Tetrahydroisoquinolines derived from catecholamines or DOPA: effects on brain tyrosine hydroxylase activity.
Explore the source record for details and available documents.
Animals↗
SEARCH · Search PubMed
Results for “TYROSINE DECARBOXYLASE”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
A site-specific mutation of tyrosine hydroxylase reduces feedback inhibition by dopamine in genetically modified cells grafted in parkinsonian rats.
Aromatic L-amino acid decarboxylase (AADC) is necessary for conversion of L-DOPA to dopamine. Therefore, AADC gene therapy has been proposed to enhance pharmacological or gene therapies delivering L-DOPA. However, addition of AADC to the grafts of genetically modified cells expressing tyrosine hydroxylase (TH) and GTP cyclohydrolase 1 (GCH1), which produce L-DOPA in parkinsonian rats, resulted in decreased production of L-DOPA and dopamine owing to feedback inhibition of TH by dopamine. End-product feedback inhibition has been shown to be mediated by the regulatory domain of TH, and site-specific mutation of serine 40 makes TH less susceptible to dopamine inhibition. Therefore, we investigated the efficacy of using TH with serine 40 mutated to leucine (mTH) in an ex vivo gene-therapy paradigm. Primary fibroblasts (PF) from Fischer 344 rats were transduced with retrovirus to express mTH or wild-type rat TH cDNA (wtTH). Both cell types were also transduced with GCH1 to provide the obligate TH cofactor, tetrahydrobiopterin. PF transfected with AADC were used as coculture and cografting partners. TH activities and L-DOPA production in culture were comparable between PFwtTHGC and PFmTHGC cells. In cocultures with PFAADC cells, PFmTHGC cells showed significant reduction in the inhibitory effect of dopamine compared with PFwtTHGC cells. In vivo microdialysis measurement showed that cografting PFAADC cells with PFmTHGC cells resulted in smaller decreases in L-DOPA and no reduction in dopamine levels compared with cografts of PFAADC cells with PFwtTHGC cells, which decreased both L-DOPA and dopamine levels. Maintenance of dopamine levels with lower levels of L-DOPA would result in more focused local delivery of dopamine and less potential side-effects arising from L-DOPA diffusion into other structures. These data support the hypothesis that mutation of serine 40 attenuates TH end-product inhibition in vivo and illustrates the importance of careful consideration of biochemical pathways and interactions between multiple genes in gene therapy.
Amino Acid Substitution↗
Specific inhibition of the synthesis of putrescine and spermidine by 1,3-diaminopropane in rat liver in vivo.
Chronic administration of 1,3-diaminopropane, a compound inhibiting mammalian ornithine decarboxylase (EC 4.1.1.17) in vivo, effectively prevented the large increases in the concentration of putrescine that normally occur during rat liver regeneration. Furthermore, repeated injections of diaminopropane depressed by more than 85% ornithine decarboxylase activity in rat kidney. Administration of diaminopropane 60 min before partial hepatectomy only marginally inhibited ornithine decarboxylase activity at 4 h after the operation. However, when the compound was given at the time of the operation (4 h before death), or any time thereafter, it virtually abolished the enhancement in ornithine decarboxylase activity in regenerating rat liver remnant. An injection of diaminopropane given 30 to 60 min after operation, but not earlier or later, depressed S-adenosyl-L-methionine decarboxylase activity (EC 4.1.1.50) 4 h after partial hepatectomy. Diaminopropane likewise inhibited ornithine decarboxylase activity during later periods of liver regeneration. In contrast to early regeneration, a total inhibition of the enzyme activity was only achieved when the injection was given not earlier than 2 to 3 h before the death of the animals. Diaminopropane also exerted an acute inhibitory effect on adenosylmethionine decarboxylase activity in 28-h regenerating liver whereas it invariably enhanced the activity of tyrosine aminotransferase (EC 2.6.1.5), used as a standard enzyme of short half-life. Treatment of the rats with diaminopropane entirely abolished the stimulation of spermidien synthesis in vivo from [14C]methionine 4 h after partial hepatectomy or after administration of porcine growth hormone. Both partial hepatectomy and the treatment with growth hormone produced a clear stimulation of hepatic RNA synthesis, the extent of which was not altered by injections of diaminopropane in doses sufficient to prevent any enhancement of ornithine decarboxylase activity and spermidine synthesis.
Adenosylmethionine Decarboxylase↗
Enzymic sulphation of dopa and tyrosine isomers by HepG2 human hepatoma cells: stereoselectivity and stimulation by Mn2+.
HepG2 human hepatoma cells, labelled with [35S]sulphate in media containing L-3,4-dihydroxyphenylalanine (L-dopa), (D-dopa), DL-m-tyrosine or D-p-tyrosine, were found to produce the [35S]sulphated forms of these compounds. Addition to the labelling media of m-hydroxybenzylhydrazine, an aromatic amino acid decarboxylase inhibitor, greatly enhanced the production of L-dopa O-[35S]sulphate and DL-m-tyrosine O-[35S]sulphate, with a concomitant decrease in the formation of dopamine O-[35S]sulphate and m-tyramine O-[35S]sulphate. With 3'-phosphoadenosine 5'-phospho[35S]sulphate as the sulphate donor., HepG2-cell cytosol was shown to contain enzymic activity catalysing the sulphation of L-dopa, D-dopa, L-m-tyrosine, D-m-tyrosine, L-p-tyrosine and D-p-tyrosine. The pH optimum of the enzyme, designated dopa/tyrosine sulphotransferase, was determined to be 8.75 with D-m-tyrosine as the substrate. The enzyme exhibited stereoselectivity for the D-form of dopa or tyrosine isomers. Addition of 10mM MnCl2 to the reaction mixture resulted in a remarkable stimulation of dopa/tyrosine sulphotransferase activity, being as high as 267.8 times with D-p-tyrosine as the substrate. Quantitative assays revealed L-dopa, D-dopa and D-m-tyrosine to be better substrates than L-p-tyrosine. When the HepG2-cell cytosol was subjected to DEAE Bio-Gel and hydroxyapatite column chromatography, dopa/tyrosine sulphotransferase was co-eluted with the thermolabile 'M-form' phenol sulphotransferase. Furthermore dopa/tyrosine sulphotransferase displayed properties similar to that of the M-form phenol sulphotransferase with respect to thermostability and sensitivity to 2,6-dichloro-4-nitrophenol. Whether the M-form phenol sulphotransferase is truly (solely) responsible for the dopa/tyrosine sulphotransferase activity present in HepG2 cells remains to be clarified.
Blood Platelets↗
GABAergic and catecholaminergic synaptic interactions in the macaque hypothalamus: double label immunostaining with peroxidase-antiperoxidase and colloidal gold.
Immunogold staining (IGS) for glutamic acid decarboxylase (GAD) was combined with the peroxidase-antiperoxidase (PAP) technique for tyrosine hydroxylase (TH) to analyze gamma-aminobutyric acid-catecholaminergic neuronal interactions in the rhesus hypothalamus. At the light-microscopic level, TH-immunoreactive (-IR) perikarya and their fibers (brown) were observed in the anterior ventral periventricular area (AVPV), the arcuate nucleus (ARC) and the adjacent periventricular zone (ARC-PVZ). GAD-IR processes (light red) were also present throughout the hypothalamus and appeared to contact some TH-IR neurons. At the electron-microscopic level, PAP was present in perikarya, dendrites, axons and axon terminals of TH-IR neurons. Colloidal gold particles (15 nm) were found only in dendrites and axon terminals of GAD-IR neurons. Labeled GAD terminals typically contained small, clear synaptic vesicles, while TH terminals contained these and sometimes one or two dense-core vesicles. In the ARC and ARC-PVZ, asymmetrical (Gray I) axodendritic synapses occurred between GAD and TH-IR profiles, with TH/GAD directionality more prevalent. Symmetrical (Gray II) synapses were less common, with either TH or GAD presynaptic in axodendritic and dendrodendritic contacts. GAD/GAD interactions were not observed, but TH/TH contacts appeared to be mostly dendrodendritic. In the AVPV, only symmetrical synapses were encountered, and their directionality was difficult to determine. GAD- and TH-IR dendrites frequently established dendrodendritic synapses, but GAD/TH dendrosomatic synapses were seldom seen. These results illustrate the complex interactions of GAD- and TH-containing elements in the neuroendocrine hypothalamus.
Animals↗
Relationship of the 37,000- and 40,000-M(r) tryptic fragments of islet antigens in insulin-dependent diabetes to the protein tyrosine phosphatase-like molecule IA-2 (ICA512).
Sera from patients with insulin-dependent diabetes immunoprecipitate 64,000-M(r) proteins, distinct from glutamate decarboxylase, that are cleaved to 37,000- and 40,000-M(r) fragments by trypsin. We investigated possible relationships between 37,000- or 40,000-M(r) fragments of antigen and the tyrosine phosphatase-like protein, IA-2 (ICA512). Antibodies from nondiabetic relatives bound differentially to 37,000- and 40,000-M(r) fragments indicating presence of distinct epitopes. Precursors of these fragments could be separated on immobilized lectins, suggesting different carbohydrate content. Levels of antibodies to 40,000-M(r) fragments were strongly associated with those to the intracellular domain of IA-2. Recombinant intracellular domain of IA-2 blocked binding of antibodies to 40,000-M(r) fragments expressed by insulinoma cells and partially blocked binding to 37,000-M(r) fragments. Furthermore, trypsinization of recombinant intracellular domain of IA-2 generated proteolytic fragments of identical M(r) to the 40,000-M(r) fragments of insulinoma antigen; 37,000-M(r) fragments were not generated. Thus, 40,000-M(r) fragments of islet autoantigen are derived from a protein similar or identical to the tyrosine phosphatase-like molecule, IA-2. The 37,000-M(r) fragments are derived from a different, although related, protein.
Animals↗
CNS monoamine cell groups projecting to pancreatic vagal motor neurons: a transneuronal labeling study using pseudorabies virus.
The CNS monoamine cell groups that project to the pancreatic parasympathetic preganglionic neurons were identified with the use of the viral retrograde transneuronal labeling method. Pseudorabies virus (PRV) was injected into the pancreas of C8 spinal rats and subsequently, transneuronally-labelled central monoamine neurons were mapped in brain tissue sections that had been stained by an immunohistochemical procedure that allowed for the visualization of PRV products and biogenic amine neurotransmitter enzymes or serotonin (5-HT) in the same neuron. The enzymes studied were tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), phenylethanolamine-N-methyltransferase (PNMT), and histidine decarboxylase. Pancreatic vagal motor neurons originate exclusively from the dorsal vagal motor nucleus and some of these may be dopamine neurons because they were TH immunopositive, but DBH and PNMT immunonegative. Transneuronally labeled aminergic neurons were found throughout the medulla oblongata. The adrenergic inputs arose from the C1, C2, and C3 cell groups. Noradrenergic inputs originated predominantly from the A5 cell group, with lesser contributions from the A1 and A2 cell groups as well as from the area postrema. None of the other CNS catecholamine cells were labeled, except for some weakly staining TH-immunoreactive neurons, presumably dopaminergic, in the paraventricular hypothalamic nucleus (PVN). The greatest number of 5-HT neurons that innervate the pancreatic vagal motor neurons come from the gigantocellular reticular nucleus, pars alpha with lesser inputs from the raphe magnus, obscurus, and pallidus nuclei. None of the CNS histaminergic cell groups were labeled.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Altered basal release and pressor effect of L-DOPA in the rostral ventrolateral medulla of spontaneously hypertensive rats.
1. Transmitter-like L-DOPA functions as a tonic to produce postsynaptic cardiopressor responses in the rostral ventrolateral medulla (RVLM) of rats. We attempted to clarify whether a transmitter-like L-DOPA system is altered in the RVLM of spontaneously hypertensive rats (SHR) to maintain the hypertension. 2. By microdialysis of left RVLM area, the basal L-DOPA release was higher in SHR than in Wistar-Kyoto (WKY) rats. This release was partially inhibited by tetrodotoxin (TTX, 1 mu mol/L) to a similar degree in both strains. TTX-sensitive L-DOPA release was higher in SHR than in WKY. 3. L-DOPA (10-600 ng) and L-glutamate (10-300 ng) microinjected into left RVLM produced dose-dependent hypertension and tachycardia. Pressor but not tachycardiac responses to L-DOPA at lower doses were slightly greater in SHR than in WKY, whereas no difference to L-glutamate was observed in either strain. 4. In RVLM regions, there was no difference of tyrosine hydroxylase activity in SHR or WKY; however, L-aromatic amino acid decarboxylase activity was lower in SHR than in WKY. 5. Enhanced presynaptic neuronal L-DOPA release, including a decrease in decarboxylation and sensitization of postsynaptic pressor sites to L-DOPA in RVLM, may be involved in the maintenance of hypertension in SHR.
Animals↗
Screening of potential chemopreventive agents using biochemical markers of carcinogenesis.
Ninety potential chemopreventive agents were screened using 6 chemoprevention-associated biochemical end points. These compounds were tested using rodent (tracheal epithelial or liver) cells and human cells [neonatal foreskin fibroblasts, bronchial epithelial cells, or human leukemic cells (HL-60)]. The effects measured were: (a) inhibition of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced tyrosine kinase activity in HL-60 cells; (b) inhibition of TPA-induced ornithine decarboxylase (ODC) activity in rat tracheal epithelial cells; (c) inhibition of poly(ADP-ribose)polymerase in propane sultone-treated primary human fibroblasts; (d) inhibition of benzo[a]pyrene(B[a]P)-DNA binding in human bronchial epithelial cells; (e) induction of reduced glutathione in Buffalo rat liver cells; and (f) inhibition of TPA-induced free radical formation in primary human fibroblasts or HL-60 cells. Fifty compounds were highly effective in inhibiting TPA-induced tyrosine kinase activity. This assay identified compounds from a wide variety of chemical classes as effective inhibitors, including all the vitamins, retinoic acid analogues, protein kinase C inhibitors, and chemicals belonging to the amino acid category. Fifty-two chemicals were classified as highly positive compounds when examined for their ability to inhibit TPA-induced ODC activity. These agents showed a dose-dependent inhibition or inhibition at all doses. Retinoids, in general, exhibited strong inhibition of ODC activity. A category of compounds showing dose-dependent inhibition were the sulfur compounds, especially the thiols and thiones. Among the natural products, terpenes were strong inhibitors of ODC. Forty-seven compounds were classified as strong inhibitors of poly(ADP-ribose)polymerase. In the carcinogen-DNA binding inhibition assay, 21 compounds were identified as strong inhibitors, which include phenolic compounds as well as sulfur compounds. Vitamins and their analogues were also good inhibitors. Testing for induced glutathione yielded 19 compounds that were good inducers. Sulfur-containing compounds and most of the phenolic compounds were also inducers of glutathione. Twenty compounds were highly positive for inhibition of TPA-induced free radical formation. A significant number of phenolic and sulfur compounds were again strong oxygen radical scavengers. Some antiinflammatory agents were also identified as free radical inhibitors. In general, retinoids were quite active in all the assays. Eight compounds were positive in all of the six assays; these were vitamin C (ascorbic acid), bismuththiol, esculetin, etoperidone, folic acid, hydrocortisone, indole-3-carbinol, and tocopherol succinate. Agents that were positive in these assays may inhibit the carcinogenesis process by similar mechanisms in humans and are identified as candidates for development as chemopreventive agents.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
The metabolism of tyrosine-O-phosphate in Drosophila.
Explore the source record for details and available documents.
Animals↗
Tyrosine hydroxylation in the rat striatum after fentanyl and droperidol in vivo.
The effects of fentanyl, droperidol, and morphine on tyrosine hydroxylation in the rat striatum was measured after inhibition of aromatic amino acid decarboxylase with R04-4602. Any direct change in the product of hydroxylation (i.e.dopa) was supposed to reflect a change in enzyme activity. Fentanyl (0.1 mg/kg intraperitoneally) first (10 min) appears to slightly depress, and later (30 min) significantly activate tyrosine hydroxylation. Morphine (10 mg/kg i.p.) activates tyrosine hydroxylation to a much lesser degree than fentanyl. Droperidol (3 mg/kg i.p.) only slightly activates the in vivo activity of the enzyme.
Animals↗
Inhibition of 3,4-dihydroxy-L-phenylalanine decarboxylase in rat striatal synaptosomes by amino acids interacting with substrate transport.
Dopamine synthesis from 3,4-dihydroxy-L-phenylalanine in rat striatal synaptosomes was inhibited by a number of amino acids with aromatic or large aliphatic side chains. Inhibition was not seen when aromatic amino acid decarboxylase activity was measured in disrupted synaptosomes. Similarly, inhibition of dopamine synthesis from tyrosine was seen in the presence of leucine. The inhibition most likely results from interactions of the amino acids with substrate transport across the synaptosome plasma membrane, rather than directly with the catalytic enzymes. The kinetic data obtained are used to infer information about the relevant transport process; they suggest the potential importance of amino acid efflux as a regulatory step.
Amino Acids↗
Poststimulation catecholamine synthesis and tyrosine hydroxylase activation in central noradrenergic neurons. I. In vivo stimulation of the locus coeruleus.
The ability of increased neuronal activity to accelerate catecholamine biosynthesis and tyrosine hydroxylase activity in the rat brain was tested. Noradrenergic neurons of the locus coeruleus (LC) were stimulated unilaterally at 20 Hz and the cortex and/or hippocampus from stimulated and contralateral (control) sides of the brain were analyzed and compared. Rats were injected with a dopa decarboxylase inhibitor and the accumulation of endogenously synthesized dopa used as an in vivo index of tyrosine hydroxylase activity. Thirty minutes after termination of 15 min of unilateral LC stimulation, dopa accumulation was 35% greater in the ipsilateral cortex + hippocampus. In untreated rats, at the end of 15 min of LC stimulation, there was an ipsilateral depletion of cortical norepinephrine (NE) which recovered within 30 min. When rats were injected with [3H]tyrosine (i.v.) during this half-hour recovery period, a poststimulation increase in [3H]catecholamine synthesis was observed in both the cortex (63%) and hippocampus (55%). In the cortex, there was more newly synthesized [3H]dopamine than [3H]NE, but LC stimulation preferentially increased the synthesis of [3H]NE. The hippocampus contained negligible amounts of [3H]dopamine and was used in subsequent studies. Tyrosine hydroxylase activity was assayed in vitro in supernatants derived from stimulated and control hippocampi. Ten minutes of LC stimulation (20 Hz) maximally activated hippocampal tyrosine hydroxylase and this activation was maintained for up to 20 min after stimulation was terminated. The results illustrate a stimulation-induced activation of NE biosynthesis and tyrosine hydroxylase activity in central NE neurons in vivo. This activation is maintained in the immediate poststimulation period and is not necessarily due to removal of end product inhibition by NE.
Animals↗
Dopamine synthesis by non-dopaminergic neurons expressing individual complementary enzymes of the dopamine synthetic pathway in the arcuate nucleus of fetal rats.
This study was aimed to test our hypothesis about dopamine (DA) synthesis by non-DAergic neurons expressing individual complementary enzymes of the DA synthetic pathway in cooperation, i.e. L-dihydroxyphenylalanine (L-DOPA) synthesized in tyrosine hydroxylase (TH)-expressing neurons is transported to aromatic L-amino acid decarboxylase (AADC)-expressing neurons for conversion to DA. The mediobasal hypothalamus of rats at the 21st embryonic day was used as an experimental model because it contains mainly monoenzymatic TH neurons and AADC neurons (>99%) whereas the fraction of bienzymatic (DAergic) neurons does not exceed 1%. The fetal substantia nigra containing DAergic neurons served as a control. DA and L-DOPA were measured by high performance liquid chromatography in: (1) cell extracts of the cell suspension prepared ex tempora; (2) cell extracts and incubation medium after the static incubation of the cell suspension with, or without exogenous L-tyrosine; (3) effluents of the incubation medium during perifusion of the cell suspension in the presence, or the absence of L-tyrosine. Total amounts of DA and L-DOPA in the incubation medium and cell extracts after the static incubation were considered as the indexes of the rates of their syntheses. L-Tyrosine administration caused the increased L-DOPA synthesis in the mediobasal hypothalamus and substantia nigra. Moreover, L-tyrosine provoked an increase of DA synthesis in the substantia nigra and its decrease in the mediobasal hypothalamus. This contradiction is most probably explained by the L-tyrosine-induced competitive inhibition of the L-DOPA transport to the monoenzymatic AADC-neurons after its release from the monoenzymatic TH neurons. Thus, this study provides convincing evidence of cooperative DA synthesis by non-DAergic neurons expressing TH or AADC in fetal rats at the end of the intrauterine development.
Animals↗
Synthesis of stereo (R and S) and geometric (E and Z) isomers of [18F]fluoro-beta-fluoromethylene-m-tyrosine derivatives: in vivo probes of central dopaminergic function.
Fluorination of pure R and S enantiomers of (E)-beta-fluoromethylene-m-tyrosine [(E)-FMMT] and its racemic geometric isomer, (Z)-beta-fluoromethylene-m-tyrosine [(Z)-FMMT] with [18F]acetyl hypofluorite ([18F]AcOF) gave a mixture of aromatic ring fluorinated products and a pair of diastereomeric products of addition across the exocyclic double bond. Semipreparative high performance liquid chromatography (HPLC) enabled a complete separation and isolation of these products, namely, 6-[18F]fluoro, 2-[18F]fluoro, and 2,6-[18F]difluoro (E)-FMMT and (Z)-FMMT derivatives. No attempt was made to isolate the individual components of the addition product. Pure racemic 4-[18F]fluoro-(E)-beta-fluoromethylene-m-tyrosine was also synthesized from a substituted (E)-FMMT precursor involving a fluorodestannylation reaction with [18F]F2. The availability of stereo (R and S) isomers of 6-[18F]fluoro and 2-[18F]fluoro (E)-FMMT and those of the racemic (Z)-FMMT along with 4-[18F]fluoro-(E)-beta-fluoromethylene-m-tyrosine would now enable a systematic investigation of the central monoamine oxidase/aromatic amino acid decarboxylase enzyme system with positron emission tomography.
Brain↗
Genetic dissection of dopamine and serotonin synthesis in the nervous system of Drosophila melanogaster.
Catecholamine- and serotonin-containing neurons were studied in pale, a Drosophila third-chromosome recessive lethal mutant. Using histofluorescent and immunocytochemical techniques, we show that this mutation only alters catecholamine levels in the CNS. Both the presence of catecholamine-neurons and the expression of serotonin are not affected by the mutation. Furthermore, we show that normal characteristics of catecholamine neurons, such as the presence of the enzyme DOPA decarboxylase and the selective uptake properties are normal in pale mutants. We suggest that pale is either the tyrosine hydroxylase structural gene, or a gene controlling tyrosine hydroxylase activity in Drosophila. The similar genetic location of the putative tyrosine hydroxylase gene and the mutation pale supports the former suggestion.
Animals↗
Tyrosine hydroxylase activation and inactivation by protein phosphorylation conditions.
Tyrosine hydroxylase, the rate-limiting enzyme in catecholamine biosynthesis, catalyzes the conversion of tyrosine to DOPA, Cyclic AMP-dependent protein phosphorylation conditions alter tyrosine hydroxylase activity in rat striatal homogenates. In agreement with other laboratories, we find that short-term pre-incubation (3 min) of extracts under phosphorylating conditions (Mg . ATP, cAMP) increases enzyme activity two- to tenfold over control as measured during a subsequent 15-min assay. We now report that preincubation under phosphorylating conditions for longer periods (30 min) results in a loss of activity to levels equal to or below that of the control enzyme. Addition of purified bovine brain protein kinase catalytic subunit and Mg . ATP enhances activation and increases the rate of inactivation. To demonstrate that inactivation is not associated with proteolytic degradation or irreversible denaturation, the inactivated form of the enzyme can be reactivated. The protein kinase inhibitor protein decreases the activation process and prevents inactivation of the enzyme to below control values. The sedimentation coefficient is not changed by phosphorylation conditions (S = 8.8 +/- 0.1). Although the apparent Km of the enzyme for the 6-methyltetrahydropterine (6-MPH4) cofactor is reduced (0.86 mM, control; 0.32 mM, activated), it is also reduced in the inactivated form (0.38 mM). The Ki for dopamine is increased from 4.5 microM for the control to 28 microM for the activated enzyme, whereas the inactivated form of the enzyme exhibits a Ki of 10 microM. Removal of catecholamines by gel filtration fails to alter activity and the apparent cofactor Km. Moreover, both the activated and the inactivated states persist following gel filtration. It therefore appears that the activation-inactivation process is not mediated solely by the modulation of enzyme feedback inhibition or changes in the Km for 6-MPH4. We also describe a coupled decarboxylase assay in which labeled dopamine is resolved from the precursors tyrosine and DOPA by low-voltage paper electrophoresis.
Adrenal Medulla↗
Neurochemical characteristics of cerebral catecholamine neurons during the postnatal development in the rat.
The development of cerebral dopamine (DA) and noradrenaline (NA) neurons was studied biochemically in 1- to 28-day-old rats. The precursor amino acid tyrosine increased to a steady state level in the whole brain during early postnatal age. After 14 days, however, there was a marked drop in the brain tyrosine level. In general, regional tyrosine levels also decreased between 4 and 28 days. DOPA accumulation was measured in whole brain and regional brain areas after inhibition of L-aromatic amino acid decarboxylase with NSD 1015. This is a well recognized method for estimating the in vivo rate of tyrosine hydroxylase activity. Whole brain DOPA accumulation after NSD 1015 increased progressively during postnatal life, and between 1 and 28 days a more than 3-fold increase was noted. Similar increases were also found in whole brain DA and NA levels during the corresponding period. Regional DA and NA levels generally increased with advancing age, with the highest DA levels in the striatum region and NA levels in the midbrain region. Regional DOPA accumulation after NSD 1015, reflecting in vivo tyrosine hydroxylase activity, increased in a similar fashion as the endogenous neurotransmitter levels. Measurements of DA and NA disappearance after alpha-methyltyrosine indicated the existence of an adult-like nerve impulse activity in DA neurons in the striatum and midbrain regions and in NA neurons in the hemispheres region of the 4-day-old rats. Gammahydroxybutyric acid (GHBA), which causes an inhibition of nerve impulse activity, was given to 4- and 28-day-old rats. This induced an initial increase in CA synthesis as measured by the NSD 1015 method at both ages. The results from this investigation show that the age-related regional increases in endogenous DA and NA levels closely follow the increase in regional tyrosine hydroxylase activity measured in vivo. Furthermore, the study also supports the view that feed-back mechanisms, resulting in an increased transmitter synthesis, exist in DA neurons after nerve impulse inhibition by GHBA at an early postnatal age.
Age Factors↗