Measures of dopamine-regulating proteins in somatodendritic regions are often used only as static indicators of neuron viability, overlooking the possible impact of somatodendritic dopamine (DA) signaling on behavior and the potential autonomy of DA regulation between somatodendritic and terminal field compartments. DA reuptake capacity is less in somatodendritic regions, possibly placing a greater burden on de novo DA biosynthesis within this compartment to maintain DA signaling. Therefore, regulation of tyrosine hydroxylase (TH) activity may be particularly critical for somatodendritic DA signaling. Phosphorylation of TH at ser31 or ser40 can increase activity, but their impact on L-DOPA biosynthesis in vivo is unknown. Thus, determining their relationship with L-DOPA tissue content could reveal a mechanism by which DA signaling is normally maintained. In Brown-Norway Fischer 344 F1 hybrid rats, we quantified TH phosphorylation versus L-DOPA accumulation. After inhibition of aromatic acid decarboxylase, L-DOPA tissue content per recovered TH protein was greatest in NAc, matched by differences in ser31, but not ser40, phosphorylation. The L-DOPA per catecholamine and DA turnover ratios were significantly greater in SN and VTA, suggesting greater reliance on de novo DA biosynthesis therein. These compartmental differences reflected an overall autonomy of DA regulation, as seen by decreased DA content in SN and VTA, but not in striatum or NAc, following short-term DA biosynthesis inhibition from local infusion of the TH inhibitor α-methyl-p-tyrosine, as well as in the long-term process of aging. Such data suggest ser31 phosphorylation plays a significant role in regulating TH activity in vivo, particularly in somatodendritic regions, which may have a greater reliance on de novo DA biosynthesis. Thus, to the extent that somatodendritic DA release affects behavior, TH regulation in the midbrain may be critical for DA bioavailability to influence behavior.
References
[1]
Nagatsu T, Levitt M, Udenfriend S (1964) Tyrosine hydroxylase. The initial step in norepinephrine biosynthesis. J Biol Chem 239: 2910–2917.
[2]
Levitt M, Spector S, Sjoerdsma A, Udenfriend S (1965) Elucidation of the rate-limiting step in norepinephrine biosynthesis in the perfused guinea-pig heart. J Pharmacol Exp Ther 148: 1–8.
[3]
Morgenroth VH, Hegstrand LR, Roth RH, Greengard P (1975) Evidence for involvement of protein kinase in the activation by adenosine 3′:5′-monophosphate of brain tyrosine 3-monooxygenase. J Biol Chem 250: 1946–1948.
[4]
Haycock JW (1990) Phosphorylation of tyrosine hydroxylase in situ at serine 8, 19, 31, and 40. J Biol Chem 265: 11682–11691.
[5]
Haycock JW, Ahn NG, Cobb MH, Krebs EG (1992) ERK1 and ERK2, two microtubule-associated protein 2 kinases, mediate the phosphorylation of tyrosine hydroxylase at serine-31 in situ. Proc Natl Acad Sci U S A 89: 2365–2369.
[6]
Vulliet PR, Langan TA, Weiner N (1980) Tyrosine hydroxylase: a substrate of cyclic AMP-dependent protein kinase. Proc Natl Acad Sci U S A 77: 92–96.
[7]
Waymire JC, Craviso GL, Lichteig K, Johnston JP, Baldwin C, et al. (1991) Vasoactive intestinal peptide stimulates catecholamine biosynthesis in isolated adrenal chromaffin cells: evidence for a cyclic AMP-dependent phosphorylation and activation of tyrosine hydroxylase. J Neurochem 57: 1313–1324.
[8]
Bobrovskaya L, Gilligan C, Bolster EK, Flaherty JJ, Dickson PW, et al. (2007) Sustained phosphorylation of tyrosine hydroxylase at serine 40: a novel mechanism for maintenance of catecholamine synthesis. J Neurochem 100: 479–489.
[9]
Haycock JW, Haycock DA (1991) Tyrosine hydroxylase in rat brain dopaminergic nerve terminals. Multiple-site phosphorylation in vivo and in synaptosomes. J Biol Chem 266: 5650–5657.
[10]
Harada , Wu J, Haycock JW, Goldstein M (1996) Regulation of L-DOPA biosynthesis by site-specific phosphorylation of tyrosine hydroxylase in AtT-20 cells expressing wild-type and serine 40-substituted enzyme. J Neurochem 67: 629–635.
[11]
Salvatore MF, Waymire JC, Haycock JW (2001) Depolarization-stimulated catecholamine biosynthesis: involvement of protein kinases and tyrosine hydroxylase phosphorylation sites in situ. J Neurochem 79: 349–360.
[12]
Salvatore MF, Garcia-Espana A, Goldstein M, Deutch AY, Haycock JW (2000) Stoichiometry of tyrosine hydroxylase phosphorylation in the nigrostriatal and mesolimbic systems in vivo: effects of acute haloperidol and related compounds. J Neurochem 75: 225–232.
[13]
Salvatore MF, Zhang JL, Large DM, Wilson PE, Gash CR, et al. (2004) Striatal GDNF administration increases tyrosine hydroxylase phosphorylation in the rat striatum and substantia nigra. J Neurochem 90: 245–254.
[14]
Salvatore MF, Pruett BS, Spann SL, Dempsey C (2009) Aging reveals a role for nigral tyrosine hydroxylase ser31 phosphorylation in locomotor activity generation. PLoS One 4: e8466.
[15]
Haycock JW, Lew JY, Garcia-Espana A, Lee KY, Harada K, et al. (1998) Role of serine-19 phosphorylation in regulating tyrosine hydroxylase studied with site- and phosphospecific antibodies and site-directed mutagenesis. J Neurochem 71: 1670–1675.
[16]
Lindgren N, Xu ZQ, Lindskog M, Herrera-Marschitz M, Goiny M, et al. (2000) Regulation of tyrosine hydroxylase activity and phosphorylation at Ser(19) and Ser(40) via activation of glutamate NMDA receptors in rat striatum. J Neurochem 74: 2470–2477.
[17]
Cragg S, Rice ME, Greenfield SA (1997) Heterogeneity of electrically evoked dopamine release and reuptake in substantia nigra, ventral tegmental area, and striatum. J Neurophysiol 77: 863–873.
[18]
Hoffman AF, Lupica CR, Gerhardt GA (1998) Dopamine transporter activity in the substantia nigra and striatum assessed by high-speed chronoamperometric recordings in brain slices. J Pharmacol Exp Ther 287: 487–496.
[19]
Hoffman AF, Gerhardt GA (1998) In vivo electrochemical studies of dopamine clearance in the rat substantia nigra: effects of locally applied uptake inhibitors and unilateral 6-hydroxydopamine lesions. J Neurochem 70: 179–189.
[20]
Chen BT, Rice ME (2001) Novel Ca2+ dependence and time course of somatodendritic dopamine release: substantia nigra versus striatum. J Neurosci 21: 7841–7847.
[21]
Rice ME, Cragg SJ (2008) Dopamine spillover after quantal release: rethinking dopamine transmission in the nigrostriatal pathway. Brain Res Rev 58: 303–313.
[22]
Ford CP, Gantz SC, Phillips PE, Williams JT (2010) Control of extracellular dopamine at dendrite and axon terminals. J Neurosci 30: 6975–6983.
[23]
Jones SR, Gainetdinov RR, Jaber M, Giros B, Wightman RM, et al. (1998) Profound neuronal plasticity in response to inactivation of the dopamine transporter. Proc Natl Acad Sci U S A 95: 4029–4034.
[24]
Keller CM, Salvatore MF, Pruett BS, Guerin GF, Goeders NE (2011) Biphasic dopamine regulation in mesoaccumbens pathway in response to non-contingent binge and escalating methamphetamine regimens in the Wistar rat. Psychopharmacology (Berl) 215: 513–526.
[25]
Dunkley PR, Bobrovskaya L, Graham ME, von Nagy-Felsobuki EI, Dickson PW (2004) Tyrosine hydroxylase phosphorylation: regulation and consequences. J Neurochem 91: 1025–1043.
[26]
Lavicky J, Dunn AJ (1993) Corticotropin-releasing factor stimulates catecholamine release in hypothalamus and prefrontal cortex in freely moving rats as assessed by microdialysis. J Neurochem 60: 602–612.
[27]
Salvatore MF, Fisher B, Surgener SP, Gerhardt GA, Rouault T (2005) Neurochemical investigations of dopamine neuronal systems in iron-regulatory protein 2 (IRP-2) knockout mice. Brain Res Mol Brain Res 139: 341–347.
[28]
Leak RK, Castro SL, Jaumotte JD, Smith AD, Zigmond MJ (2010) Assaying multiple biochemical variables from the same tissue sample. J Neurosci Methods 191: 234–238.
[29]
Bevilaqua LR, Graham ME, Dunkley PR, von Nagy-Felsobuki EI, Dickson PW (2001) Phosphorylation of Ser(19) alters the conformation of tyrosine hydroxylase to increase the rate of phosphorylation of Ser(40). J Biol Chem 276: 40411–40416.
[30]
Nissbrandt H, Sundstrom E, Jonsson G, Hjorth S, Carlsson A (1989) Synthesis and release of dopamine in rat brain: comparison between substantia nigra pars compacts, pars reticulata, and striatum. J Neurochem 52: 1170–1182.
[31]
Tachikawa E, Tank AW, Weiner DH, Mosimann WF, Yanagihara N, et al. (1987) Tyrosine hydroxylase is activated and phosphorylated on different sites in rat pheochromocytoma PC12 cells treated with phorbol ester and forskolin. J Neurochem 48: 1366–1376.
[32]
Mitchell JP, Hardie DG, Vulliet PR (1990) Site-specific phosphorylation of tyrosine hydroxylase after KCl depolarization and nerve growth factor treatment of PC12 cells. J Biol Chem 265: 22358–22364.
[33]
Hakansson K, Pozzi L, Usiello A, Haycock J, Borrelli E, et al. (2004) Regulation of striatal tyrosine hydroxylase phosphorylation by acute and chronic haloperidol. Eur J Neurosci 20: 1108–1112.
[34]
Peng X, Tehranian R, Dietrich P, Stefanis L, Perez RG (2005) Alpha-synuclein activation of protein phosphatase 2A reduces tyrosine hydroxylase phosphorylation in dopaminergic cells. J Cell Sci 118: 3523–3530.
[35]
Zhang D, Kanthasamy A, Yang Y, Anantharam V, Kanthasamy A (2007) Protein kinase C delta negatively regulates tyrosine hydroxylase activity and dopamine synthesis by enhancing protein phosphatase-2A activity in dopaminergic neurons. J Neurosci 27: 5349–5362.
[36]
Connor JR, Wang XS, Allen RP, Beard JL, Wiesinger JA, et al. (2009) Altered dopaminergic profile in the putamen and substantia nigra in restless leg syndrome. Brain 132: 2403–2412.
[37]
Lou H, Montoya SE, Alerte TN, Wang J, Wu J, et al. (2010) Serine 129 phosphorylation reduces the ability of alpha-synuclein to regulate tyrosine hydroxylase and protein phosphatase 2A in vitro and in vivo. J Biol Chem 285: 17648–17661.
[38]
Haycock JW, Wakade AR (1992) Activation and multiple-site phosphorylation of tyrosine hydroxylase in perfused rat adrenal glands. J Neurochem 58: 57–64.
[39]
Waymire JC, Johnston JP, Hummer-Lickteig K, Lloyd A, Vigny A, et al. (1988) Phosphorylation of bovine adrenal chromaffin cell tyrosine hydroxylase. Temporal correlation of acetylcholine's effect on site phosphorylation, enzyme activation, and catecholamine synthesis. J Biol Chem 263: 12439–12447.
[40]
Saraf A, Virshup DM, Strack S (2007) Differential expression of the B'beta regulatory subunit of protein phosphatase 2A modulates tyrosine hydroxylase phosphorylation and catecholamine synthesis. J Biol Chem 282: 573–580.
[41]
Sarre S, Yuan H, Jonkers N, Van HA, Ebinger G, et al. (2004) In vivo characterization of somatodendritic dopamine release in the substantia nigra of 6-hydroxydopamine-lesioned rats. J Neurochem 90: 29–39.
[42]
Bustos G, Abarca J, Bustos V, Riquelme E, Noriega V, et al. (2009) NMDA receptors mediate an early up-regulation of brain-derived neurotrophic factor expression in substantia nigra in a rat model of presymptomatic Parkinson's disease. J Neurosci Res 87: 2308–2318.
[43]
Yurek DM, Hipkens SB, Hebert MA, Gash DM, Gerhardt GA (1998) Age-related decline in striatal dopamine release and motoric function in brown Norway/Fischer 344 hybrid rats. Brain Res 791: 246–256.
[44]
Pruett BS, Salvatore MF (2010) GFR alpha-1 receptor expression in the aging nigrostriatal and mesoaccumbens pathways. J Neurochem 115: 707–715.
[45]
Cheramy A, Leviel V, Glowinski J (1981) Dendritic release of dopamine in the substantia nigra. Nature 289: 537–542.
[46]
Jackson EA, Kelly PH (1983) Role of nigral dopamine in amphetamine-induced locomotor activity. Brain Res 278: 366–369.
[47]
Robertson GS, Robertson HA (1989) Evidence that L-dopa-induced rotational behavior is dependent on both striatal and nigral mechanisms. J Neurosci 9: 3326–3331.
[48]
Trevitt JT, Carlson BB, Nowend K, Salamone JD (2001) Substantia nigra pars reticulata is a highly potent site of action for the behavioral effects of the D1 antagonist SCH 23390 in the rat. Psychopharmacology (Berl) 156: 32–41.
[49]
Bergquist F, Shahabi HN, Nissbrandt H (2003) Somatodendritic dopamine release in rat substantia nigra influences motor performance on the accelerating rod. Brain Res 973: 81–91.
[50]
Andersson DR, Nissbrandt H, Bergquist F (2006) Partial depletion of dopamine in substantia nigra impairs motor performance without altering striatal dopamine neurotransmission. Eur J Neurosci 24: 617–624.
[51]
Nimitvilai S, Brodie MS (2010) Reversal of prolonged dopamine inhibition of dopaminergic neurons of the ventral tegmental area. J Pharmacol Exp Ther 333: 555–563.
[52]
Lu L, Dempsey J, Liu SY, Bossert JM, Shaham Y (2004) A single infusion of brain-derived neurotrophic factor into the ventral tegmental area induces long-lasting potentiation of cocaine seeking after withdrawal. J Neurosci 24: 1604–1611.
[53]
Lu L, Wang X, Wu P, Xu C, Zhao M, et al. (2009) Role of ventral tegmental area glial cell line-derived neurotrophic factor in incubation of cocaine craving. Biol Psychiatry 66: 137–145.
[54]
Wang J, Carnicella S, Ahmadiantehrani S, He DY, Barak S, et al. (2010) Nucleus accumbens-derived glial cell line-derived neurotrophic factor is a retrograde enhancer of dopaminergic tone in the mesocorticolimbic system. J Neurosci 30: 14502–14512.
[55]
Ai Y, Markesbery W, Zhang Z, Grondin R, Elseberry D, et al. (2003) Intraputamenal infusion of GDNF in aged rhesus monkeys: distribution and dopaminergic effects. J Comp Neurol 461: 250–261.
[56]
Gash DM, Zhang Z, Ai Y, Grondin R, Coffey R, et al. (2005) Trophic factor distribution predicts functional recovery in parkinsonian monkeys. Ann Neurol 58: 224–233.
[57]
Salvatore MF, Gerhardt GA, Dayton RD, Klein RL, Stanford JA (2009) Bilateral Effects of Unilateral GDNF Administration on Dopamine- and GABA-regulating Proteins in the Rat Nigrostriatal System. Exp Neurol 219: 197–207.
[58]
Hoffer BJ, Hoffman A, Bowenkamp K, Huettl P, Hudson J, et al. (1994) Glial cell line-derived neurotrophic factor reverses toxin-induced injury to midbrain dopaminergic neurons in vivo. Neurosci Lett 182: 107–111.
[59]
Gash DM, Zhang Z, Ovadia A, Cass WA, Yi A, et al. (1996) Functional recovery in parkinsonian monkeys treated with GDNF. Nature 380: 252–255.
[60]
Grondin R, Cass WA, Zhang Z, Stanford JA, Gash DM, et al. (2003) Glial cell line-derived neurotrophic factor increases stimulus-evoked dopamine release and motor speed in aged rhesus monkeys. J Neurosci 23: 1974–1980.
