The determination of histamine in the Drosophila head

Borycz, J; Vohra, M. M.; Tokarczyk, G; Meinertzhagen, Ian A.

doi:10.1016/s0165-0270(00)00259-4

Cited by 35 publications

(54 citation statements)

References 39 publications

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“…Our initial observations began when we used our methods (Borycz et al, 2000) to undertake routine determinations of head histamine in various Drosophila mutants that had been isolated in a genetic background containing the mutant white gene as a marker. These loss-of-function white-eyed mutants lack filled pigment granules in the eye but were thought otherwise to be normal.…”

Section: Resultsmentioning

confidence: 99%

“…For determinations of brain biogenic amines, flies were collected, frozen at -80°C, shaken to decapitate them, and were then processed either for histamine determinations using high-performance liquid chromatography (HPLC) with electrochemical detection, as previously reported (Borycz et al, 2000) or, in parallel samples, for determinations of both dopamine and 5-HT. We used samples from: 50 heads (Drosophila), five heads (Musca) or one head (Calliphora and Sarcophaga) and between six and 12 samples per reported mean.…”

Section: High-performance Liquid Chromatographymentioning

confidence: 99%

“…The tan peak with the shorter retention time (4 min) is an unknown metabolite. For a detailed comment see Borycz et al (Borycz et al, 2000).…”

Section: Dopamine and Serotonin Content In The Drosophila Headmentioning

confidence: 99%

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DrosophilaABC transporter mutantswhite,brownandscarlethave altered contents and distribution of biogenic amines in the brain

Borycz

Kubów

Lloyd

et al. 2008

Journal of Experimental Biology

147

169

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SUMMARYMonoamines such as dopamine, histamine and serotonin (5-HT) are widely distributed throughout the brain of the fruit fly Drosophila melanogaster, where many of their actions have been investigated. For example, histamine is released from photoreceptor synapses in the lamina neuropile of the visual system. Mutations of the genes white, an important eye pigmentation marker in fly genetics that encodes an ABC transporter, and its binding partner brown, cause neural phenotypes not readily reconciled solely with actions in eye pigmentation. We find that flies mutant for these genes, and another binding partner, scarlet, have about half the wild-type amount of histamine in the head, as well as reduced 5-HT and dopamine. These differences parallel reductions in immunoreactivity to the corresponding biogenic amines. They also correlate with the amine content of fractions after differential centrifugation of head homogenates. Thus, most of the amine is found in the vesicle-rich fraction of wild-type head homogenates, whereas it is found in the supernatant fractions from white, brown and scarlet flies. White co-expresses in lamina epithelial glia with Ebony, which conjugates histamine to β-alanine. Histamine is then released when the conjugate is hydrolyzed in photoreceptors, by Tan. Mutant white ameliorates the effects of tan on head histamine whereas it exacerbates the effects of ebony. Our results are consistent with the proposal that histamine uptake by the epithelial glia may be white dependent. Behavioral abnormalities in white, brown and scarlet mutants could arise because aminergic neurons in the Drosophila brain have reduced amine for release.

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Section: Resultsmentioning

confidence: 99%

Section: High-performance Liquid Chromatographymentioning

confidence: 99%

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DrosophilaABC transporter mutantswhite,brownandscarlethave altered contents and distribution of biogenic amines in the brain

Borycz

Kubów

Lloyd

et al. 2008

Journal of Experimental Biology

147

169

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“…For determinations of brain -alanine and histamine, flies were collected, frozen at -80°C, shaken to decapitate them, and the heads then processed using HPLC with electrochemical detection, all as previously reported (Borycz et al, 2000). In this method, -alanine is clearly separated from neighbouring peaks and has a retention time that is approximately 2min shorter than that for histamine.…”

Section: High-performance Liquid Chromatography (Hplc)mentioning

confidence: 99%

The metabolism of histamine in theDrosophilaoptic lobe involves an ommatidial pathway: β-alanine recycles through the retina

Borycz

Edwards

Boulianne

et al. 2012

Journal of Experimental Biology

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SUMMARYFlies recycle the photoreceptor neurotransmitter histamine by conjugating it to -alanine to form -alanyl-histamine (carcinine). The conjugation is regulated by Ebony, while Tan hydrolyses carcinine, releasing histamine and -alanine. In Drosophila, -alanine synthesis occurs either from uracil or from the decarboxylation of aspartate but detailed roles for the enzymes responsible remain unclear. Immunohistochemically detected -alanine is present throughout the flyʼs entire brain, and is enhanced in the retina especially in the pseudocone, pigment and photoreceptor cells of the ommatidia. HPLC determinations reveal 10.7ng of -alanine in the wild-type head, roughly five times more than histamine. When wild-type flies drink uracil their head -alanine increases more than after drinking L-aspartic acid, indicating the effectiveness of the uracil pathway. Mutants of black, which lack aspartate decarboxylase, cannot synthesize -alanine from L-aspartate but can still synthesize it efficiently from uracil. Our findings demonstrate a novel function for pigment cells, which not only screen ommatidia from stray light but also store and transport -alanine and carcinine. This role is consistent with a -alanine-dependent histamine recycling pathway occurring not only in the photoreceptor terminals in the lamina neuropile, where carcinine occurs in marginal glia, but vertically via a long pathway that involves the retina. The laminaʼs marginal glia are also a hub involved in the storage and/or disposal of carcinine and -alanine.

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“…In all cases, histamine determinations were performed on groups of 20-50 isolated heads using HPLC with electrochemical detection as reported for D. melanogaster [3,13,46], and values are reported for the means of 3-12 such samples.…”

Section: Methodsmentioning

confidence: 99%

Drosophila tan encodes a novel hydrolase required in pigmentation and vision

et al. 2005

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Many proteins are used repeatedly in development, but usually the function of the protein is similar in the different contexts. Here we report that the classical Drosophila melanogaster locus tan encodes a novel enzyme required for two very different cellular functions: hydrolysis of N-b-alanyl dopamine (NBAD) to dopamine during cuticular melanization, and hydrolysis of carcinine to histamine in the metabolism of photoreceptor neurotransmitter. We characterized two tan-like P-element insertions that failed to complement classical tan mutations. Both are inserted in the 59 untranslated region of the previously uncharacterized gene CG12120, a putative homolog of fungal isopenicillin-N N-acyltransferase (EC 2.3.1.164). Both P insertions showed abnormally low transcription of the CG12120 mRNA. Ectopic CG12120 expression rescued tan mutant pigmentation phenotypes and caused the production of striking black melanin patterns. Electroretinogram and head histamine assays indicated that CG12120 is required for hydrolysis of carcinine to histamine, which is required for histaminergic neurotransmission. Recombinant CG12120 protein efficiently hydrolyzed both NBAD to dopamine and carcinine to histamine. We conclude that D. melanogaster CG12120 corresponds to tan. This is, to our knowledge, the first molecular genetic characterization of NBAD hydrolase and carcinine hydrolase activity in any organism and is central to the understanding of pigmentation and photoreceptor function.

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The determination of histamine in the Drosophila head

Cited by 35 publications

References 39 publications

DrosophilaABC transporter mutantswhite,brownandscarlethave altered contents and distribution of biogenic amines in the brain

DrosophilaABC transporter mutantswhite,brownandscarlethave altered contents and distribution of biogenic amines in the brain

The metabolism of histamine in theDrosophilaoptic lobe involves an ommatidial pathway: β-alanine recycles through the retina

Drosophila tan encodes a novel hydrolase required in pigmentation and vision

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