The potential for egg-guarding care in females of the damselfish,Dascyllus reticulatus,in the absence of uniparental male care

Male damselfish typically demonstrate uniparental egg-guarding care in nature. Potential plasticity in sexual behavior has recently been reported in various teleost fish. To examine behavioral plasticity in parental care, we conducted aquarium experiments to explore the potential for egg-guarding care in the female damselfish, Dascyllus reticulatus. After initial caretaking, males were removed from the mating nests, and cohabiting females frequently exhibited egg predation on the same day. However, we confirmed that females showed significantly decreased egg-predation frequencies on the following day and showed egg-caring behaviors. All experimental females guarded their eggs until they hatched. Females subsequently spawned eggs as females even after performing parental care behaviors, indicating no progression of sex change into males. Molecular analysis of select pituitary gland hormones indicated that egg-caring females and males showed high expression levels of prolactin, suggesting its involvement in the development of parental care behaviors. The cryptic possession of caretaking ability in females may be a tactical response to the need for temporary replacement of the care roles in cases where caretaking males are removed, for example, through predation, in damselfish species living in sexually cohabiting groups.     

A knock-in mouse model of N-terminal R420W mutation of cardiac ryanodine receptor exhibits arrhythmogenesis with abnormal calcium dynamics in cardiomyocytes

Cardiac ryanodine receptor gene (RyR2) mutations cause fatal arrhythmogenic diseases such as catecholaminergic polymorphic ventricular tachycardia and arrhythmogenic right ventricular cardiomyopathy. The N-terminal region of RyR2 is one of the hot spots for mutations. In this study, we investigated cardiac phenotypes of a knock-in mouse model carrying R420W mutation of RyR2. The N-terminal R420W mutation has already been found in juvenile sudden death cadavers of unrelated families. The depolarization-induced Ca²⁺ transient amplitude was significantly lower in cardiomyocytes from RyR2^R420W/R420W mice compared with wild-type mice. The time to peak of the Ca²⁺ transient was significantly increased in RyR2^R420W/R420W mice. Furthermore, the prolonged decay time from the peak of the Ca²⁺ transient was detected in RyR2^R420W/R420W mice. ECG telemetry revealed that various types of arrhythmias were induced in RyR2^R420W/R420W mice in response to administration of caffeine and adrenaline. The mutant mice showed high occurrences of arrhythmias in response to heart stimulants compared with wild-type mice. These findings suggest that R420W mutation impairs depolarization-induced Ca²⁺ oscillation in cardiomyocytes, which possibly results in sudden death due to stress-induced arrhythmias.     

Nest thermoregulation in Polybia scutellaris (White) (Hymenoptera: Vespidae)

Polybia scutellaris (White) builds large nests characterized by numerous spiny projections on the surface. In order to determine whether or not the nest temperature is maintained because of homeothermic conditions of the nest individuals or otherwise, we investigated the thermal conditions within the nests built by P. scutellaris. We measured the temperature within active and abandoned nests. The temperature in the active nest was almost stable at 27°C during data collection, whereas the temperature in the abandoned nest varied with changes in ambient temperature. These results suggest that nest temperature was maintained by the thermogenesis of the individuals of the colony. This is the first report of nest incubation caused by thermogenesis of species of Polybia wasps.     

A part of the Japan Sea form of the threespine stickleback, Gasterosteus aculeatus, spawns in the seawater tidal pools of western Hokkaido Island, Japan

The Japan sea form of the threespine stickleback, Gasterosteus aculeatus, was found to spawn in the seawater tidal pool of the Cape Benkei, western Hokkaido Island, Japan, in 2001 and 2003. Nest-guarding males, adult females and juveniles of this form were also observed in four tidal pools of the Capes Benkei, Kabuto and Kamui, in 2001–2003. Otolith Sr:Ca ratios in sticklebacks collected from these tidal pools were relative high from the core to the edge, suggesting they lived in seawater environments throughout its life cycle. These findings suggest that a part of the Japan Sea formȁ9s individuals in western Hokkaido Island have the potential to breed in the coastal sea and the life history style has evolved in dependence on the seawater environment.     

Determination of endogenous peptides in the porcine brain: possible construction of peptidome,a fact database for endogenous peptides

Peptides play crucial roles in many physiological events. However, a database for endogenous peptides has not yet been developed, because the peptides are easily degraded by proteolytic enzymes during extraction and purification. In this study, we demonstrated that the data for endogenous peptides could be collected by minimizing the proteolytic degradation. We separated porcine brain peptides into 5250 fractions by 2-dimensional chromatography (first ion-exchange and second reversed-phase high-performance liquid chromatography), and 75 fractions of average peptide contents were analyzed in detail by mass spectrometers and a protein sequencer. Based on the analysis data obtained in this study, more than 10000 peptides were deduced to be detected, and more than 1000 peptides to be identified starting from 2 g of brain tissue. Thus, we deduce that it is possible to construct a database for endogenous peptides starting from a gram level of tissue by using 2-dimensional high-performance liquid chromatography coupled with a mass spectrometer.     

Intranuclear verrucomicrobial symbionts and evidence of lateral gene transfer to the host protist in the termite gut

In 1944, Harold Kirby described microorganisms living within nuclei of the protists Trichonympha in guts of termites; however, their taxonomic assignment remains to be accomplished. Here, we identified intranuclear symbionts of Trichonympha agilis in the gut of the termite Reticulitermes speratus. We isolated single nuclei of T. agilis, performed whole-genome amplification, and obtained bacterial 16S rRNA genes by PCR. Unexpectedly, however, all of the analyzed clones were from pseudogenes of 16S rRNA with large deletions and numerous sequence variations even within a single-nucleus sample. Authentic 16S rRNA gene sequences were finally recovered by digesting the nuclear DNA; these pseudogenes were present on the host Trichonympha genome. The authentic sequences represented two distinct bacterial species belonging to the phylum Verrucomicrobia, and the pseudogenes have originated from each of the two species. Fluorescence in situ hybridization confirmed that both species are specifically localized, and occasionally co-localized, within nuclei of T. agilis. Transmission electron microscopy revealed that they are distorted cocci with characteristic electron-dense and lucent regions, which resemble the intranuclear symbionts illustrated by Kirby. For these symbionts, we propose a novel genus and species, ‘Candidatus Nucleococcus trichonymphae'' and ‘Candidatus Nucleococcus kirbyi''. These formed a termite-specific cluster with database sequences, other members of which were also detected within nuclei of various gut protists, including both parabasalids and oxymonads. We suggest that this group is widely distributed as intranuclear symbionts of diverse protists in termite guts and that they might have affected the evolution of the host genome through lateral gene transfer.     

Structure of cytochrome b5 unique to tardigrades

Cytochrome b₅ is an essential electron transfer protein, which is ubiquitously found in living systems and involved in wide variety of biological processes. Tardigrades (also known as water bears), some of which are famous for desiccation resistance, have many proteins unique to them. Here, we report spectroscopic and structural characterization of a cytochrome b₅ like protein from one of the desiccation‐tolerant tardigrades, Ramazzottius varieornatus strain YOKOZUNA‐1 (RvCytb₅). A 1.4 Å resolution crystal structure revealed that RvCytb₅ is a new cytochrome b₅ protein specific to tardigrades.     

Efficient production of Japanese encephalitis virus-like particles by recombinant lepidopteran insect cells

The production of Japanese encephalitis (JE) virus-like particles (VLPs) in stably transformed lepidopteran insect cells was investigated. The DNA fragment encoding the JE virus (JEV) prM signal peptide, the precursor (prM) of the viral membrane protein (M), and the envelope glycoprotein (E) was cloned into the plasmid vector pIHAbla. The pIHAbla contained the Bombyx mori actin promoter downstream of the B. mori nucleopolyhedrovirus (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression, together with a blasticidin resistance gene for use as a selectable marker. DNA encoding a form of prM with a pr/M cleavage site mutation was used to suppress the cell-fusion activity of VLPs. After transfection with the resultant plasmid, Trichoplusia ni BTI-TN-5B1-4 (High Five) cells were incubated with blasticidin, and cells resistant to the antibiotic were obtained. Western blot analysis and enzyme-linked immunosorbent assay of a culture supernatant showed that transfected High Five cells secreted an E antigen equivalent to the authentic JEV E. Sucrose density-gradient sedimentation analysis of the culture supernatant from recombinant High Five cells indicated that secreted E antigen molecules were produced in a particulate form. VLPs recovered from the supernatant successfully induced neutralizing antibodies in mice, particularly when adsorbed to alum adjuvant. High yields (≈30 μg/ml) of E antigen were achieved in shake-flask cultures. These results indicate that recombinant insect cells may offer a novel approach for efficient VLP production.     

New 2-Acylcyclohexane-1,3-diones: Kairomone Components against a Parasitic Wasp,Venturia canescens,from Feces of the Almond Moth,Cadra cautella,and the Indian Meal Moth,Plodia interpunctella

As new kairomone components, 2-palmitoyl- (I) and 2-stearoylcyclohexane-1,3-dione (II) were identified for the parasitic wasp Venturia canescens from feces of the Indian meal moth larva, Plodia interpunctella. One more component, 2-arachidoylcyclohexane-1,3-dione (III), together with the afore mentioned two compounds were newly obtained from feces of the almond moth larva Cadra cautella.

Purified kairomone fractions from both species indicated similar activity to synthetic 2- oleoylcyclohexane-1,3-dione (active at 0.1 mg/ml). 2-Palmitoylcyclohexane-1,3-dione (I) was the most active (at 1 mg/ml) among the synthetic homologues with saturated 2-acyl groups.

No traces of 2-acylcyclohexane-1,3-dione were found in the rearing medium of the host; therefore, the kairomone was suggested to have been biosynthesized.