Recent ecological and biogeochemical changes in alpine lakes of Rocky Mountain National Park (Colorado, USA): a response to anthropogenic nitrogen deposition

Dated sediment cores from five alpine lakes (>3200 m asl) in Rocky Mountain National Park (Colorado Front Range, USA) record near‐synchronous stratigraphic changes that are believed to reflect ecological and biogeochemical responses to enhanced nitrogen deposition from anthropogenic sources. Changes in sediment proxies include progressive increases in the frequencies of mesotrophic planktonic diatom taxa and diatom concentrations, coupled with depletions of sediment δ¹⁵N and C : N values. These trends are especially pronounced since approximately 1950. The most conspicuous diatoms to expand in recent decades are Asterionella formosa and Fragilaria crotonensis. Down‐core species changes are corroborated by a year‐long sediment trap experiment from one of the lakes, which reveals high frequencies of these two taxa during autumn and winter months, the interval of peak annual limnetic []. Although all lakes record recent changes, the amplitude of stratigraphic shifts is greater in lakes east of the Continental Divide relative to those on the western slope, implying that most nitrogen enrichment originates from urban, industrial and agricultural sources east of the Rocky Mountains. Deviations from natural trajectories of lake ontogeny are illustrated by canonical correspondence analysis, which constrains the diatom record as a response to changes in nitrogen biogeochemistry. These results indicate that modest rates of anthropogenic nitrogen deposition are fully capable of inducing directional biological and biogeochemical shifts in relatively pristine ecosystems.     

Potential role for ADAM15 in pathological neovascularization in mice

ADAM15 (named for a disintegrin and metalloprotease 15, metargidin) is a membrane-anchored glycoprotein that has been implicated in cell-cell or cell-matrix interactions and in the proteolysis of molecules on the cell surface or extracellular matrix. To characterize the potential roles of ADAM15 during development and in adult mice, we analyzed its expression pattern by mRNA in situ hybridization and generated mice carrying a targeted deletion of ADAM15 (adam15(-/-) mice). A high level of expression of ADAM15 was found in vascular cells, the endocardium, hypertrophic cells in developing bone, and specific areas of the hippocampus and cerebellum. However, despite the pronounced expression of ADAM15 in these tissues, no major developmental defects or pathological phenotypes were evident in adam15(-/-) mice. The elevated levels of ADAM15 in endothelial cells prompted an evaluation of its role in neovascularization. In a mouse model for retinopathy of prematurity, adam15(-/-) mice had a major reduction in neovascularization compared to wild-type controls. Furthermore, the size of tumors resulting from implanted B16F0 mouse melanoma cells was significantly smaller in adam15(-/-) mice than in wild-type controls. Since ADAM15 does not appear to be required for developmental angiogenesis or for adult homeostasis, it may represent a novel target for the design of inhibitors of pathological neovascularization.     

Smooth muscle length-dependent PI(4,5)P2 synthesis and paxillin tyrosine phosphorylation

We studiedeffects of increasing the length of porcine trachealis muscle on 5.5 µM carbachol (CCh)-evoked phosphatidylinositol 4,5-bisphosphate[PI(4,5)P₂] synthesis and other parametersof phosphatidylinositol (PI) turnover.PI(4,5)P₂ resynthesis rates in muscle held at1.0 optimal length (L_o), measured over the first 6 min of CCh stimulation, were 140 ± 12 and 227 ± 14% ofvalues found in muscle held at 0.5 L_o and infree-floating muscle, respectively. Time-dependent changes in cellularmasses of PI(4,5)P₂, PI, and phosphatidicacid, and PI resynthesis rates, were also altered by the muscle lengthat which contraction occurred. In free-floating muscle, CCh did notevoke increases in tyrosine-phosphorylated paxillin (PTyr-paxillin), anindex of ₁-integrin signaling; however, there wereprogressive increases in PTyr-paxillin in muscle held at 0.5 and 1.0 L_o during contraction, which correlated withincreases in PI(4,5)P₂ synthesis rates. Thesedata indicate that PI(4,5)P₂ synthesis ratesand other parameters of CCh-stimulated inositol phospholipid turnoverare muscle length-dependent and provide evidence that supports thehypothesis that length-dependent ₁-integrin signals mayexert control on CCh-activated PI(4,5)P₂ synthesis.

     

Mouse smad8 phosphorylation downstream of BMP receptors ALK-2, ALK-3, and ALK-6 induces its association with Smad4 and transcriptional activity

Smads are intracellular signaling mediators for TGF-beta superfamily. Smad1 and Smad5 are activated by BMP receptors. Here, we have cloned mouse Smad8 and functionally characterized its ability to transduce signals from BMP receptors. Constitutively active BMP type I receptors, ALK-3 and ALK-6, as well as ALK-2, were phosphorylated Smad8 and induced Smad8 interaction with Smad4. Nuclear translocation of Smad8 was stimulated by constitutively active BMP type I receptors. In contrast, constitutively active TGF-beta type I receptor, ALK-5, did not exhibit any action on Smad8. Smad8 and Smad4 cooperatively induced the promoter of Xvent2, a homeobox gene that responds specifically to BMP signaling. Dominant-negative Smad8 was shown to inhibit the increase of alkaline phosphatase activity induced by BMP-2 on pluripotent mesenchymal C3H10T1/2 and myoblastic C2C12 cell lines. The presence of Smad8 mRNA in mouse calvaria cells and osteoblasts suggests a role of Smad8 in the osteoblast differentiation and maturation.     

DIP: the database of interacting proteins

The Database of Interacting Proteins (DIP; http://dip.doe-mbi.ucla.edu) is a database that documents experimentally determined protein-protein interactions. This database is intended to provide the scientific community with a comprehensive and integrated tool for browsing and efficiently extracting information about protein interactions and interaction networks in biological processes. Beyond cataloging details of protein-protein interactions, the DIP is useful for understanding protein function and protein-protein relationships, studying the properties of networks of interacting proteins, benchmarking predictions of protein-protein interactions, and studying the evolution of protein-protein interactions.     

A functional link between dynamin and the actin cytoskeleton at podosomes

Cell transformation by Rous sarcoma virus results in a dramatic change of adhesion structures with the substratum. Adhesion plaques are replaced by dot-like attachment sites called podosomes. Podosomes are also found constitutively in motile nontransformed cells such as leukocytes, macrophages, and osteoclasts. They are represented by columnar arrays of actin which are perpendicular to the substratum and contain tubular invaginations of the plasma membrane. Given the similarity of these tubules to those generated by dynamin around a variety of membrane templates, we investigated whether dynamin is present at podosomes. Immunoreactivities for dynamin 2 and for the dynamin 2-binding protein endophilin 2 (SH3P8) were detected at podosomes of transformed cells and osteoclasts. Furthermore, GFP wild-type dynamin 2aa was targeted to podosomes. As shown by fluorescence recovery after photobleaching, GFP-dynamin 2aa and GFP-actin had a very rapid and similar turnover at podosomes. Expression of the GFP-dynamin 2aa(G273D) abolished podosomes while GFP-dynamin(K44A) was targeted to podosomes but delayed actin turnover. These data demonstrate a functional link between a member of the dynamin family and actin at attachment sites between cells and the substratum.     

Genetic characterization of the Drosophila melanogaster Suppressor of deltex gene: A regulator of notch signaling.

The Notch receptor signaling pathway regulates cell differentiation during the development of multicellular organisms. A number of genes are known to be components of the pathway or regulators of the Notch signal. One candidate for a modifier of Notch function is the Drosophila Suppressor of deltex gene [Su(dx)]. We have isolated four new alleles of Su(dx) and mapped the gene between 22B4 and 22C2. Loss-of-function Su(dx) mutations were found to suppress phenotypes resulting from loss-of-function of Notch signaling and to enhance gain-of-function Notch mutations. Hairless, a mutation in a known negative regulator of the Notch pathway, was also enhanced by Su(dx). Phenotypes were identified for Su(dx) in wing vein development, and a role was demonstrated for the gene between 20 and 30 hr after puparium formation. This corresponds to the period when the Notch protein is involved in refining the vein competent territories. Taken together, our data indicate a role for Su(dx) as a negative regulator of Notch function.