Honey bee kin recognition: learning self and nestmate phenotypes

It is known that there is a genetic basis to the labelling of individuals for kin recognition in the honey bee, Apis mellifera. This study shows that individual workers reared in total isolation are able to discriminate between their full sisters and maternal half sisters. When individuals were reared with a half sister, recognition of their own patritype persisted, together with a comparable awareness of the patritype of their half sisters. Workers, when reared in mixed patritype groups of 10, showed no tendency to discriminate between full and half sisters, i.e. they appeared to learn both nestmate patritypes equally well. However, the labelling phenotypes of individuals reared together became more uniform, possibly through the transfer of substances during trophallaxis and mutual grooming. Workers exposed to 10 patritypes from their full sister patriline were more likely to accept an unfamiliar full sister than workers exposed to only five. Finally, workers reared in the hive appeared to retain an ability to discriminate their own patritype; i.e., even though the hive consisted of two worker patritypes, they discriminated between unfamiliar full and half sisters that had been reared under the same controlled conditions in an incubator.     

Wax moth,Galleria mellonella,high density lipophorin receptor: alternative splicing,tissue-specific expression,and developmental regulation

A lipophorin (Lp) receptor cDNA from the fat body of Galleria mellonella (Lepidoptera) was cloned and sequenced. This is the first result in this order, Lepidoptera. It showed the pattern of the VLDL receptor belonging to the LDL receptor family. Sequence homology with other Lp receptors in insects, Locusta migratoria and Aedes aegypti, was 70 and 61%, respectively and each domain was highly conserved. Polyclonal anti-Lp receptor antibody prepared against expressed Lp receptor fragment between ligand binding domain and EGF-precursor homology domain (R305-D549 of amino acid residues) specifically detected the Lp receptor. Through immuno-blotting, the Lp receptor of larval fat body has an approximate molecular mass of about 97 and 110 kDa under non-reducing and reducing conditions, respectively. This result was in agreement with that of the ligand-blotting. The variant Lp receptors were expressed in the fat body of G. mellonella; one is an Lp receptor which lacks 84 bp of O-linked sugar domain and the other is a full length form of the Lp receptor. Both forms were detected by the polyclonal anti-Lp receptor antibody. The Lp receptor from the fat body of G. mellonella was differently expressed depending on the tissue and the developmental stage with specific abundance in prepupal stage. A steroid hormone, 20-hydroxyecdysone (20-HE) plays a crucial role in insect development. With regards to this conception, day 1-2 last instar larvae were treated with 20-HE and drastic induction of the Lp receptor was observed 48 h after treatment. It was also observed that cholesterol caused an induction of the Lp receptor.     

Structure and tissue-specific developmental expression of a sea urchin arylsulfatase gene

Arylsulfatases are a group of enzymes that remove sulfate moieties from a diverse set of substrates including glycoproteins, steroids, and cerebrosides. We have isolated recombinant cDNA clones corresponding to an arylsulfatase (SpARS) message that encodes an abundant protein of pluteus larvae of the sea urchin Strongylocentrotus purpuratus. Although vertebrate arylsulfatases have broad tissue distributions, in situ hybridization with a probe for SpARS shows that the sea urchin message accumulates in the embryo only in the single cell type of aboral ectoderm and its precursors. The message is first detectable by RNase protection assays around hatching blastula stage and accumulates through pluteus larva stage. The open reading frame of cDNA clones is 1701 nt long and encodes a deduced protein with a predicted molecular mass of 61 kDa. Analysis of corresponding genomic DNA clones reveals that the pre-mRNA contains six exons. Consistent with the fact that arylsulfatase enzyme activity is extracellular, this polypeptide has a hydrophobic leader sequence and three potential glycosylation sites. Furthermore, hybridization in situ shows that in blastulae arylsulfatase message is preferentially concentrated around nuclei at the basal sides of cells. The S. purpuratus sequence is very similar to that recently reported for the same enzyme from Hemicentrotus pulcherrimus and 30% of the amino acid residues are also identical to those of both human arylsulfatase C (steroid sulfatase) and arylsulfatase A. Sequence relationships among these four mRNAs suggest that, assuming equal rates of evolution, the duplication separating the human genes occurred at about the time of separation of the echinoderm and vertebrate lineages.     

Contrasting developmental and tissue-specific expression of alpha and beta thyroid hormone receptor genes.

Thyroid hormones and their receptors (TRs) have critical functions in development. Here we show that a chicken TR beta cDNA clone encodes a receptor with a novel, short N-terminal domain. In vitro-expressed TR beta protein bound thyroid hormone with similar affinity as the chicken TR alpha. Comparison of expression of TR alpha and TR beta mRNAs throughout chicken development until 3 weeks post-hatching revealed ubiquitous expression of TR alpha mRNAs (in 14 different tissues) with some variations in levels, from early embryonic stages. In contast, expression of TR beta mRNA was restricted, occurring notably in brain, eye, lung, yolk sac and kidney, and was subject to striking developmental control, especially in brain where levels increased 30-fold upon hatching. Levels also sharply increased in late embryonic lung, but were relatively high earlier in embryonic eye and yolk sac. RNase protection analyses detected no obvious mRNAs for alpha and beta TRs with variant C-termini as demonstrated previously for the rat TR alpha gene. The data suggest a general role for TR alpha and specific developmental functions for TR beta, and that thyroid-dependent development involves temporal and tissue-specific expression of the TR beta gene.     

Mapping tissue-specific genes correlated with age-dependent changes in protein stability and function

Biophysical measurements indicative of protein stability and function were performed on crude extracts from liver, muscle, and lens of a genetically heterogeneous mouse population. Genetic information was used to search for quantitative trait loci (QTL) that influenced the biophysical traits, with emphasis on phenotypes that previously have been shown to be altered in aged animals. Spectroscopic and enzymatic assays of crude liver and muscle tissue extracts from approximately 600 18-month-old mice, the progeny of (BALB/cJxC57BL/6J)F1 females and (C3H/HeJxDBA/2J)F1 males, were used to measure the susceptibility of a ubiquitous glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), to thermal denaturation. The rate constant for thermal inactivation of GAPDH correlated with markers on chromosome 5 (D5Mit79 and D5Mit251) for muscle lysates and chromosome 15 (D15Mit63 and D15Mit100) for liver tissue. The degree of variability of inactivation rate constants, a measure of the heterogeneity of muscle GAPDH in tissue extracts, was also associated with markers on chromosome 5 (D5Mit79 and D5Mit205). In addition, spectroscopic characteristics of extracted eye lens proteins were evaluated for their susceptibility to photooxidative stress. Absorbance and fluorescence emission characteristics of the lens proteins were mapped to QTL on chromosomes 5 and 15 (D5Mit25 and D15Mit171) while the degree of heterogeneity in photochemical oxidation kinetics was associated with a marker on the chromosome 8 (D8Mit42). Recent work has shown that GAPDH possesses a number of non-glycolytic functions including DNA/RNA binding and regulation of protein expression. Tissue specific differences in GAPDH stability may have significant consequences to these alternate functions during aging.     

When to bee social: interactions among environmental constraints, incentives, guarding, and relatedness in a facultatively social carpenter bee

In the facultatively social carpenter bee, Xylocopa pubescens,foundresses usually establish nests solitarily. However, nestsmay become social if a second foundress (referred to as alpha)successfully usurps the nest, with the original foundress (referredto as beta) remaining as a guard. Reproductive skew theory predictsthat beta foundresses should remain as helpers only if alphausurpers allow them a share of reproduction. Because alpha femalesdestroy much of beta's brood and beta females do not lay eggsafter takeovers, studies have concluded that usurpers offerno staying incentives or concessions in return for helping behavior.This conclusion is paradoxical, and we suggest that by refrainingfrom destroying all of beta's brood, alpha females do indeedoffer concessions to beta females. We constructed a model toexamine the conditions under which social nesting is favoredby both alpha and beta females. Female preference for socialversus solitary nesting is proportional to expected fitnessin either setting and is affected by current environmental conditions,the value of guarding behavior in protecting brood from pollenrobbery, the size of the concession offered by alpha, and thedegree of genetic relatedness between the foundresses. Our modelshows that at a minimum, establishing sociality after unrelatedusurpations always requires a concession, whereas in relatedusurpations, a concession is not always required. Generally,agreement between alpha and beta is difficult because alpharequires a much higher premium in pollen robbery protectionthan beta in order for sociality to be advantageous. Alpha femalesprefer social nesting only under the most severe environmentalconditions because usually they gain less by the presence ofa guard than by replacing beta brood with their own. In contrast,beta females always strongly prefer social nesting because thechances of successful renesting elsewhere are low and rarelyoutweigh the advantages of guarding their own brood that surviveusurpation. Effects of relatedness between foundresses on preferencefor social nesting are nonintuitive: first, alpha's preferenceincreases with relatedness, whereas beta's preference declines,and second, unrelated beta females prefer sociality more stronglythan related ones. This is because replacement of beta's offspringwith related alpha offspring partially compensates her for theloss of her own brood, even should she leave the nest.     

Rates of protein evolution are positively correlated with developmental timing of expression during mouse spermatogenesis

Male reproductive genes often evolve very rapidly, and sexual selection is thought to be a primary force driving this divergence. We investigated the molecular evolution of 987 genes expressed at different times during mouse spermatogenesis to determine if the rate of evolution and the intensity of positive selection vary across stages of male gamete development. Using mouse-rat orthologs, we found that rates of protein evolution were positively correlated with the developmental timing of expression. Genes expressed early in spermatogenesis had rates of divergence similar to the genome median, while genes expressed after the onset of meiosis were found to evolve much more quickly. Rates of protein evolution were fastest for genes expressed during the dramatic morphogenesis of round spermatids into spermatozoa. Late-expressed genes were also more likely to be specific to the male germline. To test for evidence of positive selection, we analyzed the ratio of nonsynonymous to synonymous changes using a maximum likelihood framework in comparisons among mouse, rat, and human. Many genes showed evidence of positive selection, and most of these genes were expressed late in spermatogenesis and were testis specific. Overall, these data suggest that the intensity of positive selection associated with the evolution of male gametes varies considerably across development and acts primarily on phenotypes that develop late in spermatogenesis.     

Analysis of the tissue-specific expression, developmental regulation, and linkage relationships of a rodent gene encoding heart fatty acid binding protein

The rat contains at least three homologous cytosolic proteins that bind long chain fatty acids, termed liver (L-), intestinal (I-), and heart (H-) fatty acid binding protein (FABP). I-FABP mRNA is confined to the gastrointestinal tract while L-FABP mRNA is abundantly represented in hepatocytes as well as enterocytes. We have isolated a rat heart FABP cDNA clone and determined the pattern of H-FABP mRNA accumulation in a wide variety of tissues harvested from late fetal, suckling, weaning, and adult rats. RNA blot hybridizations and primer extension analysis disclosed that the distribution of H-FABP mRNA in adult rat tissues is different from that of I- or L-FABP mRNA. H-FABP mRNA is most abundant in adult heart. This mRNA was also present in an adult slow twitch (type I) skeletal muscle (soleus, 63% of the concentration in heart), testes (28%), a fast twitch skeletal muscle (psoas, 17%), brain (10%), kidney (5%), and adrenal gland (5%). H-FABP mRNA was not detected in adult small intestine, colon, spleen, lung, or liver RNA. Distinct patterns of developmental change in H-FABP mRNA accumulation were documented in heart, placenta, brain, kidney, and testes. Myocardial H-FABP mRNA levels rise rapidly during the 48 h prior to and after birth, reaching peak levels by the early weaning period. The postnatal increase in myocardial H-FABP mRNA concentration and its relative distribution in adult fast and slow twitch skeletal muscle are consistent with its previously proposed function in facilitating mitochondrial beta-oxidation of fatty acids. However, the presence of H-FABP mRNA in brain, a tissue which does not normally significantly oxidize fatty acids in late postnatal life, suggests that H-FABP may play a wider role in fatty acid metabolism than previously realized. Mouse-hamster somatic cell hybrids were utilized to map H-FABP. Using stringencies which did not produce cross-hybridization between L-, I-, and H-FABP DNA sequences, we found at least three loci in the mouse genome, each located on different chromosomes, which reacted with our cloned H-FABP cDNA. None of these H-FABP-related loci were linked to the gene which specifies a highly homologous adipocyte-specific protein termed aP2 or to genes encoding two other members of this protein family, cellular retinol binding protein and cellular retinol binding protein II.(ABSTRACT TRUNCATED AT 400 WORDS)     

Cryptic castes,social context and colony defence in a social bee,Tetragonula carbonaria

Division of labour in social insect colonies is facilitated in two ways: through temporal sharing of tasks or by morphologically specialised castes. In casteless species, colony defence is maintained by morphologically indistinct workers, who lack the obvious defensive specialisation of polymorphic species. Discrimination of intruders is carried out via antenna, which also detects defensive social cues such as alarm pheromones. Despite their functional importance however, antennal morphology is rarely considered in studies of nestmate recognition. We investigated antennal morphology and the necessity of social cues in mediating defensive behaviour across differentially tasked workers of a casteless social bee, Tetragonula carbonaria. Our results suggest that the current understanding of division of labour in casteless worker species remains poorly understood, with differences in antennal morphology and aggression creating morphologically and behaviourally distinct ‘cryptic castes’. Further, we found that defensive behaviour was only elicited near nest odours, highlighting the importance of mediating aggression among workers.