The expansion of GPCR transactivation-dependent signalling to include serine/threonine kinase receptors represents a new cell signalling frontier

G protein-coupled receptor (GPCR) signalling is mediated through transactivation-independent signalling pathways or the transactivation of protein tyrosine kinase receptors and the recently reported activation of the serine/threonine kinase receptors, most notably the transforming growth factor-β receptor family. Since the original observation of GPCR transactivation of protein tyrosine kinase receptors, there has been considerable work on the mechanism of transactivation and several pathways are prominent. These pathways include the “triple membrane bypass” pathway and the generation of reactive oxygen species. The recent recognition of GPCR transactivation of serine/threonine kinase receptors enormously broadens the GPCR signalling paradigm. It may be predicted that the transactivation of serine/threonine kinase receptors would have mechanistic similarities with transactivation of tyrosine kinase pathways; however, initial studies suggest that these two transactivation pathways are mechanistically distinct. Important questions are the relative importance of tyrosine and serine/threonine transactivation pathways, the contribution of transactivation to overall GPCR signalling, mechanisms of transactivation and the range of cell types in which this phenomenon occurs. The ultimate significance of transactivation-dependent signalling remains to be defined but it appears to be prominent and if so will represent a new cell signalling frontier.     

Protease-activated-receptor-2 affects protease-activated-receptor-1-driven breast cancer

Mammalian protease-activated-receptor-1 and -2 (PAR₁ and PAR₂) are activated by proteases found in the flexible microenvironment of a tumor and play a central role in breast cancer. We propose in the present study that PAR₁ and PAR₂ act together as a functional unit during malignant and physiological invasion processes. This notion is supported by assessing pro-tumor functions in the presence of short hairpin; shRNA knocked-down hPar2 or by the use of a truncated PAR₂ devoid of the entire cytoplasmic tail. Silencing of hPar2 by shRNA-attenuated thrombin induced PAR₁ signaling as recapitulated by inhibiting the assembly of Etk/Bmx or Akt onto PAR₁-C-tail, by thrombin-instigated colony formation and invasion. Strikingly, shRNA-hPar2 also inhibited the TFLLRN selective PAR₁ pro-tumor functions. In addition, while evaluating the physiological invasion process of placenta extravillous trophoblast (EVT) organ culture, we observed inhibition of both thrombin or the selective PAR₁ ligand; TFLLRNPNDK induced EVT invasion by shRNA-hPar2 but not by scrambled shRNA-hPar2. In parallel, when a truncated PAR₂ was utilized in a xenograft mouse model, it inhibited PAR₁–PAR₂-driven tumor growth in vivo. Similarly, it also attenuated the interaction of Etk/Bmx with the PAR₁-C-tail in vitro and decreased markedly selective PAR₁-induced Matrigel invasion. Confocal images demonstrated co-localization of PAR₁ and PAR₂ in HEK293T cells over-expressing YFP-hPar2 and HA-hPar1. Co-immuno-precipitation analyses revealed PAR₁-PAR₂ complex formation but no PAR₁-CXCR4 complex was formed. Taken together, our observations show that PAR₁ and PAR₂ act as a functional unit in tumor development and placenta-uterus interactions. This conclusion may have significant consequences on future breast cancer therapeutic modalities and improved late pregnancy outcome.     

Development and pathologies of the arterial wall

Arteries consist of an inner single layer of endothelial cells surrounded by layers of smooth muscle and an outer adventitia. The majority of vascular developmental studies focus on the construction of endothelial networks through the process of angiogenesis. Although many devastating vascular diseases involve abnormalities in components of the smooth muscle and adventitia (i.e., the vascular wall), the morphogenesis of these layers has received relatively less attention. Here, we briefly review key elements underlying endothelial layer formation and then focus on vascular wall development, specifically on smooth muscle cell origins and differentiation, patterning of the vascular wall, and the role of extracellular matrix and adventitial progenitor cells. Finally, we discuss select human diseases characterized by marked vascular wall abnormalities. We propose that continuing to apply approaches from developmental biology to the study of vascular disease will stimulate important advancements in elucidating disease mechanism and devising novel therapeutic strategies.     

Indicators of red squirrel (Tamiasciurus hudsonicus) abundance in the whitebark pine zone

We investigated occupied squirrel middens and squirrel sightings and vocalizations as indicators of red squirrel ( Tamiasciurus hudsonicus ) abundance in high-elevation whitebark pine ( Pinus albicaulis ) zone. Data were collected 1984-1989 from line transects located on 2 study sites in the Yellowstone ecosystem. We evaluated the performance of each measure on the basis of precision and biological considerations. We concluded that, of the 3 measures, active middens were the best indicator of red squirrel abundance. We also observed that the density of active middens dropped by 48%-66% between 1987 and 1989, following a severe drought and extensive wildfires that burned one of the study sites during 1988.     

Black-tailed prairie dog populations one year after treatment with rodenticides

Three rodenticide treatments, zinc phosphide with prebait, strychnine with prebait, and strychnine without prebait, were applied to black-tailed prairie dog Cynomys ludovicianus colonies in west central South Dakota. Results were compared immediately posttreatment and for one year after application. Zinc phosphide was the most effective for reducing prairie dog numbers immediately. When burrow activity levels of prairie dogs were initially reduced by 45% with strychnine only, they returned to untreated levels within ten months. When initial reductions were 95% with zinc phosphide, however, the number of active burrows was still reduced 77% in September the following year. Strychnine with prebait treatment showed initial reductions of 83% in burrow activity. Bait consumption by prairie dogs was highest for zinc phosphide.     

Field study of plant survival as affected by amendments to bentonite spoil

Efforts to reclaim amended and raw bentonite spoils with six plant species (two forbs, three shrubs, and one tree) were evaluated over a 4-year period. Plant species included fourwing saltbush ( Atriplex canescens [Pursh] Nutt.), big sagebrush ( Artemisia tridentata tridentata Nutt.), Rocky Mountain Juniper ( Juniperus scopulorum Sarg.), Russian olive ( Elaeagnus angustifolia L.) common yarrow ( Achillea millifolium L.) and scarlet globemallow ( Sphaeralcea coccinea [Pursh] Rydb.). Spoil treatments included addition of gypsum, sawdust, perlite, straw, and vermiculite; the control treatment was amended. Fourwing saltbush had 52% survival across all spoil treatments, with greatest survival occurring on perlite-treated spoil (80%), followed by gypsum (70%) and vermiculite amendments (70%). Survival of other plant species ranged from 0 to 2% averaged across all treatments after 4 years. No differences in plant survival occurred among amendments when all species were considered.     

Association of black-tailed prairie dog colonies with cattle point attractants in the Northern Great Plains

In October 1991 we recorded all black-tailed prairie dog ( Cynomys ludovicianus ) colonies and cattle points in a 1248-km 2 study area in southwest North Dakota and southeast Montana. Cattle point attractants were defined as fabricated water tanks and long-term supplemental feed sites. We found that a signivicant number of prairie dog colonies encompassed or adjoined cattle point attractants ( p < .001). Prairie dog colonies associated with cattle point attractants were a mean distance of 1.0 km from the next nearest town. The existence of cattle point attractants may encourage prairie dog colonization. Conversely, refraining from using long-term cattle point attractants can discourage prairie dog colonization.     

Cartilage biology, pathology, and repair

Osteoarthritis is one of the most common forms of musculoskeletal disease and the most prominent type of arthritis encountered in all countries. Although great efforts have been made to investigate cartilage biology and osteoarthritis pathology, the treatment has lagged behind that of other arthritides, as there is a lack of effective disease-modifying therapies. Numerous approaches for dealing with cartilage degradation have been tried, but enjoyed very little success to develop approved OA treatments with not only symptomatic improvement but also structure-modifying effect. In this review we discuss the most recent findings regarding the regulation of cartilage biology and pathology and highlight their potential therapeutic values.     

Response of understory species to changes in ponderosa pine stocking levels in the Black Hills

The objective of this study was to test the hypothesis that there are no differences in understory production, by species, due to stocking levels of Pinus ponderosa (ponderosa pine). Understory production was estimated, by species, on 3 replicates each of 8 growing stock levels, ranging from clearcuts to unthinned stands, in both sapling- and pole-sized pine stands (48 plots) over 3 nonconsecutive years. All stands were approximately 70 yr old when thinning treatments were applied. Production of many herbaceous species, especially Agropyron spp. (wheatgrasses) and Carex spp. (sedges), declined as growing stock levels (measured in terms of basal area) of ponderosa pine increased. While trends in total production were similar, there were specific differences between sapling and pole stands. Sedges and Oryzopsis asperfolia (roughleaf ricegrass) produced more in sapling stands, whereas Danthonia intermedia (timber oatgrass) was more abundant in pole stands. Shrub production, dominated by Acrtostaphylos uva-ursi (bearberry), was relatively consistent across all stocking levels except unthinned. Although the total number of species declined as pine basal area increased, a few species, such as Linnaea borealis (twinflower) and Shepherdia canadensis (buffaloberry), were found only under relatively dense pine canopies. While floristic species richness was greater at lower stocking levels of ponderosa pine, the total number of species would be greater if all stocking levels were present.