Abstract
The influence of age, postmortem delay and freezing storage period on receptor-mediated G-protein activity was quantified in cortical membranes from 34 healthy subjects. Concentration-response curves of the [35S]GTPγS binding stimulation by agonists for α2-adrenoceptors (UK14304), μ-opioid (DAMGO), 5-HT1A (8-OH-DPAT), GABAB (baclofen) and muscarinic (carbachol) receptors were analyzed. Immunoreactivities of Gα-protein subunits were also determined. Basal binding and UK14304, 8-OH-DPAT, and baclofen potency to stimulate [35S]GTPγS binding decreased with aging (1–88 years) without changes of efficacy. DAMGO-mediated stimulation increased both in potency and efficacy with aging. A negative correlation between age and immunoreactivity was observed for Gαi1/2-, but not for Gαi3-, Gαo-,and Gαs-proteins. Neither [35S]GTPγS binding nor Gα-proteins changed with the postmortem delay (8–92 h). Basal [35S]GTPγS binding decreased with the sample storage period (1–85 months). A careful match between cases and controls should be taken into account when designing signal transduction studies in human disorders, specially for variables such as age and storage period.
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References
Arranz B, Eriksson A, Mellerup E, Plenge P, Marcusson J . (1993): Effect of aging in human cortical pre- and postsynaptic serotonin binding sites. Brain Res 620: 163–166
Birnbaumer L, Abramowitz J, Brown AM . (1990): Receptor-effector coupling by G proteins. Biochim Biophys Acta 1031: 163–224
Bradford MM . (1976): A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254
Burke RE, Greenbaum D . (1987): Effect of postmortem factors on muscarinic receptor subtypes in rat brain. J Neurochem 49: 592–596
Burnet PW, Eastwood SL, Harrison PJ . (1994): Detection and quantitation of 5-HT1A and 5-HT2A receptor mRNAs in human hippocampus using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique and their correlation with binding site densities and age. Neurosci Lett 178: 85–89
Busquets X, Ventayol P, Sastre M, García-Sevilla JA . (1996): Age-dependent increases in protein kinase C-αβ immunoreactivity and activity in the human brain: possible in vivo modulatory effects on guanine nucleotide Gi proteins. Brain Res 710: 28–34
Calne DB . (1994): Is idiopathic parkinsonism the consequence of an event or a process? Neurology 44: 5–10
Cerione RA, Regan JW, Nakata H, Codina J, Benovic JL, Gierschik P, Somers RL, Spiegel AM, Birnbaumer L, Lefkowitz RJ, Caron MG . (1986): Fuctional reconstitution of the α2-adrenergic receptor with guanine nucleotide regulatory proteins in phospholipid vesicles. J Biol Chem 261: 3901–3909
Cowburn RF, O'Neill C, Ravid R, Alafuzoff I, Winblad B, Fowler CJ . (1992): Adenylyl cyclase activity in postmortem human brain: evidence of altered G-protein mediation in Alzheimer's disease. J Neurochem 58: 1409–1419
Cowburn RF, Marcusson JO, Eriksson A, Wiehager B, O'Neill C . (1994): Adenylyl cyclase activity and G-protein subunit levels in postmortem frontal cortex of suicide victims. Brain Res 633: 297–304
Cowburn RF, Wiehager B, Ravid R, Winblad B . (1996): Acetylcholine muscarinic M2 receptor stimulated [35S]GTPγS binding shows regional selective changes in Alzheimer's disease postmortem brain. Neurodegeneration 5: 19–26
Cutler R, Joseph JA, Yamagami K, Villalobos-Molina R, Roth GS . (1994): Area specific alterations in muscarinic stimulated low Km GTPase activity in aging and Alzheimer's disease: implications for altered signal transduction. Brain Res 644: 54–60
Dewey SL, Volkow ND, Logan J, MacGregor RR, Fowler JS, Schyler DJ, Bendriem B . (1990): Age-related decreases in muscarinic cholinergic receptor binding in the human brain measured with positron emission tomography (PET). J Neurosci Res 27: 569–575
Dillon KA, Gross-Isseroff R, Israeli M, Biegon A . (1991): Autoradiographic analysis of serotonin 5-HT1A receptor binding in the human brain postmortem: effects of age and alcohol. Brain Res 554: 56–64
Dowlatshahi D, MacQueen GM, Wang J-F, Reiach JS, Young LT . (1999): G protein-coupled cyclic AMP signaling in postmortem brain of subjects with mood disorders: effects of diagnosis, suicide, and treatment at the time of death. J Neurochem 73: 1121–1126
Dupuis DS, Pauwels PJ, Radu D, Hall H . (1999): Autoradiographic studies of 5-HT1A-receptor-stimulated [35S]GTPγS-binding responses in the human and monkey brain. Eur J Neurosci 11: 1809–1817
Elliot J, Reynolds GP . (1999): Agonist-stimulated GTPγ[35S]binding to 5-HT1A receptors in human post-mortem brain. Eur J Pharmacol 386: 313–315
Escribá PV, Sastre M, García-Sevilla JA . (1994): Increased density of guanine nucleotide-binding proteins in the postmortem brains of heroin addicts. Arch Gen Psychiatry 51: 494–501
Florio VA, Sternweis PC . (1989): Mechanisms of muscarinic receptor action on Go in reconstituted phospholipid vesicles. J Biol Chem 264: 3909–3915
Friedman E, Wang H-Y . (1996): Receptor-mediated activation of G proteins is increased in postmortem brains of bipolar affective disordes subjects. J Neurochem 67: 1145–1152
Fülöp T Jr, Seres I . (1994): Age-related changes in signal transduction. Implications for neuronal transmission and potential for drug intervetion. Drugs Aging 5: 366–390
Gabilondo AM, Meana JJ, García-Sevilla JA . (1995): Increased density of μ-opioid receptors in the postmortem brain of suicide victims. Brain Res 682: 245–250
Gessi S, Campi F, Varani K, Borea PA . (1999): α2-Adrenergic agonist modulation of [35S]GTPγS binding to guanine-nucleotide-binding-proteins in human platelet membranes. Life Sci 64: 1403–1413
Giacobini E, DeSarno P, Clark B, Mcllhany M . (1989): The cholinergic receptor system of the human brain: neurochemical and pharmacological aspects in aging and Alzheimer. Prog Brain Res 79: 335–343
Giacobini E . (1990): Cholinergic receptors in human brain: effects of aging and Alzheimer disease. J Neurosci Res 27: 548–560
Gilman GA . (1987): G proteins: transducers of receptor-generated signals. Annu Rev Biochem 56: 615–649
Greenwood AF, Powers RE, Jope RS . (1995): Phosphoinositide hydrolysis, Gαq, phospholipase C, and protein kinase C in post mortem human brain: effects of post mortem interval, subject age and Alzheimer's disease. Neuroscience 69: 125–138
González-Maeso J, Rodríguez-Puertas R, Gabilondo AM, Meana JJ . (2000): Characterization of receptor-mediated [35S]GTPγS binding to cortical membranes from postmortem human brain. Eur J Pharmacol 390: 25–36
Grijalba B, Callado LF, Meana JJ, García-Sevilla JA, Pazos A . (1996): α2-Adrenoceptor subtypes in the human brain: a pharmacological delineation of [3H]RX-821002 binding to membranes and tissue sections. Eur J Pharmacol 310: 83–93
Gudermann T, Schöneberg T, Schultz G . (1997): Functional and structural complexity of signal transduction via G-protein-coupled receptors. Annu Rev Neurosci 20: 399–427
Happe HK, Bylund DB, Murrin LC . (1999): Alpha-2 adrenergic receptor functional coupling to G proteins in rat brain during postnatal development. J Pharmacol Exp Ther 288: 1134–1142
Hilf G, Gierschik P, Jakobs KH . (1989): Muscarinic acetylcholine receptor-stimulated binding of guanosine 5′-O-(3-thiotriphosphate) to guanine-nucleotide-binding proteins in cardiac membranes. Eur J Biochem 186: 725–731
Jope RS, Song L, Grimes CA, Pacheco MA, Dilley GE, Li X, Meltzer HY, Overholser JC, Stockmeier CA . (1998): Selective increases in phosphoinositide signaling activity and G protein levels in postmortem brain from subjects with schizophrenia or alcohol dependence. J Neurochem 70: 763–771
Lazareno S, Birdsall NJM . (1993): Pharmacological characterization of acetylcholine-stimulated [35S]-GTPγS binding mediated by human muscarinic m1-m4 receptors: antagonists studies. Br J Pharmacol 109: 1120–1127
Lee KS, Frey KA, Koeppe RA, Buck A, Mulholland GK, Kuhl DE . (1996): In vivo quantification of cerebral muscarinic receptors in normal human aging using positron emission tomography and [11C]tropanyl benzilate. J Cereb Blood Flow Metab 16: 303–310
Li X, Greenwood AF, Powers R, Jope RS . (1996): Effects of postmortem interval, age, and Alzheimer's disease on G-proteins in human brain. Neurobiol Aging 17: 115–122
Manji HK . (1992): G proteins: implications for psychiatry. Am J Psychiatry 149: 746–760
Mann DMA, Yates PO, Hawks J . (1983): Pathology of the human locus coeruleus. Clin Neuropathol 2: 1–7
Meana JJ, Barturen F, Garro MA, García-Sevilla JA, Fontán A, Zarranz JJ . (1992): Decreased density of presynaptic α2-adrenoceptors in postmortem brains of patients with Alzheimer's disease. J Neurochem 58: 1896–1904
Meana JJ, González-Maeso J, García-Sevilla JA, Guimón J . (2000): μ-Opioid receptor and α2-adrenoceptor agonist stimulation of [35S]GTPγS to G-proteins in postmortem brains of opioid addicts. Mol Psychiatry 5: 308–315
Milligan G . (1988): Techniques used in the identification and analysis of function of pertussis toxin-sensitive guanine nucleotide binding proteins. Biochem J 255: 1–13
Newman-Tancredi A, Conte C, Chaput C, Verrièle L, Millan MJ . (1997): Agonist and inverse agonist efficacy at human recombinant serotonin 5-HT1A receptors as function of receptor: G-protein stoichiometry. Neuropharmacology 36: 451–459
Nordberg A, Alafuzoff I, Winblad B . (1992): Nicotinic and muscarinic subtypes in the human brain: changes with aging and dementia. J Neurosci Res 31: 103–111
Odagaki Y, Nishi N, Ozawa H, Saito T, Takahata N, Riederer P, Koyama T . (1998): Measurement of receptor-mediated functional activation of G proteins in postmortem human brain membranes. Brain Res 789: 84–91
Olianas MC, Onali P . (1996): Stimulation of guanosine 5′-O-(3-[35S]thiotriphosphate) binding by cholinergic muscarinic receptors in membranes of rat olfactory bulb. J Neurochem 67: 2549–2556
Ozawa H, Ukai W, Kornhuber J, Yamaguchi T, Froelich L, Ikeda H, Saito T, Riederer P . (1999): Postnatal ontogeny of GTP binding protein in the human frontal cortex. Life Sci 65: 2315–2323
Pacheco MA, Stockmeier C, Meltzer HY, Overholser JC, Dilley GE, Jope RS . (1996): Alterations in phosphoinositide signalling and G-protein levels in depressed suicide brain. Brain Res 723: 37–45
Palego L, Giromella A, Marazziti D, Borsini F, Naccarato AG, Giannaccini G, Lucacchini A, Cassano GB, Mazzoni MR . (1999): Effects of postmortem delay on serotonin and (+)8-OH-DPAT-mediated inhibition of adenylyl cyclase activity in rat and human tissues. Brain Res 816: 165–174
Pascual J, del Arco C, González AM, Díaz A, del Olmo E, Pazos A . (1991): Regionally specific age-dependent decline in α2-adrenoceptors: an autoradiographic study in human brain. Neurosci Lett 133: 279–283
Paul D, Gauthier CA, Minor LD, Gonzales GR . (1997): The effects of postmortem delay on mu, delta and kappa opioid receptor subtypes in rat brain and guinea pig cerebellum evaluated by radioligand receptor binding. Life Sci 61: 1993–1998
Peltonen JM, Pihlavisto M, Scheinin M . (1998): Subtype-specific stimulation of [35S]GTPγS binding by recombinant α2-adrenoceptors. Eur J Pharmacol 355: 275–279
Perry EK, Perry RH . (1983): Human brain neurochemistry. Some postmortem problems. Life Sci 33: 1733–1743
Rinne JO . (1987): Muscarinic and dopaminergic receptors in the aging human brain. Brain Res 404: 162–168
Rodríguez-Puertas R, Pascual J, Pazos A . (1996): Effects of freezing storage time on the density of muscarinic receptors in the human postmortem brain: an autoradiographic study in control and Alzheimer's disease brain tissues. Brain Res 728: 65–71
Rodríguez-Puertas R, González-Maeso J, Meana JJ, Pazos A . (2000): Autoradiography of receptor-activated G-proteins in postmortem human brain. Neuroscience 96: 169–180
Sastre M, García-Sevilla JA . (1994): Density of alpha-2 adrenoceptors and Gi proteins in the human brain: ratio of high-affinity agonist sites to antagonist sites and effect of age. J Pharmacol Exp Ther 269: 1062–1072
Sastre M, Guimón J, García-Sevilla JA . (2001): Relationship between β- and α2-adrenoceptors and G coupling proteins in the human brain: Effects of age and suicide. Brain Res 898: 242–255
Scherman D, Desnos C, Darchen F, Pollak P, Javoy-Agid F, Agid Y . (1989): Striatal dopamine deficiency in Parkinson's disease: role of aging. Ann Neurol 26: 551–557
Sim LJ, Selley DE, Childers SR . (1995): In vitro autoradiography of receptor-activated G-proteins in rat brain by agonist-stimulated guanylyl 5′-[γ-[35S]thio]-triphosphate binding. Proc Natl Acad Sci USA 92: 7242–7246
Simon MI, Strathmann MP, Gautam N . (1991): Diversity of G proteins in signal transduction. Science 252: 802–808
Spiegel AM . (1996): Defects in G protein-coupled signal transduction in human disease. Annu Rev Physiol 58: 143–170
Turgeon SM, Albin RL . (1994): Postnatal ontogeny of GABAB binding in rat brain. Neuroscience 62: 601–613
Vijayashankar N, Brody H . (1979): A quantitative study of the pigmented neurons in the nuclei locus coeruleus and subcoeruleus in man as related to aging. J Comp Neurol 38: 490–497
Wang H-Y, Friedman E . (1994): Receptor-mediated activation of G proteins is reduced in postmortem brains from Alzheimer's disease patients. Neurosci Lett 173: 37–39
Whitehouse PJ, Lynch D, Kuhar MJ . (1984): Effects of postmortem delay and temperature on neurotransmitter receptor binding in a rat model of the human autopsy process. J Neurochem 43: 553–559
Young LT, Warsh JJ, Li PP, Siu KP, Becker L, Gilbert J, Hornykiewicz O, Kish SJ . (1991): Maturational and aging effects on guanine nucleotide binding protein immunoreactivity in human brain. Dev Brain Res 61: 243–248
Young LT, Li PP, Kish SJ, Siu KP, Kamble A, Hornykiewicz O, Warsh JJ . (1993): Cerebral cortex Gsα protein levels and forskolin-stimulated cyclic AMP formation are increased in bipolar affective disorder. J Neurochem 61: 890–898
Zubieta JK, Dannals RF, Frost JJ . (1999): Gender and age related influences of human brain mu-opioid receptor binding measured by PET. Am J Psychiatry 156: 842–848
Acknowledgements
This study was supported by grants from FIS (01/0358), the Basque Government (PI98/8) and the University of the Basque Country (G13/98), Spain to JJM, and from FNSRS (32.57066.99), Switzerland to JAG-S. JG-M was recipient of a predoctoral fellowship from MEC, Spain. The authors wish to thank the staff members of the Basque Institute of Legal Medicine, Bilbao, and the Department of Forensic Medicine, Geneva, for their cooperation in the study. JAG-S is a member of the Institut d'Estudis Catalans (Barcelona, Spain).
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González-Maeso, J., Torre, I., Rodríguez-Puertas, R. et al. Effects of Age, Postmortem Delay and Storage Time on Receptor-mediated Activation of G-proteins in Human Brain. Neuropsychopharmacol 26, 468–478 (2002). https://doi.org/10.1016/S0893-133X(01)00342-6
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DOI: https://doi.org/10.1016/S0893-133X(01)00342-6
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