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At least 523 records · Page 29Linked to original sources

PubMed2007

Towards abolition of immunogenic structures in insect cells: characterization of a honey-bee (Apis mellifera) multi-gene family reveals both an allergy-related core alpha1,3-fucosyltransferase and the first insect Lewis-histo-blood-group-related antigen-synthesizing enzyme.

Glycoproteins from honey-bee (Apis mellifera), such as phospholipase A2 and hyaluronidase, are well-known major bee-venom allergens. They carry N-linked oligosaccharide structures with two types of alpha1,3-fucosylation: the modification by alpha1,3-fucose of the innermost core GlcNAc, which constitutes an epitope recognized by IgE from some bee-venom-allergic patients, and an antennal Lewis-like GalNAcbeta1,4(Fucalpha1,3)GlcNAc moiety. We now report the cloning and expression of two cDNAs encoding the relevant active alpha1,3-FucTs (alpha1,3-fucosyltransferases). The first sequence, closest to that of fruitfly (Drosophila melanogaster) FucTA, was found to be a core alpha1,3-FucT (EC 2.4.1.214), as judged by several enzyme and biochemical assays. The second cDNA encoded an enzyme, most related to Drosophila FucTC, that was shown to be capable of generating the Le(x) [Galbeta1-4(Fucalpha1-3)GlcNAc] epitope in vitro and is the first Lewis-type alpha1,3-FucT (EC 2.4.1.152) to be described in insects. The transcription levels of these two genes in various tissues were examined: FucTA was found to be predominantly expressed in the brain tissue and venom glands, whereas FucTC transcripts were detected at highest levels in venom and hypopharyngeal glands. Very low expression of a third homologue of unknown function, FucTB, was also observed in various tissues. The characterization of these honey-bee gene products not only accounts for the observed alpha1,3-fucosylation of bee-venom glycoproteins, but is expected to aid the identification and subsequent down-regulation of the FucTs in insect cell lines of biotechnological importance.

Allergens↗

PubMed1992

The antigenicity of the carbohydrate moiety of an insect glycoprotein, honey-bee (Apis mellifera) venom phospholipase A2. The role of alpha 1,3-fucosylation of the asparagine-bound N-acetylglucosamine.

A rabbit polyclonal antiserum raised against honey-bee (Apis mellifera) venom phospholipase A2 (PLA2) contains antibodies that react exclusively with its glycosylated variants and cross-react with plant glycoproteins. The interaction of anti-(horseradish peroxidase) antiserum with PLA2 suggests the existence of a carbohydrate determinant common to both glycoproteins. E.l.i.s.a. binding and inhibition experiments, employing glycoproteins and glycopeptides of plant and animal origin with known N-glycan structures, in combination with chemical and enzymic deglycosylation, identified alpha 1,3-fucosylation of the asparagine-bound N-acetylglucosamine as the antigenic determinant. This fucose residue is present in the N-glycan of PLA2 and is frequently found in plant glycoproteins, whereas mammalian glycoproteins lack this modification.

Acetylglucosamine↗

PubMed1999

Phylogeographical autocorrelation of phenotypic evolution in honey bees (Apis mellifera L.).

The analysis of phenotypic divergence among local populations within a species has been traditionally performed in a spatial context, although advances in genetic analysis using mtDNA have permitted a simultaneous evaluation of geographical and historical patterns of variation, so-called phylogeographical analysis. In this paper, we combine these two dimensions of variation (geographical space and phylogenetic history) to evaluate patterns of phenotypic evolution in honey bees (Apis mellifera L.). Data on 39 phenotypic traits, derived from 417 colonies grouped into 14 subspecies, were analysed using autocorrelation methods. Mantel tests indicated that the relationship between phenotypic divergence, estimated by Euclidean distances among subspecies' morphological centroids, was significant both when compared to geographical distance (r=0.371; P < 0.01) and to genetic distance (estimated as sequence divergence (%) in a mtDNA region encompassing part of the NADH dehydrogenase subunit 2 and isoleucine transfer RNA (r=0.329; P < 0.01)). For the analysis of each trait, the effects of the geographical co-ordinates (latitude and longitude of subspecies geographical range) and of the phylogenetic patterns (defined by eigenvectors of the genetic distance matrix) on phenotypic variation were simultaneously analysed using an extension of a recently developed model, called Phylogenetic Eigenvector Regression (PVR). In general terms, the partial regression slopes indicated that the variation in the characters traditionally associated with adaptive processes, such as body and wing size, were better explained by geographical position. However, characters usually thought to be neutral, such as wing venation angle, were more associated with phylogeny. This is expected because PVR can be interpreted as a partition model, in which adaptive variation tends to be independent of phylogeny (and, in this case, associated with geography). In addition, the first principal component derived from the expected values of the model for each trait, which can be interpreted as the phenotypic variation predicted by phylogeny, is more structured in a north-south cline than are the original data, supporting an adaptive interpretation. The phylogeographical autocorrelation analyses performed in this study show that different traits are more related to one of the two dimensions of variation (geography and phylogeny), and these patterns can furnish insights into the nature of phenotypic evolution in these organisms.

Animals↗

PubMed2000

MtDNA variation in Apis cerana populations from the Philippines.

The cavity-nesting honeybee Apis cerana occurs in Asia, from Afghanistan to China and from Japan to southern Indonesia. Based on morphometric values, this species can be grouped into four subspecies: A. c. cerana, A. c. indica, A. c. japonica and A. c. himalaya. In order to analyse the geographical variability of A. c. indica from the Philippine Islands, 47 colonies from different locations in three of the larger islands (Mindanao, Luzon and Palawan) and four of the Visayan Islands (Panay, Negros, Cebu and Leyte) were studied. Genetic variation was estimated by restriction and sequence analysis of PCR-amplified fragments of the tRNAleu-COII region. We found four different haplotypes, Ce1, Ce2, Ce3 and Ce4, that discriminate among the bee populations from different islands. The Ce1 haplotype is present in Mindanao and Visayan Islands, Ce2 is restricted to Luzon, and both Ce3 and Ce4 are only present in Palawan. Phylogenetic analysis of the sequences shows a great intraspecific variability, is in accordance with the geological history of these islands and partially agrees with some previous morphological and molecular studies.

Animals↗

PubMed2000

Sperm-mediated transformation of the honey bee, Apis mellifera.

Our primary objective was to identify techniques to transform the genome of the honey bee (Apis mellifera) with foreign DNA constructs. The strategy we adopted was to linearize foreign DNA and introduce it with sperm during the instrumental insemination of virgin queen honey bees. We analysed extracts from larvae within the same cohort and isolated the predicted fragment by means of PCR amplification of genomic DNA. Larvae that carried the construct also expressed the introduced DNA. We propagated several transgenic lines for up to three generations, which demonstrates its heritability. Once carried by a queen, the construct can be detected in that queen's larvae over several months. However, there was no evidence of integration of the construct, at least as determined by genomic Southern analysis. Nevertheless, this demonstrates the general viability of the technique for introduction of DNA, and it should be augmented by further use of transposable elements that enhance integration.

Animals↗

PubMed2003

Identification and localization of the nicotinic acetylcholine receptor alpha3 mRNA in the brain of the honeybee, Apis mellifera.

The nicotinic acetylcholine receptors are ligand-gated ion channels responsible for rapid neurotransmission and are target sites for pesticides in insects. In the honeybee Apis mellifera, pharmacological and electrophysiological studies have shown that different nicotinic acetylcholine receptor subtypes may exist in the brain. Here, we have identified a honeybee cDNA that encodes a 537 amino acid protein with features typical of nicotinic acetylcholine receptor alpha subunit, and sequence homology to human alpha3. In situ hybridization on cryosections shows that the Apisalpha3 mRNA is differently expressed in larvae and adult. In larvae, Apisalpha3 mRNA expression is restricted to the suboesophageal ganglia. In adult, it is further expressed in the optic lobes, the dorsal lobes, the antennal lobes and the calyces of mushroom bodies. Together our results suggest that Apisalpha3 shows a controlled expression pattern during development.

Amino Acid Sequence↗

PubMed2003

Antibacterial activity of honey and propolis from Apis mellifera and Tetragonisca angustula against Staphylococcus aureus.

AIMS: The antibacterial activity against Staphylococcus aureus of honey and propolis produced by Apis mellifera and Tetragonisca angustula was evaluated. Secondary aims included the study of the chemical composition of propolis and honey samples and its relationship with antibacterial activity against S. aureus. METHODS AND RESULTS: The antibacterial activity of honey and propolis was determined by the method of macrodilution. The minimum inhibitory concentration (MICs) of A. mellifera honey ranged from 126.23 to 185.70 mg ml(-1) and of T. angustula from 142.87 to 214.33 mg ml(-1). For propolis, the MIC ranged from 0.36 to 3.65 mg ml(-1) (A. mellifera) and from 0.44 to 2.01 mg ml(-1) (T. angustula). Honey and propolis were evaluated by high-performance liquid chromatography. Some typical compounds of Brazilian propolis were also identified in honey samples. Principal component analysis revealed that the chemical composition of honey and propolis samples was distinct based on the geographical location of the samples. CONCLUSIONS: Propolis samples had higher antibacterial activity against S. aureus when compared with honey. However, both propolis and honey samples had antibacterial against S. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: These antimicrobial properties would warrant further studies on the clinical applications of propolis and honey against S. aureus.

Animals↗

PubMed1998

Mitochondrial DNA variability in the Canary Islands honeybees (Apis mellifera L.).

The mitochondrial DNA (mtDNA) of individuals from 79 colonies of Apis mellifera from five Canary Islands was studied using the DraI test based on the restriction of PCR products of the tRNA(leu)-COII intergenic region. Five haplotypes of the African (A) lineage and one of the west European (C) lineage were found. The haplotypes A14 and A15 are described for the first time. These haplotypes have a new P sequence named P1. The wide distribution and high frequency of haplotype A15 suggest that it is characteristic of the Canarian Archipelago. Sources of haplotype variability of honeybee mtDNA in the Canary Islands (waves of colonization from Africa, queen importations, habitat diversification) are discussed.

Animals↗

PubMed2000

Hybrid origins of honeybees from italy (Apis mellifera ligustica) and sicily (A. m. sicula).

The genetic variability of honeybee populations Apis mellifera ligustica, in continental Italy, and of A. m. sicula, in Sicily, was investigated using nuclear (microsatellite) and mitochondrial markers. Six populations (236 individual bees) and 17 populations (664 colonies) were, respectively, analysed using eight microsatellite loci and DraI restriction fragment length polymorphism (RFLP) of the cytochrome oxidase I (COI)-cytochrome oxidase II (COII) region. Microsatellite loci globally confirmed the southeastern European heritage of both subspecies (evolutionary branch C). However, A. m. ligustica mitochondrial DNA (mtDNA) appeared to be a composite of the two European (M and C) lineages over most of the Italian peninsula, and only mitotypes from the African (A) lineage were found in A. m. sicula samples. This demonstrates a hybrid origin for both subspecies. For A. m. ligustica, the most widely exported subspecies, this hybrid origin has long been obscured by the fact that in the main area of queen production (from which most of the previous ligustica bee samples originated) the M mitochondrial lineage is absent, whereas it is present almost everywhere else in Italy. This presents a new view of the evolutionary history of European honeybees. For instance, the Iberian peninsula was considered as the unique refuge for the M branch during the quaternary ice periods. Our results show that the Apennine peninsula played a similar role. The differential distribution of nuclear and mitochondrial markers observed in Italy seems to be a general feature of introgressed honeybee populations. Presumably, it stems from the social nature of the species in which both genome compartments are differentially affected by the two (individual and colonial) reproduction levels.

Alleles↗

PubMed2001

Origin of honeybees (Apis mellifera L.) from the Yucatan peninsula inferred from mitochondrial DNA analysis.

Honeybees (Apis mellifera L.) sampled at sites in Europe, Africa and South America were analysed using a mitochondrial DNA restriction fragment length polymorphism (RFLP) marker. These samples were used to provide baseline information for a detailed analysis of the process of Africanization of bees from the neotropical Yucatan peninsula of Mexico. Radical changes in mitochondrial haplotype (mitotype) frequencies were found to have occurred in the 13-year period studied. Prior to the arrival of Africanized bees (1986) the original inhabitants of the Yucatan peninsula appear to have been essentially of southeastern European origin with a smaller proportion having northwestern European ancestry. Three years after the migration of Africanized bees into the area (1989), only very low levels of maternal gene flow from Africanized populations into the resident European populations had occurred. By 1998, however, there was a sizeable increase in the proportion of African mitotypes in domestic populations (61%) with feral populations having 87% of mitotypes classified as African derived. The results suggest that the early stages of Africanization did not involve a rapid replacement of European with African mitotypes and that earlier studies probably overestimated the prevalence of African mitotypes.

Africa↗

PubMed2001

Genetic structure and distinctness of Apis mellifera L. populations from the Canary Islands.

The genetic structure of Apis mellifera populations from the Canary Islands has been assessed by mitochondrial (restriction fragment length polymorphisms of the intergenic transfer RNAleu-COII region) and nuclear (microsatellites) studies. These populations show a low level of genetic variation in terms of average number of alleles and degree of heterozygosity. Significant differences in the distribution of alleles were found in both data sets, confirming the genetic differentiation among some of the islands but not within them. Two mitochondrial haplotypes characteristic of the Canary Islands are found at high frequencies, although populations are introgressed by imported honeybees of eastern European C lineage. This introgression is rather high on Tenerife and El Hierro and low on Gran Canaria and La Gomera, whereas on La Palma it has not been recorded. The finding of microsatellite alleles characteristic of the eastern European lineage corroborates the genetic introgression. Phylogenetic analyses indicate that the Canarian honeybees are differentiated from other lineages and provide genetic evidence of their African origin.

Alleles↗

PubMed2003

Male fitness of honeybee colonies (Apis mellifera L.).

Honeybees (Apis mellifera L.) have an extreme polyandrous mating system. Worker offspring of 19 naturally mated queens was genotyped with DNA microsatellites, to estimate male reproductive success of 16 drone producing colonies. This allowed for estimating the male mating success on both the colony level and the level of individual drones. The experiment was conducted in a closed population on an isolated island to exclude interferences of drones from unknown colonies. Although all colonies had produced similar numbers of drones, differences among the colonies in male mating success exceeded one order of magnitude. These differences were enhanced by the siring success of individual drones within the offspring of mated queens. The siring success of individual drones was correlated with the mating frequency at the colony level. Thus more successful colonies not only produced drones with a higher chance of mating, but also with a significantly higher proportion of offspring sired than drones from less successful colonies. Although the life cycle of honeybee colonies is very female centred, the male reproductive success appears to be a major driver of natural selection in honeybees.

Animals↗

PubMed1999

Primary structure and properties of the cathepsin G/chymotrypsin inhibitor from the larval hemolymph of Apis mellifera.

A member of the Ascaris inhibitor family exhibiting anti-cathepsin G and anti-chymotrypsin activity was purified from the larval hemolymph of the honey bee (Apis mellifera). Three forms of the inhibitor, designated AMCI 1-3, were isolated using gel filtration and anion-exchange chromatographies followed by reverse-phase HPLC. The amino-acid analyses indicated that AMCI-1 and AMCI-2 have an identical composition whereas AMCI-3 is shorter by two residues (Thr, Arg). All three forms contain as many as 10 cysteine residues and lack tryptophan, tyrosine, and histidine. The sequence of the isoinhibitors showed that the major form (AMCI-1) consisting of 56 amino-acid residues was a single-chain protein of molecular mass 5972 Da, whereas the other two forms were two-chain proteins with a very high residue identity. The AMCI-2 appeared to be derived from AMCI-1, as a result of the Lys24-Thr25 peptide bond splitting, while AMCI-3 was truncated at its N-terminus by the dipeptide Thr25-Arg26. The association constants for the binding of bovine alpha-chymotrypsin to all purified forms of the inhibitor were high and nearly identical, ranging from 4.8 x 10(10) M-1 for AMCI-1 to 2.7 x 10(9) M-1 for AMCI-3. The sensitivity of cathepsin G to inhibition by each inhibitor was different. Only the association constant for the interaction of this enzyme with AMCI-1 was high (2 x 10(8) M-1) whereas those for AMCI-2 and AMCI-3 were significantly lower, and appeared to be 3.7 x 10(7) M-1 and 4.5 x 10(6) M-1, respectively. The reactive site of the inhibitor, as identified by cathepsin G degradation and chemical modification, was found to be at Met30-Gln31. A search in the Protein Sequence Swiss-Prot databank revealed a significant degree of identity (44%) between the primary structure of AMCI and the trypsin isoinhibitor from Ascaris sp (ATI). On the basis of the cysteine residues alignment, the position of the reactive site as well as some sequence homology, the cathepsin G/chymotrypsin inhibitor from larval hemolymph of the honey bee may be considered to be a member of the Ascaris inhibitor family.

Amino Acid Sequence↗

PubMed2003

The expression of genes encoding visual components is regulated by a circadian clock, light environment and age in the honeybee (Apis mellifera).

The honeybee, Apis mellifera, has been used as a model to study the development of the visual system and adult bee behaviour. However, the regulation of the levels of visual component genes has never been addressed in this organism. We isolated honeybee cDNAs encoding green-sensitive opsin and visual arrestin and then measured their mRNA levels in honeybee workers. Both mRNAs fluctuate on a daily cycle that depends on a pacemaker that functions separately from the pacemaker which controls rhythmic locomotor activity. The cycling-patterns of opsin and arrestin mRNAs are different from each other and are modified by light. Furthermore, light exposure can increase the absolute levels of both mRNAs and the arrestin mRNA level is also dependent on age. Consistent with these results, both mRNA levels are higher in foragers than in in-hive bees under natural conditions. This study thus shows that the expression of genes encoding visual components is regulated by multiple factors and is adjusted to the honeybees' need for vision during the day, and throughout their lives. Comparison of data obtained with honeybees and other organisms indicates that there is a link between the regulation of phototransduction components and vision-related animal behaviour.

Aging↗

PubMed2000

Amtyr1: characterization of a gene from honeybee (Apis mellifera) brain encoding a functional tyramine receptor.

Biogenic amine receptors are involved in the regulation and modulation of various physiological and behavioral processes in both vertebrates and invertebrates. We have cloned a member of this gene family from the CNS of the honeybee, Apis mellifera. The deduced amino acid sequence is homologous to tyramine receptors cloned from Locusta migratoria and Drosophila melanogaster as well as to an octopamine receptor cloned from Heliothis virescens. Functional properties of the honeybee receptor were studied in stably transfected human embryonic kidney 293 cells. Tyramine reduced forskolin-induced cyclic AMP production in a dose-dependent manner with an EC50 of approximately 130 nM. A similar effect of tyramine was observed in membrane homogenates of honeybee brains. Octopamine also reduced cyclic AMP production in the transfected cell line but was both less potent (EC50 of approximately 3 microM) and less efficacious than tyramine. Receptor-encoding mRNA has a wide-spread distribution in the brain and subesophageal ganglion of the honeybee, suggesting that this tyramine receptor is involved in sensory signal processing as well as in higher-order brain functions.

Amino Acid Sequence↗

PubMed2003

Molecular and functional characterization of an octopamine receptor from honeybee (Apis mellifera) brain.

Biogenic amines and their receptors regulate and modulate many physiological and behavioural processes in animals. In vertebrates, octopamine is only found in trace amounts and its function as a true neurotransmitter is unclear. In protostomes, however, octopamine can act as neurotransmitter, neuromodulator and neurohormone. In the honeybee, octopamine acts as a neuromodulator and is involved in learning and memory formation. The identification of potential octopamine receptors is decisive for an understanding of the cellular pathways involved in mediating the effects of octopamine. Here we report the cloning and functional characterization of the first octopamine receptor from the honeybee, Apis mellifera. The gene was isolated from a brain-specific cDNA library. It encodes a protein most closely related to octopamine receptors from Drosophila melanogaster and Lymnea stagnalis. Signalling properties of the cloned receptor were studied in transiently transfected human embryonic kidney (HEK) 293 cells. Nanomolar to micromolar concentrations of octopamine induced oscillatory increases in the intracellular Ca2+ concentration. In contrast to octopamine, tyramine only elicited Ca2+ responses at micromolar concentrations. The gene is abundantly expressed in many somata of the honeybee brain, suggesting that this octopamine receptor is involved in the processing of sensory inputs, antennal motor outputs and higher-order brain functions.

Amino Acid Sequence↗

PubMed2000

A veterinary approach to the European honey bee (Apis mellifera)

The European honey bee (Apis mellifera) has the unusual status of being an inherently wild species from which a natural foodstuff (honey) is derived by manipulating its behaviour to deposit this in man-made wooden frames. Bees also produce propolis and Royal Jelly which can be harvested but their most important effect is one not immediately obvious as an economic product: that of pollination. Bee diseases are predominantly infectious and parasitic conditions accentuated by the close confinement in which they congregate, either in man-made hives or in colonies in a natural cavity. Treatment or at least control of some of these conditions can be attempted. In some cases natural bee behavioural traits limit the effect of the disease while in others, such as the notifiable disease American foulbrood, destruction of the colony is the only method of control. The mite Varroa jacobsoni can be controlled by the synthetic pyrethroids flumethrin and tau-fluvalinate. The introduction of these products has heightened veterinary interest in this important invertebrate species.

Animal Diseases↗

PubMed2000

Sperm ultrastructure of the honey bee (Apis mellifera) (L) (Hymenoptera, Apidae) with emphasis on the nucleus-flagellum transition region.

The flagellum of Apis mellifera (Hymenoptera, Apidae) consists of two mitochondrial derivatives, an axoneme and two accessory bodies. The mitochondrial derivatives are of unequal size and lie parallel to the axoneme. In the larger derivative four regions can be distinguished while in the smaller, only three. The region occurring only in the larger derivative consists of paracystalline material. The smaller mitochondrial derivative terminates anterior to the larger one. An extremely long centriolar adjunct is observed between the nucleus and the smaller mitochondrial derivative. This adjunct is compact, very electron dense and gradually tapers from base toward apex, finishing at the anterior extremity of the axonemal microtubules. In this flagellar region, there is only one accessory body present between the larger mitochondrial derivative and the axoneme. Anteriorly, the tips of the axonemal microtubules are inserted in a well developed mass of granular appearance. This material surrounds the nuclear base, separating it from the anterior end of the larger mitochondrial derivative. We believe that the structure identified here as a centriolar adjunct is homologous to that observed in Formicidae, Ichneumonoidea and Symphyta. Therefore, very probably, it is common to most Hymenoptera.

Animals↗