Summary of the morphological and ecological traits of Central European dung beetles

Ecological, morphological and life‐history traits have been increasingly used in community ecology during the last decade. Dung beetles represent a model group of insects frequently used in studies of landscape ecology and grassland management. Their body sizes and nesting behavioral traits are regularly used to help understand ecological processes at the community level. However, information on their seasonal activity, wing morphometry and dung specialization is sparse in published reports, or is simply not available yet. We thus compiled a comprehensive list of the morphological and ecological traits of Central European dung beetles (Geotrupidae, Scarabaeidae and Aphodiidae). We gathered information from published works and, for the first time, took morphometric measurements of wings. We provide a database of 12 traits for all 100 dung beetle species occurring in Central Europe. Most species are not restricted to one specific dung type, and the most frequently used dung types are sheep/goat, cattle and horse dung, which are almost equally exploited by 90, 89 and 87 species, respectively. More than one‐third of all species are active in winter, and the number of active species is the highest in June. The wing morphometry shows a high variation and is largely determined by the family identity; the ratio of elytron length to wing area is the largest in Aphodiidae but the smallest in Geotrupidae. Our database is the first standardized set of information for Central European dung beetles and can be used in future trait‐based studies focusing on the ecology and conservation of these beetles.     

Acetylcholine Receptor (AChR) Clustering Is Regulated Both by Glycogen Synthase Kinase 3β (GSK3β)-dependent Phosphorylation and the Level of CLIP-associated Protein 2 (CLASP2) Mediating the Capture of Microtubule Plus-ends

The postsynaptic apparatus of the neuromuscular junction (NMJ) traps and anchors acetylcholine receptors (AChRs) at high density at the synapse. We have previously shown that microtubule (MT) capture by CLASP2, a MT plus-end-tracking protein (+TIP), increases the size and receptor density of AChR clusters at the NMJ through the delivery of AChRs and that this is regulated by a pathway involving neuronal agrin and several postsynaptic kinases, including GSK3. Phosphorylation by GSK3 has been shown to cause CLASP2 dissociation from MT ends, and nine potential phosphorylation sites for GSK3 have been mapped on CLASP2. How CLASP2 phosphorylation regulates MT capture at the NMJ and how this controls the size of AChR clusters are not yet understood. To examine this, we used myotubes cultured on agrin patches that induce AChR clustering in a two-dimensional manner. We show that expression of a CLASP2 mutant, in which the nine GSK3 target serines are mutated to alanine (CLASP2–9XS/9XA) and are resistant to GSK3β-dependent phosphorylation, promotes MT capture at clusters and increases AChR cluster size, compared with myotubes that express similar levels of wild type CLASP2 or that are noninfected. Conversely, myotubes expressing a phosphomimetic form of CLASP2 (CLASP2–8XS/D) show enrichment of immobile mutant CLASP2 in clusters, but MT capture and AChR cluster size are reduced. Taken together, our data suggest that both GSK3β-dependent phosphorylation and the level of CLASP2 play a role in the maintenance of AChR cluster size through the regulated capture and release of MT plus-ends.     

Delaying insect access alters community composition on small carrion: a quantitative approach

We present a study of interactions in the highly competitive insect communities inhabiting the carrion of small mammals. Via manipulation in a fully quantitative design, we delayed community development by excluding insect colonization in mouse and rat carcasses for 3 days, to study the role of early competitively dominant colonizers [burying beetles (Nicrophorus spp.; Coleoptera: Silphidae) and blowfly larvae (Diptera: Calliphoridae)] in the course of heterotrophic succession on small cadavers. Earlier studies demonstrated that in the case of large mammalian carrion, exclusion of insects’ access to the carcass in the early stages of decomposition altered the successional trajectory and species assemblages. However, the effect of such manipulation in easy monopolizable small vertebrate carrion remained unknown. Our results demonstrate that delaying insect access to carrion significantly lowered blowfly larvae abundances, while it simultaneously had no effect on colonization and carrion burial by burying beetles. Higher abundances of blowfly larvae seem to deter necrophagous beetles, whereas they are not harmful to the larvae of flesh flies, at least in larger rat carcasses. Predatory beetle species preferred the lower abundances of blowfly larvae, presumably due to better accessibility of their prey. Our results therefore suggest that in the course of the entire season, larvae of blowflies are the dominant competitors in small carcasses, and significantly affect the assembly of other insect groups, whereas burying beetles may exhibit a more temporal pattern of dominance.     

Temporal segregations in the surface community of an ephemeral habitat: Time separates the potential competitors of coprophilous Diptera

Temporal separations among species greatly enhance the species' coexistence, especially in insect communities inhabiting temporally unstable, yet resource‐rich, ephemeral habitats like dung or carrion. The insect communities inhabiting ephemeral habitats consist of two components, the internal community dwelling within the substrate (mostly Coleoptera), and the surface community inhabiting the habitat's outer rim (mostly adult Diptera). In contrast to the internal community, the surface community is very rarely studied. We present here the first quantitative study of temporal trends in the surface community of coprophilous dipteran adults. Using artificially created 1.5 L cow dung pats, we studied the succession and seasonality in the surface community during six sampling periods in 2011 and 2012. In total, we sampled 13579 adults of dung‐visiting Diptera. Both the abundance and species richness decreased rapidly throughout the succession, and were highest during summer. Along the successional gradient, the community was separated into two main groups (early and late) and four subgroups: (i) species occurring during the first few hours (mainly Calyptratae: Diptera); (ii) species occurring between the first and second days; (iii) species occurring between the second and third days (mainly Acalyptratae: Diptera); and (iv) species with optima after the third day of dung pat existence (mainly Nematocera). The earliest and latest successional groups, occurring mainly during spring–autumn, were seasonally separated from the two mid‐successional groups, occurring during summer. The ecologically similar species displayed detectable seasonal micro‐optima, which likely facilitate their coexistence. There was a high overall similarity in temporal patterns between dung and carrion surface communities.     

CLASP2-dependent microtubule capture at the neuromuscular junction membrane requires LL5β and actin for focal delivery of acetylcholine receptor vesicles

A hallmark of the neuromuscular junction (NMJ) is the high density of acetylcholine receptors (AChRs) in the postsynaptic muscle membrane. The postsynaptic apparatus of the NMJ is organized by agrin secreted from motor neurons. The mechanisms that underlie the focal delivery of AChRs to the adult NMJ are not yet understood in detail. We previously showed that microtubule (MT) capture by the plus end–tracking protein CLASP2 regulates AChR density at agrin-induced AChR clusters in cultured myotubes via PI3 kinase acting through GSK3β. Here we show that knockdown of the CLASP2-interaction partner LL5β by RNAi and forced expression of a CLASP2 fragment blocking the CLASP2/LL5β interaction inhibit microtubule capture. The same treatments impair focal vesicle delivery to the clusters. Consistent with these findings, knockdown of LL5β at the NMJ in vivo reduces the density and insertion of AChRs into the postsynaptic membrane. MT capture and focal vesicle delivery to agrin-induced AChR clusters are also inhibited by microtubule- and actin-depolymerizing drugs, invoking both cytoskeletal systems in MT capture and in the fusion of AChR vesicles with the cluster membrane. Combined our data identify a transport system, organized by agrin through PI3 kinase, GSK3β, CLASP2, and LL5β, for precise delivery of AChR vesicles from the subsynaptic nuclei to the overlying synaptic membrane.     

Agrin regulates CLASP2-mediated capture of microtubules at the neuromuscular junction synaptic membrane

Transforming growth factor β (TGF-β)-stimulated epithelial-mesenchymal transition (EMT) is an important developmental process that has also been implicated in increased cell invasion and metastatic potential of cancer cells. Expression of the focal adhesion protein Hic-5 has been shown to be up-regulated in epithelial cells in response to TGF-β. Herein, we demonstrate that TGF-β-induced Hic-5 up-regulation or ectopic expression of Hic-5 in normal MCF10A cells promoted increased extracellular matrix degradation and invasion through the formation of invadopodia. Hic-5 was tyrosine phosphorylated in an Src-dependent manner after TGF-β stimulation, and inhibition of Src activity or overexpression of a Y38/60F nonphosphorylatable mutant of Hic-5 inhibited matrix degradation and invasion. RhoC, but not RhoA, was also required for TGF-β- and Hic-5-induced matrix degradation. Hic-5 also induced matrix degradation, cell migration, and invasion in the absence of TGF-β via Rac1 regulation of p38 MAPK. These data identify Hic-5 as a critical mediator of TGF-β-stimulated invadopodia formation, cell migration, and invasion.