Abstract
The microdialysis technique for the measurement of brain extracellular levels of monoamines has become very popular over the last 2–3 decades, particularly in laboratories involved in neuropharmacology studies. Nevertheless, microdialysis of monoamines is a challenging technique for several reasons and to get reliable results it is necessary to develop sufficient skill and be ready to solve technical and analytical problems that may occur. This chapter is intended to help obtain reliable and reproducible results by describing in detail practical aspects of probe construction and implantation into the rat brain and measurement of noradrenaline, dopamine, and serotonin with HPLC coupled to electrochemical detection, set up, optimized, and validated in this author’s laboratory.
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References
Dahlstroem A, Fuxe K (1964) Evidence for the existence of monoamine-containing neurons in the central nervous system. I. Demonstration of monoamines in the cell bodies of brain stem neurons. Acta Physiol Scand 62(p. SUPPL 232):1–55
Sarna GS, Hutson PH, Tricklebank MD, Curzon G (1983) Determination of brain 5-hydroxytryptamine turnover in freely moving rats using repeated sampling of cerebrovascular fluid. J Neurochem 40(2):383–388
Gaddum JH (1961) Push-pull cannulae. J Physiol 155:1–2P
Glowinski J (1981) In vivo release of transmitters in the cat basal ganglia. Fed Proc 40(2):135–141
Philippu A (1984) Use of push-pull cannulae to determine the release of endogenous neurotransmitters in distinct brain areas of anaesthetized and freely moving animals. In: Marsden CA (ed) Measurement of neurotransmitters release in vivo. Wiley, Chichester, UK, pp 3–37
Besson M, Cheramy A, Feltz P, Glowinski J (1971) Dopamine: spontaneous and drug-induced release from the caudate nucleus in the cat. Brain Res 32(2):407–424
Gonon F, Buda M, Cespuglio R, Jouvet M, Pujol JF (1980) In vivo electrochemical detection of catechols in the neostriatum of anaesthetized rats: dopamine or DOPAC? Nature 286(5776):902–904
Stamford JA (1985) In vivo voltammetry: promise and perspective. Brain Res 357(2):119–135
Ungerstedt U, Pycock C (1974) Functional correlates of dopamine neurotransmission. Bull Schweiz Akad Med Wiss 30(1–3):44–55
Kissinger PT, Refshuage CJ, Dreiling R, Blank L, Freeman R, Adams RN (1973) An electrochemical detector for liquid chromatography with picograms sensitivity. Analytical Letters 6:465–477
Mefford IN (1981) Application of high performance liquid chromatography with electrochemical detection to neurochemical analysis: measurement of catecholamines, serotonin and metabolites in rat brain. J Neurosci Methods 3(3):207–224
Imperato A, Di Chiara G (1984) Trans-striatal dialysis coupled to reverse phase high performance liquid chromatography with electrochemical detection: a new method for the study of the in vivo release of endogenous dopamine and metabolites. J Neurosci 4(4):966–977
Ungerstedt U, Herrera-Marschitz M, Zetterstrom T (1982) Dopamine neurotransmission and brain function. Prog Brain Res 55:41–49
Zetterstrom T, Sharp T, Marsden CA, Ungerstedt U (1983) In vivo measurement of dopamine and its metabolites by intracerebral dialysis: changes after d-amphetamine. J Neurochem 41(6):1769–1773
L’Heureux R, Dennis T, Curet O, Scatton B (1986) Measurement of endogenous noradrenaline release in the rat cerebral cortex in vivo by transcortical dialysis: effects of drugs affecting noradrenergic transmission. J Neurochem 46(6):1794–1801
Carboni E, Di Chiara G (1989) Serotonin release estimated by transcortical dialysis in freely-moving rats. Neuroscience 32(3):637–645
Hernandez L, Lee F, Hoebel BG (1987) Simultaneous microdialysis and amphetamine infusion in the nucleus accumbens and striatum of freely moving rats: increase in extracellular dopamine and serotonin. Brain Res Bull 19(6):623–628
Kalen P, Strecker RE, Rosengren E, Bjorklund A (1988) Endogenous release of neuronal serotonin and 5-hydroxyindoleacetic acid in the caudate-putamen of the rat as revealed by intracerebral dialysis coupled to high-performance liquid chromatography with fluorimetric detection. J Neurochem 51(5):1422–1435
Calcagno E, Carli M, Baviera M, Invernizzi RW (2009) Endogenous serotonin and serotonin2C receptors are involved in the ability of M100907 to suppress cortical glutamate release induced by NMDA receptor blockade. J Neurochem 108(2):521–532
Calcagno E, Carli M, Invernizzi RW (2006) The 5-HT(1A) receptor agonist 8-OH-DPAT prevents prefrontocortical glutamate and serotonin release in response to blockade of cortical NMDA receptors. J Neurochem 96(3):853–860
Calcagno E, Guzzetti S, Canetta A, Fracasso C, Caccia S, Cervo L, Invernizzi RW (2009) Enhancement of cortical extracellular 5-HT by 5-HT1A and 5-HT2C receptor blockade restores the antidepressant-like effect of citalopram in non-responder mice. Int J Neuropsychopharmacol 12:793–803
Calcagno E, Invernizzi RW (2010) Strain-dependent serotonin neuron feedback control: role of serotonin 2 C receptors. J Neurochem 114(6):1701–1710
Ceglia I, Acconcia S, Fracasso C, Colovic M, Caccia S, Invernizzi RW (2004) Effects of chronic treatment with escitalopram or citalopram on extracellular 5-HT in the prefrontal cortex of rats: role of 5-HT1A receptors. Br J Pharmacol 142(3):469–478
Ceglia I, Carli M, Baviera M, Renoldi G, Calcagno E, Invernizzi RW (2004) The 5-HT receptor antagonist M100,907 prevents extracellular glutamate rising in response to NMDA receptor blockade in the mPFC. J Neurochem 91(1):189–199
Invernizzi R, Belli S, Samanin R (1992) Citalopram’s ability to increase the extracellular concentrations of serotonin in the dorsal raphe prevents the drug’s effect in the frontal cortex. Brain Res 584(1–2):322–324
Invernizzi R, Bramante M, Samanin R (1994) Chronic treatment with citalopram facilitates the effect of a challenge dose on cortical serotonin output: role of presynaptic 5-HT1A receptors. Eur J Pharmacol 260(2–3):243–246
Invernizzi R, Bramante M, Samanin R (1995) Extracellular concentrations of serotonin in the dorsal hippocampus after acute and chronic treatment with citalopram. Brain Res 696(1–2):62–66
Invernizzi R, Bramante M, Samanin R (1996) Role of 5-HT1A receptors in the effects of acute chronic fluoxetine on extracellular serotonin in the frontal cortex. Pharmacol Biochem Behav 54(1):143–147
Invernizzi R, Morali F, Pozzi L, Samanin R (1990) Effects of acute and chronic clozapine on dopamine release and metabolism in the striatum and nucleus accumbens of conscious rats. Br J Pharmacol 100(4):774–778
Invernizzi R, Pozzi L, Samanin R (1995) Selective reduction of extracellular dopamine in the rat nucleus accumbens following chronic treatment with DAU 6215, a 5-HT3 receptor antagonist. Neuropharmacology 34(2):211–215
Invernizzi R, Velasco C, Bramante M, Longo A, Samanin R (1997) Effect of 5-HT1A receptor antagonists on citalopram-induced increase in extracellular serotonin in the frontal cortex, striatum and dorsal hippocampus. Neuropharmacology 36(4–5):467–473
Invernizzi RW, Garattini S (2004) Role of presynaptic alpha2-adrenoceptors in antidepressant action: recent findings from microdialysis studies. Prog Neuropsychopharmacol Biol Psychiatry 28(5):819–827
Invernizzi RW, Garavaglia C, Samanin R (2003) The alpha 2-adrenoceptor antagonist idazoxan reverses catalepsy induced by haloperidol in rats independent of striatal dopamine release: role of serotonergic mechanisms. Neuropsychopharmacology 28(5):872–879
Invernizzi RW, Parini S, Sacchetti G, Fracasso C, Caccia S, Annoni K, Samanin R (2001) Chronic treatment with reboxetine by osmotic pumps facilitates its effect on extracellular noradrenaline and may desensitize alpha(2)- adrenoceptors in the prefrontal cortex. Br J Pharmacol 132(1):183–188
Invernizzi RW, Pierucci M, Calcagno E, Di Giovanni G, Di Matteo V, Benigno A, Esposito E (2007) Selective activation of 5-HT(2 C) receptors stimulates GABA-ergic function in the rat substantia nigra pars reticulata: a combined in vivo electrophysiological and neurochemical study. Neuroscience 144(4):1523–1535
Invernizzi RW, Sacchetti G, Parini S, Acconcia S, Samanin R (2003) Flibanserin, a potential antidepressant drug, lowers 5-HT and raises dopamine and noradrenaline in the rat prefrontal cortex dialysate: role of 5-HT(1A) receptors. Br J Pharmacol 139(7):1281–1288
Parini S, Renoldi G, Battaglia A, Invernizzi RW (2005) Chronic reboxetine desensitizes terminal but not somatodendritic alpha2-adrenoceptors controlling noradrenaline release in the rat dorsal hippocampus. Neuropsychopharmacology 30(6):1048–1055
Pozzi L, Acconcia S, Ceglia I, Invernizzi RW, Samanin R (2002) Stimulation of 5-hydroxytryptamine (5-HT(2 C) ) receptors in the ventrotegmental area inhibits stress-induced but not basal dopamine release in the rat prefrontal cortex. J Neurochem 82(1):93–100
Pozzi L, Invernizzi R, Cervo L, Vallebuona F, Samanin R (1994) Evidence that extracellular concentrations of dopamine are regulated by noradrenergic neurons in the frontal cortex of rats. J Neurochem 63(1):195–200
Pozzi L, Invernizzi R, Garavaglia C, Samanin R (1999) Fluoxetine increases extracellular dopamine in the prefrontal cortex by a mechanism not dependent on serotonin: a comparison with citalopram. J Neurochem 73(3):1051–1057
Renoldi G, Calcagno E, Borsini F, Invernizzi RW (2007) Stimulation of group I mGlu receptors in the ventrotegmental area enhances extracellular dopamine in the rat medial prefrontal cortex. J Neurochem 100(6):1658–1666
Renoldi G, Invernizzi RW (2006) Blockade of tachykinin NK1 receptors attenuates stress-induced rise of extracellular noradrenaline and dopamine in the rat and gerbil medial prefrontal cortex. J Neurosci Res 84(5):961–968
Sacchetti G, Bernini M, Bianchetti A, Parini S, Invernizzi RW, Samanin R (1999) Studies on the acute and chronic effects of reboxetine on extracellular noradrenaline and other monoamines in the rat brain. Br J Pharmacol 128(6):1332–1338
Chefer VI, Thompson AC, Zapata A, Shippenberg TS (2009) Overview of brain microdialysis. Curr Protoc Neurosci Chapter 7: p. Unit 7 1
Zapata A, Chefer VI, Shippenberg TS (2009) Microdialysis in rodents. Curr Protoc Neurosci Chapter 7: p. Unit 7 2
Zapata A, Chefer VI, Shippenberg TS, Denoroy L (2009) Detection and quantification of neurotransmitters in dialysates. Curr Protoc Neurosci Chapter 7: p. Unit 7 4 1–30
Di Chiara G (1991) Brain dialysis of monoamines. In: Robinson TE, Justice JB Jr (eds) Microdialysis in the neurosciences. Elsevier, Amsterdam, pp 175–185
Sharp T, Zetterstrom T (1991) In vivo measurement of monoamine neurotransmitter release using brain microdialysis. In: Stamford JA (ed) Monitoring neuronal activity: a practical approach. IRL, Oxford, pp 147–179
Santiago M, Westerink BH (1990) Characterization of the in vivo release of dopamine as recorded by different types of intracerebral microdialysis probes. Naunyn Schmiedebergs Arch Pharmacol 342(4):407–414
Benveniste H, Huttemeier PC (1990) Microdialysis–theory and application. Prog Neurobiol 35(3):195–215
Moghaddam B, Bunney BS (1989) Ionic composition of microdialysis perfusing solution alters the pharmacological responsiveness and basal outflow of striatal dopamine. J Neurochem 53(2):652–654
Silver IA, Erecinska M (1990) Intracellular and extracellular changes of (Ca2+) in hypoxia and ischemia in rat brain in vivo. J Gen Physiol 95(5):837–866
Nicholson C (1980) Modulation of extracellular calcium and its functional implications. Fed Proc 39(5):1519–1523
Invernizzi R, Pozzi L, Vallebuona F, Bonini I, Sacchetti G, Samanin R (1992) Effect of amineptine on regional extracellular concentrations of dopamine and noradrenaline in the rat brain. J Pharmacol Exp Ther 262(2):769–774
Lehmann J, Valentino R, Robine V (1992) Cortical norepinephrine release elicited in situ by N-methyl-D-aspartate (NMDA) receptor stimulation: a microdialysis study. Brain Res 599(1):171–174
Sharp T, Bramwell SR, Grahame-Smith DG (1989) 5-HT1 agonists reduce 5-hydroxytryptamine release in rat hippocampus in vivo as determined by brain microdialysis. Br J Pharmacol 96(2):283–290
Gundlah C, Martin KF, Heal DJ, Auerbach SB (1997) In vivo criteria to differentiate monoamine reuptake inhibitors from releasing agents: sibutramine is a reuptake inhibitor. J Pharmacol Exp Ther 283(2):581–591
Nakahara D, Ozaki N, Kapoor V, Nagatsu T (1989) The effect of uptake inhibition on dopamine release from the nucleus accumbens of rats during self- or forced stimulation of the medial forebrain bundle: a microdialysis study. Neurosci Lett 104(1–2):136–140
Romero L, Artigas F (1997) Preferential potentiation of the effects of serotonin uptake inhibitors by 5-HT1A receptor antagonists in the dorsal raphe pathway: role of somatodendritic autoreceptors. J Neurochem 68(6):2593–2603
Paxinos G, Watson C (2005) The rat brain in stereotaxic coordinates. Elsevier Academic Press, Sidney
Franklin KBJ, Paxinos G (1997) The mouse brain in stereotaxic coordinates. Academic Press, San Diego
Auerbach SB, Minzenberg MJ, Wilkinson LO (1989) Extracellular serotonin and 5-hydroxyindoleacetic acid in hypothalamus of the unanesthetized rat measured by in vivo dialysis coupled to high-performance liquid chromatography with electrochemical detection: dialysate serotonin reflects neuronal release. Brain Res 499(2):281–290
Sharp T, Bramwell SR, Clark D, Grahame-Smith DG (1989) In vivo measurement of extracellular 5-hydroxytryptamine in hippocampus of the anaesthetized rat using microdialysis: changes in relation to 5-hydroxytryptaminergic neuronal activity. J Neurochem 53(1):234–240
Deacon RM, Rawlins JN (1996) Equithesin without chloral hydrate as an anaesthetic for rats. Psychopharmacology (Berl) 124(3):288–290
Waynforth HB, Flecknell PA (1992) Experimental and surgical techniques in the rat, 2nd edn. Academic, London
Tao R, Hjorth S (1992) Differences in the in vitro and in vivo 5-hydroxytryptamine extraction performance among three common microdialysis membranes. J Neurochem 59(5):1778–1785
Dolan JW (2010) Where did that peak come from? LC-GC, 23(7):358–361
Cosford RJ, Vinson AP, Kukoyi S, Justice JB Jr (1996) Quantitative microdialysis of serotonin and norepinephrine: pharmacological influences on in vivo extraction fraction. J Neurosci Methods 68(1):39–47
Parsons LH, Justice JB Jr (1994) Quantitative approaches to in vivo brain microdialysis. Crit Rev Neurobiol 8(3):189–220
Shippenberg TS, He M, Chefer V (1999) The use of microdialysis in the mouse: conventional versus quantitative techniques. Psychopharmacology (Berl) 147(1):33–34
Invernizzi R, Garavaglia C, Samanin R (2000) JL13, a pyridobenzoxazepine compound with potential atypical antipsychotic activity, increases extracellular dopamine in the prefrontal cortex, but not in the striatum and the nucleus accumbens of rats. Naunyn Schmiedebergs Arch Pharmacol 361(3):298–302
Calcagno E, Canetta A, Guzzetti S, Cervo L, Invernizzi RW (2007) Strain differences in basal and post-citalopram extracellular 5-HT in the mouse medial prefrontal cortex and dorsal hippocampus: relation with tryptophan hydroxylase-2 activity. J Neurochem 103(3):1111–1120
Acknowledgements
I wish to thank all my coworkers that have contributed to set up, optimize, and validate the microdialysis technique applied to monoamine measurement in the rat, mouse, and gerbil brain as currently used in this author’s laboratory. This Chapter is dedicated to the memory of my mentor, Dr. R. Samanin, who passed away in 2001.
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Appendix
Appendix
List of main suppliers and Web page address
Supplier | Product | Web page address |
Beckman | Collection vials | |
Carlo Erba Reagenti | Chemicals | |
Clark Electromedical | Tungsten wires | |
CMA/Microdialysis | Microinfusion pump | |
Coopers Needle Work | Stainless steel tubing | |
Datalys | Chromatographic software | |
David Kopf Instruments | Stereotaxic | |
Eicom | Swivels | |
Eppendorf | Tubes | |
ESA | Electrochemical detector | |
Fluka | Monoamine standards | |
Harvard Apparatus | Microinfusion pump | |
Hospal | Microdialysis fibers | |
Instech | Swivels | |
Leica | Stereomicroscope | |
Merck | Chemicals | |
Microbiotech | FEP tubing | |
Millipore | HPLC-grade water | |
Polymicro Technologies | Silica tubing | |
Portex | PE tubing | |
Schleicher & Schuell | Disc filters | |
Shimadzu | HPLC pumps | |
Shiseido | HPLC columns | |
Sorin Biomedica | Microdialysis fibers | |
Spark Holland | Autosamplers | |
SSI-LabAlliance | Pulse dampener | |
Supelco | HPLC columns | |
Univentor | Fraction collectors |
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Invernizzi, R.W. (2013). Monitoring Extracellular Monoamines with In Vivo Microdialysis in Awake Rats: A Practical Approach. In: Di Giovanni, G., Di Matteo, V. (eds) Microdialysis Techniques in Neuroscience. Neuromethods, vol 75. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-173-8_9
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DOI: https://doi.org/10.1007/978-1-62703-173-8_9
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