In holometabolous insects, the larval body is almost completely decomposed and reconstructed into the adult body during the pupal-pharate adult stages. Therefore, the total energetic cost of this process is a key thermodynamic quantity necessary for evaluating the benefit of their life history. Here, we measured whole-body thermal dissipation of single pupae of the fruit fly, Drosophila melanogaster, during the period from puparium formation to adult eclosion as a function of age, using a high-precision isothermal calorimeter at T = 298 K. The mass-specific energy consumption during the period from the onset of larval-pupal apolysis to adult eclosion was determined to be 2.3 kJ/g for an individual of mass (adult) = 1.0 mg, while it was observed to follow Kleiber's law for individuals smaller than mass (adult) = 1.0 mg. During the pupal-pharate adult period, in addition to the U-shaped variation, several characteristic thermal dissipations related to various events, including somatic muscle contractions, ecdyses, pulsatile hormone secretion in a pharate adult, and vaporization of the exuvial fluid, were observed. The periodic bursts in the pharate adult stage grew exponentially, suggesting that the positive feedback in the metabolic system synchronized with the progression of development, making the energy consumption in this stage more efficient. The present study showed that high-precision calorimetry is a powerful and credible method for measuring not only the total energy spent during development but also the energy spent during every specific developmental event in an organism.

Some anguillid eels migrate thousands of kilometers from their spawning grounds, dispersing across vast geographic areas to fresh and brackish water habitats, where they settle and grow. Japanese eels (Anguilla japonica) and giant mottled eels (A. marmorata) are both found in Japan, although their distributions differ, and their exact distributions are poorly known. We assumed that topographic distribution patterns of Japanese and giant mottled eels must differ among and within rivers along the northwest coast of Kyushu, Japan. Environmental DNA (eDNA) analysis was conducted at 87 sites in 23 rivers. Japanese eel eDNA was detected in 19 rivers (82.6%) and that of giant mottled eels was detected in eight (34.8%). We detected giant mottled eel eDNA in five rivers where they were previously unknown. eDNA for Japanese eels was detected at six of nine sites in the north (66.7%), 13 of 23 sites in Omura (56.5%), and 37 of 55 sites in the south (67.3%). In contrast, giant mottled eel eDNA was detected at one of nine sites in the north (11.1%), no sites in Omura, and 15 of 55 sites in the south (27.3%). There was no correlation between eDNA concentrations of the two species at 10 sites in the five rivers where eDNA of both species was detected. These findings suggest differences in the distribution of the two eel species and the northern distributional limit of giant mottled eels in the area facing the East China Sea.

A thorough understanding of the development of complex plumages in birds necessitates the acquisition of genetic data pertaining to the mechanism underlying this phenomenon from various avian species. The oriental honey-buzzard (Pernis ptilorhynchus orientalis), a tropical summer migrant to Northeast Asia, including Japan, exemplifies this aspect owing to the diversity of its ventral coloration and intra-feather barring patterns. However, genetic polymorphism responsible for this diversity has not been identified yet. This study aimed to investigate the link between dark-plumed phenotypes of this subspecies and haplotypes of the melanocortin-1-receptor (MC1R) gene. A draft sequence of MC1R was constructed using next generation sequencing and subsequently amplified using designed polymerase chain reaction (PCR) primers. The genome sequences of 32 honey-buzzard individuals were determined using PCR, and 12 MC1R haplotype sequences were obtained. Among these haplotypes, we found that unique haplotypes with nine non-synonymous substitutions and four or five synonymous substitutions in the coding region had a perfect correlation with the dark-plumed phenotype. The lack of correlation between the genotype of ASIP coding region and plumage phenotype reiterated that the dark morph is attributable to specific MC1R haplotypes. The absence of a correlation between genetic polymorphisms of MC1R and the intra-feather barring patterns, as well as the diversity observed within lighter ground color classes (pale and intermediate), implies the involvement of alternative molecular mechanisms in the manifestation of the aforementioned phenotypes.

Praesagittifera naikaiensis is an acoel flatworm that inhabits the sandy beaches in the intertidal zone of the Seto Inland Sea. This species carries Tetraselmis sp., a green unicellular chlorophyte, as a symbiont in its body, and depends on algal photosynthetic products to survive. However, the eggs of P. naikaiensis contain no symbiotic algae, and juvenile P. naikaiensis acquire symbionts from the surrounding environment through horizontal transfer after hatching, thereby establishing new symbiotic relationships in each generation. Other acoel species, Symsagittifera spp., also inhabit the Seto Inland Sea shores and acquire symbiotic green algae via horizontal transfers. To characterize their symbionts, these acoels were collected from a wide area of the Seto Inland Sea and partial nucleotide sequences of the chloroplast ribulose diphosphate carboxylase large subunit (rbcL) of the symbiotic algae were determined and used for molecular phylogenetic analysis. Symbionts of both P. naikaiensis and Symsagittifera spp. belonged to the genus Tetraselmis but were phylogenetically distant, and both species established symbiotic relationships with different symbionts even when they were sympatric. To test whether each species selects specific algae in the environment for symbiosis, we established algal strains from P. naikaiensis and Symsagittifera sp. symbionts and conducted uptake experiments on aposymbiotic juveniles of P. naikaiensis. The results suggest that symbiotic algae from Symsagittifera could be taken up by P. naikaiensis juveniles, but were unable to establish a normal symbiotic relationship with the juveniles.

The circadian system comprises multiple clocks, including central and peripheral clocks. The central clock generally governs peripheral clocks to synchronize circadian rhythms throughout the animal body. However, whether the peripheral clock influences the central clock is unclear. This issue can be addressed through a system comprising a peripheral clock (compound eye clock [CE clock]) and central clock (the optic lobe [OL] clock) in the cricket Gryllus bimaculatus. We previously found that the compound eye regulates the free-running period (τ) and the stability of locomotor rhythms driven by the OL clock, as measured by the daily deviation of τ at 30°C. However, the role of the CE clock in this regulation remains unexplored. In this study, we investigated the importance of the CE clock in this regulation using RNA interference (RNAi) of the period (per) gene localized to the compound eye (per^CE-RNAi). The per^CE-RNAi abolished the compound eye rhythms of the electroretinogram (ERG) amplitude and clock gene expression but the locomotor rhythm driven by the OL clock was maintained. The locomotor rhythm of the tested crickets showed a significantly longer τ and greater daily variation of τ than those of control crickets treated with dsDsRed2. The variation of τ was comparable with that of crickets with the optic nerve severed. The τ was considerably longer but was comparable with that of crickets with the optic nerve severed. These results suggest that the CE clock regulates the OL clock to maintain and stabilize τ.

Enucleated erythrocytes are characteristic of adult mammals. In contrast, fish, amphibians, reptiles, birds, and fetal mammals possess nucleated erythrocytes in their circulation. Erythroid maturation is regulated by erythropoietin (EPO) and its receptor (EPOR), which are conserved among vertebrates. In mammals, EPOR on the erythroid progenitor membrane disappears after terminal differentiation. However, in western clawed frog, Xenopus tropicalis, mature erythrocytes maintain EPOR expression, suggesting that they have non-canonical functions of the EPO-EPOR axis rather than proliferation and differentiation. In this study, we investigated the non-canonical functions of EPOR in Xenopus mature erythrocytes. EPO stimulation of peripheral erythrocytes did not induce proliferation but induced phosphorylation of intracellular proteins, including signal transducer and activator of transcription 5 (STAT5). RNA-Seq analysis of EPO-stimulated peripheral erythrocytes identified 45 differentially expressed genes (DEGs), including cytokine inducible SH2 containing protein gene (cish) and suppressor of cytokine signaling 3 gene (socs3), negative regulators of the EPOR-Janus kinase (JAK)-STAT pathway. These phosphorylation studies and pathway analysis demonstrated the activation of the JAK-STAT pathway through EPO-EPOR signaling in erythrocytes. Through comparison with EPO-responsive genes in mouse erythroid progenitors obtained from a public database, we identified 31 novel EPO-responsive genes indicating non-canonical functions. Among these, we focused on ornithine decarboxylase 1 gene (odc1), which is the rate-limiting enzyme in polyamine synthesis and affects hematopoietic progenitor differentiation and the endothelial cell response to hypoxic stress. An EPO-supplemented culture of erythrocytes showed increased odc1 expression followed by a decrease in polyamine-rich erythrocytes, suggesting EPO-responsive polyamine excretion. These findings will advance our knowledge of the unknown regulatory systems under the EPO-EPOR axis and functional differences between vertebrates' nucleated and enucleated erythrocytes.