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The isolation of an o-diphenal oxidase from third-instar larvae of the blowfly Calliphora erythrocephala.

ABSTRACT: 1. The isolation of an o-diphenol oxidase from an acetone-dried powder of late-third-instar larvae of Calliphora erythrocephala was investigated. An insoluble and micro-crystalline fraction containing the enzyme activity was obtained after fractionating extracts of the acetone-dried powder with (NH4)2SO4 and acetone. 2. This fraction can be solubilized in 0.1% sodium dodecyl sulphate without loss of activity. 3. Polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate shows that the o-diphenol oxidase is a minor component of the extracts from the acetone-dried powder. 4. The o-diphenol oxidase was purified by zonal centrifugation on a sucrose density gradient in the presence of sodium dodecyl sulphate. 5. The amino acid composition of the purified enzyme resembles that of some other o-diphenol oxidases. 6. The subunit composition of the o-diphenol oxidase is discussed.

SUBMITTER: Pau RN

PROVIDER: S-EPMC1165677 | biostudies-other | 1975 Sep

REPOSITORIES: biostudies-other

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The hydroxylation of tyrosine by an enzyme from third-instar larvae of the blowfly Calliphora erythrocephala.

Project description:1. Two pro-(phenol oxidase) were distinguished when the blood of late-third-instar larvae of Calliphora erythrocephala was electrophoresed in polyacrylamide gels with Tris-glycine buffer, pH 8.3. One pro-(phenol oxidase), after activation by an enzyme readily catalyses the oxidation of both L-tyrosine and L-3,4-dihydroxyphenylalanine (L-dopa). The second enzyme catalyses the oxidation of L-dopa but not of L-tyrosoine. 2. One of the pro-(phenol oxidases) was purified over 2000-fold from homogenates of whole larvae. This enzyme, after activation, catalyses the oxidation of both dopa and tyrosine. On electrophoresis in polyacrylamide gels with Tris-glycine buffer, pH 8.3, it has the same mobility as the enzyme in the blood which catalyses the oxidation of both tyrosine and dopa. 3. The pro-(phenol oxidase)-activating enzyme was purified over 100-fold from homogenates of whole larvae. 4. The oxidation of L-tyrosine, in the presence of the activated purified phenol oxidase, reached a steady maximum rate after a lag period that was directly related to tyrosine concentration and inversely related to enzyme concentration. 5. The effect of the addition of electron donors on the lag period was studied. Dopa, dopamine (3,4-dihydroxyphenethylamine) and 2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine are the most effective hydrogen donors. 3,4-Dihydroxybenzoic acid, the oxidation of which was not catalysed by the activated pro-(phenol oxidase), did not affect the lag period.

| S-EPMC1165484 | biostudies-other

Chemosensory genes identified in the antennal transcriptome of the blowfly Calliphora stygia.

Project description:BackgroundBlowflies have relevance in areas of forensic science, agriculture, and medicine, primarily due to the ability of their larvae to develop on flesh. While it is widely accepted that blowflies rely heavily on olfaction for identifying and locating hosts, there is limited research regarding the underlying molecular mechanisms. Using next generation sequencing (Illumina), this research examined the antennal transcriptome of Calliphora stygia (Fabricius) (Diptera: Calliphoridae) to identify members of the major chemosensory gene families necessary for olfaction.ResultsRepresentative proteins from all chemosensory gene families essential in insect olfaction were identified in the antennae of the blowfly C. stygia, including 50 odorant receptors, 22 ionotropic receptors, 21 gustatory receptors, 28 odorant binding proteins, 4 chemosensory proteins, and 3 sensory neuron membrane proteins. A total of 97 candidate cytochrome P450s and 39 esterases, some of which may act as odorant degrading enzymes, were also identified. Importantly, co-receptors necessary for the proper function of ligand-binding receptors were identified. Putative orthologues for the conserved antennal ionotropic receptors and candidate gustatory receptors for carbon dioxide detection were also amongst the identified proteins.ConclusionsThis research provides a comprehensive novel resource that will be fundamental for future studies regarding blowfly olfaction. Such information presents potential benefits to the forensic, pest control, and medical areas, and could assist in the understanding of insecticide resistance and targeted control through cross-species comparisons.

| S-EPMC4392625 | biostudies-literature

16S rRNA Gene Amplicon Profiling of the New Zealand Parasitic Blowfly Calliphora vicina.

Project description:Here, we present a 16S rRNA gene amplicon sequence data set and profiles demonstrating the bacterial diversity of larval and adult Calliphora vicina, collected from Ashhurst, New Zealand (May 2020). The three dominant genera among the adult male and female C. vicina blowflies were Serratia and Morganella (phylum Proteobacteria) and Carnobacterium (phylum Firmicutes), while the larvae were also dominated by the genera Lactobacillus (phylum Firmicutes).

| S-EPMC8103871 | biostudies-literature

The complete mitochondrial genome of a blowfly <i>Calliphora nigribarbis</i> (Vollenhoven, 1863) (Insecta: Diptera: Calliphoridae).

Project description:In the present study, the complete mitochondrial genome of a blowfly Calliphora nigribarbis has been sequenced and analyzed. The length of complete the mitochondrial genome is 16,279?bp, with 39.50% A, 13.20% C, 9.30% G, and 38.0% T nucleotide distribution. The complete mitochondrial genome consists of 13 protein-coding genes, 22 transfer RNAs, and 2 ribosomal RNAs likewise the most metazoan mitochondrial genomes. Furthermore, phylogenetic relationships of C. nigribarbis in the subfamily Calliphorinae investigated. The results suggested that C. vomitoria is the most related species to C. nigribarbis and the genus Calliphora is not monophyletic. This study provides the first complete mitochondrial genome sequence for the species.

| S-EPMC7687389 | biostudies-literature

Impact of walking speed and motion adaptation on optokinetic nystagmus-like head movements in the blowfly Calliphora.

Project description:The optokinetic nystagmus is a gaze-stabilizing mechanism reducing motion blur by rapid eye rotations against the direction of visual motion, followed by slower syndirectional eye movements minimizing retinal slip speed. Flies control their gaze through head turns controlled by neck motor neurons receiving input directly, or via descending neurons, from well-characterized directional-selective interneurons sensitive to visual wide-field motion. Locomotion increases the gain and speed sensitivity of these interneurons, while visual motion adaptation in walking animals has the opposite effects. To find out whether flies perform an optokinetic nystagmus, and how it may be affected by locomotion and visual motion adaptation, we recorded head movements of blowflies on a trackball stimulated by progressive and rotational visual motion. Flies flexibly responded to rotational stimuli with optokinetic nystagmus-like head movements, independent of their locomotor state. The temporal frequency tuning of these movements, though matching that of the upstream directional-selective interneurons, was only mildly modulated by walking speed or visual motion adaptation. Our results suggest flies flexibly control their gaze to compensate for rotational wide-field motion by a mechanism similar to an optokinetic nystagmus. Surprisingly, the mechanism is less state-dependent than the response properties of directional-selective interneurons providing input to the neck motor system.

| S-EPMC9262929 | biostudies-literature

Isolation of material displaying insulin-like immunological biological activity from the brain of the blowfly Calliphora vomitoria.

Project description:An insulin-like material from the brain of the blowfly Calliphora vomitoria was partially purified by acid alcohol extraction, gel filtration and ion-exchange cellulose chromatography. In addition, the RF value on polyacrylamide-gel electrophoresis was determined. The material was characterized by its ability to cross-react with bovine insulin antibody and by displaying diminished immunoreactivity on dilution. It displaced specifically bound 125I-labelled insulin from rat liver plasma membrane insulin receptors and displayed insulin-like biological activity on the isolated rat fat-cell. Within 30 min of injection into Calliphora, made hypertrehalocaemic and hyperglucaemic as a result of median neurosecretory cell removal, it caused the concentrations of both sugars to return to normal. The hypothesis is put forward that the median neurosecretory cells are the source of the material.

| S-EPMC1161755 | biostudies-other

Isolation and partial characterization of pancreatic polypeptide-like material in the brain of the blowfly Calliphora vomitoria.

Project description:Using 10(6) flies (5 kg of heads) a pancreatic polypeptide-like material has been partially purified from the blowfly Calliphora vomitoria. The isolated material was eluted on Sephadex G-50 similarly to bovine pancreatic polypeptide and had an RF on polyacrylamide-gel electrophoresis that was identical with that of the bovine hormone. The material diluted linearly and showed parallelism with bovine standards in a bovine pancreatic polypeptide immunoassay. In specificity controls the immunoreactivity was not abolished by trasylol and no cross-reactivity was discerned in assay for glucagon, proangiotensin and cyclic AMP. These data suggest that the pancreatic polypeptide material in the brain of the blowfly has close structural similarity to the mammalian hormone.

| S-EPMC1163195 | biostudies-other

Characterization of the complete mitochondrial genome of the New Zealand parasitic blowfly <i>Calliphora vicina</i> (Insecta: Diptera: Calliphoridae).

Project description:In this study, the complete mitochondrial (mt) genome of the New Zealand parasitic blowfly Calliphora vicina (blue bottle blowfly) field strain NZ_CalVic_NP was generated using next-generation sequencing technology and annotated. The 16,518 bp mt genome consists of 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and a 1689 bp non-coding region, similar to the two other available C. vicina and most metazoan mt genomes. Phylogenetic analysis showed that C. vicina NZ_CalVic_NP forms a monophyletic cluster with the remaining three Calliphorinae species. The complete mt genome sequence of C. vicina NZ_CalVic_NP is a resource to facilitate future species- and strain-level identification research and investigations into the evolutionary provenance within the Calliphoridae.

| S-EPMC8008873 | biostudies-literature

Calliphora vicina (blue bottle blowfly) and Lucilia sericata

Project description:Calliphora vicina (blue bottle blowfly) and Lucilia sericata (common green bottle fly) genome sequencing

| PRJNA667961 | ENA

Molecular and pharmacological characterization of serotonin 5-HT2? and 5-HT7 receptors in the salivary glands of the blowfly Calliphora vicina.

Project description:Secretion in blowfly (Calliphora vicina) salivary glands is stimulated by the biogenic amine serotonin (5-hydroxytryptamine, 5-HT), which activates both inositol 1,4,5-trisphosphate (InsP(3))/Ca(2+) and cyclic adenosine 3',5'-monophosphate (cAMP) signalling pathways in the secretory cells. In order to characterize the signal-inducing 5-HT receptors, we cloned two cDNAs (Cv5-ht2?, Cv5-ht7) that share high similarity with mammalian 5-HT(2) and 5-HT(7) receptor genes, respectively. RT-PCR demonstrated that both receptors are expressed in the salivary glands and brain. Stimulation of Cv5-ht2?-transfected mammalian cells with 5-HT elevates cytosolic [Ca(2+)] in a dose-dependent manner (EC(50) = 24 nM). In Cv5-ht7-transfected cells, 5-HT produces a dose-dependent increase in [cAMP](i) (EC(50) = 4 nM). We studied the pharmacological profile for both receptors. Substances that appear to act as specific ligands of either Cv5-HT(2?) or Cv5-HT(7) in the heterologous expression system were also tested in intact blowfly salivary gland preparations. We observed that 5-methoxytryptamine (100 nM) activates only the Cv5-HT(2?) receptor, 5-carboxamidotryptamine (300 nM) activates only the Cv5-HT(7) receptor, and clozapine (1 µM) antagonizes the effects of 5-HT via Cv5-HT(7) in blowfly salivary glands, providing means for the selective activation of each of the two 5-HT receptor subtypes. This study represents the first comprehensive molecular and pharmacological characterization of two 5-HT receptors in the blowfly and permits the analysis of the physiological role of these receptors, even when co-expressed in cells, and of the modes of interaction between the Ca(2+)- and cAMP-signalling cascades.

| S-EPMC3493529 | biostudies-literature

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