Many lepidopteran larvae produce silk secretions to build feeding tubes and cocoons that play important protective roles in their lives. Recent research on the silk of bombycoid and pyralid moths has shown that it contains several highly abundant silk components with remarkable mechanical properties. It was also found to contain a number of other proteins of which the functions have yet to be identified. To gain an overview of the silk composition in more primitive lepidopteran species and to identify the core silk components common to most species, we analyzed the cocoon proteins of Tineola bisselliella, which belongs to the basal ditrysian moth line. Using de novo transcriptome sequencing combined with mass spectrometry (MS)-based proteomics, we detected more than 100 secretory proteins in the silk cocoons. Fibroin, sericins, and protease inhibitors were found to be the most abundant proteins, along with several novel candidate silk components. We also verified the tissue and developmental stage specificity of the silk protein expression and characterized the morphology of both the silk glands and silk in T. bisselliella. Our study provides a detailed analysis of silk in the primitive moth, expands the known set of silk-specific genes in Lepidoptera, and helps to elucidate their evolutionary relationships.
- MeSH
- biologická evoluce * MeSH
- fibroiny metabolismus MeSH
- hedvábí * chemie genetika metabolismus MeSH
- hmyzí proteiny genetika metabolismus MeSH
- inhibitory proteas metabolismus MeSH
- larva genetika metabolismus fyziologie MeSH
- můry * genetika metabolismus fyziologie MeSH
- proteomika metody MeSH
- sericiny metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The late 5th instar caterpillar of the cecropia silk moth (Hyalophora cecropia) spins a silken cocoon with a distinct, multilayered architecture. The cocoon construction program, first described by the seminal work of Van der Kloot and Williams, consists of a highly ordered sequence of events. We perform behavioral experiments to re-evaluate the original cecropia work, which hypothesized that the length of silk that passes through the spinneret controls the orderly execution of each of the discrete events of cocoon spinning. We confirm and extend by three-dimensional scanning and quantitative measurements of silk weights that if cocoon construction is interrupted, upon re-spinning, the caterpillar continues the cocoon program from where it left off. We also confirm and extend by quantitative measurements of silk weights that cecropia caterpillars will not bypass any of the sections of the cocoon during the construction process, even if presented with a pre-spun section of a cocoon spun by another caterpillar. Blocking silk output inhibits caterpillars from performing normal spinning behaviors used for cocoon construction. Surprisingly, unblocking silk output 24-hr later did not restart the cocoon construction program, suggesting the involvement of a temporally-defined interval timer. We confirm with surgical reductions of the silk glands that it is the length of silk itself that matters, rather than the total amount of silk extracted by individuals. We used scanning electron microscopy to directly show that either mono- or dual-filament silk (i.e., equal silk lengths but which vary in their total amount of silk extracted) can be used to construct equivalent cocoons of normal size and that contain the relevant layers. We propose that our findings, taken together with the results of prior studies, strongly support the hypothesis that the caterpillar uses a silk "odometer" to measure the length of silk extracted during cocoon construction but does so in a temporally regulated manner. We further postulate that our examination of the anatomy of the silk spinning apparatus and ablating spinneret sensory output provides evidence that silk length measurement occurs upstream of output from the spinneret.
- MeSH
- biobehaviorální přístup MeSH
- biologická proměna fyziologie MeSH
- bourec anatomie a histologie fyziologie MeSH
- chování zvířat fyziologie MeSH
- čití, cítění fyziologie MeSH
- hedvábí analýza chemie metabolismus MeSH
- kukla fyziologie MeSH
- Manduca anatomie a histologie fyziologie MeSH
- mikroskopie elektronová rastrovací MeSH
- senzorická zpětná vazba fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Lepidopteran silk is a complex assembly of proteins produced by a pair of highly specialized labial glands called silk glands. Silk composition has been examined only in a handful of species. Here we report on the analysis of silk gland-specific transcriptomes from three developmental stages of the greater wax moth, Galleria mellonella, combined with proteomics, Edman microsequencing and northern blot analysis. In addition to the genes known earlier, we identified twenty seven candidate cDNAs predicted to encode secretory proteins, which may represent novel silk components. Eight were verified by proteomic analysis or microsequencing, and several others were confirmed by similarity with known silk genes and their expression patterns. Our results revealed that most candidates encode abundant secreted proteins produced by middle silk glands including ten sericins, two seroins, one or more mucins, and several sequences without apparent similarity to known proteins. We did not detect any novel PSG-specific protein, confirming that there are only three fibroin subunits. Our data not only show that the number of sericin genes in the greater wax moth is higher than in other species thus far examined, but also the total content of soluble proteins in silk is twice as high in G. mellonella than in B. mori or A. yamamai. Our data will serve as a foundation for future identification and evolutionary analysis of silk proteins in the Lepidoptera.
- MeSH
- fylogeneze MeSH
- glykoproteiny chemie genetika metabolismus MeSH
- hedvábí genetika metabolismus MeSH
- hmyzí proteiny chemie genetika metabolismus MeSH
- larva genetika růst a vývoj metabolismus MeSH
- muciny chemie genetika metabolismus MeSH
- můry genetika růst a vývoj metabolismus MeSH
- proteom * MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční seřazení MeSH
- sericiny chemie genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Agroecosystems are subject to regular disturbances that cause extinction or migration of much of their fauna, followed by recolonization from surrounding refuges. In small-sized aeronaut spiders, such recolonization is potentiated by their ability to rappel and balloon. These are complex behaviors that we hypothesized to be affected by neurotoxins, namely, neonicotinoids. We tested this hypothesis using two common farmland spider species, Oedothorax apicatus (Linyphiidae) and Phylloneta impressa (Theridiidae). The spiders were topically exposed by dorsal wet application or tarsal dry exposure to commercial neonicotinoid formulations Actara 25 WG, Biscaya 240 OD, Mospilan 20 SP and Confidor 200 OD at concentrations that are recommended for application in agriculture. Contact exposure to neonicotinoids suppressed the ability of spiders to produce the major ampullate fiber and anchor it to the substratum by piriform fibrils. Contact exposure to neonicotinoids also suppressed the ballooning behavior that was manifested by climbing to elevated places, adopting a tiptoe position and producing silk gossamer in the wind. Impaired ability of affected common farmland spiders to quickly recolonize disturbed agroecosystems by silk-mediated dispersal may explain their decline in multiple farmland ecosystems, in which neonicotinoids are applied.
- MeSH
- ekosystém * MeSH
- farmy * MeSH
- hedvábí metabolismus MeSH
- insekticidy farmakologie MeSH
- neonikotinoidy farmakologie MeSH
- pavouci metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Seroins are small lepidopteran silk proteins known to possess antimicrobial activities. Several seroin paralogs and isoforms were identified in studied lepidopteran species and their classification required detailed phylogenetic analysis based on complete and verified cDNA sequences. We sequenced silk gland-specific cDNA libraries from ten species and identified 52 novel seroin cDNAs. The results of this targeted research, combined with data retrieved from available databases, form a dataset representing the major clades of Lepidoptera. The analysis of deduced seroin proteins distinguished three seroin classes (sn1-sn3), which are composed of modules: A (includes the signal peptide), B (rich in charged amino acids) and C (highly variable linker containing proline). The similarities within and between the classes were 31-50% and 22.5-25%, respectively. All species express one, and in exceptional cases two, genes per class, and alternative splicing further enhances seroin diversity. Seroins occur in long versions with the full set of modules (AB1C1B2C2B3) and/or in short versions that lack parts or the entire B and C modules. The classes and the modular structure of seroins probably evolved prior to the split between Trichoptera and Lepidoptera. The diversity of seroins is reflected in proposed nomenclature.
