Vitis pollen dispersal in and from organic vineyards: I. Pollen trap and soil pollen data

The recent discovery of Roman vineyards in Britain [Antiquity 75 (2001) 745] based on both archaeological and palynological data has raised questions concerning the palynological record of viticulture in NW Europe. This paper presents data collected from 30 pollen traps and 29 soil pollen samples from within, and around, three organic vineyards in England. The soil trap data were collected over one flowering season. Within-vineyard pollen accumulation is compared with off-site accumulation and the soil pollen data. Although there is extremely high intravineyard variation in Vitis accumulation, the data confirm the role of stemflow transport and the exponential decrease in Vitis pollen accumulation with distance away from the vineyard. Pollen trap values of over 2% total pollen (TP) Vitis and soils with over 0.2% TP (based on 1000+ pollen sum) are likely to indicate the on-site presence of a vineyard. This is in accordance with the values recorded from known Roman vineyards in the Nene Valley. A compilation of occasional finds of Vitis pollen in England reveals typical values of 0.2% or less; although higher values are recorded, they are generally an artefact of a low total pollen count. However, the clustering of these occasional finds into the Roman and high Medieval periods suggests that long-distance Vitis influx increased during periods of widespread viticulture in NW Europe including England. The monitoring of Vitis pollen dispersal suggests that where encountered as part of routine pollen counting, the total pollen sum should be increased to ca. 1000 TP in order to establish a better estimate of the frequency of occurrence. This will allow palynology to play a less ambiguous part in the mapping of ancient viticulture.     

Pollen tube cell walls of wild and domesticated tomatoes contain arabinosylated and fucosylated xyloglucan

Background and Aims In flowering plants, fertilization relies on the delivery of the sperm cells carried by the pollen tube to the ovule. During the tip growth of the pollen tube, proper assembly of the cell wall polymers is required to maintain the mechanical properties of the cell wall. Xyloglucan (XyG) is a cell wall polymer known for maintaining the wall integrity and thus allowing cell expansion. In most angiosperms, the XyG of somatic cells is fucosylated, except in the Asterid clade (including the Solanaceae), where the fucosyl residues are replaced by arabinose, presumably due to an adaptive and/or selective diversification. However, it has been shown recently that XyG of Nicotiana alata pollen tubes is mostly fucosylated. The objective of the present work was to determine whether such structural differences between somatic and gametophytic cells are a common feature of Nicotiana and Solanum (more precisely tomato) genera.Methods XyGs of pollen tubes of domesticated (Solanum lycopersicum var. cerasiforme and var. Saint-Pierre) and wild (S. pimpinellifolium and S. peruvianum) tomatoes and tobacco (Nicotiana tabacum) were analysed by immunolabelling, oligosaccharide mass profiling and GC-MS analyses.Key Results Pollen tubes from all the species were labelled with the mAb CCRC-M1, a monoclonal antibody that recognizes epitopes associated with fucosylated XyG motifs. Analyses of the cell wall did not highlight major structural differences between previously studied N. alata and N. tabacum XyG. In contrast, XyG of tomato pollen tubes contained fucosylated and arabinosylated motifs. The highest levels of fucosylated XyG were found in pollen tubes from the wild species.Conclusions The results clearly indicate that the male gametophyte (pollen tube) and the sporophyte have structurally different XyG. This suggests that fucosylated XyG may have an important role in the tip growth of pollen tubes, and that they must have a specific set of functional XyG fucosyltransferases, which are yet to be characterized.     

A male cone of Pseudofrenelopsis dalatzensis with in situ pollen grains from the Lower Cretaceous of Northeast China

A Classopollis-containing male cone found in close association with Pseudofrenelopsis dalatzensis from the Lower Cretaceous Dalazi Formation of Yanji Basin, eastern Jilin, China was studied using scanning electron microscopy and described as Classostrobus dalatzensis sp. nov. The small oval cone, borne on the top of a stalk (fertile short shoot), consists of helically arranged microsporophylls, each with a rhomboid and expanded distal head. The short shoot is divided into indistinct nodes and internodes with fine longitudinal striations. Cuticles of the microsporophyll and the internode of the short shoot are generally similar to Pseudofrenelopsis dalatzensis internode cuticles, all of them having papillate outer surfaces. Stomata are similar in structure and arrangement, with papillae of subsidiary cells overhanging the stomatal pit. Epidermal cells are rectangular or isodiametrical, each with a robust papilla on the periclinal wall. Hypodermal cells are present between the stomatal files. The internal structure of the in situ Classopollis pollen grains was studied in detail. Pollen grain is small, with thin sexine, poorly developed nexine and may be immature. This is the first record of in situ pollen grains Classopollis in China of which the internal structure is known in detail.     

Delirium subsindrómico en pacientes ancianos: revisión sistemática

In this systemic review, the articles published between1990 and November 2012 on subsyndromal delirium (SSD), and specifically those with reference to geriatric patients, were analysed. In SSD, symptoms from the three nuclear domains of delirium (cognitive, circadian and higher order thinking) are simultaneously present, with mild to moderate severity. Although the search for these clinical characteristics is a useful guide, there are no universally accepted diagnostic criteria for SSD. Regardless of the criteria used for diagnosis, SSD is persistently associated with poor functional and cognitive outcome, longer hospital stay, institutionalisation, and increased mortality. Studies are needed on the physiopathology, treatment and prevention in the field of SSD, which is a particularly important clinical condition in geriatric patients.     

Ultrastructural study of pollen and anther development in Luehea divaricata (Malvaceae,Grewioideae) and its systematic implications: Role of tapetal transfer cells,orbicules and male germ unit

Developmental processes of microsporogenesis and microgametogenesis in Luehea divaricata (Malvaceae, Grewioideae) were analyzed with transmission electron microscopy. The species studied has perfect flowers. The young anthers are bithecal and tetrasporangiate; microspore mother cells undergo simultaneous meiosis, forming tetrads with a tetrahedral arrangement. The development of the anther wall conforms to the basic type and the tapetum is secretory. The results highlight the presence of multinucleate tapetal cells, which acquire ultrastructural features characteristic of transfer cells at the young pollen grain stage, and which are associated with the presence of orbicules. Tapetal transfer cells have not been previously investigated in detail for other species of angiosperms; their function during pollen development is discussed. The present work is the first contribution to the knowledge of ultrastructural pollen development in the genus Luehea, as well as in the subfamily Grewioideae.     

S-RNase-based self-incompatibility in Petunia inflata

Background

For the Solanaceae-type self-incompatibility, also possessed by Rosaceae and Plantaginaceae, the specificity of self/non-self interactions between pollen and pistil is controlled by two polymorphic genes at the S-locus: the S-locus F-box gene (SLF or SFB) controls pollen specificity and the S-RNase gene controls pistil specificity.

Scope

This review focuses on the work from the authors'' laboratory using Petunia inflata (Solanaceae) as a model. Here, recent results on the identification and functional studies of S-RNase and SLF are summarized and a protein-degradation model is proposed to explain the biochemical mechanism for specific rejection of self-pollen tubes by the pistil.

Conclusions

The protein-degradation model invokes specific degradation of non-self S-RNases in the pollen tube mediated by an SLF, and can explain compatible versus incompatible pollination and the phenomenon of competitive interaction, where SI breaks down in pollen carrying two different S-alleles. In Solanaceae, Plantaginaceae and subfamily Maloideae of Rosaceae, there also exist multiple S-locus-linked SLF/SFB-like genes that potentially function as the pollen S-gene. To date, only three such genes, all in P. inflata, have been examined, and they do not function as the pollen S-gene in the S-genotype backgrounds tested. Interestingly, subfamily Prunoideae of Rosaceae appears to possess only a single SLF/SFB gene, and competitive interaction, observed in Solanaceae, Plantaginaceae and subfamily Maloideae, has not been observed. Thus, although the cytotoxic function of S-RNase is an integral part of SI in Solanaceae, Plantaginaceae and Rosaceae, the function of SLF/SFB may have diverged. This highlights the complexity of the S-RNase-based SI mechanism. The review concludes by discussing some key experiments that will further advance our understanding of this self/non-self discrimination mechanism.     

Thank you for not flowering: conservation genetics and gene flow analysis of native and non-native populations of Fraxinus (Oleaceae) in Ireland

The risks of gene flow between interfertile native and introduced plant populations are greatest when there is no spatial isolation of pollen clouds and phenological patterns overlap completely. Moreover, invasion probabilities are further increased if introduced populations are capable of producing seeds by selfing. Here we investigated the mating system and patterns of pollen-mediated gene flow among populations of native ash (Fraxinus excelsior) and mixed plantations of non-native ash (F. angustifolia and F. excelsior) as well as hybrid ash (F. excelsior × F. angustifolia) in Ireland. We analysed the flowering phenology of the mother trees and genotyped with six microsatellite loci in progeny arrays from 132 native and plantation trees (1493 seeds) and 444 potential parents. Paternity analyses suggested that plantation and native trees were pollinated by both native and introduced trees. No signs of significant selfing in the introduced trees were observed and no evidence of higher male reproductive success was found for introduced trees compared with native ones either. A small but significant genetic structure was found (φ_ft=0.05) and did not correspond to an isolation-by-distance pattern. However, we observed a significant temporal genetic structure related to the different phenological groups, especially with early and late flowering native trees; each phenological group was pollinated with distinctive pollen sources. Implications of these results are discussed in relation to the conservation and invasiveness of ash and the spread of resistance genes against pathogens such as the fungus Chalara fraxinea that is destroying common ash forests in Europe.     

Effects at Nearctic north-temperate latitudes of indoor versus outdoor overwintering on the microsporidium Nosema ceranae and western honey bees (Apis mellifera)

In northern temperate climates, western honey bee (Apis mellifera) colonies can be wintered outdoors exposed to ambient conditions, or indoors in a controlled setting. Because very little is known about how this affects the recently-detected microsporidium Nosema ceranae, we investigated effects of indoor versus outdoor overwintering on spring N. ceranae intensity (spores per bee), and on winter and spring colony mortality. For colonies medicated with Fumagilin-B® to control N. ceranae, overwintering treatment did not affect N. ceranae intensity, despite outdoor-wintered colonies having significantly greater mortality. These findings suggest that N. ceranae may not always pose the most significant threat to western honey bees, and that indoor-wintering may ensure that a greater number of colonies are available for honey production and pollination services during the summer.     

Crop Pollination Exposes Honey Bees to Pesticides Which Alters Their Susceptibility to the Gut Pathogen Nosema ceranae

Recent declines in honey bee populations and increasing demand for insect-pollinated crops raise concerns about pollinator shortages. Pesticide exposure and pathogens may interact to have strong negative effects on managed honey bee colonies. Such findings are of great concern given the large numbers and high levels of pesticides found in honey bee colonies. Thus it is crucial to determine how field-relevant combinations and loads of pesticides affect bee health. We collected pollen from bee hives in seven major crops to determine 1) what types of pesticides bees are exposed to when rented for pollination of various crops and 2) how field-relevant pesticide blends affect bees’ susceptibility to the gut parasite Nosema ceranae. Our samples represent pollen collected by foragers for use by the colony, and do not necessarily indicate foragers’ roles as pollinators. In blueberry, cranberry, cucumber, pumpkin and watermelon bees collected pollen almost exclusively from weeds and wildflowers during our sampling. Thus more attention must be paid to how honey bees are exposed to pesticides outside of the field in which they are placed. We detected 35 different pesticides in the sampled pollen, and found high fungicide loads. The insecticides esfenvalerate and phosmet were at a concentration higher than their median lethal dose in at least one pollen sample. While fungicides are typically seen as fairly safe for honey bees, we found an increased probability of Nosema infection in bees that consumed pollen with a higher fungicide load. Our results highlight a need for research on sub-lethal effects of fungicides and other chemicals that bees placed in an agricultural setting are exposed to.     

Going to great lengths: selection for long corolla tubes in an extremely specialized bat–flower mutualism

In a hypothesis that has remained controversial since its inception, Darwin suggested that long-tubed flowers and long-tongued pollinators evolved together in a coevolutionary race, with each selecting for increasing length in the other. Although the selective pressures that flowers impose on tongue length are relatively straightforward, in that longer tongues allow access to more nectar, selective pressures that pollinators impose on flower length are less clear. Here, we test for such selective pressures in the highly specialized mutualism between the nectar bat Anoura fistulata, which can extend its tongue twice as far as other nectar bats, and Centropogon nigricans, which has flowers of a similar length (8–9 cm). We used flight cage experiments to examine the effects of artificially manipulated flower lengths on (i) bat behaviour and (ii) pollen transfer. Increased length produced longer visits, but did not affect the force bats applied during visits. In the second experiment, flower length increased both the male and female components of flower function: long male flowers delivered more pollen grains and long female flowers received more pollen grains. However, pollen transfer was not correlated with visit duration, so the mechanism behind differences in pollen transfer remains unclear. By demonstrating that bats select for increasing flower length, these results are consistent with the hypothesis that A. fistulata evolved its remarkable tongue in a coevolutionary race with long-tubed flowers similar to that envisioned by Darwin.     

F-actin forms mobile and unwinding ring-shaped structures in germinating Arabidopsis pollen expressing Lifeact

The flowering plant pollen tube is the fastest elongating plant cell and transports the sperm cells for double fertilization. The highly dynamic formation and reorganization of the actin cytoskeleton is essential for pollen germination and pollen tube growth. To drive pollen-specific expression of fluorescent marker proteins, commonly the strong Lat52 promoter is used. Here we show by quantitative fluorescent analysis that the gametophyte-specific ARO1 promoter from Arabidopsis drives an about 3.5 times weaker transgene expression than the Lat52 promoter. In one third of the pollen of F-actin-labeled ARO1p:tagRFP-T-Lifeact transgenic lines we observed mobile ring-shaped actin structures in pollen grains and pollen tubes. Pollen tube growth, transgene transmission and seed production were not affected by tagRFP-T-Lifeact expression. F-actin rings were able to integrate into emerging actin filaments and they may reflect a particular physiological state of the pollen or a readily available storage form provided for rapid actin network remodeling.     

Mating system and early viability resistance to habitat fragmentation in a bird-pollinated eucalypt

Habitat fragmentation has been shown to disrupt ecosystem processes such as plant-pollinator mutualisms. Consequently, mating patterns in remnant tree populations are expected to shift towards increased inbreeding and reduced pollen diversity, with fitness consequences for future generations. However, mating patterns and phenotypic assessments of open-pollinated progeny have rarely been combined in a single study. Here, we collected seeds from 37 Eucalyptus incrassata trees from contrasting stand densities following recent clearance in a single South Australian population (intact woodland=12.6 trees ha⁻¹; isolated pasture=1.7 trees ha⁻¹; population area=10 km²). 649 progeny from these trees were genotyped at eight microsatellite loci. We estimated genetic diversity, spatial genetic structure, indirect contemporary pollen flow and mating patterns for adults older than the clearance events and open-pollinated progeny sired post-clearance. A proxy of early stage progeny viability was assessed in a common garden experiment. Density had no impact on mating patterns, adult and progeny genetic diversity or progeny growth, but was associated with increased mean pollen dispersal. Weak spatial genetic structure among adults suggests high historical gene flow. We observed preliminary evidence for inbreeding depression related to stress caused by fungal infection, but which was not associated with density. Higher observed heterozygosities in adults compared with progeny may relate to weak selection on progeny and lifetime-accumulated mortality of inbred adults. E. incrassata appears to be resistant to the negative mating pattern and fitness changes expected within fragmented landscapes. This pattern is likely explained by strong outcrossing and regular long-distance pollen flow.     

Pollen morphology of Chinese Curcuma L. and Boesenbergia Kuntz (Zingiberaceae): Taxonomic implications

In order to find new non-molecular evidence to support the phylogenetic and taxonomic position, pollen grains of 20 populations of 16 species of Chinese Curcuma L. and Boesenbergia Kuntz (Zingiberaceae) were investigated under SEM and TEM. The pollen grains are spherical and ovoid, nonaperturate. The pollen wall is composed of a very thin exine and a thick intine. The exine is psilate or echinate. The intine consists of two layers, i.e., a thick, channeled layer (exintine) and an inner homogenous layer (endintine). The results reveal morphological congruence between the pollen grains of species of Curcuma, which according to DNA sequence data appears to be a polyphyletic genus. However the uniform pollen morphology in Curcuma provides no evidence to divide this genus into separate taxonomic entities. Our results on pollen morphology also do not provide any additional evidence to either unite or segregate Boesenbergia albomaculata and Curcumorpha longiflora in the same genus and demonstrate that more taxonomic data on the genus Boesenbergia and its relatives are needed before a final decision can be made.     

In vivo imaging of the S-locus receptor kinase,the female specificity determinant of self-incompatibility,in transgenic self-incompatible Arabidopsis thaliana

Background and Aims The S-locus receptor kinase (SRK), which is expressed in stigma epidermal cells, is responsible for the recognition and inhibition of ‘self’ pollen in the self-incompatibility (SI) response of the Brassicaceae. The allele-specific interaction of SRK with its cognate pollen coat-localized ligand, the S-locus cysteine-rich (SCR) protein, is thought to trigger a signalling cascade within the stigma epidermal cell that leads to the arrest of ‘self’ pollen at the stigma surface. In addition to the full-length signalling SRK receptor, stigma epidermal cells express two other SRK protein species that lack the kinase domain and whose role in the SI response is not understood: a soluble version of the SRK ectodomain designated eSRK and a membrane-tethered form designated tSRK. The goal of this study was to describe the sub-cellular distribution of the various SRK protein species in stigma epidermal cells as a prelude to visualizing receptor dynamics in response to SCR binding.Methods The Arabidopsis lyrata SRKb variant was tagged with the Citrine variant of yellow fluorescent protein (cYFP) and expressed in A. thaliana plants of the C24 accession, which had been shown to exhibit a robust SI response upon transformation with the SRKb–SCRb gene pair. The transgenes used in this study were designed for differential production and visualization of the three SRK protein species in stigma epidermal cells. Transgenic stigmas were analysed by pollination assays and confocal microscopy.Key Results and Conclusions Pollination assays demonstrated that the cYFP-tagged SRK proteins are functional and that the eSRK is not required for SI. Confocal microscopic analysis of cYFP-tagged SRK proteins in live stigma epidermal cells revealed the differential sub-cellular localization of the three SRK protein species but showed no evidence for redistribution of these proteins subsequent to incompatible pollination.     

Responses to damage in an arborescent bryozoan: Effects of injury location

A common consequence of the predation or physical disturbance of sessile modular animals is the removal of part of the colony. This damage is often sub-lethal, and leaves the remainder of the colony to repair and regenerate lost tissue. Damage may, however, have lasting effects on the life history characteristics of a colony. We examined the effects of damage on growth and reproduction in the arborescent bryozoan Bugula neritina (Linnaeus), with a particular focus on the effects of the location of that damage. We predicted that a damaged colony would initially allocate more energy to damage repair and subsequent growth, increasing in size to decrease the risk of mortality, at the expense of its short-term reproductive potential. A short-term experiment assessed the immediate regenerative capacities of colonies, while a longer-term experiment measured overall colony responses to damage at different colony locations. In this experiment, zooids were removed in three colony regions; all on one colony branch, spread evenly across all growing tips, and a third, intermediate pattern. A third experiment tested the effects of predator-induced damage versus damage alone, by measuring the recovery of colonies subjected to either a predation event by the nudibranch Polycera hedgpethi, or to an imitation predation event, where an equivalent section of the colony was removed manually. Regeneration was rapid, with budding resuming a few days after damage occurred. Removing around 20% of the colony did not consistently alter colony growth rates in any experiment, but the onset of reproduction was delayed by 7 to 20 days, and subsequent colony fecundity reduced by as much as 70% in one damage treatment. The removal of the same amount of tissue by nudibranch predators, or manually removed in a treatment that mimicked this predation, had no detrimental effect on the onset of reproduction or the eventual reproductive output. Nudibranchs removed tissue by consuming whole colony branches, and the experimental treatment that removed one branch also had the least effect of three damage treatments. These results suggest the severity of long-term effects depends on the location of the injury, and that B. neritina does not modify its growth or reproductive patterns to minimise these effects.     

Effects of Parietaria judaica pollen extract on human microvascular endothelial cells

Pollinosis from Parietaria judaica is one of the main causes of allergy in the Mediterranean area. The present study is designed to assess if P. judaica pollens contain bioactive compounds able to elicit a functional response in endothelial cells.We have demonstrated that addition of pollen extract to human lung microvascular endothelial cells (HMVEC-L) induces a modification of cell morphology, actin cytoskeletal rearrangements and an increase in endothelial cell permeability. We further showed that the treatment of endothelial cells with pollen extract causes an increase of E-selectin and VCAM-1 protein levels as well as an increase of IL-8 production. The stimulation of cell-cell adhesion molecules was paralleled by a dose-dependent increase of adhesion of polymorphonuclear cells (PMNs) to HMVEC-L monolayer. Our results suggest for the first time that pollen affect directly endothelial cells (EC) modulating critical functions related to the inflammatory response.     

Colony pollen reserves affect body size,sperm production and sexual development in males of the stingless bee <Emphasis Type="Italic">Melipona beecheii</Emphasis>

The production of male sexual offspring by social insect colonies is often strongly seasonal or resource-dependent. In stingless bees, males are produced in smaller numbers under conditions of low colony food reserves; whether such males are negatively affected in traits related to reproductive success is not known. We compared body size, sperm production and sexual maturity in Melipona beecheii males reared with experimentally supplemented or reduced pollen reserves, but with otherwise equal numbers of workers and equal quantities of honey reserves. We also studied the same traits in males collected from non-manipulated colonies with pollen reserves intermediate between the supplemented or reduced groups but with more workers and honey reserves. Males reared under experimentally reduced pollen reserves had significantly smaller bodies and lower sperm counts compared to those reared in colonies with experimentally supplemented pollen reserves. There was also a significantly positive relationship between the number of sperm and body size in males across all colony treatments. The maximum number of sperm in seminal vesicles was recorded 2 days later in males from colonies with reduced pollen compared to males from colonies with supplementary pollen. Males from non-manipulated colonies were intermediate in size, sperm count and speed of maturation. Our study documents for the first time the existence of large size variation in males of stingless bees that is related with the amount of pollen reserves in their natal colony. We conclude that a colony’s pollen reserves have a major impact on male body size, sperm production and speed of sexual maturity in this stingless bee, which may be the case in other social insects. Stingless bees are a good model system to study the balance between colony-level selection and individual-level selection on male sexually selected traits such as body size.     

Counting pollen grains using readily available, free image processing and analysis software

Background and Aims

Although many methods exist for quantifying the number of pollen grains in a sample, there are few standard methods that are user-friendly, inexpensive and reliable. The present contribution describes a new method of counting pollen using readily available, free image processing and analysis software.

Methods

Pollen was collected from anthers of two species, Carduus acanthoides and C. nutans (Asteraceae), then illuminated on slides and digitally photographed through a stereomicroscope. Using ImageJ (NIH), these digital images were processed to remove noise and sharpen individual pollen grains, then analysed to obtain a reliable total count of the number of grains present in the image. A macro was developed to analyse multiple images together. To assess the accuracy and consistency of pollen counting by ImageJ analysis, counts were compared with those made by the human eye.

Key Results and Conclusions

Image analysis produced pollen counts in 60 s or less per image, considerably faster than counting with the human eye (5–68 min). In addition, counts produced with the ImageJ procedure were similar to those obtained by eye. Because count parameters are adjustable, this image analysis protocol may be used for many other plant species. Thus, the method provides a quick, inexpensive and reliable solution to counting pollen from digital images, not only reducing the chance of error but also substantially lowering labour requirements.     

The within-nest behaviour of honeybee pollen foragers in colonies with a high or low need for pollen

Numerous studies have documented that honeybee colonies can rapidly adjust the number of foragers collecting pollen in response to changes in quantities of brood, pollen and nectar in the nest. However, few studies have examined the behaviour of individual pollen foragers while in the nest between trips. Thus, little is known about how a pollen forager actually assesses its colony's needs. To understand this process better, we observed the behaviour of 319 pollen foragers while in their nests between foraging trips. We observed foragers in two types of nest environments: one with a relatively high need for pollen and one with a relatively low need for pollen. Foragers performed as many as 14 activities during two phases demarcated by the unloading of pollen loads. They inspected empty cells and cells with pollen, and performed the waggle dance at higher relative frequencies before unloading (P≤ 0.0004 each act). They antennated nestmates, autogroomed and received trophallaxis at higher relative frequencies after unloading (P≤ 0.0004 each act). All acts were performed both before and after unloading, but not always by each bee. Pollen foragers discriminated among cells based on cell contents in two contexts. First, they inspected cells already containing pollen more often than expected by chance. Second, their pattern of inspecting cells with different contents differed from their pattern of unloading pollen loads in those cells. Pollen foragers performed 42.3±4.6% (least square mean±SE) of preunloading inspection events on empty cells, but unloaded in them only 19.9±4.6% of the time. They performed 42.2±4.6% of preunloading inspection events on cells already containing pollen, but unloaded in them 79.4±4.6% of the time (P< 0.0001). Our data show that pollen foragers can determine the contents of cells in the nest and suggest that the regulation of pollen collection involves direct assessment of colony need by pollen foragers. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Fibronectin modulates thymocyte-thymic epithelial cell interactions following Trypanosoma cruzi infection

Developing thymocytes interact with thymic epithelial cells (TECs) through cell-cell interactions, TEC-derived secretory moieties and extracellular matrix (ECM)-mediated interactions. These physiological interactions are crucial for normal thymocyte differentiation, but can be disrupted in pathological situations. Indeed, there is severe thymic atrophy in animals acutely infected with Trypanosoma cruzi due to CD4+CD8+ thymocyte depletion secondary to caspase-mediated apoptosis, together with changes in ECM deposition and thymocyte migration. We studied an in vitro model of TEC infection by T. cruzi and found that infected TEC cultures show a reduced number of cells, which was likely associated with decreased proliferative capacity, but not with increased cell death, as demonstrated by bromodeoxyuridine and annexin-V labelling. The infected TEC cultures exhibited increased expression of fibronectin (FN), laminin (LM) and type IV collagen. Importantly, treatment with FN increased the relative number of infected cells, whereas treatment with anti-FN or anti-LM antibodies resulted in lower infection rates. Consistent with these data, we observed increased thymocyte adhesion to infected TEC cultures. Overall, these results suggest that ECM molecules, particularly FN, facilitate infection of the thymic epithelium and that the consequent enhancement of ECM expression might be associated with changes in TEC-thymocyte interactions.