Microarchitectural cues drive aligned fibrillar collagen deposition in vivo and in biomaterial scaffolds, but the cell-signaling events that underlie this process are not well understood. Utilizing a multicellular patterning model system that allows for observation of intracellular signaling events during collagen matrix assembly, we investigated the role of calcium (Ca2+) signaling in human mesenchymal stem cells (MSCs) during this process. We observed spontaneous Ca2+oscillations in MSCs during fibrillar collagen assembly, and hypothesized that the transient receptor potential vanilloid 4 (TRPV4) ion channel, a mechanosensitive Ca2+-permeable channel, may regulate this signaling. Inhibition of TRPV4 nearly abolished Ca2+signaling at initial stages of collagen matrix assembly, while at later times had reduced but significant effects. Importantly, blocking TRPV4 activity dramatically reduced aligned collagen fibril assembly; conversely, activating TRPV4 accelerated aligned collagen formation. TRPV4-dependent Ca2+oscillations were found to be independent of pattern shape or subpattern cell location, suggesting this signaling mechanism is necessary for aligned collagen formation but not sufficient in the absence of physical (microarchitectural) cues that force multicellular alignment. As cell-generated mechanical forces are known to be critical to the matrix assembly process, we examined the role of TRPV4-mediated Ca2+signaling in force generated across the load-bearing focal adhesion protein vinculin within MSCs using an FRET-based tension sensor. Inhibiting TRPV4 decreased tensile force across vinculin, whereas TRPV4 activation caused a dynamic unloading and reloading of vinculin. Together, these findings suggest TRPV4 activity regulates forces at cell-matrix adhesions and is critical to aligned collagen matrix assembly by MSCs.

We present the first evidence for isolation by distance in a foliar fungal endophyte that is horizontally transmitted. Cryptic species may be common among microbial symbionts and are important to delimit when exploring their genetic structure and microevolutionary processes. The hyperdiversity of endophytic fungi may be explained in part by cryptic species without apparent ecological and morphological differences as well as genetic diversification within rare fungal species across large spatial scales.

Identifying the influence of environmental variables on the spatio-temporal distribution of organisms remains a central goal of ecology. Although environmental limits to the geographic range of species have been investigated, less information is available regarding the influence of environmental variables limiting species to discrete patches within their geographic range. Identifying environmental variables of importance to the species in question is further complicated where the patches appear to be distributed across a variety of outwardly dissimilar habitats. We assessed the influence of environmental variables on the patchy distribution of Macrozamia lucida L.A.S. Johnson (Zamiaceae) cycads endemic to south-east Queensland, Australia. Here we show that patches of M. lucida are consistently associated with elevated moisture, despite the patches being located in contrasting habitats and topographic positions. Our survey results across five field sites demonstrate that both cycad abundance and adult cycad condition (estimated from measures of adult cycad leaf production and reproductive output) are positively correlated with elevated moisture, as measured by mean minimum relative humidity at the plant–soil interface. The historical biogeography of the genus Macrozamia may be intimately linked with the contraction and fragmentation of Australia’s mesic biomes during the Miocene. It is plausible that current distribution pattern of M. lucida, restricted to discrete patches that meet the species’ environmental requirements, reflects a contraction and fragmentation within its range from a more regular historical distribution under more favourable climatic regimes.