Mandible microwear texture analysis of crickets raised on diets of different abrasiveness reveals universality of diet-induced wear.

Animals have evolved diverse comminuting tools. While vertebrates possess mineralized teeth, insect mandibles often bear metal-inclusion-hardened serrated cusps. Microscopic dental enamel wear (microwear) is known to be caused by contact with ingesta. To test if insect mandible microwear is also diet-dependent, we kept newly moulted adult two-spotted crickets (Gryllus bimaculatus) for four weeks on alfalfa-based rodent pellets with and without added mineral abrasives (loess, quartz, volcanic ash). Six crickets per diet were examined after 1, 3, 7, 14, 21 and 28 days. All diets induced progressive mandible wear, affecting specific locations along the distal tooth cusps differently. The depth of furrows increased on most abrasive-containing diets until day 21, while wear mark complexity increased from day 1 to 3 and 14 to 21. After 28 days, these parameter values for large volcanic ash and large quartz diets significantly exceeded those for the control diet. These results are comparable to observations from guinea pig feeding experiments with the same diets. Cricket mandible wear was affected by all abrasives. Notably, large volcanic ash and large quartz induced the deepest, most complex lesions, akin to observations in guinea pigs. This suggests a universal wear process, supporting that microwear analyses are suitable for inferring invertebrate diets.

Best Practices for Comprehensive Annotation of Neuropeptides of Gryllus bimaculatus.

Genome annotation is critically important data that can support research. Draft genome annotations cover representative genes; however, they often do not include genes that are expressed only in limited tissues and stages, or genes with low expression levels. Neuropeptides are responsible for regulation of various physiological and biological processes. A recent study disclosed the genome draft of the two-spotted cricket Gryllus bimaculatus, which was utilized to understand the intriguing physiology and biology of crickets. Thus far, only two of the nine reported neuropeptides in G. bimaculatus were annotated in the draft genome. Even though de novo assembly using transcriptomic analyses can comprehensively identify neuropeptides, this method does not follow those annotations on the genome locus. In this study, we performed the annotations based on the reference mapping, de novo transcriptome assembly, and manual curation. Consequently, we identified 41 neuropeptides out of 43 neuropeptides, which were reported in the insects. Further, 32 of the identified neuropeptides on the genomic loci in G. bimaculatus were annotated. The present annotation methods can be applicable for the neuropeptide annotation of other insects. Furthermore, the methods will help to generate useful infrastructures for studies relevant to neuropeptides.

Different transcriptional levels of Corazonin, Elevenin, and PDF according to the body color of the two-spotted cricket, Gryllus bimaculatus.

Body color in insects changes according to the living environment and physiological stresses possibly involved in endocrine factors. To date, 3 predominant bioactive peptides, Corazonin, Elevenin, and pigment-dispersing factor (PDF), have been illuminated to be involved in the body color in insects and crustaceans. Here, we examined the possibilities that these 3 factors would contribute to body color changes via melanization in the two-spotted cricket, Gryllus bimaculatus, whose body color changes according to population density drastically. Quantitative analyses revealed the higher transcriptional levels of Corazonin and Elevenin in the crowded-conditioned crickets, whereas the transcriptional level of PDF was higher in the isolated-conditioned crickets. However, the body color was not changed by knockdown of Corazonin, Elevenin, and PDF by RNA interference. The present data indicated that coloration mechanisms in G. bimaculatus is differently controlled from the previous observation in Locusta migratoria, a closely related orthopteran species.

Function of myosuppressin in regulating digestive function in the two-spotted cricket, Gryllus bimaculatus.

Myosuppressin is one of essential peptides controlling biological processes including feeding behavior. Here we identified and characterized the cDNAs that encode myosuppressin precursor and its receptor in the two-spotted cricket Gryllus bimaculatus. The presence of the mature peptide (Grybi-MS) was confirmed by direct measurement of adult brain. RT-PCR revealed the tissue distribution of these transcripts; myosuppressin is expressed predominantly in the brain and central nervous system, whereas its receptor is ubiquitously expressed in the cricket body. To address the function of Grybi-MS, we performed several bioassays to test concerning feeding behavior and digestive function upon exposure to Grybi-MS. Administration of synthetic Grybi-MS resulted in increased feeding motivation, accompanied by an increase in food intake. Meanwhile, the hemolymph lipid and carbohydrate titers were both elevated after Grybi-MS injection. As the intestinal contraction is significantly inhibited by the exposure to Grybi-MS, the upregulating feeding index might be complicated in the cricket body. The current data indicate that Grybi-MS modulates feeding behavior to control the physiological processes in the cricket.

Antagonistic Effect of Short Neuropeptide F on Allatotropin-Inhibited Feeding Motivation of the Silkworm Larva, Bombyx mori.

Here, we demonstrated an antagonistic effect of short neuropeptide F (sNPF) in modulating feeding motivation in the silkworm Bombyx mori; sNPF reduced the feeding-delaying effects caused by administration of an inhibitory peptide, allatotropin (AT). In situ hybridization and MALDI-TOF MS analysis revealed the presence of three subtypes of sNPFs (sNPF-1, -2, and -3) in the midgut enteroendocrine cells. Ca2+-imaging analyses revealed that three subtypes of sNPF receptors (sNPFRs) (BNGR-A7, -A10, and -A11) showed different affinities with the three subtypes of sNPFs. In addition, sNPF activated its signaling via ERK phosphorylation in the midgut, while mixture of sNPF and AT reduced the phosphorylation level, agreeing with the results of behavioral assay. Together, our current findings suggest that intestinal sNPF positively modulates the feeding motivation by reducing the inhibitory effects by AT within the midgut.