Differential Regulation of Drosophila Tyrosine Hydroxylase Isoforms by Dopamine Binding and cAMP-dependent Phosphorylation

Vié, Agnès; Cigna, Mireille; Toci, René; Birman, Serge

doi:10.1074/jbc.274.24.16788

Cited by 53 publications

(30 citation statements)

References 59 publications

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“…S3). Since the DTH protein is subject to feedback inhibition by DA (Vié et al 1999), low level expression of DTH1 in a DA-deficient neuron should result in a highly active/phosphorylated DTH1 protein that would remain fully active until intracellular DA levels approach normal. We tested if DTH phosphorylation, potentially modifying the epitope recognized by the antibody, could be responsible for the lack of TH IR in TH-GAL4/DDC-Gal4:DTH1m rescued flies, by increasing brain DA levels with L-DOPA/carbidopa feeding (Fig S3).…”

Section: Resultsmentioning

confidence: 99%

A new brain dopamine‐deficient Drosophila and its pharmacological and genetic rescue

Cichewicz

Garren

Adiele

et al. 2016

Genes Brain and Behavior

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Dopamine (DA) is a neurotransmitter with conserved behavioral roles between invertebrate and vertebrate animals. In addition to its neural functions, in insects DA is a critical substrate for cuticle pigmentation and hardening. Drosophila tyrosine hydroxylase (DTH) is the rate limiting enzyme for DA biosynthesis. Viable brain DA deficient flies were previously generated using tissue selective GAL4-UAS binary expression rescue of a DTH null mutation and these flies show specific behavioral impairments. To circumvent the limitations of rescue via binary expression, here we achieve rescue utilizing genomically integrated mutant DTH. As expected, our DA deficient flies have no detectable DTH or DA in the brain, and show reduced locomotor activity. This deficit can be rescued by L-DOPA/carbidopa feeding, similar to human Parkinson’s disease treatment. Genetic rescue via GAL4/UAS-DTH was also successful, although this required the generation of a new UAS-DTH1 transgene devoid of most untranslated regions, since existing UAS-DTH transgenes express in the brain without a Gal4 driver via endogenous regulatory elements. A surprising finding of our newly constructed UAS-DTH1m is that it expresses DTH at an undetectable level when regulated by dopaminergic GAL4 drivers even when fully rescuing DA, indicating that DTH immunostaining is not necessarily a valid marker for DA expression. This finding necessitated optimizing DA immunohistochemistry, revealing details of DA innervation to the mushroom body and the central complex. When DA rescue is limited to specific DA neurons, DA does not diffuse beyond the DTH-expressing terminals, such that DA signaling can be limited to very specific brain regions.

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Section: Resultsmentioning

confidence: 99%

A new brain dopamine‐deficient Drosophila and its pharmacological and genetic rescue

Cichewicz

Garren

Adiele

et al. 2016

Genes Brain and Behavior

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“…Considering all our results and the findings in Drosophila, we suggest that BmTh is not only a lethal gene but also a temperature-regulated gene. In Drosophila, both isoforms of TH have their highest enzymatic activity at 25 to 29°C and are only 30% active at 37°C (40). Candidate heat-shock protein (HSP) binding sites were predicted within the 70-kb upstream region of BmTh in the wild-type strain p50T using TFSEARCH (41).…”

Section: Discussionmentioning

confidence: 99%

Repression of tyrosine hydroxylase is responsible for the sex-linked chocolate mutation of the silkworm, Bombyx mori

Lei

Yamamoto

Cheng

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Pigmentation patterning has long interested biologists, integrating topics in ecology, development, genetics, and physiology. Wild-type neonatal larvae of the silkworm, Bombyx mori , are completely black. By contrast, the epidermis and head of larvae of the homozygous recessive sex-linked chocolate ( sch ) mutant are reddish brown. When incubated at 30 °C, mutants with the sch allele fail to hatch; moreover, homozygous mutants carrying the allele sch lethal ( sch l ) do not hatch even at room temperature (25 °C). By positional cloning, we narrowed a region containing sch to 239,622 bp on chromosome 1 using 4,501 backcross (BC1) individuals. Based on expression analyses, the best sch candidate gene was shown to be tyrosine hydroxylase ( BmTh ). BmTh coding sequences were identical among sch , sch l , and wild-type. However, in sch the ∼70-kb sequence was replaced with ∼4.6 kb of a Tc1-mariner type transposon located ∼6 kb upstream of BmTh , and in sch l , a large fragment of an L1Bm retrotransposon was inserted just in front of the transcription start site of BmTh . In both cases, we observed a drastic reduction of BmTh expression. Use of RNAi with BmTh prevented pigmentation and hatching, and feeding of a tyrosine hydroxylase inhibitor also suppressed larval pigmentation in the wild-type strain, pnd + and in a pS (black-striped) heterozygote. Feeding L-dopa to sch neonate larvae rescued the mutant phenotype from chocolate to black. Our results indicate the BmTh gene is responsible for the sch mutation, which plays an important role in melanin synthesis producing neonatal larval color.

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“…Alternative splicing of Drosophila TH (DTH) produces two enzyme isoforms, DTH1 and DTH2 (20,21) (Fig. S1B).…”

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confidence: 99%

“…S1A). Because DA is also required in Drosophila as a precursor substrate for cuticle sclerotization and melanization, inactivating mutations of the genomic TH, alias pale (ple) locus, results in unpigmented cuticle and late embryonic lethality (19).Alternative splicing of Drosophila TH (DTH) produces two enzyme isoforms, DTH1 and DTH2 (20,21) (Fig. S1B).…”

mentioning

confidence: 99%

Behavioral consequences of dopamine deficiency in the Drosophila central nervous system

Riemensperger

Isabel

Coulom

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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The neuromodulatory function of dopamine (DA) is an inherent feature of nervous systems of all animals. To learn more about the function of neural DA in Drosophila, we generated mutant flies that lack tyrosine hydroxylase, and thus DA biosynthesis, selectively in the nervous system. We found that DA is absent or below detection limits in the adult brain of these flies. Despite this, they have a lifespan similar to WT flies. These mutants show reduced activity, extended sleep time, locomotor deficits that increase with age, and they are hypophagic. Whereas odor and electrical shock avoidance are not affected, aversive olfactory learning is abolished. Instead, DA-deficient flies have an apparently "masochistic" tendency to prefer the shock-associated odor 2 h after conditioning. Similarly, sugar preference is absent, whereas sugar stimulation of foreleg taste neurons induces normal proboscis extension. Feeding the DA precursor L-DOPA to adults substantially rescues the learning deficit as well as other impaired behaviors that were tested. DA-deficient flies are also defective in positive phototaxis, without alteration in visual perception and optomotor response. Surprisingly, visual tracking is largely maintained, and these mutants still possess an efficient spatial orientation memory. Our findings show that flies can perform complex brain functions in the absence of neural DA, whereas specific behaviors involving, in particular, arousal and choice require normal levels of this neuromodulator.neurotransmitters | locomotor activity | memory formation | choice behavior | feeding behavior A n important challenge in neuroscience is to understand the roles of specific neurotransmitter systems on brain homeostasis and functioning. Dopamine (DA), a biogenic amine biosynthesized from tyrosine, is an essential neuromodulator in the mammalian central nervous system that is involved in attention, movement control, motivation, and cognition. Studies in Drosophila melanogaster indicate that DA also plays central regulatory roles in insects, specifically in the neural networks controlling locomotor activity and stereotypical behaviors (1-3), sleep and arousal (4-7), registration of salient stimuli (4,8,9), and associative olfactory learning (10-15). Some of these studies were based on genetic inactivation or overactivation of dopaminergic neurons. Dopaminergic neurons can corelease other neuroactive agents, such as neuropeptides, however. Therefore, one must ensure that the behavioral phenotypes observed specifically result from the lack of DA release to draw firm conclusions on brain DA function.Nearly all neuropil regions of the insect CNS receive dense dopaminergic innervation. In particular, the Drosophila adult brain contains six paired clusters of dopaminergic neurons, some of which specifically project to higher brain centers, such as the central complex and the mushroom bodies (1,10,12,13,(16)(17)(18). Tyrosine hydroxylase (TH) catalyzes the first and rate-limiting step in DA biosynthesis (Fig. S1A). Because DA is also ...

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Differential Regulation of Drosophila Tyrosine Hydroxylase Isoforms by Dopamine Binding and cAMP-dependent Phosphorylation

Cited by 53 publications

References 59 publications

A new brain dopamine‐deficient Drosophila and its pharmacological and genetic rescue

A new brain dopamine‐deficient Drosophila and its pharmacological and genetic rescue

Repression of tyrosine hydroxylase is responsible for the sex-linked chocolate mutation of the silkworm, Bombyx mori

Behavioral consequences of dopamine deficiency in the Drosophila central nervous system

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