energy expenditure, particularly in hypoxic-ischemic injury. However, data about clonidine effects on the functional parameters regulating brain energy metabolism are lacking. In this study, the effects of acute clonidine treatment (5 μg×kg−1 i.p., 30 min) were evaluated on the catalytic activity of regulatory energy-linked enzymes of Krebs’ cycle, Electron Transport Chain and glutamate metabolism of temporal cerebral cortex of 3-month-old male Sprague–Dawley rats. Enzyme activities were assayed on non-synaptic “free” mitochondria (FM) of neuronal perikaryon and partly of glial cells, and on intra-synaptic “light” (LM) and “heavy” mitochondria (HM), localized within synaptic terminals. This subcellular analysis differentiates clonidine effects on post-synaptic and pre-synaptic neuronal compartments. The results showed that clonidine increased citrate synthase, cytochrome oxidase and glutamate–oxaloacetate transaminase activities of FM. In LM, citrate synthase activity was decreased, while cytochrome oxidase and glutamate–oxaloacetate transaminase activities were increased; on the contrary, citrate synthase, cytochrome oxidase and glutamate dehydrogenase were all decreased in HM. Therefore, clonidine exerted different effects with respect to brain mitochondria, coherently with the in vivo energy requirements of each synaptic compartment: the drug increased energy-linked enzyme activities in post-synaptic compartment, while the metabolic variations were complex in the pre-synaptic one, being enzyme activities heterogeneously modified in LM and decreased in HM. This study highlights the relationships existing between the clonidine-induced neuroreceptorial effects and the energy metabolism in pre- and post- synaptic bioenergetics.
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The data that support the findings of this study are available from the corresponding author upon reasonable request.
References
Wang X, Li G, Abdel-Rahman AA (2005) Site-dependent inhibition of neuronal c-jun in the brainstem elicited by imidazoline I1 receptor activation: role in rilmenidine-evoked hypotension. Eur J Pharmacol 514:191–199
Huwyler J, Fricker G, Török M, Schneider M, Drewe J (1997) Transport of clonidine across cultured brain microvessel endothelial cells. J Pharmacol Exp Ther 282:81–85
Hall DL, Tatakis DN, Walters JD, Rezvan E (2006) Oral clonidine pre-treatment and diazepam/meperidine sedation. J Dent Res 85:854–858
Schneider JS, Tinker JP, Decamp E (2010) Clonidine improves attentional and memory components of delayed response performance in a model of early Parkinsonism. Behav Brain Res 211:236–239
Bardgett ME, Points M, Ramsey-Faulkner C, Topmiller J, Roflow J, McDaniel T, Lamontagne T, Griffith MS (2008) The effects of clonidine on discrete-trial delayed spatial alternation in two rat models of memory loss. Neuropsychopharmacology 33:1980–1991
Gupta S, Sharma B (2014) Pharmacological modulation of I1-imidazoline and α2-adrenoceptors in sub-acute brain ischemia induced vascular dementia. Eur J Pharmacol 723:80–90
Jellish WS, Murdoch J, Kindel G, Zhang X, White FA (2005) The effect of clonidine on cell survival, glutamate, and aspartate release in normo- and hyperglycemic rats after near complete forebrain ischemia. Exp Brain Res 167:526–534
Milot MR, Plamondon H (2011) Changes in HPA reactivity and noradrenergic functions regulate spatial memory impairments at delayed time intervals following cerebral ischemia. Horm Behav 59:594–604
Zhang Y (2004) Clonidine preconditioning decreases infarct size and improves neurological outcome from transient forebrain ischemia in the rat. Neuroscience 125:625–631
Chao HM, Chidlow G, Melena J, Wood JP, Osborne NN (2000) An investigation into the potential mechanisms underlying the neuroprotective effect of clonidine in the retina. Brain Res 877:47–57
Gorini A, Villa RF (2001) Effect of in vivo treatment of clonidine on ATP-ase’s enzyme systems of synaptic plasma membranes from rat cerebral cortex. Neurochem Res 26:821–827
Jiménez-Rivera CA, Figueroa J, Vázquez-Torres R, Vélez-Hernandez ME, Schwarz D, Velásquez-Martinez MC, Arencibia-Albite F (2012) Presynaptic inhibition of glutamate transmission by α2 receptors in the VTA. Eur J Neurosci 35:1406–1415
Karmen NB (2003) Oxidative modification of erythrocyte membranes in the acute stage of severe craniocerebral trauma and its correction with clonidine. Bull Exp Biol Med 136:362–365
Moretti A, Ferrari F, Villa RF (2015) Neuroprotection for ischaemic stroke: current status and challenges. Pharmacol Ther 146:23–34
Villa RF, Gorini A, Ferrari F, Hoyer S (2013) Energy metabolism of cerebral mitochondria during aging, ischemia and post-ischemic recovery assessed by functional proteomics of enzymes. Neurochem Int 63:765–781
Hoffman WE, Cheng MA, Thomas C, Baughman VL, Albrecht RF (1991) Clonidine decreases plasma catecholamines and improves outcome from incomplete ischemia in the rat. Anesth Analg 73:460–464
Schmidt K, Philipsenburg C, Zivkovic A, Hofer S (2013) Effects of clonidine on microcirculatory alterations during endotoxemia. Crit Care 17(S2):P388
Ståhl N, Ungerstedt U, Nordström CH (2001) Brain energy metabolism during controlled reduction of cerebral perfusion pressure in severe head injuries. Intensive Care Med 27:1215–1223
Taittonen M, Kirvelä O, Aantaa R, Kanto J (1997) Cardiovascular and metabolic responses to clonidine and midazolam premedication. Eur J Anaesthesiol 14:190–196
Quintin L, Viale JP, Annat G, Hoen JP, Butin E, Cottet-Emard JM, Levron JC, Bussery D, Motin J (1991) Oxygen uptake after major abdominal surgery: effect of clonidine. Anesthesiology 74:236–241
Villa RF, Ferrari F, Gorini A (2013) Functional proteomics related to energy metabolism of synaptosomes from different neuronal systems of rat hippocampus during aging. J Proteome Res 12:5422–5435
Battino M, Quiles JL, Huertas JR, Mataix JF, Villa RF, Gorini A (2000) Cerebral cortex synaptic heavy mitochondria may represent the oldest synaptic mitochondrial population: biochemical heterogeneity and effects of L-acetylcarnitine. J Bioenerg Biomembr 32:163–173
Battino M, Ferreiro MS, Littarru G, Quiles JL, Ramírez-Tortosa MC, Huertas JR, Mataix J, Villa RF, Gorini A (2002) Structural damage induced by peroxidation may account for functional impairment of heavy synaptic mitochondria. Free Radic Res 36:479–484
Lai JCK, Walsh JM, Dennis SC, Clark JB (1977) Synaptic and non-synaptic mitochondria from rat brain: isolation and characterization. J Neurochem 28:625–631
Villa RF, Gorini A, Hoyer S (2006) Differentiated effect of ageing on the enzymes of Krebs’ cycle, electron transfer complexes and glutamate metabolism of non-synaptic and intra-synaptic mitochondria from cerebral cortex. J Neural Transm 113:1659–1570
Villa RF, Gorini A, Hoyer S (2009) Effect of ageing and ischemia on enzymatic activities linked to Krebs’ cycle, electron transfer chain, glutamate and aminoacids metabolism of free and intrasynaptic mitochondria of cerebral cortex. Neurochem Res 34:2102–2116
Villa RF, Ferrari F, Gorini A (2012) Effect of CDP-choline on age-dependent modifications of energy- and glutamate-linked enzyme activities in synaptic and non-synaptic mitochondria from rat cerebral cortex. Neurochem Int 61:1424–1432
Villa RF, Ferrari F, Gorini A, Brunello N, Tascedda F (2016) Effect of desipramine and fluoxetine on energy metabolism of cerebral mitochondria. Neuroscience 330:326–334
Villa RF, Ferrari F, Bagini L, Gorini A, Brunello N, Tascedda F (2017) Mitochondrial energy metabolism of rat hippocampus after treatment with the antidepressants desipramine and fluoxetine. Neuropharmacology 121:30–38
Fisher RA, Yates F (1963) Statistical tables for biological, agricultural and medical research. Edinburgh, Oliver and Boyd
Benzi G, Gorini A, Ghigini B, Arnaboldi R, Villa RF (1994) Modifications by hypoxia and drug treatment of cerebral ATPase plasticity. Neurochem Res 19:517–524
Conway EL, Jarrott B (1982) Tissue pharmacokinetics of clonidine in rats. J Pharmacokinet Biopharm 10:187–200
Paalzow LK, Edlund PO (1979) Pharmacokinetics of clonidine in the rat and cat. J Pharmacokinet Biopharm 7:481–494
Claessens AJ, Risler LJ, Eyal S, Shen DD, Easterling TR, Hebert MF (2010) CYP2D6 mediates 4-hydroxylation of clonidine in vitro: implication for pregnancy-induced changes in clonidine clearance. Drug Metab Dispos 38:1393–1396
Khan ZP, Ferguson CN, Jones RM (1999) alpha-2 and imidazoline receptor agonists their pharmacology and therapeutic role. Anaesthesia 54:146–165
Villa RF, Gorini A, LoFaro A, Dell’Orbo C (1989) A critique on the preparation and enzymatic characterization of synaptic and non-synaptic mitochondria from hippocampus. Cell Mol Neurobiol 9:247–262
Glowinsky J, Iversen LL (1966) Regional studies of catecholamines in the rat brain - I. The disposition of 3H-norepinephrine, 3H-dopamine and 3H-DOPA in various regions of the brain. J Neurochem 13:655–669
Alosco ML, Gunstad J, Xu X, Clark US, Labbe DR, Riskin-Jones HH, Terrero G, Schwarz NF, Walsh EG, Poppas A, Cohen RA, Sweet LH (2014) The impact of hypertension on cerebral perfusion and cortical thickness in older adults. J Am Soc Hypertens 8:561–570
Jennings JR, Mendelson DN, Muldoon MF, Ryan CM, Gianaros PJ, Raz N, Aizenstein H (2012) Regional grey matter shrinks in hypertensive individuals despite successful lowering of blood pressure. J Hum Hypertens 26:295–305
Sugden PH, Newsholme EA (1975) Activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenase, glutamate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in nervous tissues from vertebrates and invertebrates. Biochem J 150:105–111
Ackrell BAC, Kearney EB, Singer TP (1978) Mammalian succinate dehydrogenase. In: Estabrook RW, Pullman ME (eds) Methods in Enzymology, vol 23. Academic Press. New York, London, pp 446–483
Battino M, Bertoli E, Formaggini G, Sassi S, Gorini A, Villa RF, Lenaz G (1991) Structural and functional aspects of the respiratory chain of synaptic and nonsynaptic mitochondria derived from selected brain regions. J Bioenerg Biomembr 23:245–263
Smith L (1955) Spectrophotometric assay of cytochrome c oxidase. In: Glick D (ed) Methods of Biochemical Analysis, vol 2. Wiley-Interscience, New York, pp 427–434
Smith L, Davies HC, Nava M (1974) Oxidation and reduction of soluble cytochrome c by membrane-bound oxidase and reductase system. J Biol Chem 249:2904–1910
Wharton DC, Tzagoloff A (1967) Cytochrome oxidase from beef heart mitochondria. In: Estabrook RW, Pullman ME (eds) Methods in Enzymology, vol 10. Academic Press. New York, London, pp 245–250
Wong-Riley MT (1989) Cytochrome oxidase: an endogenous metabolic marker for neuronal activity. Trends Neurosci 12:94–101
Bergmeyer HU, Bernt E (1974) Glutamate-pyruvate transaminase: UV-assay, manual method. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 752–758
Cooper AJ, Jeitner TM (2016) Central role of glutamate metabolism in the maintenance of nitrogen homeostasis in normal and hyperammonemic brain. Biomolecules 6:16
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Shapiro S, Wilk MB (1965) An analysis of variance test for normality (complete samples). Biometrika 52:591–611
Kang CW, Kvam PH (2012) Shewhart Control Charts. Basic statistical tools for improving quality. John Wiley & Sons, New York, pp 97–124
Ferrari F, Gorini A, Hoyer S, Villa RF (2018) Glutamate metabolism in cerebral mitochondria after ischemia and post-ischemic recovery during aging: relationships with brain energy metabolism. J Neurochem 146:416–428
Wong-Riley MT (2012) Bigenomic regulation of cytochrome c oxidase in neurons and the tight coupling between neuronal activity and energy metabolism. Adv Exp Med Biol 748:283–304
Smith TJ, Stanley CA (2008) Untangling the glutamate dehydrogenase allosteric nightmare. Trends Biochem Sci 33:557–564
Dienel GA (2019) Brain glucose metabolism: integration of energetics with function. Physiol Rev 99:949–1045
Andersen JV, Markussen KH, Jakobsen E, Schousboe A, Waagepetersen HS, Rosenberg PA, Aldana BI (2021) Glutamate metabolism and recycling at the excitatory synapse in health and neurodegeneration. Neuropharmacology 196:108719
Hohnholt MC, Andersen VH, Andersen JV, Christensen SK, Karaca M, Maechler P, Waagepetersen HS (2018) Glutamate dehydrogenase is essential to sustain neuronal oxidative energy metabolism during stimulation. J Cerebr Blood Flow Metabol 38:1754–1768
Andersen JV, Jakobsen E, Waagepetersen HS, Aldana BI (2019) Distinct differences in rates of oxygen consumption and ATP synthesis of regionally isolated non-synaptic mouse brain mitochondria. J Neurosci Res 97:961–974
Lindblom P, Rafter I, Copley C, Andersson U, Hedberg JJ, Berg AL, Samuelsson A, Hellmold H, Cotgreave I, Glinghammar B (2007) Isoforms of alanine aminotransferases in human tissues and serum-differential tissue expression using novel antibodies. Arch Biochem Biophys 466:66–77
Ouyang Q, Nakayama T, Baytas O, Davidson SM, Yang C, Schmidt M, Lizarraga SB, Mishra S, Ei-Quessny M, Niaz S, Gul Butt M, Imran Murtaza S, Javed A, Chaudhry HR, Vaughan DJ, Hill RS, Partlow JN, Yoo SY, Lam AT, Nasir R, Al-Saffar M, Barkovich AJ, Schwede M, Nagpal S, Rajab A, DeBerardinis RJ, Housman DE, Mochida GH, Morrow EM (2016) Mutations in mitochondrial enzyme GPT2 cause metabolic dysfunction and neurological disease with developmental and progressive features. Proc Natl Acad Sci USA 113:E5598–E5607
Weinberg FF, Hamanaka R, Wheaton WW, Weinberg S, Joseph J, Lopez M, Kalyanaraman B, Mutlu GM, Budinger GR, Chandel NS (2010) Mitochondrial metabolism and ROS generation are essential for Kras-mediated tumorigenicity. Proc Natl Acad Sci USA 107:8788–8793
Attwell D, Laughlin SB (2001) An energy budget for signaling in the grey matter of the brain. J Cereb Blood Flow Metab 21:1133–1145
Rangaraju V, Calloway N, Ryan TA (2014) Activity-driven local ATP synthesis is required for synaptic function. Cell 156:825–835
Harris JJ, Jolivet R, Attwell D (2012) Synaptic energy use and supply. Neuron 75:762–777
Villa RF, Gorini A, Hoyer S (2002) ATPases of synaptic plasma membranes from hippocampus after ischemia and recovery during ageing. Neurochem Res 27:861–870
Nicholls DG (2004) Mitochondrial membrane potential and aging. Aging Cell 3:35–40
Saxena U (2012) Bioenergetics failure in neurodegenerative diseases: back to the future. Expert Opin Ther Targets 16:351–354
Small SA, Schobel SA, Buxton RB, Witter MP, Barnes CA (2011) A pathophysiological framework of hippocampal dysfunction in ageing and disease. Nat Rev Neurosci 12:585–601
Villa RF, Ferrari F, Gorini A (2012) Energy metabolism of rat cerebral cortex, hypothalamus and hypophysis during ageing. Neuroscience 227:55–66
Gorini A, D’Angelo A, Villa RF (1998) Action of L-acetylcarnitine on different cerebral mitochondrial populations from cerebral cortex. Neurochem Res 23:1485–1491
Dennis SC, Clark JB (1977) The pathway of glutamate metabolism in rat brain mitochondria. Biochem J 168:521–527
McKenna MC, Stridh MH, McNair LF, Sonnewald U, Waagepetersen HS, Schousboe A (2016) Glutamate oxidation in astrocytes: roles of glutamate dehydrogenase and aminotransferases. J Neurosci Res 94:1561–1571
Schousboe A, Scafidi S, Bak LK, Waagepetersen HS, McKenna MC (2014) Glutamate metabolism in the brain focusing on astrocytes. Adv Neurobiol 11:13–30
Gorini A, Villa RF (1983) Action of dihydroergocristine on enzyme activities related to energy transduction. II Farmaco 38:191–199
Borst P (2020) The malate-aspartate shuttle (Borst cycle): How it started and developed into a major metabolic pathway. IUBMB Life 72:2241–2259
Fonnum F (1968) The distribution of glutamate decarboxylase and aspartate transaminase in subcellular fractions of rat and Guinea-pig brain. Biochem J 106:401–412
Nik Yusoff NS, Mustapha Z, Govindasamy C, Sirajudeen KN (2013) Effect of Clonidine (an antihypertensive drug) treatment on oxidative stress markers in the heart of spontaneously hypertensive rats. Oxid Med Cell Longev 2013:927214
Filos KS, Panteli ES, Fligou F, Papamichail C, Papapostolou I, Zervoudakis G, Spiliopoulou I, Georgiou C (2012) Clonidine pre-treatment prevents hemorrhagic shock-induced endotoxemia and oxidative stress in the gut, liver, and lungs of the rat. Redox Rep 17:246–251
Fedorovich SV, Waseem TV, Puchkova LV (2017) Biogenetic and morphofunctional heterogeneity of mitochondria: the case of synaptic mitochondria. Rev Neurosci 28:363–373
Naga KK, Sullivan PG, Geddes JW (2007) High cyclophilin D content of synaptic mitochondria results in increased vulnerability to permeability transition. J Neurosci 27:7469–7475
Hill RL, Kulbe JR, Singh IN, Wang JA, Hall ED (2018) Synaptic mitochondria are more susceptible to traumatic brain injury-induced oxidative damage and respiratory dysfunction than non-synaptic mitochondria. Neuroscience 386:265–283
Gurd JW, Jones IR, Mahler HR, Moore WJ (1974) Isolation and partial characterization of rat brain synaptic plasma membranes. J Neurochem 22:281–290
Gorini A, Canosi U, Devecchi E, Geroldi D, Villa RF (2002) ATPases enzyme activities during ageing in different types of somatic and synaptic plasma membranes from rat frontal cerebral cortex. Prog Neuropsychopharmacol Biol Psychiatry 26:81–90
Villa RF, Arnaboldi R, Gorini A, Geroldi D (1989) Action of piracetam and clonidine on different mitochondrial populations from hippocampus Il. Farmaco 44:215–226
Lakshminarayan K, Anderson DC, Borbas C, Duval S, Luepker RV (2007) Blood pressure management in acute ischemic stroke. J Clin Hypertens 9:444–453
Burns TW, Langley PE, Terry BE, Bylund DB, Forte LR (1982) Alpha-2 adrenergic activation inhibits forskolin-stimulated adenylate cyclase activity and lipolysis in human adipocytes. Life Sci 31:815–821
Okada M, Mine K, Fujiwara M (1989) Relationship of calcium and adenylate cyclase messenger systems in rat brain synaptosomes. Brain Res 501:23–31
De Langen CDJ, Mulder AH (1980) On the role of calcium ions in the presynaptic alpha-receptors mediated inhibition of [3H]noradrenaline release from rat brain cortex synaptosomes. Brain Res 185:399–408
Adamson P, Brammer MJ, Campbell IC (1988) Cyclic AMP analogues potentiate k-opiate and α2-adrenoceptor agonist effects on intrasynaptosomal free calcium. J Neurochem 51:542–547
Adamson P, McWilliam JR, Campbell IC (1988) Mutual antagonism of κ-opiate and α2-adrenoceptor agonist effect on intra-synaptosomal free [Ca2+]i. J Neurochem 50:65–68
Ouyang H, Bai X, Huang W, Chen D, Dohi S, Zeng W (2012) The antinociceptive activity of intrathecally administered amiloride and its interactions with morphine and clonidine in rats. J Pain 13:41–48
Denton RM (2009) Regulation of mitochondrial dehydrogenases by calcium ions. Biochim Biophys Acta 1787:1309–1316
Nicholls DG (2017) Brain mitochondrial calcium transport: Origins of the set-point concept and its application to physiology and pathology. Neurochem Int 109:5–12
Madison DV, Nicoll RA (1986) Actions of noradrenaline recorded intracellulary in rat hippocampal pyramidal neurons, in vitro. J Physiol 372:221–244
Dodt HU, Pawelzik H, Zieglgansberger W (1991) Actions of noradrenaline on neocortical neurons in vitro. Brain Res 545:307–311
Mori-Okamoto J, Namii Y, Tatsuno J (1991) Subtypes of adrenergic receptors and intracellular mechanism involved in modulatory effects of noradrenaline on glutamate. Brain Res 539:67–75
Kamisaki Y, Hamahashi T, Hamada T, Maeda K, Itoh T (1992) Presynaptic inhibition by clonidine of neurotransmitter aminoacid release in various brain regions. Eur J Pharmacol 217:57–63
Ferrari F, Gorini A, Villa RF (2015) Functional proteomics of synaptic plasma membrane ATP-ases of rat hippocampus: effect of L-acetylcarnitine and relationships with dementia and depression pathophysiology. Eur J Pharmacol 756:67–74
Ferrari F, Viscardi P, Gorini A, Villa RF (2019) Synaptic ATPases system of rat frontal cerebral cortex during aging. Neurosci Lett 694:74–79
Villa RF, Ferrari F, Gorini A (2011) Effect of in vivo L-acetylcarnitine administration on ATP-ases enzyme systems of synaptic plasma membranes from rat cerebral cortex. Neurochem Res 36:1372–1382
Villa RF, Ferrari F, Gorini A (2013) ATP-ases of synaptic plasma membranes in striatum: enzymatic systems for synapses functionality by in vivo administration of L-acetylcarnitine in relation to Parkinson’s Disease. Neuroscience 248:414–426
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This Study was supported by: Ministero dell'Università e della Ricerca (MUR), Rome, Italy; Catholic Universitary Center (Centro Universitario Cattolico) of the “Conferenza Episcopale Italiana” (CEI), Rome, Italy.
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Conceptualization: RFV; Methodology: RFV and AG; Formal analysis and investigation: AG and FF; Writing—original draft preparation: FF; Writing—review and editing: FF and RFV; Supervision: RFV.
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Villa, R.F., Gorini, A. & Ferrari, F. Clonidine and Brain Mitochondrial Energy Metabolism: Pharmacodynamic Insights Beyond Receptorial Effects. Neurochem Res 47, 1429–1441 (2022). https://doi.org/10.1007/s11064-022-03541-z
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DOI: https://doi.org/10.1007/s11064-022-03541-z