Vasoactive intestinal peptide induces both tyrosine hydroxylase activity and tetrahydrobiopterin biosynthesis in PC12 cells

doi:10.1016/S0306-4522(97)00611-8

Neuroscience

Volume 86, Issue 1, 21 May 1998, Pages 179-189

https://doi.org/10.1016/S0306-4522(97)00611-8 Get rights and content

Abstract

Vasoactive intestinal peptide plays an important role in the trans-synaptic activation of tyrosine hydroxylase in sympathoadrenal tissues in response to physiological stress. Since tyrosine hydroxylase is thought to be subsaturated with its cofactor, tetrahydrobiopterin, we tested the hypothesis that up-regulation of tyrosine hydroxylase gene expression following vasoactive intestinal peptide treatment is accompanied by a concomitant elevation of intracellular tetrahydrobiopterin biosynthesis. We also investigated the second messenger systems involved in vasoactive intestinal peptide's effects on tetrahydrobiopterin metabolism. Our results demonstrate that treatment of PC12 cells for 24 h with vasoactive intestinal peptide induced intracellular tetrahydrobiopterin levels 3.5-fold. This increase was due to increased expression of the gene encoding GTP cyclohydrolase, the initial and rate-limiting enzyme in tetrahydrobiopterin biosynthesis, which was blocked by the transcriptional inhibitor, actinomycin D. Activation of tyrosine hydroxylase and GTP cyclohydrolase by vasoactive intestinal peptide was mediated by cyclic-AMP. Furthermore, stimulation of cyclic-AMP-mediated responses or protein kinase C activity induced the maximal in vitro activities of both tyrosine hydroxylase and GTP cyclohydrolase; the responses were additive when both treatments were combined. Induction of sphingolipid metabolism had no effect on the activation of tyrosine hydroxylase, while it induced GTP cyclohydrolase in a protein kinase C-independent manner.

Our results support the hypothesis that intracellular tetrahydrobiopterin levels are tightly linked to tyrosine hydroxylation and that tetrahydrobiopterin bioavailability modulates catecholamine synthesis.

Section snippets

Experimental procedures

PC12 cells were maintained in tissue culture flasks (150 cm²; Falcon), as reported previously.[5]Cells were harvested by mechanical dislodgment, collected by centrifugation at room temperature (7 min, 500×g) and resuspended in Dulbecco's modified Eagle's medium, as described previously.[5]Cell viability was determined by Trypan Blue (0.2%) exclusion, and cells were re-plated in 25-cm² flasks (12×10⁴ cells/cm²). After 24 h, cells were incubated in test conditions for various times at 37°C. VIP was

Results

Incubation of PC12 cells for 24 h with 10 μM VIP increased the in vitro maximal activity of TH two-fold (Fig. 1). VIP treatment also induced both the activity of GTP cyclohydrolase (3.7-fold) and intracellular BH₄ levels (3.5-fold). In contrast, the activity of sepiapterin reductase, the final enzyme in BH₄ biosynthesis, was not altered (Fig. 1). Total catecholamine levels were also increased 2.7-fold (Fig. 1).

Fig. 2 shows that VIP induces the content of GTP cyclohydrolase mRNA to 220% of control

Discussion

As indicated by our results, VIP induces GTP cyclohydrolase gene expression and this induction in turn causes an increase in intracellular BH₄ levels. The activity of sepiapterin reductase was not altered by VIP, suggesting that the induction of the initial enzyme in BH₄ synthesis is the rate-limiting step in PC12 cell BH₄ biosynthesis. Increased intracellular BH₄ is essential for the physiological action of TH in inducing catecholamine synthesis following long-term stimulation, since TH is

Conclusion

Our results demonstrate that the enhancement of TH gene expression is coupled under most conditions to the elevation of intracellular BH₄ biosynthesis. Such a coordinate induction has physiological relevance, since TH is believed to be subsaturated with BH₄ in vivo40, 43, 54, 63, 69and in PC12 cells.5, 14The elevation of BH₄ biosynthesis may be especially important at early stages of TH long-term induction before the newly synthesized TH molecules are phosphorylated (phosphorylation decreases K_m

References (85)

P.Z Anastasiadis et al.
Regulation of tyrosine hydroxylase and tetrahydrobiopterin biosynthetic enzymes in PC12 cells by NGF, EGF and IFN-gamma
Brain Res.
(1996)
P.Z Anastasiadis et al.
Tetrahydrobiopterin uptake in rat brain synaptosomes, cultured PC12 cells, and rat striatum
Brain Res.
(1994)
S Audigier et al.
Vasoactive intestinal polypeptide increases inositol phospholipid breakdown in the rat superior cervical ganglion
Brain Res.
(1986)
N Blau et al.
Guanosine triphosphate cyclohydrolase I assay in human and rat liver using high-performance liquid chromatography of neopterin phosphates and guanine nucleotides
Analyt. Biochem.
(1983)
S Boularand et al.
Characterization of the human tryptophan hydroxylase gene promoter. Transcriptional regulation by cAMP requires a new motif distinct from the cAMP-responsive element
J. biol. Chem.
(1995)
M.M Bradford
A rapid method for the quantification of microgram quantities of protein using the principle of dye binding
Analyt. Biochem.
(1976)
A.W Burg et al.
The biosynthesis of folic acid. 8. Purification and properties of the enzyme that catalyses the production of formate from carbon atom 8 of guanosine triphosphate
J. biol. Chem.
(1968)
E Costa et al.
Do cyclic nucleotides promote the transsynaptic induction of tyrosine hydroxylase?
Life Sci.
(1974)
N.N Desai et al.
Sphingosine-1-phosphate, a metabolite of sphingosine, increases phosphatidic acid levels by phospholipase D activation
J. biol. Chem.
(1992)
P.J Deutsch et al.
The 38-amino acid form of pituitary adenylate cyclase-activating polypeptide stimulates dual signaling cascades in PC12 cells and promotes neurite outgrowth
J. biol. Chem.
(1992)

N.J Dun et al.

Pituitary adenylate cyclase activating polypeptide-immunoreactive sensory neurons innervate rat adrenal medulla

Brain Res.

(1996)

S.J Fluharty et al.

Short- and long-term changes in adrenal tyrosine hydroxylase activity during insulin-induced hypoglycemia and cold stress

Brain Res.

(1983)

T Fukushima et al.

Analysis of reduced forms of biopterin in biological tissues and fluids

Analyt. Biochem.

(1980)

A Guidotti et al.

Commentary: trans-synaptic regulation of tyrosine 3-mono-oxygenase biosynthesis in rat adrenal medulla

Biochem. Pharmac.

(1977)

K Hatakeyama et al.

Cloning and sequencing of cDNA encoding rat GTP cyclohydrolase I

J. biol. Chem.

(1991)

B Hiremagalur et al.

Nicotine increases expression of tyrosine hydroxylase gene

J. biol. Chem.

(1993)

H Ichinose et al.

Characterization of mouse and human GTP cyclohydrolase I genes. Mutations in patients with GTP cyclohydrolase I deficiency

J. biol. Chem.

(1995)

K Isobe et al.

Pituitary adenylate cyclase-activating polypeptide induces gene expression of the catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine beta hydroxylase, through 3′,5′-cyclic adenosine monophosphate- and protein kinase C-dependent mechanisms in cultured porcine adrenal medullary chromaffin cells

Neuropeptides

(1996)

D Katsaros et al.

Site-selective cyclic AMP analogs provide a new approach in the control of cancer cell growth

Fedn Eur. biochem. Socs Lett.

(1987)

T.L Krukoff

Segmental distribution of corticotropin-releasing factor-like and vasoactive intestinal peptide-like immunoreactivities in presumptive sympathetic preganglionic neurons of the cat

Brain Res.

(1986)

E.J Lewis et al.

Regulation of tyrosine hydroxylase mRNA by glucocorticoid and cyclic AMP in a rat pheochromocytoma cell line

J. biol. Chem.

(1983)

R.K Malhotra et al.

Vasoactive intestinal polypeptide and muscarine mobilize intracellular Ca²⁺ through breakdown of phosphoinositides to induce catecholamine secretion: role of IP3 in exocytosis

J. biol. Chem.

(1988)

P.D Marley et al.

Activation of tyrosine hydroxylase by pituitary adenylate cyclase-activating polypeptide (PACAP-27) in bovine adrenal chromaffin cells

J. auton. nerv. Syst.

(1996)

B Mayer et al.

Kinetics and mechanism of tetrahydrobiopterin-induced oxidation of nitric oxide

J. biol. Chem.

(1995)

M Misbahuddin et al.

Stimulatory effect of vasoactive intestinal polypeptide on catecholamine secretion from isolated guinea pig adrenal chromaffin cells

Neurosci. Lett.

(1988)

S Miwa et al.

6R-l-Erythro-5,6,7,8-tetrahydrobiopterin as a regulator of dopamine and serotonin biosynthesis in the rat brain

Archs Biochem. Biophys.

(1985)

A Miyata et al.

Isolation of a novel 38-residue hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells

Biochem. biophys. Res. Commun.

(1989)

A Miyata et al.

Isolation of a neuropeptide corresponding to the N-terminal 27 residues of the pituitary adenylate cyclase activating polypeptide with 38 residues (PACAP38)

Biochem. biophys. Res. Commun.

(1990)

K Moller et al.

Expression of pituitary adenylate cyclase activating peptide (PACAP) and PACAP type I receptors in the rat adrenal medulla

Regul. Pept.

(1996)

R.A Mueller et al.

Inhibition of neuronally induced tyrosine hydroxylase by nicotinic receptor blockade

Eur. J. Pharmac.

(1970)

R.A Rius et al.

Pituitary adenylate cyclase activating polypeptide (PACAP) potently enhances tyrosine hydroxylase (TH) expression in adrenal chromaffin cells

Life Sci.

(1994)

Y Shiotani et al.

Immunohistochemical localization of pituitary adenylate cyclase-activating polypeptide (PACAP) in the adrenal medulla of the rat

Peptides

(1995)

G.K Smith

On the role of sepiapterin reductase in the biosynthesis of tetrahydrobiopterin

Archs Biochem. Biophys.

(1987)

R Strong et al.

GABA receptor modulation of tyrosine hydroxylase gene expression in the rat adrenal gland

Neurosci. Lett.

(1990)

S Suzuki et al.

Decrease in tetrahydrobiopterin content and neurotransmitter amine biosythesis in rat brain by an inhibitor of guanosine triphosphate cyclohydrolase

Brain Res.

(1988)

H.J Tae et al.

cAMP activation of CAAT enhancer-binding protein-beta gene expression and promoter I of acetyl-CoA carboxylase

J. biol. Chem.

(1995)

M Wessels-Reiker et al.

Vasoactive intestinal polypeptide induces tyrosine hydroxylase in PC12 cells

J. biol. Chem.

(1991)

K Witter et al.

Cloning, sequencing and functional studies of the gene encoding human GTP cyclohydrolase I

Gene

(1996)

M Zhu et al.

Regulation of tetrahydrobiopterin biosynthesis in cultured dopamine neurons by depolarization and cAMP

J. biol. Chem.

(1994)

M.M Abou-Donia et al.

Tetrahydrobiopterin increases in adrenal medulla and cortex: a factor in the regulation of tyrosine hydroxylase

Proc. natn. Acad. Sci. U.S.A.

(1981)

M.M Abou-Donia et al.

Regulation of guanosine triphosphate cyclohydrolase and tetrahydrobiopterin levels and the role of the cofactor in tyrosine hydroxylation in primary cultures of adrenomedullary chromaffin cells

J. Neurochem.

(1986)

P.Z Anastasiadis et al.

Tetrahydrobiopterin as obligatory mediator of PC12 cell proliferation induced by EGF and NGF

Molec. Pharmac.

(1996)

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¹: Present address: Department of Cell Biology, Medical Center North, Rm no. 2310, Vanderbilt University, Nashville, TN 37232-2175, U.S.A.

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