An air puff stimulus evoked the swimming of an intact cricket, Gryllus bimaculatus, placed on a water surface. When only the forelegs were intact, swimming was initiated frequently, but flying was never initiated. On the other hand, flying was initiated when only the middle legs or hindlegs were intact. Therefore, the sensory inputs from the forelegs are important in the initiation of swimming and for the inhibition of flying when on the water surface. After bilateral ablation of the middle legs and hindlegs, the bilateral segments of the remaining forelegs were sequentially ablated from the distal area to the proximal area of the legs. After bilateral ablation of all tarsomeres, the relative occurrence of swimming decreased and that of flying increased. After the following ablation of the bilateral tibiae, most insects responded to an air puff stimulus by flying. Experiments performed after coating the leg surface with enamel resulted in almost the same behavioral change as that observed in the ablation experiments. These results suggest that the sensory receptors responsible for the initiation of swimming and the inhibition of flying are mainly located on the surface of the tibia and the tarsus of the forelegs. The behavioral change between swimming and walking was also studied using methylcellulose solutions of various viscosities. On the methylcellulose solution, the relative occurrence of walking in the crickets increased with an increase in the viscosity of the solution.

To identify the sensory organs that are sensitive to water stimuli in the cricket Gryllus bimaculatus, cuticular structures on the legs and the number of sensory neurons innervating them were studied. Some small hair sensilla on the legs were innervated by 2–5 sensory neurons. All such sensilla had a tiny pore at the tip of their hairs. The diameter of the pore was approximately 0.2 μm. These findings suggest that these are chemosensitive hairs (LCS: leg chemosensillum). Of the three pairs of legs, the anterior legs (forelegs) possessed the largest number of LCSs. Of the five leg segments (i.e., coxa, trochanter, femur, tibia and tarsus), the tarsus possessed the largest number of LCSs on each leg. Electrophysiological investigation by tip recording revealed that some of the LCSs contained water-receptor cells. Because the basitarsus possessed a larger number of LCSs than the other tarsomeres, the distribution of water-receptor-containing LCSs in the basitarsus of a foreleg was investigated morphologically and electrophysiologically. LCSs that contained water-receptor cells were mainly distributed on the ventral surface of the basitarsus. There were two types of water receptor that showed different response patterns to a stimulus, that is, phasic- and tonic-type water receptors. From the distribution of LCSs on the legs, the roles of these different types of water receptors in behavioral selection, that is, the initiation of swimming and the inhibition of flying, will be discussed.

Interleukin (IL)-1α is one of the important cytokines involved in regulating immunological reactions in the mouse skin. However, it is not known whether IL-1α regulates the proliferation and differentiation of mouse epidermal melanocytes. In this study, to investigate the role of IL-1α in the regulation of the proliferation and differentiation of mouse epidermal melanocytes, IL-1α was supplemented to serum-free primary cultures of epidermal cell suspensions from the initiation of the primary culture (keratinocytes and melanoblasts-melanocytes) as well as to pure cultures of melanoblasts-melanocytes (keratinocyte-depleted cultures, after 14 days), and its effect was tested. IL-1α inhibited the proliferation of undifferentiated melanoblasts irrespective of the presence or absence of keratinocytes, whereas the cytokine inhibited the proliferation of differentiated melanocytes only in the presence of keratinocytes. Moreover, IL-1α induced the differentiation of melanocytes and, in addition, stimulated tyrosinase activity, melanin synthesis, and dendritogenesis of melanocytes irrespective of the presence or absence of keratinocytes. These results suggest that IL-1α is involved in inhibiting the proliferation of neonatal murine epidermal melanoblasts and in stimulating the differentiation, melanogenesis, and dendritogenesis of melanocytes. The results also suggest that IL-1α inhibits the proliferation of differentiated melanocytes in cooperation with keratinocyte-derived factors.

We previously provided preliminary evidence for the presence of a putative membrane ecdysone receptor (mEcR) anchored in the plasma membranes of anterior silk glands (ASGs) in Bombyx mori. This receptor may act in concert with the conventional EcR in 20E-dependent programmed cell death of these glands. We report here, for the first time, the solubilization of mEcR from ASG membranes using the zwitterionic detergent CHAPS in the presence of NaCl. Our results show by ligand binding assay that mEcR solubilized this way is functionally active and retains 75% of its native binding activity. We also defined experimental conditions that yielded protein/detergent complexes with partial binding activity, which makes it possible to purify the membrane-bound ecdysone binding protein.

Den sites are a conspicuous feature of Eurasian badgers, Meles meles, and in many environments include large communal burrows used by several group members. In Serra de Grândola, southwest Portugal, nine badgers from three social groups were captured and radio collared from 2000 to 2004. A total of 1,787 locations of badgers in their resting sites were registered along with a brief description of the type of site and weather conditions. Resting sites were grouped according to structure (burrows, shrubs, rocks, hollow trees and man-made structures) and function (main, secondary and occasional). Although main setts were the most frequently used shelter (62.25%), an average of 14 (SD 7.55) resting sites were used in each territory. The pattern of use varied seasonally, showing differences according to sex and social group. Overall, females used more than twice as many occasional resting sites as did males. Generally burrows, predominantly main setts, were most frequently used during winter and autumn, whilst non-burrow shelters were preferred during spring and summer, when the weather was hot, dry and not windy. Proximity to food patches had no apparent influence on the location of resting sites. Our results offered no support for the foraging-related hypotheses that multiple resting sites are a means of conserving energy or of maintaining proximity to rich food patches. We suggest that other factors such as thermoregulation needs, disturbance, and reproductive status, could be influencing the observed pattern of resting-site use by badgers in Serra de Grândola.

We isolated cDNA clones of two estrogen receptors (ER1 and ER2) from testis of the Japanese common goby (Acanthogobius flavimanus). The cDNAs of ER1 contained 3,796 nucleotides, including an open reading frame encoding 564 amino acids (M_r: 61.9 kDa); the cDNA for ER2 was 3,274 nucleotides long, with an open reading frame of 567 amino acids (M_r: 63.5 kDa). The deduced aminoacid sequences of ER1 and ER2 each had a characteristic ER structure consisting of five domains (A/B, DNA-binding, D, ligand-binding, and F) and were homologous to ERα and ERβ of other vertebrates. We therefore named ER1 and ER2 as goby ERα (gERα) and goby ERβ (gERβ), respectively. The DNA- and ligand-binding domains in each gER showed high similarity to the corresponding domains of other vertebrates. Analysis of the distribution of gERα and gERβ mRNAs in tissue by reverse transcription–polymerase chain reaction (RT-PCR) analysis revealed that gERα was expressed at high levels in brain and testis, and gERβ was expressed most strongly in testis. In situ hybridization showed that the mRNA of each gER was expressed mainly in the Sertoli cells of goby testis.

In medaka, we found a C16orf35-like gene in the region within 1 Kbp 3′ downstream of the ψβ end of the 36-Kbp embryonic globin gene cluster (^5′α0^3′–^3′β1^5′–^5′α1^3′–^5′β2^3′–^5′α2^3′–^3′α3^5′–^5′β3^3′–^3′β4^5′–^5′α4^3′–^3′ψα^5′–^5′ψβ^3′). Intron 5 of the gene contained a region having NF-E2 binding sites located between GATA boxes. The region was homologous to human HS-40 in terms of the existence and structure of characteristic transcription-factor binding sites and was named Ol-HS-40. Injection of the fusion gene construct Ol-HS-40-α0(up-2)GFP, consisting of Ol-HS-40, a 5′ upstream 200-bp minimum promoter for α0, and green fluorescent protein (GFP), showed that Ol-HS-40, as in human HS-40, had the ability to strongly enhance GFP expression in erythroid cells of embryos. Further analysis using transgenic technology revealed that Ol-HS-40 had the ability to change the type of the GFP expression from embryo-to-young fish to embryo-to-adult. In addition, the results suggest that Ol-HS-40, although its natural function remains unclear, has strong enhancer activity for the expression of not only the α-globin gene but also the β-globin gene.

The gross anatomy of the mastication system of the giant anteater (Myrmecophaga tridactyla) was examined by means of three-dimensional image analysis. The anteater rotates the mandibles medially and laterally to control its tongue when it is elongated and to house it when it is relaxed. Three-dimensional CT image analysis demonstrated that the shape and size of the oral cavity changes drastically when the mandibles are rotated. The oral cavity expands bilaterally when the dorsal part of the mandibles bend medially. Macroscopic observations and muscle-weight data supported the observation that the superficial temporal and medial pterygoid muscles act as the main medial and lateral rotators of the mandible, respectively. The low height of the mandibular ramus and the incomplete zygomatic arch in this species represent adaptations for the rotational movement of the mandibles, since they both contribute to the medially oriented transmission of force from the temporal muscles and to preventing collision between the mandibles and the cranium during the rotational movement.

Extant and fossil Australian chelonioid turtles were examined for 57 osteological morphometric variables. Data were analysed using principal components analysis and canonical variates analysis, after Burnaby isometric ‘size’ removal, as well as multivariate allometry. Results indicate that Lepidochelys spp. are proportionately differentiated from other Cheloniidae. The majority of chelonioid osteological variables scale isometrically, with less than a third exhibiting positive or negative allometry. Skull length and width, postero-ventral skull, mandible length, scapular and pubic variables are found to be useful for differentiating between extant and extinct chelonioids. Skull length and width, mandible height, jaw symphysis length, premaxilla height, femoral length, scapular, pelvic, plastral and rib variables are established as useful differentiators of cheloniids. Australian fossil protostegids are morphometrically more similar to cheloniids than dermochelyids, and no cranial morphometric evidence could be found for the presence of more than one protostegid species. Some osteological allometric variables may be valuable for use in determining the relationships of chelonioids; however these should be examined in conjunction with morphology-based cladistic analyses to test established phylogenies.

The species- and situation-specific sound production of grasshoppers can be stimulated by focal application of both nicotinic and muscarinic receptor agonists into the central body complex of the protocerebrum. Pressure injection of the intrinsic transmitter acetylcholine only elicits fast and short-lived responses related to nicotinic receptor-mediated excitation. Prolonged sound production that includes complex song patterns requires muscarinic receptor-mediated excitation. In addition, basal muscarinic excitation in the central body neuropil seems to determine the general motivation of a grasshopper to stridulate. To demonstrate that endogenous acetylcholinesterase limits the activation of muscarinic receptors by synaptically released acetylcholine in the central body of Chorthippus biguttulus, we investigated both its presence in the brain and effects on sound production resulting from inhibition of esterase activity. Acetylcholinesterase activity was detected in the upper and lower division of the central body. Both these neuropils known to be involved in the cephalic control of stridulation were also shown to contain muscarinic acetylcholine receptors expressed by columnar neurons suggested to serve as output neurons of the central complex. Pressure injection of the acetylcholinesterase inhibitor eserine into protocerebral control circuits of restrained male grasshoppers stimulated long-lasting stridulation that depended on scopolamine-sensitive muscarinic receptors. In restrained males, eserine released the typical response song by potentiating the stimulatory effect of the conspecific female song. Eserine-mediated inhibition of acetylcholinesterase in the central body prolongs the presence of synaptically released acetylcholine at its postsynaptic receptors and increases its potency to activate muscarinic receptor-initiated signaling pathways acting to promote grasshopper sound production.

Dichotomous spermatogenesis was examined in relation to diapause in the sweet potato hornworm, Agrius convolvuli. In non-diapause individuals, eupyrene metaphase began during the fifth larval instar and eupyrene spermatids appeared in wandering larvae. Bundles of mature sperm were found after pupation. Apyrene spermatocytes also appeared during the fifth larval instar, but meiotic divisions occurred irregularly and their nuclei were discarded from the cells during spermiogenesis. Morphometric analyses of flagellar axonemes showed a variable sperm number in apyrene bundles. The variation ranging from 125 to 256 sperm per bundle indicated abnormal divisions or the elimination of apyrene spermatocytes. In diapause-induced hornworms, spermatogenesis progressed similarly during the larval stages. The cessation of spermatogenesis during diapause is characterized by 1) secondary spermatocytes and sperm bundles degenerating gradually as the diapause period lengthens, and 2) spermatogonia or primary spermatocytes appearing throughout diapause. A TUNEL (TdT-mediated dUTP-biotin nick end-labeling) assay revealed that DNA fragmentation occurred in the nuclei of secondary spermatocytes and early spermatids. Aggregates of heterochromatin along the nuclear membrane indicated the onset of apoptosis, and condensed chromatin was confirmed by electron microscopy to be the apoptotic body. These results show that the degenerative changes in spermatogenic cells during pupal diapause were controlled by apoptosis.

Camptandriid crabs collected in the Kumanoe River Estuary, Kyushu, Japan were studied on the basis of morphological characters and molecular analysis. As a result, a new species, Deiratonotus kaoriae, was recognized. These crabs were found mainly in a creek of the sandy tidal flat within the Kumanoe River Estuary. The new species shares a very diagnostic character, the presence of a transverse ridge on the carapace, with D. cristatus (de Man, 1895) and differs markedly from the other congeners that lack this feature. The new species, however, differs from D. cristatus in the absence of harpoon-shaped setae on the subdistal end of the first gonopod and the presence of an extremely reduced second abdominal segment. According to a molecular analysis based on 12S 16S mitochondrial rRNA gene sequences, with Cleistostoma dilatatum (de Haan, 1833) and Camptandrium sexdentatum Stimpson, 1858 as outgroups, Deiratonotus kaoriae is more closely related to D. cristatus than to D. japonicus (Sakai, 1934).

A new species in the monotypic genus Symbioribates, S. aokii sp. nov., is described on the basis of adult material collected from canopy habitats and wind traps in the Ryukyu Archipelago, southwestern Japan. Symbioribates aokii sp. nov. expresses different dimorphism in the octotaxic system from that of the type species, S. papuensis Aoki, 1966. The characteristics of this genus and its relationships to others of the superfamily Oripodoidea are discussed and a revised diagnosis of the family is given.