Integrin expression in developing human salivary glands

The development and complete differentiation of salivary glands is a complex process that involves a large number of co-ordinated events. Little is known about the molecular basis for salivary gland development. However, we have reported previously that integrins appear to play a role. Integrins are heterodimeric transmembrane receptors consisting of one α and one β subunit that play a pivotal role in the interaction of cells with the extracellular matrix. Such interactions regulate the organisation of cells of tissues and organs during development as well as cell proliferation and differentiation. Using immunohistochemistry and Western and Northern blot analysis, we mapped the localisation and expression of integrins β1, β3 and β4 in human salivary glands obtained from foetuses ranging from weeks 4–24 of gestation and compared it with adult salivary glands. Integrin β1 first appeared during the canalisation stage and during the differentiation stage. A message first appeared at week 6 of development. The expression of β4 integrin protein and message was observed only in the late stage of differentiation. Integrin β3 was not detected in the developing glands; however, integrins β1, β3 and β4 were all expressed in adult salivary gland tissues. The data suggest that integrins, particularly β1, have a role to play in salivary gland development and differentiation.     

Molecular evidence for host specificity of parasitic nematode microfilariae in some African rainforest birds

Here we describe, determine the prevalence, and examine the host-specificity of some parasitic nematode microfilariae in selected bird species from West and Central Africa. We used microscopy to determine the prevalence of microfilariae in 969 host individuals representing 121 rainforest bird species from Cameroon, Côte d’Ivoire and Equatorial Guinea. Thirteen (11%) of these potential host species harboured microfilariae, and 35 individuals (3.6%) were infected. From the 35 infected individuals, we identified eight distinct morphological microfilarial forms. Sixteen of the 35 infected individuals were of one host species, the Fire-crested Alethe (Alethe diademata), at a prevalence rate of 62%. To examine host and geographical specificity, we sequenced a portion of the LSU rDNA gene from representative microfilariae drawn from different hosts and collecting locations. Identical sequences of the nematode LSU rDNA gene were found in A. diademata collected from locations in Côte d’Ivoire and Equatorial Guinea, locations separated by the Dahomey Gap and associated with different hypothesized refugial areas. In contrast, several other bird species collected at the same sites harboured different microfilaria lineages. We sequenced the mitochondrial ATP synthase genes of the host species A. diademata, and found a 5.4% sequence divergence between the birds sampled in Côte d’Ivoire, and those from Cameroon. Thus, despite this split between the two populations, they harbour microfilariae with identical lineages. These data provide evidence that the microfilariae found in A. diademata may be highly host specific. This apparent specificity may have important implications for the evolutionary and ecological interactions between parasitic nematodes and their avian hosts.     

Kin interactions and changing social structure during a population outbreak of feral house mice

Populations of feral house mice (Mus domesticus L.) in Australia undergo multiannual fluctuations in density, and these outbreaks may be partly driven by some change in behavioural self-regulation. In other vertebrate populations with multiannual fluctuations, changes in kin structure have been proposed as a causal mechanism for changes in spacing behaviour, which consequently result in density fluctuations. We tested the predictions of two alternative conceptual models based on kin selection in a population of house mice during such an outbreak. Both published models (Charnov & Finerty 1980; Lambin & Krebs 1991) propose that the level of relatedness between interacting individuals affects their behavioural response and that this changes with population density, though the nature of this relationship differs between the two models. Neither of the models was consistent with all observed changes in relatedness between interacting female mice; however, our results suggested that changes in kin structure still have potential for explaining why mouse outbreaks begin. Therefore, we have developed a variant of one of these conceptual models suggesting that the maintenance of female kin groups through the preceding winter significantly improves recruitment during the subsequent breeding season, and is therefore necessary for mouse outbreaks. We provide six testable predictions to falsify this hypothesis.     

Three-dimensional solution structure of a unique S100 protein

S100A13 is a homodimeric protein that belongs to the S100 subfamily of EF-hand Ca2+-binding proteins. S100A13 exhibits unique physical and functional properties not observed in other members of the S100 family. S100A13 is crucial for the non-classical export of acidic fibroblast growth factors (FGFs-1), which lack signal peptide at their N-terminal end. In the present study, we report the three-dimensional solution structure of Ca2+-bound S100A13 using a variety of 3D NMR experiments. The structure of S100A13 is globular with four helices and an antiparallel beta-sheet in each subunit. The dimer interface is formed mainly by an antiparallel arrangement of helices H1, H1', H4, and H4'. Isothermal titration calorimetry (ITC) experiments show that S100A13 binds non-cooperatively to four calcium ions. Prominent differences exist between the three-dimensional structures of S100A13 and other S100 proteins. The hydrophobic pocket that largely contributes to protein-protein interactions in other S100 proteins is absent in S100A13. The structure of S100A13 is characterized by a large patch of negatively charged residues flanked by dense cationic clusters contributed largely by the positively charged residues located at the C-terminal end. Results of ITC experiments reveal that S100A13 lacking the C-terminal segment (residues 88-98) fails to bind FGF-1. The three-dimensional structure of S100A13 not only provides useful clues on its role in the non-classical export of signal peptide-less proteins such as FGF-1 but also paves the way for rational design of drugs against FGF-induced tumors.     

Extracellular matrix protein 1 interacts with the domain III of fibulin-1C and 1D variants through its central tandem repeat 2

Extracellular matrix protein 1 (ECM1), a widely expressed glycoprotein, has been shown to harbor mutations in lipoid proteinosis (LP), an autosomal recessive disorder characterized by profound alterations in the extracellular matrix of connective tissue. The biological function of ECM1 and its role in the pathomechanisms of LP are unknown. Fibulins comprise a family of extracellular matrix components, and the prototype of this family, fibulin-1, is expressed in various connective tissues and plays a role in developmental and pathologic processes. In this study, we demonstrate that ECM1, and specifically the second tandem repeat domain which is alternatively spliced, interacts with the C-terminal segments of fibulins 1C and 1D splice variants which differ in their C-terminal domain III. The interactions were detected by yeast two-hybrid genetic system and confirmed by co-immunoprecipitations. Kinetics of the binding between ECM1 and fibulin-1D, measured by biosensor assay, revealed a K(d) of 5.71 x 10(-8) M, indicating a strong protein-protein interaction. Since distinct splice variants of ECM1 and fibulin-1 have been shown to be co-expressed in tissues affected in LP, we propose that altered ECM1/fibulin-1 interactions may play a role in the pathogenesis of this disease as well as in a number of processes involving the extracellular matrix of connective tissues.     

TGF-beta, c-Cbl, and PDGFR-alpha the in mammary stroma

Transforming growth factor-beta (TGF-beta) is thought to regulate ductal and lobuloalveolar development as well as involution in the mammary gland. In an attempt to understand the role TGF-beta plays during normal mammary gland development, and ultimately cancer, we previously generated transgenic mice that express a dominant-negative TGF-beta type II receptor under control of the metallothionine promoter (MT-DNIIR). Upon stimulation with zinc sulfate, the transgene was expressed in the mammary stroma and resulted in an increase in ductal side branching. In this study, mammary gland transplantation experiments confirm that the increase in side branching observed was due to DNIIR activity in the stroma. Development during puberty through the end buds was also accelerated. Cbl is a multifunctional intracellular adaptor protein that regulates receptor tyrosine kinase ubiquitination and downregulation. Mice with a targeted disruption of the c-Cbl gene displayed increased side branching similar to that observed in MT-DNIIR mice; however, end bud development during puberty was normal. Transplantation experiments showed that the mammary stroma was responsible for the increased side branching observed in Cbl-null mice. Cbl expression was reduced in mammary glands from DNIIR mice compared to controls and TGF-beta stimulated expression of Cbl in cultures of primary mammary fibroblasts. In addition, both TGF-beta and Cbl regulated platelet-derived growth factor receptor-alpha (PDGFR alpha) expression in vivo and in isolated mammary fibroblasts. The hypothesis that TGF-beta mediates the levels of PDGFR alpha protein via regulation of c-Cbl was tested. We conclude that TGF-beta regulates PDGFR alpha in the mammary stroma via a c-Cbl-independent mechanism. Finally, the effects of PDGF-AA on branching were determined. Treatment in vivo with PDGF-AA did not affect branching making a functional interaction between TGF-beta and PDGF unlikely.     

Immunolocalization of retinoic acid biosynthesis systems in selected sites in rat

Vitamin A deficiency leads to focal metaplasia of numerous epithelial tissues with altered differentiation from columnar (in general) to stratified squamous cells. This process can be reversed with vitamin A repletion. Previously, we described a system of retinoic acid (RA) synthesis in the cycling rat uterus consisting of cellular retinol binding protein (Crbp), epithelial retinol dehydrogenase (eRoldh), retinal dehydrogenase 2 (Aldh1a2), and cellular retinoic acid binding protein type II (Crabp2). Western blot analysis, RT-PCR, and immunohistochemistry were performed to test whether this retinoic acid synthesis system was also present in other vitamin A sensitive tissues. We found that combinations of Crbp, eRoldh, Aldh1a2 or Aldh1a3, and Crabp2 were present in all vitamin A sensitive tissues examined. In the ureter, while eRoldh was present, another short chain alcohol dehydrogenase reductase (possibly Roldh 1, 2, or 3) was in higher concentration in the transitional epithelia. In several tissues, Crbp, Aldh1a2, and/or Aldh1a3 localized to mesenchyme and/or epithelial cells, while eRoldh and Crabp2 were expressed only in epithelial cells. This suggests that mesenchymal-epithelial interactions may be as important in the adult as they are during development and that local synthesis of RA is important in maintenance of these tissues.     

Dimethylsulfoxide gold-thallium complexes. Effects of the metal-metal interactions in the luminescence

The heteropolynuclear complexes [AuTl(C₆X₅)₂]_n (X = F, Cl) react with dimethylsulfoxide (DMSO) in different molar ratios leading to products of stoichiometry [Tl₂{Au(C₆F₅)₂}₂{μ-DMSO}₃]_n (1), and [Tl₂{Au(C₆Cl₅)₂}₂{μ-DMSO}₂]_n (2). These complexes have been structurally characterized and can be viewed as extended linear chains built with Tl-Au-Tl units in which the thallium atoms are bridged by the oxygen atoms of DMSO ligands. Additional [Au(C₆X₅)₂]⁻ fragments interact with one or two thallium centres, respectively, giving rise to two different types of metal-metal interactions in each molecule. Both of them show a strong luminescence in solid state and complex 2 also in solution. The thallium-thallium interaction in this complex is considered to be the responsible of its luminescence, which remains in solution.     

An overview of arbuscular mycorrhizal fungi–nematode interactions

Plant parasitic nematodes and arbuscular mycorrhizal fungi (AMF) share plant roots as a resource for food and space. The interest in AMF-nematode interactions lies in the possibility of enhanced resistance or tolerance of AMF-infected plants to nematodes, and the potential value of this for control of crop pests. Data collated from previous studies revealed a great diversity of AMF-nematode responses and we seek to generalise from these by evaluating and discussing interactions involving three groups of nematodes distinguished by their mode of parasitism: (i) ectoparasites; (ii) sedentary endoparasites; and (iii) migratory endoparasites. Based on proximity in tissue, we expected that the interactions between endoparasites and AMF would be stronger, i.e. more reciprocal effects of endoparasitic nematodes on AMF, than those between ectoparasites and AMF. Contrary to this hypothesis, we found that, relative to AMF-free plants, AMF-infected plants were damaged more by ectoparasites than by endoparasites. Of the sedentary endoparasites, numbers of root-knot nematodes were reduced more by mycorrhizal infection than were those of cyst nematodes. The reduction in nematode damage by AMF was not different for root-knot or cyst nematode infested plants. Migratory endoparasitic nematodes were the only group whose numbers were greater on AMF-infected plants. However, the experiments involving migratory nematodes were characterised by relatively high levels of AMF infection and little nematode damage compared to the other feeding types. The outcomes of the AMF-nematode interactions are determined by many factors during the interactions between organisms and their physical, physiological and temporal environments. Assessing effects by recording plant sizes and total nematode or AMF populations at the end of experiments gives very little information on the mechanisms of the interactions. It is time to stop doing studies of black boxes and time to start observing processes, directly by using microscopy and indirectly by application of molecular genetics.     

Community context and the influence of non-indigenous species on juvenile salmon survival in a Columbia River reservoir

Non-indigenous species (NIS) have been called biological pollutants, which implies that reducing their numbers should reduce negative impacts. To test this hypothesis, we used food web models, parameterized with data from field studies, to ask how reducing the number of NIS co-occurring with endangered salmon would affect salmon mortality. Our analyses indicate that predation on Upper Columbia River spring chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss juveniles was affected very little by NIS reduction. The effects of removing NIS were partly or totally offset by indirect food web interactions, and were subtle compared to effects of native predator management. We predict that the most effective way of reducing predation on salmon smolts will involve managing native predators and targeted removals of specific NIS. Minimizing impact of established NIS thus entails not only reducing NIS prevalence, but also considering background management practices and community context.