Cellular migration and contractility are fundamental processes that are regulated by a variety of concerted mechanisms such as cytoskeleton rearrangements, focal adhesion turnover, and Ca2+ oscillations. TRPM4 is a Ca2+-activated non-selective cationic channel (Ca2+-NSCC) that conducts monovalent but not divalent cations. Here, we used a mass spectrometry-based proteomics approach to identify putative TRPM4-associated proteins. Interestingly, the largest group of these proteins has actin cytoskeleton-related functions, and among these nine are specifically annotated as focal adhesion-related proteins. Consistent with these results, we found that TRPM4 localizes to focal adhesions in cells from different cellular lineages. We show that suppression of TRPM4 in MEFs impacts turnover of focal adhesions, serum-induced Ca2+ influx, focal adhesion kinase (FAK) and Rac activities, and results in reduced cellular spreading, migration and contractile behavior. Finally, we demonstrate that the inhibition of TRPM4 activity alters cellular contractility in vivo, affecting cutaneous wound healing. Together, these findings provide the first evidence, to our knowledge, for a TRP channel specifically localized to focal adhesions, where it performs a central role in modulating cellular migration and contractility.

Biomineralization is a process in which specialized cells secrete and deliver inorganic ions into confined spaces limited by organic matrices or scaffolds. Chicken eggshell is the fastest biomineralization system on earth, and therefore, it is a good experimental model for the study of biomineralization. Eggshell mineralization starts on specialized dispersed sites of the soft fibrillar eggshell membranes referred to as negatively charged keratan sulfate mammillae. However, the rest of the fibrillar eggshell membranes never mineralizes, although 21% of their amino acids are acidic. We hypothesized that, relative to the mammillae, the negatively charged amino acids of the fibrillar eggshell membranes are not competitive enough to promote calcite nucleation and growth. To test this hypothesis, we experimentally increased the number of negatively charged carboxylate groups on the eggshell membrane fibers and compared it with in vitro calcite deposition of isolated intact eggshell membranes. We conclude that the addition of poly-carboxylated groups onto eggshell membranes increases the number of surface nucleation sites but not the crystal size.

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This study examines the new fossiliferous site Huapilacuy II of Mid-Holocene age (7,344±51-6,865±58 cal years BP.) located in the northwestern coast of the Isla Grande de Chiloé. This area was not affected by the successive Pleistocene glaciations, and therefore it presents a biogeographic relevance as a potential area of refugia and stability for the vegetation. The presence of plant macrofossils contained in a sedimentary sequence of ca. 300 cm thick, confers a special interest to the site, due to the scarce information available on this type of indicator in paleoenvironmental studies of southern Chile. Additionally, several pollen-based reconstructions from the southern Lake District of Chile (40-44˚ S), document the Holocene sequence of recolonization by the different temperate rainforests types that today occupy this region, although there are non-Holocene records for the Pacific coast of the region. The aim of this study is to reconstruct the local environmental conditions and paleoecology based on the stratigraphic context and the analysis of plant macrofossils at the site Huapilacuy II. In addition, based on the pollen analysis of the deposit, we provide new information to reconstruct the regional characteristics of the vegetation during the Middle-Holocene. In particular, the plant macrofossil record of marsh species contained in the sediments of the lower section of the studied sedimentary sequence, together with the pollen analysis of the same sequence, document a first phase of plant colonization at 7,344±51 cal yrs. BP, with predominance of Poaceae, ferns, and trees with regeneration capacity in open areas, such as Embothrium coccineum and Drimys winteri. The analysis of leaf macrofossils and palynomorphs recovered from several intercalated layers, from the middle section of the sedimentary sequence, show the local and regional development of dense and very humid forests dominated by Aextoxicon punctatum, associated with several species of Myrtaceae. The presence of soil moisture indicator species, such as Luma chequen, Myrceugenia sp. and Myrtaceae Blepharocalyx-type is consistent with the sedimentary environment and the local development of swamp or riverine forests. This hygrophile forest environment is also consistent with the assemblage of fossil mosses, dominated by species that grow today in dense closed-canopy forests, such as Weymouthia, Ptychomnium, Rigodium, Porothamnium and Eucamptodon. The regional correlation of the pollen spectra from Huapilacuy II and other records from the Lake District allows us to establish latitudinal and longitudinal differences of tree composition in the temperate-rainforests that expanded during the Early to Mid-Holocene. In particular, this study established for the northwestern coast of the Isla Grande de Chiloé the presence of the coastal association of the valdivian forest (As. Lapagerio-Aextoxiconetum), currently distributed along the Chilean coastline between 30˚- 43˚S. In contrast, the Valdivian associations recorded in other areas of the region exhibit the dominance of Eucryphia cordifolia, Caldcluvia paniculata, Weinmannia trichosperma and different species of Nothofagus. Despite the differences in tree composition, the fossil bryophyte species recorded in several of the sites compared are common with those reported for Huapilacuy II, thus showing the wide ecological range of Chilean bryophytes associated with closed-canopy temperate-rainforests.