Long-term exposure to muscarinic agonists decreases expression of contractile proteins and responsiveness of rabbit tracheal smooth muscle cells

Background: Chronic airway diseases, like asthma or COPD, are characterized by excessive acetylcholine release and airway remodeling. The aim of this study was to investigate the long-term effect of muscarinic agonists on the phenotype and proliferation of rabbit tracheal airway smooth muscle cells (ASMCs). Methods: ASMCs were serum starved before treatment with muscarinic agonists. Cell phenotype was studied by optical microscopy and indirect immunofluorescence, using smooth muscle a-actin, desmin and SM-Myosin Heavy Chain (SM-MHC) antibodies. [N-methyl-H-3] scopolamine binding studies were performed in order to assess M-3 muscarinic receptor expression on isolated cell membranes. Contractility studies were performed on isolated ASMCs treated with muscarinic agonists. Proliferation was estimated using methyl-[3H] thymidine incorporation, MTT or cell counting methods. Involvement of PI3K and MAPK signalling pathways was studied by cell incubation with the pathway inhibitors LY294002 and PD98059 respectively. Results: Prolonged culture of ASMCs with acetylcholine, carbachol or FBS, reduced the expression of a-actin, desmin and SM-MHC compared to cells cultured in serum free medium. Treatment of ASMCs with muscarinic agonists for 3-15 days decreased muscarinic receptor expression and their responsiveness to muscarinic stimulation. Acetylcholine and carbachol induced DNA synthesis and increased cell number, of ASMCs that had acquired a contractile phenotype by 7 day serum starvation. This effect was mediated via a PI3K and MAPK dependent mechanism. Conclusions: Prolonged exposure of rabbit ASMCs to muscarinic agonists decreases the expression of smooth muscle specific marker proteins, down-regulates muscarinic receptors and decreases ASMC contractile responsiveness. Muscarinic agonists are mitogenic, via the PI3K and MAPK signalling pathways

Sunlight and Soil-Litter Mixing: Drivers of Litter Decomposition in Drylands

Decomposition of leaf litter is a key component of biogeochemical cycles but the mechanisms driving it in arid and semiarid ecosystems (drylands) remain unresolved. Here, we review recent findings that demonstrate dual roles of solar radiation (ultraviolet and photosynthetically active radiation) and soil–litter mixing as drivers of decomposition in drylands. We focus on the known and potential mechanisms by which these factors influence leaf litter decomposition, explore how the importance of these two drivers may shift over time, and propose possible avenues by which these factors may interact. Special attention is given to UV in sunlight, as this radiation is known to have multiple roles in influencing decomposition and has received considerable recent research attention. We also identify important uncertainties and challenges and offer a generalized conceptual model to guide future research aimed at enhancing our mechanistic understanding and quantitative modeling of the processes by which soil deposition and solar radiation together influence leaf litter decomposition rates in globally extensive dryland ecosystems