BackgroundFree-living flatworms, in both marine and freshwater environments, are able to adhere to and release from a substrate several times within a second. This reversible adhesion relies on adhesive organs comprised of three cell types: an adhesive gland cell, a releasing gland cell, and an anchor cell, which is a modified epidermal cell responsible for structural support. However, nothing is currently known about the molecules that are involved in this adhesion process.ResultsIn this study we present the detailed morphology of the adhesive organs of the free-living marine flatworm Macrostomum lignano. About 130 adhesive organs are located in a horse-shoe-shaped arc along the ventral side of the tail plate. Each organ consists of exactly three cells, an adhesive gland cell, a releasing gland cell, and an anchor cell. The necks of the two gland cells penetrate the anchor cell through a common pore. Modified microvilli of the anchor cell form a collar surrounding the necks of the adhesive- and releasing glands, jointly forming the papilla, the outer visible part of the adhesive organs. Next, we identified an intermediate filament (IF) gene, macif1, which is expressed in the anchor cells. RNA interference mediated knock-down resulted in the first experimentally induced non-adhesion phenotype in any marine animal. Specifically, the absence of intermediate filaments in the anchor cells led to papillae with open tips, a reduction of the cytoskeleton network, a decline in hemidesmosomal connections, and to shortened microvilli containing less actin.ConclusionOur findings reveal an elaborate biological adhesion system in a free-living flatworm, which permits impressively rapid temporary adhesion-release performance in the marine environment. We demonstrate that the structural integrity of the supportive cell, the anchor cell, is essential for this adhesion process: the knock-down of the anchor cell-specific intermediate filament gene resulted in the inability of the animals to adhere. The RNAi mediated changes of the anchor cell morphology are comparable to situations observed in human gut epithelia. Therefore, our current findings and future investigations using this powerful flatworm model system might contribute to a better understanding of the function of intermediate filaments and their associated human diseases.

Two new genera (Streptosarcina and Streptofilum) and three new species (Streptosarcina arenaria, S. costaricana and Streptofilum capillatum) of streptophyte algae were detected in cultures isolated from terrestrial habitats of Europe and Central America and described using an integrative approach. Additionally, a strain isolated from soil in North America was identified as Hormidiella parvula and proposed as an epitype of this species. The molecular phylogeny based on 18S rRNA and rbcL genes, secondary structure of ITS-2, as well as the morphology of vegetative and reproductive stages, cell ultrastructure, ecology and distribution of the investigated strains were assessed. The new genus Streptosarcina forms a sister lineage to the genus Hormidiella (Klebsormidiophyceae). Streptosarcina is characterized by packet-like (sarcinoid) and filamentous thalli with true branching and a cell organization typical for Klebsormidiophyceae. Streptofilum forms a separate lineage within Streptophyta. This genus represents an easily disintegrating filamentous alga which exhibits a cell coverage of unique structure: layers of submicroscopic scales of piliform shape covering the plasmalemma and exfoliate inside the mucilage envelope surrounding cells. The implications of the discovery of the new taxa for understanding evolutionary tendencies in the Streptophyta, a group of great evolutionary interest, are discussed.

IntroductionCochlear micromechanics and frequency tuning depend on the macromolecular organization of the basilar membrane (BM), which is still unclear in man. Novel techniques in cochlear implantation (CI) motivate further analyses of the BM.Materials and methodsNormal cochleae from patients undergoing removal of life-threatening petro-clival meningioma and an autopsy specimen from a normal human were used. Laser-confocal microscopy, high resolution scanning (SEM) and transmission electron microscopy (TEM) were carried out in combination. In addition, one human temporal bone was decellularized and investigated by SEM.ResultsThe human BM consisted in four separate layers: (1) epithelial basement membrane positive for laminin-β2 and collagen IV, (2) BM “proper” composed of radial fibers expressing collagen II and XI, (3) layer of collagen IV and (4) tympanic covering layer (TCL) expressing collagen IV, fibronectin and integrin. BM thickness varied both radially and longitudinally (mean 0.55–1.16 μm). BM was thinnest near the OHC region and laterally.ConclusionsThere are several important similarities and differences between the morphology of the BM in humans and animals. Unlike in animals, it does not contain a distinct pars tecta (arcuate) and pectinata. Its width increases and thickness decreases as it travels apically in the cochlea. Findings show that the human BM is thinnest and probably most vibration-sensitive at the outer pillar feet/Deiter cells at the OHCs. The inner pillar and IHCs seem situated on a fairly rigid part of the BM. The gradient design of the BM suggests that its vulnerability increases apical wards when performing hearing preservation CI surgery.Electronic supplementary materialThe online version of this article (doi:10.1007/s00441-014-2098-z) contains supplementary material, which is available to authorized users.

Green algae of the genus Zygnema form extensive mats and produce large amounts of biomass in shallow freshwater habitats. Environmental stresses including freezing may perturb these mats, which usually have only annual character. To estimate the limits of survival at subzero temperatures, freezing resistance of young Zygnema sp. (strain MP2011Skan) cells and pre-akinetes was investigated. Young, 2-week-old cultures were exposed to temperatures of 0 to – 14 °C at 2-K steps, whereas 8-month-old cultures were frozen from – 10 to – 70 °C at 10-K intervals. Cell viability after freezing was determined by 0.1% auramine O vital fluorescence staining and measurements of the effective quantum yield of photosystem II (ФPSII). At – 8 °C, the young vegetative cells were unable to recover from severe frost damage. But temperatures even slightly below zero (– 2 °C) negatively affected the cells’ physiology. Single pre-akinetes could survive even at – 70 °C, but their LT50 value was – 26.2 °C. Severe freezing cytorrhysis was observed via cryo-microscopy at – 10 °C, a temperature found to be lethal for young cells. The ultrastructure of young cells appeared unchanged at – 2 °C, but severe damage to biomembranes and formation of small foamy vacuoles was observed at – 10 °C. Pre-akinetes did not show ultrastructural changes at – 20 °C; however, vacuolization increased, and gas bubbles appeared at – 70 °C. Our results demonstrate that the formation of pre-akinetes increases freezing resistance. This adaptation is crucial for surviving the harsh temperature conditions prevailing in the High Arctic in winter and a key feature in seasonal dynamics of Zygnema sp.