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Fibronectin (Fn1) fibrils have long been viewed as continuous fibers composed of extended, periodically aligned Fn1 molecules. However, our live-imaging and single-molecule localization microscopy data are inconsistent with this traditional view and show that Fn1 fibrils are composed of roughly spherical nanodomains containing six to eleven Fn1 dimers. As they move toward the cell center, Fn1 nanodomains become organized into linear arrays, in which nanodomains are spaced with an average periodicity of 105±17 nm. Periodical Fn1 nanodomain arrays can be visualized between cells in culture and within tissues; they are resistant to deoxycholate treatment and retain nanodomain periodicity in the absence of cells. The nanodomain periodicity in fibrils remained constant when probed with antibodies recognizing distinct Fn1 epitopes or combinations of antibodies recognizing epitopes spanning the length of Fn1. Treatment with FUD, a peptide that binds the Fn1 N-terminus and disrupts Fn1 fibrillogenesis, blocked the organization of Fn1 nanodomains into periodical arrays. These studies establish a new paradigm of Fn1 fibrillogenesis. This article has an associated First Person interview with the first author of the paper., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Abundance on and access time to the host plant are the pivotal factors in Xylella fastidiosa transmission to olive by the meadow spittlebug Philaenus spumarius. Therefore, olive varieties suitable for the vectors, i.e., plants providing all the necessary cues to the insect for their location, settling and acceptance, and devoid of antixenotic defenses, could be more susceptible to infection than varieties non- or less suitable for the vector. Here we evaluated whether a bacterium-susceptible olive variety, Ogliarola Salentina, could be a more suitable host for P. spumarius than the two resistant varieties Leccino and FS-17. We carried out: (i) an evaluation of between-hosts and within-host preference; (ii) an insect survival analysis; (iii) an Electrical Penetration Graph (EPG)-assisted analysis of the probing behavior; (iv) light microscopy of the tissues the spittlebugs had access to; (v) an analysis of the xylem sap primary metabolites. In choice tests, the insect exhibited a significant preference for Ogliarola Salentina. In addition, spittlebugs displayed longer xylem sap ingestion bouts on the bacterium-susceptible variety compared to resistant genotypes, possibly because of differences in the xylem sap chemical profile rather than xylem anatomy. Spittlebugs preference for Ogliarola over both Leccino and FS-17 could be a relevant and so far overlooked component of the low disease prevalence in these two olive varieties reported in Southern Italian olive orchards. Overall, our data point toward the importance of incorporating studies on vector-plant interaction and host traits of resistance to the vector in research on genotypes resistant to X. fastidiosa. [ABSTRACT FROM AUTHOR]

Many cardiac diseases have been associated with increased fibrosis and changes in the organization of fibrillar collagen. The degree of fibrosis is routinely analyzed with invasive histological and immunohistochemical methods, giving a limited and qualitative understanding of the tissue's morphological adaptation to disease. Our aim is to quantitatively evaluate the increase in fibrosis by three-dimensional imaging of the collagen network in the myocardium using the non-linear optical microscopy techniques Two-Photon Excitation microscopy (TPE) and Second Harmonic signal Generation (SHG). No sample staining is needed because numerous endogenous fluorophores are excited by a two-photon mechanism and highly non-centrosymmetric structures such as collagen generate strong second harmonic signals. We propose for the first time a 3D quantitative analysis to carefully evaluate the increased fibrosis in tissue from a rat model of heart failure post myocardial infarction. We show how to measure changes in fibrosis from the backward SHG (BSHG) alone, as only backward-propagating SHG is accessible for true in vivo applications. A 5-fold increase in collagen I fibrosis is detected in the remote surviving myocardium measured 20 weeks after infarction. The spatial distribution is also shown to change markedly, providing insight into the morphology of disease progression. [ABSTRACT FROM AUTHOR]