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Chimpanzee societies generally show male philopatry and female dispersal. However, demographic data on wild chimpanzee societies from long-term study sites have revealed that some females give birth in their natal group (i.e., “remaining females”). Here, we report two remaining females in the M group in Mahale, Tanzania, and compare their cases with previous reports to explore the social and ecological factors that lead to females remaining in their natal group. The results revealed that neither the social traits of the remaining females nor the ecological factors they experienced showed a coherent trend. However, we found multiple, non-mutually exclusive potential factors that may influence the decision by females to remain in their natal group: a decrease in indirect feeding competition, support from mothers or allomothers in the care of offspring and in aggressive interactions with other individuals, close relationships with the other remaining females, and a short adolescent infertility period. Additionally, we observed a natal female copulating with her older brother, which was the first observation of brother–sister incest in Mahale. Although DNA analysis revealed that her infant was not a product of inbreeding, the pair copulated frequently in the latter half of her estrus period, suggesting that they did not avoid incest behaviorally to avoid inbreeding. Furthermore, there was no hard evidence that the remaining female avoided mating with her maternal brother, suggesting that incest avoidance may not be a proximate factor responsible for female dispersal., 出自集団で出産するメスは「例外」ではないことを解明 --チンパンジー父系社会でメスが出自集団に居残る要因の検討--. 京都大学プレスリリース. 2021-01-15.

Parkinson's disease (PD) is a common neurodegenerative disorder with motor symptoms linked to the loss of dopaminergic neurons in the brain. α-Synuclein is an aggregation-prone neural protein that plays a role in the pathogenesis of PD. In our previous paper, we found that saffron; the stigma of Crocus sativus Linne (Iridaceae), and its constituents (crocin and crocetin) suppressed aggregation of α-synuclein and promoted the dissociation of α-synuclein fibrils in vitro. In this study, we investigated the effect of dietary saffron and its constituent, crocetin, in vivo on a fly PD model overexpressing several mutant α-synuclein in a tissue-specific manner. Saffron and crocetin significantly suppressed the decrease of climbing ability in the Drosophila overexpressing A30P (A30P fly PD model) or G51D (G51D fly PD model) mutated α-synuclein in neurons. Saffron and crocetin extended the life span in the G51D fly PD model. Saffron suppressed the rough-eyed phenotype and the dispersion of the size histogram of the ocular long axis in the eye of A30P fly PD model. Saffron had a cytoprotective effect on a human neuronal cell line with α-synuclein fibrils. These data showed that saffron and its constituent crocetin have protective effects on the progression of PD disease in animals in vivo and suggest that saffron and crocetin can be used to treat PD.

Parkinson’s disease (PD) is a common neurodegenerative disorder with motor symptoms linked to the loss of dopaminergic neurons in the brain. α-Synuclein is an aggregation-prone neural protein that plays a role in the pathogenesis of PD. In our previous paper, we found that saffron; the stigma of Crocus sativus Linné (Iridaceae), and its constituents (crocin and crocetin) suppressed aggregation of α-synuclein and promoted the dissociation of α-synuclein fibrils in vitro. In this study, we investigated the effect of dietary saffron and its constituent, crocetin, in vivo on a fly PD model overexpressing several mutant α-synuclein in a tissue-specific manner. Saffron and crocetin significantly suppressed the decrease of climbing ability in the Drosophila overexpressing A30P (A30P fly PD model) or G51D (G51D fly PD model) mutated α-synuclein in neurons. Saffron and crocetin extended the life span in the G51D fly PD model. Saffron suppressed the rough-eyed phenotype and the dispersion of the size histogram of the ocular long axis in A30P fly PD model in eye. Saffron had a cytoprotective effect on a human neuronal cell line with α-synuclein fibrils. These data showed that saffron and its constituent crocetin have protective effects on the progression of PD disease in animals in vivo and suggest that saffron and crocetin can be used to treat PD.

The reproductive success of male primates is not always associated with dominance status. For example, even though male orangutans exhibit intra-sexual dimorphism and clear dominance relationships exist among males, previous studies have reported that both morphs are able to sire offspring. The present study aimed to compare the reproductive success of two male morphs, and to determine whether unflanged males sired offspring in a free-ranging population of Bornean orangutans, using 12 microsatellite loci to determine the paternity of eight infants. A single flanged male sired most of the offspring from parous females, and an unflanged male sired a firstborn. This is consistent with our observation that the dominant flanged male showed little interest in nulliparous females, whereas the unflanged males frequently mated with them. This suggests that the dominant flanged male monopolizes the fertilization of parous females and that unflanged males take advantage of any mating opportunities that arise in the absence of the flanged male, even though the conception probability of nulliparous females is relatively low.

Bread has been a staple of the human diet since antiquity. Many Saccharomyces cerevisiae strains have been used in raising the dough during bread-making. However, the genetic relationships and characteristics of these strains are not well understood. In this study, we analyzed several S. cerevisiae strains using microsatellite polymorphism. First, we extracted DNA from cooked bread (baked, steamed, and fried) and analyzed S. cerevisiae polymorphisms using six microsatellite loci, which is half of the subset reported previously. To the best of our knowledge, this is the first report that demonstrates the possibility of analyzing S. cerevisiae microsatellite polymorphisms using cooked bread as a source of yeast DNA.

Bocaparvoviruses have been studied extensively owing to their ability to cause respiratory illness or gastroenteritis in humans. Some bocaparvoviruses have been detected in non-human primates (gorillas and chimpanzees), but the diversity and evolution of these viruses are not fully understood. In this study, we collected 107 fecal samples from wild western lowland gorillas in Moukalaba-Doudou National Park in Gabon to investigate the presence of bocaparvoviruses. Using a combination of pan-bocaparvovirus PCR and individual identification by microsatellite genotyping, we found that two samples from two apparently healthy infant gorillas were positive for bocaparvovirus. Sequencing and phylogenetic analyses revealed that the two gorilla bocaparvovirus strains are nearly identical and are closely related to viruses in the species Primate bocaparvovirus 2 (with 86.0% nucleotide identity to a human bocavirus 2 isolate). To our knowledge, this is the first report showing the presence of a non-human primate bocaparovirus within Primate bocaparvovirus 2. Our findings provide novel insights into the diversity and evolution of bocaparvoviruses and highlight the importance of surveying these viruses for the safe management of gorilla-based ecotourism.

Mutations in RNA-binding proteins (RBPs) such as TAR DNA-binding protein 43 (TDP-43) and fused in sarcoma (FUS) are associated with amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Recent evidence suggests that RNA dysregulation mediated by aberrant RBPs may play a critical role in neurodegeneration, but the underlying molecular mechanisms are largely unknown. In this study, we performed whole transcriptome profiling of various brain tissues of a transgenic (Tg) mouse model of ALS/FTD overexpressing the exogenous nuclear localization signal deletion mutant of human FUS (ΔNLS-FUS) to investigate changes associated with the early stages of ALS/FTD. Although there were not many differences in expression profiles between wild-type and Tg mice, we found that Sema3g was significantly upregulated in the frontal cortex and hippocampus of Tg mice. Interestingly, analysis of alternative splicing events identified widespread exons that were differentially regulated in Tg mice in a tissue-specific manner. Our study thus identified aberrant splicing regulation mediated by mutant FUS during the early stages of ALS/FTD. Targeting this aberrant splicing regulation represents a potential therapeutic strategy for ALS/FTD.

Extraction of DNA from non-invasive samples (feces) has been used increasingly in genetic research on wildlife. For effective and reliable genetic analyses, knowledge about which samples should be selected in the field is essential. For this reason, we examined the process of DNA degradation in feces of deer. We collected fresh fecal pellets from three wild deer living in a warm temperate forest. We then assessed the effects of time (3, 5, and 10 days) under three environmental conditions (on the forest floor, on exposed ground, and inside the laboratory) on the rates of correct genotyping (CG), amplification failure (NA), genotyping error among positive amplification (ER), false alleles (FA), and allelic dropout (AD) of 15 microsatellite loci. The rate of CG significantly decreased, and those of NA and FA increased with increasing lapse of time. Rates of CG tended to be highest and those of NA, ER, FA, and AD to be lowest in feces kept inside, followed by those on the forest floor. Suitability of samples for DNA extraction was lowest in fecal pellets left on exposed ground, and we suspect that rain may hasten DNA degradation. NA rate could serve as a reliable indicator of the quality of fecal pellets because it was significantly positively correlated with ER rate. For efficient genetic analyses using deer feces in warm temperate zones, we recommend collecting fecal pellets within 3 days of defecation, during periods without rainfall and from under the cover of trees.

Genetic sex identification and efficient microsatellite genotyping using non-invasive samples are important for assessing genetic diversity and structure of wild mammals. We extracted DNA from fecal samples of endangered proboscis monkeys (Nasalis larvatus) at the Yokohama Zoological Gardens, and successfully identified their sex using a DEAD-box gene marker. Using the information of 31 published microsatellite loci developed for proboscis monkeys, we redesigned the primers for 18 loci and selected microsatellite loci with high heterozygosities and then developed a multiplex PCR system, in which 19 polymorphic microsatellite loci and a sex identification marker were amplified in three multiplex sets. In our developed method, genotyping errors rarely happened using DNA extracted from feces, and we could identify individuals and estimate parent—offspring relationship with high confidence. Our developed method is applicable for genetic analysis of wild proboscis monkeys and will provide us the info...

We investigated the relative abundance of four duiker species and the environmental effect including human impacts on their abundance in the forest of Moukalaba, Gabon. We counted and collected fresh faeces from duikers in thirteen 2-km line transects established in different types of vegetation. Genetic species identification showed that blue duiker (Philantomba monticola), Ogilby’s duiker (Cephalophus ogilbyi, which might include the sister species Peter’s duiker Cephalophus callipygus), bay duiker (Cephalophus dorsalis), and yellow-backed duiker (Cephalophus silvicultor) coexisted. Genetic analyses successfully identified Ogilby’s and bay duiker, which are difficult to discriminate using other methods. The blue, Ogilby’s, and yellow-backed duikers were found in all transects. The dung abundance of bay duiker was quite low compared to other duiker species. This result suggests the importance of accumulating information regarding the abundance of each duiker species for updating their conservation status. The dung count method with genetic species identification may be useful for this purpose. The dung abundance of Ogilby’s duikers and blue duikers was affected by human impacts, while that of yellow-backed duikers was not affected by human impacts. The dung abundance of Ogilby’s duikers was lower in young secondary forests, which are regenerating after significant human disturbance. In contrast, generalized liner mixed model showed that the dung abundance of blue duikers was not affected by vegetation type and was lower in transects nearer to the villages, which might be due to past hunting activity. The difference in human impacts among duiker species should be considered for their conservation management.

A small chimpanzee habitat in the montane forest of Kahuzi-Biega National Park, Democratic Republic of Congo, is connected with the lowland forest of this park through a corridor, which is affected by human encroachment. To assess the conservation status of the chimpanzee population in this small habitat, we estimated the size of the community and evaluated its genetic diversity by using 279 fecal samples collected in the montane forest of Kahuzi. Using autosomal microsatellite (or short tandem repeat, STR) loci, we identified 32 individuals, comprising 19 females and 13 males. Samples from 24 individuals were collected at least twice and a genetic mark-recapture analysis estimated that the community size was 36 (range: 32-42). Data on nest site sharing confirmed that all the samples belonged to the same community. Nest site sharing information may be useful in population studies of unhabituated chimpanzees. The genetic structure and diversity of the 32 genotyped individuals was assessed using Y-chromosomal short tandem repeat (Y-STR) loci and mitochondrial D-loops. One dominant Y-STR haplotype was found, whereas there was no dominant haplotype in the mitochondrial region, reflecting a female-biased dispersal pattern, which is typical of chimpanzees. The genetic diversity for three markers in Kahuzi chimpanzees was comparable to that in other eastern chimpanzee populations. A relatively high heterozygosity and negative inbreeding coefficient (FIS ) for STR loci suggests that the study community belongs to an outbreeding chimpanzee population. These findings suggest that individuals of the study community may have reproductive contact with other chimpanzee individuals from neighboring communities in Kahuzi-Biega National Park, at least in the recent past. Am. J. Primatol. 77:1015-1025, 2015. © 2015 Wiley Periodicals, Inc.

Fecal DNA-based 16S ribosomal RNA (rRNA) gene sequencing using next-generation sequencers allows us to understand the dynamic gut microbiome adaptation of animals to their specific habitats. Conventional techniques of fecal microbiome analysis have been developed within the broad contexts defined by human biology; hence, many of these techniques are not immediately applicable to wild nonhuman primates. In order to establish a standard experimental protocol for the analysis of the gut microbiomes of wild animals, we selected the Japanese macaques (Macaca fuscata yakui) on Yakushima Island. We tested different protocols for each stage of fecal sample processing: storage, DNA extraction, and choice of the sequencing region in the bacterial 16S rRNA gene. We also analyzed the gut microbiome of captive Japanese macaques as the control. The comparison of samples obtained from identical macaques but subjected to different protocols showed that the tested storage methods (RNAlater and lysis buffer) produced effectively the same composition of bacterial operational taxonomic units (OTUs) as the standard frozen storage method, although the relative abundance of each OTU was quantitatively affected. Taxonomic assignment of the detected bacterial groups was also significantly affected by the region being sequenced, indicating that sequencing regions and the corresponding polymerase chain reaction (PCR) primer pairs for the 16S rRNA gene should be carefully selected. This study improves the current standard methods for microbiome analysis in wild nonhuman primates. Japanese macaques were shown to be a suitable model for understanding microbiome adaptation to various environments.

Ecological genetic analyses have recently been applied to the field of biological invasions to describe the genetic background of the invasive species. The raccoon (Procyon lotor) is a medium-sized mammal that is native to North and Central America and has been naturalized into several countries worldwide. In the Boso Peninsula, Japan, raccoons were estimated to have been introduced during the 1990s. We collected DNA samples from 139 raccoons in seven administrative districts and sequenced a part of the mitochondrial D-loop region. We identified two haplotypes of the mitochondrial DNA, which differed by three single nucleotide polymorphisms; therefore, the raccoon population in Boso was founded by at least two females. Although the two haplotypes were identified in all districts, genetic differentiation analysis showed that the northern population was genetically different from the southern population. This genetic gap might not be affected by the landscape, but instead by the expansion history of raccoons. One of the probable hypotheses suggests that the expansion of raccoons may have occurred twice in Boso, a second expansion occurring after raccoons had already spread across Boso. Although this is consistent with historical distribution records, further studies using nuclear microsatellite markers are necessary to validate this hypothesis.

Objectives: This study was conducted to clarify the analgesic effect of toad cake and toad-cake-containing herbal drugs. Methods: We counted the writhing response of mice after the intraperitoneal administration of acetic acid as a nociceptive pain model and the withdrawal response after the plantar surface stimulation of the hind paw induced by partial sciatic nerve ligation of the mice as a neuropathic pain model to investigate the analgesic effect of toad cake and toad-cake-containing herbal drugs. A co-treatment study with serotonin biosynthesis inhibitory drug 4-chloro-DL-phenylalanine methyl ester hydrochloride (PCPA), the catecholamine biosynthesis inhibitory drug α-methyl-DL-tyrosine methyl ester hydrochloride (AMPT) or the opioid receptor antagonist naloxone hydrochloride was also conducted. Results: Analgesic effects in a mouse model of no ciceptive pain and neuropathic pain were shown by oral administration of toad cake and toad-cake-containing herbal drugs. The effects of toad cake and toad-cake-containing herbal drugs disappeared upon co-treatment with PCPA, but not with AMPT or naloxone in the nociceptive pain model; the analgesic effect of toad-cake-containing herbal drugs also dis appeared upon co-treatment with PCPA in the neuropathic pain model. Conclusion: Toad cake and toad-cake-containing herbal drugs have potential for the treatments of nociceptive pain and of neuropathic pain, such as post-herpetic neuralgia, trigeminal neuralgia, diabetic neuralgia, and postoperative or posttraumatic pain, by activation of the central serotonin nervous system.

We isolated and characterized 11 microsatellite markers for Ovophis okinavensis, a viperid species endemic to the Ryukyu Archipelago. These markers showed polymorphism among 25 individuals from Okinawajima Island. The number of alleles per locus was 3–11, and observed and expected heterozygosity were 0.240–0.960 and 0.218–0.853, respectively. Cross-amplification confirmed that at least seven out of the 11 microsatellite markers were applicable to a putative relative, Trimeresurus gracilis. The markers provided here are expected to be valuable for population- and/or individual-level genetic studies of not only O. okinavenesis, but also its relatives.

東アフリカのチンパンジーが同所的に生息するヒョウに食べられている最初の証拠を発見. 京都大学プレスリリース. 2013-05-21.

The male dispersal patterns of western lowland gorillas (WLGs, Gorilla gorilla gorilla) are not well understood. To determine whether most silverbacks stay close to their relatives, we analyzed autosomal and Y-chromosomal microsatellites (STRs) in wild WLGs at Moukalaba, Gabon. We obtained STR genotypes for 38 individuals, including eight silverbacks and 12 adult females in an approximately 40 km2 area. Among them, 20 individuals were members of one identified group (Group Gentil; GG), including one silverback and six adult females. The silverback sired all 13 of the offspring in GG and no Y-STR polymorphism within GG was found, as expected in a one-male group structure. Over all silverbacks sampled, Y-STR diversity was high considering the limited sampling area, and silverbacks with similar Y-STR haplotypes were not always located in nearby areas. Although the misclassification rate of kinship estimates in this study was not negligible, there were no kin dyads among all silverbacks sampled. These results suggest that silverbacks born in the same group do not stay close to each other after maturation. The Y-STR diversity in this study was similar to that of a previous study conducted in an area that was approximately 150 times larger than our study area. Similarity of WLG Y-STR diversity between studies at different sampling scales suggests that male gene flow may not be geographically limited. These results suggest that WLG males normally disperse from their natal areas after maturation, at least, in Moukalaba. Am J Phys Anthropol 151:583–588, 2013. © 2013 Wiley Periodicals, Inc.

Using long-term maternal pedigree data, microsatellite analysis, and behavioral tests, we examined whether personality differences in wild Japanese macaques (Macaca fuscata) are associated with additive genetic effects, maternal influences, or belonging to a particular social group. Behaviors elicited by novel-object tests were defined by a component related to caution around novel-objects (Ob-PC1) and behaviors elicited by novel food-tests were defined by correlated components related to consummatory responses (Fo-PC1) and caution around novel foods (Fo-PC2). The repeatability of Ob-PC1 was modest and not significant; the repeatabilities of Fo-PC1 and Fo-PC2 were moderate and significant. Linear mixed effects models found that sex, age, sex × age, provisioning, trial number, date, time of day, season, and distance to the closest monkey were not related to personality. Linear mixed effects models of females older than 2 years found that high rank was associated with greater caution around novel objects. Linear models were used to determine whether sex, age, group membership, maternal kinship, or relatedness had independent effects on the personality similarity of dyads. These analyses found that pairs of macaques that lived in the same group were less similar in their caution around novel objects, more closely related pairs of macaques were more similar in their tendency to eat novel food, and that pairs of macaques in the same group were more similar in how cautious they were around novel foods. Together, these findings suggest that personality in this population of wild monkeys was driven by rank, genetic effects, and group effects, the latter possibly including the need to exploit different niches in the environment. Electronic supplementary material The online version of this article (doi:10.1007/s10329-016-0572-9) contains supplementary material, which is available to authorized users.

Tanzania is located at the southeastern end of the eastern chimpanzee (Pan troglodytes schweinfurthii) distribution. Except for two national parks, their habitats have been degraded due to human activities. To clarify the gene flow and genetic diversity of chimpanzees in Tanzania, we analyzed the mitochondrial sequences of chimpanzees in six sites (Lwazi, Wansisi, Mahale, Karobwa, Ugalla-Masito, and Gombe), some of which are now isolated. The southernmost habitat (Lwazi) was about 150 km away from the nearest habitat but, considering the geographic distance, the genetic distance of the chimpanzees between Lwazi and the other habitats was not high. In contrast, the genetic distance between the chimpanzees in the northernmost habitat (Gombe), and the other habitats was relatively high considering the geographic distance. The results suggest that the Malagarasi River, which runs between Gombe and the southern habitats, limits gene flow. The genetic difference analyses also suggest that the habitats...

The grasscutter (Thryonomys swinderianus) is a hystricomorph rodent that inhabits mainly Sub-Saharan Africa and its meat is highly relished by most people in this region. The animal is hunted in the wild using techniques that are sometimes detrimental to the environment. It is therefore being domesticated to make the meat readily available to forestall the negative consequences of hunting. There are currently no genetic markers available and we report here, 33 novel microsatellite markers developed using next-generation sequencing technology. The number of alleles ranged from 3 to 11 (mean 6.4) while the observed and expected heterozygosities ranged from 0.188 to 1.000 (mean 0.591) and 0.322–0.873 (mean 0.713), respectively. Cumulative probability of identity (PI) for all loci was very low (PI = 3.1 × 10−33), indicating that these markers can be used for individual identification in the domestic setting. They will also be useful to study the genetic diversity of the grasscutter in the wild for population monitoring and conservation.

Loss of synapses is associated with cognitive impairment in Alzheimer's disease (AD). However, the molecular mechanism underlying this synaptic impairment is not well understood. EphA4 is a substrate of γ-secretase, and the γ-secretase-cleaved EphA4 intracellular domain (EICD) is known to enhance the formation of dendritic spines via activation of the Rac signaling pathway. Here, we show that the amount of Rac1 is significantly reduced, and correlated with the level of EICD in the frontal lobes of AD patients. Biochemical analyses revealed that the amount of membrane-associated EICD was decreased and strongly correlated with the level of membrane-associated Rac1, which is considered to be active Rac1. The synaptic scaffolding protein, postsynaptic density (PSD)-95, was specifically decreased in AD, and the amount of PSD-95 correlated with the level of Rac1. Moreover, the amounts of Rac1 and PSD-95 were negatively correlated with the extent of tau phosphorylation, which is crucial for neurofibrillary tangle formation. These results suggest that attenuation of the EICD-mediated Rac signaling pathway is involved in the synaptic pathogenesis of AD.

In this study, 21 microsatellite markers were used to genotype 196 Egyptian local chickens obtained from Fayoumi (n = 35), Dandarawy (n = 30), Baladi (n = 29), Sinai (n = 30), El-Salam (n = 36), and Golden Montazah (n = 36) strains. The results were compared to two pure commercial chicken populations reared in Japan-White Leghorn (n = 42) and Rhode Island Red (n = 43). A total of 162 alleles were observed, with an average of 7.7 alleles per locus. The average expected heterozygosity for the Egyptian chickens was 0.595. The closest pairwise Nei’s genetic distance was recorded between Sinai and Golden Montazah (0.038) and the smallest pairwise FST value (0.006) was observed between Baladi and Sinai. The most probable structure clustering of the eight studied populations was at K = 6. Baladi, Sinai and Golden Montazah strains were clustered together forming admixed mosaic cluster. Dandarawy ranked firstly and contributed the most to aggregate genetic diversity based on two prioritization methods. The information resulting from this study may be used as an initial guide to design further investigations for development of sustainable genetic improvement and conservation programs for the Egyptian chicken genetic resources.

The planar cell polarity (PCP) protein, Prickle (Pk), is conserved in invertebrates and vertebrates, and regulates cellular morphogenesis and movement. Vertebrate Pk consists of at least two family members, Pk1 and Pk2, both of which are expressed in the brain; however, their localization and function at synapses remain elusive. Here, we show that Pk2 is expressed mainly in the adult brain and is tightly associated with the postsynaptic density (PSD) fraction obtained by subcellular fractionation. In primary cultured rat hippocampal neurons, Pk2 is colocalized with PSD-95 and synaptophysin at synapses. Moreover, immunoelectron microcopy shows that Pk2 is localized at the PSD of asymmetric synapses in the hippocampal CA1 region. Biochemical assays identified that Pk2 forms a complex with PSD proteins including PSD-95 and NMDA receptor subunits via the direct binding to the C-terminal guanylate kinase domain of PSD-95. These results indicate that Pk2 is a novel PSD protein that interacts with PSD-95 and NMDA receptors through complex formations in the brain.

Characterization of the genetic diversity of indigenous animal populations is a prerequisite for providing needed information for the conservation of useful genotypes against future uncertainties in the face of daunting global challenges such as climate change, emerging diseases, population growth, and rising consumer demands. In this study, a total of 232 helmeted guineafowls (Numida meleagris) sampled from three populations in Ghana, one population in Benin and two populations in Japan were genotyped across six autosomal microsatellite loci. Three vulturine guineafowls (Acryllium vulturinum) were included as outgroup. A total of 66 alleles were observed with an average of 11.0 alleles per locus. The indigenous West African populations (Ghana and Benin) were more genetically diverse (Na=9.8; Ho=0.457) but less differentiated (FST=0.162) compared to the non-indigenous populations in Japan (Na=4.2; Ho=0.236; FST=0.389). The information from this study would be useful for selection and improvement programs necessary for the sustainable exploitation of this agriculturally and commercially important species as a suitable alternative to chicken.

The “Shiba inu” is the most kept and popular among the six Japanese dog breeds (Shiba, Kishu, Shikoku, Hokkaido, Kai, and Akita) officially recognized in Japan, and includes three varieties (two lines of Shinshu-Shiba, San'in-Shiba, and Mino-Shiba) showing different shapes and characters. In the preceding studies on the Japanese dog breeds, only a few studies distinguished these three varieties. Genetic backgrounds of the three varieties are still unclear. In the present study, we revealed genetic diversity and relationship among the three varieties of Shiba by microsatellite markers.We analysed genetic constitutions of the two lines of Shinshu-Shiba (Shinshu-ShibalNippo and Shinshu-Shiba/Shibaho), San'in-Shiba, and Mino-Shiba using 17 microsatellite markers. Shinshu-Shiba was composed of two different societies of preservation, “Nihonken hozonkai” (Nippo) and “Tennenkinenbutsu Shibainu hozonkai” (Shibaho) . In addition, seven dog breeds (Hokkaido, Akita, Shikoku, Satsuma, Ryukyu, Japanese Spitz, and Labrador Retriever) were also analysed by way of comparison. The total number of dogs analysed in a total of 11 populations was 430. Average expected heterozygosities over 17 microsatellite loci of Shinshu-ShibalNippo, Shinshu-Shiba/Shibaho, San'in-Shiba, and Mino-Shiba were 0.641, 0.480, 0.508, and 0.588, respectively. These estimated values were similar as the average (0.603) of the above the seven dog breeds. In addition, the degree of genetic differentiation among the three varieties (including two lines in Shinshu-shiba) of Shiba was highly significant (P

CAST and ELKS are major components of the presynaptic active zones of neurons in the central nervous system, but it remains elusive whether CAST and ELKS are also components of synapses in the peripheral nervous system. Here, we have attempted to examine their expression and localization at the synapses of neuromuscular junctions. Immunoreactivity for ELKS is partly colocalized with that for the major neuromuscular junctions marker alpha-bungarotoxin, which binds to acetylcholine receptors. Moreover, another active zone protein, Piccolo, is also present at neuromuscular junctions, together with ELKS, whereas CAST is not found. These results suggest that at least ELKS and Piccolo, but not CAST, are components of neuromuscular junction synapses in the peripheral nervous system.

The active zone protein CAST binds directly to the other active zone proteins RIM, Bassoon and Piccolo, and it has been suggested that these protein-protein interactions play an important role in neurotransmitter release. To further elucidate the molecular mechanism, we attempted to examine the function of CAST using PC12 cells as a model system. Although PC12 cells do not express CAST, they do express ELKS, a protein structurally related to CAST. Endogenous and exogenously expressed ELKS, RIM2 and Bassoon were colocalized in punctate signals in PC12 cells. Over-expression of full-length ELKS resulted in a significant increase in stimulated exocytosis of human growth hormone (hGH) from PC12 cells, similar to the effect of full-length RIM2. This increase was not observed following over-expression of deletion constructs of ELKS that lacked either the last three amino acids (IWA) required for binding to RIM2 or a central region necessary for binding to Bassoon. Moreover, over-expression of the NH(2)-terminal RIM2-binding domain of Munc13-1, which is known to inhibit the binding between RIM and Munc13-1, inhibited the stimulated increase in hGH secretion by full-length RIM2. Furthermore, this construct also inhibited the stimulated increase in hGH secretion induced by full-length ELKS. These results suggest that ELKS is involved in Ca(2+)-dependent exocytosis from PC12 cells at least partly via the RIM2-Munc13-1 pathway.

We developed nine microsatellite markers for duikers by next-generation sequencing using a muscle tissue sample of blue duiker (Cephalophus monticola). For blue duikers, the number of alleles ranged from 4 to 14 (mean 9.4), and the observed and expected heterozygosities were 0.333–0.833 (mean 0.716) and 0.625–0.878 (mean 0.817), respectively. Cumulative probability of identity was low (1.5 × 10−12), indicating that these markers are enough for individual discrimination. All nine markers were successfully amplified in three other duikers (C. dorsalis, C. ogilbyi and C. sylvicultor), and almost all loci were polymorphic. These markers will be useful for clarifying genetic structure of duikers.

We found lymphocyte proliferating substances in the water-soluble and non-dialyzable fraction prepared from the crude drug Senso (Chan Su). The effect of this fraction was increased by affinity chromatography using the concanavalin A-agarose. By analyzing the fraction using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and lectin blotting, we estimated that one of the active substances of this fraction is a glycoprotein that has about 13 kDa of molecular weight and D-mannose within the molecule. The purified fraction increased the IL-2 and the IL-12 level in the supernatant of spleen cell culture, and increased the natural killer activity of spleen lymphocyte in C3H/HeN mice. These results show that Senso contains immunopotentiating substances that may serve as an immunomodulator in an organism.

We have recently isolated a novel cytomatrix at the active zone (CAZ)–associated protein, CAST, and found it directly binds another CAZ protein RIM1 and indirectly binds Munc13-1 through RIM1; RIM1 and Munc13-1 directly bind to each other and are implicated in priming of synaptic vesicles. Here, we show that all the CAZ proteins thus far known form a large molecular complex in the brain, including CAST, RIM1, Munc13-1, Bassoon, and Piccolo. RIM1 and Bassoon directly bind to the COOH terminus and central region of CAST, respectively, forming a ternary complex. Piccolo, which is structurally related to Bassoon, also binds to the Bassoon-binding region of CAST. Moreover, the microinjected RIM1- or Bassoon-binding region of CAST impairs synaptic transmission in cultured superior cervical ganglion neurons. Furthermore, the CAST-binding domain of RIM1 or Bassoon also impairs synaptic transmission in the cultured neurons. These results indicate that CAST serves as a key component of the CAZ structure and is involved in neurotransmitter release by binding these CAZ proteins.

The cytomatrix at the active zone (CAZ) has been implicated in defining the site of Ca2+-dependent exocytosis of neurotransmitter. We have identified here a novel CAZ protein of ∼120 kD from rat brain and named it CAST (CAZ-associated structural protein). CAST had no transmembrane segment, but had four coiled-coil domains and a putative COOH-terminal consensus motif for binding to PDZ domains. CAST was localized at the CAZ of conventional synapses of mouse brain. CAST bound directly RIM1 and indirectly Munc13-1, presumably through RIM1, forming a ternary complex. RIM1 and Munc13-1 are CAZ proteins implicated in Ca2+-dependent exocytosis of neurotansmitters. Bassoon, another CAZ protein, was also associated with this ternary complex. These results suggest that a network of protein–protein interactions among the CAZ proteins exists at the CAZ. At the early stages of synapse formation, CAST was expressed and partly colocalized with bassoon in the axon shaft and the growth cone. The vesicles immunoisolated by antibassoon antibody–coupled beads contained not only bassoon but also CAST and RIM1. These results suggest that these CAZ proteins are at least partly transported on the same vesicles during synapse formation.

Background Van Gogh-like (Vangl) 2 is a planar cell polarity (PCP) protein that regulates the induction of polarized cellular and tissue morphology during animal development. In the nervous system, the core PCP signaling proteins have been identified to regulate neuronal maturation. In axonal growth cones, the antagonistic interaction of PCP components makes the tips of filopodia sensitive to guidance cues. However, the molecular mechanism by which the PCP signaling regulates spine and dendritic development remains obscure. Findings Here we explored the finding that a loss of function of Vangl2 results in a significant reduction in spine density and complexity of dendritic branching. In spite of a previous report, in which the Vangl2 C-terminal TSV motif was shown to be required for the interaction with PSD-95 and the C-terminal intracellular domain was shown to associate with N-cadherin, overexpression of deletion mutants (Vangl2-∆TSV and Vangl2-∆C) had little effect on spine density. However, when an N-terminal region deletion mutant was overexpressed, spine density was slightly down-regulated. Intriguingly, the deletion mutants had a more potent effect on dendritic branching, such that the deletion of the N-terminal region reduced dendritic branching, whereas deletion of the C-terminal region increased it. Conclusions Based on these results, Vangl2, a core PCP signaling pathway component, appears to have a functional role in neural complex formation. Especially in the case of dendritic branching, Vangl2 serves as a molecular hub to regulate neural morphology in opposite directions.

Membrane-associated guanylate kinase (MAGUK) with inverted orientation (MAGI)-1/brain angiogenesis inhibitor 1-associated protein (BAP1), is a member of the MAGUK family that has multiple PDZ domains and interacts with many transmembrane proteins, including receptors and channels, through these domains. MAGI-1/BAP1 is ubiquitously expressed and localized at tight junctions in epithelial cells. It is an isoform of the neurone-specific synaptic scaffolding molecule (S-SCAM), which is known to interact with NMDA receptors and neuroligins. We have recently found that S-SCAM also interacts with a signalling molecule, a GDP/GTP exchange protein (GEP) that is specific for Rap1 small G protein, Rap GEP, which has also recently been referred to as RA-GEF/PDZ-GEFI/CNras-GEF. In this study, we have examined whether MAGI-1/BAP1 also interacts with and serves as a scaffolding molecule for Rap GEP at tight junctions in epithelial cells. MAGI-1/BAP1 similarly interacted with Rap GEP in cell-free and intact cell systems. A Northern blot analysis revealed that Rap GEP was expressed in most tissues examined. However, neither postsynaptic density (PSD)-95/synapse-associated protein (SAP) 90 (another member of the MAGUK family) nor SAP97/human discs-large tumour suppressor gene product (another ubiquitously expressed MAGUK localizing to adherens junctions in epithelial cells and the isoform of PSD-95/SAP90) interacted with Rap GEP. These results indicate that MAGI-1/BAP1 serves as a scaffolding molecule for Rap GEP at tight junctions in epithelial cells.

Background Frabin is an actin filament (F-actin)-binding protein that shows GDP/GTP exchange activity for Cdc42 small G protein (Cdc42). Frabin furthermore induces indirect activation of Rac small G protein (Rac) in intact cells. We have recently shown that in nonepithelial cells, frabin induces the formation of both filopodia- and lamellipodia-like processes through the activation of Cdc42 and Rac, respectively. In epithelial cells such as MDCK cells, Cdc42 and Rac regulate cell-cell adherens junctions (AJs) via the accumulation of F-actin and E-cadherin, although neither Cdc42 nor Rac induces the formation of filopodia or lamellipodia. In this study, we have examined the effects of frabin on the reorganization of the actin cytoskeleton in MDCK cells. Results Frabin induces the formation of microspikes at the basal area of the lateral membranes through the activation of Cdc42 and Rac in MDCK cells, although a dominant active mutant of Cdc42 or Rac alone, or both, did not induce the formation of microspikes. Furthermore, frabin weakly increased the accumulation of F-actin and E-cadherin at cell-cell AJs and the formation of stress fibres through the activation of Cdc42 and Rac, under conditions where the dominant active mutant of Cdc42 or Rac markedly showed these effects. The Cdc42- and Rac-induced formation of stress fibres was dependent on the activation of Rho small G protein. Conclusion These results indicate that the frabin-dependent spatial activation of Cdc42 and Rac is important for the formation of microspikes.

Proteins containing formin homology domains, FH1 and FH2, are involved in cytokinesis or establishment of cell polarity in a variety of organisms. Bni1p and Bnr1p are FH proteins and potential targets of the Rho family small GTP-binding proteins in S. cerevisiae. We have shown that Bnr1p is localized at the bud neck to interact with Hof1p, involved in cytokinesis. We report here that the overexpression of BNR1 causes a cytokinesis deficiency which is similar to the phenotypes of the septin mutants, including cdc3, cdc10, cdc11, and cdc12. The region required for the septin mutant phenotypes was mapped to Bnr1p (35 – 500), which coincided with the region required for the bud-neck localization. To further isolate a gene interacting with BNI1 or BNR1, a multicopy suppressor of the bni1 bnr1 mutant was isolated. This gene encoded Smy1p, a kinesin-related protein. Bnr1p, but not Bni1p, directly interacted with the C-terminal region of Smy1p. The Smy1p-interacting region of Bnr1p was mapped to a region containing the FH2 domain. Bnr1p also directly interacted with Bud6p, a novel actin-binding protein. Bnr1p is thus a multi-functional protein which interacts with the septin system, a microtubule-dependent motor protein, and the actin system, to regulate cytoskeletal functions in S. cerevisiae.

Telomeres in eukaryotic somatic cells are destined to the age-dependent shortening, which has not been demonstrated to correlate to direct lesion of telomeric DNA by reactive oxygen intermediates (ROI); still less explicable is the inhibitory effect of ROI-scavenging on telomere shortening. Here, we succeeded in artificial slowdown of age-dependent telomere shortening to 52–62% of the untreated control, in human vascular endothelial cells, by addition of the oxidation-resistant type of ascorbic acid (Asc), Asc-2-O-phosphate (Asc2P), which concurrently achieved both extension of cellular life-span and prevention of cell size enlargement indicative of cellular senescence. The results are attributable to a 3.9-fold more marked enrichment of intracellular Asc (Ascin) by addition of Asc2P, subsequently dephosphorylated before or during transmembrane influx, than by addition of Asc itself, and also attributed to diminution of intracellular ROI to 53% of the control level by Asc2P; telomerase activity was at a trace level and underwent an age-dependent decline, which was significantly decelerated by Asc2P. Thus, age-dependent telomere-shortening can be decelerated by suppression of intracellular oxidative stress and/or by telomerase retention, both of which are achieved by enriched Ascin but not by extracellular Asc overwhelmingly more abundant than Ascin.

We report the first 18 observed cases of fruit (Treculia africana) transfer among western lowland gorillas (Gorilla gorilla gorilla) in Moukalaba-Doudou National Park, Gabon. The fruit transfer occurred during our observations of a habituated group of gorillas in 2010 and 2013. Pieces of the fruits were transferred among adults and immatures, and three cases involved a silverback male. Once an individual picked up a fallen fruit of Treculia africana, other members of the group approached the possessor, who laid pieces of the fruits nearby and tolerated the others getting them. Agonistic interaction was rarely observed between the possessor and the non-possessor. Only the silverback male seemed to force another gorilla, a subadult male, to relinquish the fruit on the ground. He tolerated an adult female taking a piece of fruit on his leg and copulated with her on the following days. From these preliminary observations, most interactions over the fruit of Treculia africana among western gorillas in Moukalaba were not active transfer by the possessor but probably passive sharing. They were not only interpreted as a means of acquiring foraging skills by immatures (Nowell and Fletcher 2006) but also similar to behaviors observed in chimpanzees and bonobos in various social contexts.

Information on the distribution and abundance of sympatric great apes (Pan troglodytes troglodytes and Gorilla gorilla gorilla) are important for effective conservation and management. Although much research has been done to improve the precision of nest-surveys, trade-offs between data-reliability and research-efficiency have not been solved. In this study, we used different approaches to assess the landscape-scale distribution patterns of great apes. We conducted a conventional nest survey and a camera-trap survey concurrently, and checked the consistency of the estimates. We divided the study area (ca. 500 km2), containing various types of vegetation and topography, into thirty 16-km2 grids (4 km × 4 km) and performed both methods along 2-km transects centered in each grid. We determined the nest creator species according to the definitions by Tutin & Fernandez [Tutin & Fernandez, 1984, Am J Primatol 6:313–336] and estimated nest-site densities of each species by using the conventional distance-sampling approach. We calculated the mean capture rate of 3 camera traps left for 3 months at each grid as the abundance index. Our analyses showed that both methods provided roughly consistent results for the distribution patterns of the species; chimpanzee groups (parties) were more abundant in the montane forest, and gorilla groups were relatively homogeneously distributed across vegetation types. The line-transect survey also showed that the number of nests per nest site did not vary among vegetation types for either species. These spatial patterns seemed to reflect the ecological and sociological features of each species. Although the consistent results may be largely dependent on site-specific conditions (e.g., high density of each species, distinct distribution pattern between the two species), conventional nest-surveys and a subsequent check of their consistency with independent estimates may be a reasonable approach to obtain certain information on the species distribution patterns. Further analytical improvement is necessary for camera-traps to be considered a stand-alone method. Am. J. Primatol. 75:1220–1230, 2013. © 2013 Wiley Periodicals, Inc.

Mycophagy (fungus-feeding) by Japanese macaques (Macaca fuscata yakui) in Yakushima has been observed by many researchers, but no detailed information is available on this behavior, including which fungal species are consumed. To provide a general description of mycophagy and to understand how and whether macaques avoid poisonous fungi, we conducted behavioral observation of wild Japanese macaques in Yakushima and used molecular techniques to identify fungal species. The results indicate that the diet of the macaques contains a large variety of fungal species (67 possible species in 31 genera), although they compose a very small portion of the total diet (2.2 % of annual feeding time). Fungi which were eaten by macaques immediately after they were picked up were less likely to be poisonous than those which were examined (sniffed, nibbled, carefully handled) by macaques. However, such examining behaviors did not appear to increase the macaques’ abilities to detect poisonous fungi. Fungi that were only partially consumed included more poisonous species than those fully consumed with/without examining behavior, yet this was not significant. Taste, therefore, might also play an important role in discriminating poisonous from non-poisonous.

In regards to the error in Figure 2, the authors write: "In this study, we generated and analyzed 51 mitochondrial genome (mtgenome) sequences of gibbons to reconstruct phylogenetic trees and estimate the divergence times in gibbon evolutionary history. We became recently aware that the sequences of two individuals (J24 and J17) in our dataset of 51 gibbon mtgenomes are partially incorrect and hence the corresponding Genbank records of these two sequences (accession numbers {"type":"entrez-nucleotide","attrs":{"text":"HQ622760","term_id":"315142051","term_text":"HQ622760"}}HQ622760 and {"type":"entrez-nucleotide","attrs":{"text":"HQ622786","term_id":"315142412","term_text":"HQ622786"}}HQ622786) have been removed from the GenBank database. Here we show the consequences of removing those two sequences from the analysis by examining newly generated phylogenetic trees and Bayesian estimates of divergence times. Using the 49 mtgenome sequences, we reconstructed the phylogenetic trees of gibbons using Bayesian and maximum likelihood methods with three partitions (rRNA, tRNA and protein). The revised information on size and characteristics of the partitions are slightly different from those in the previous table (Table 4). While the branching pattern of the gibbon genera remain unchanged, the corrected results place H. pileatus in the basal position in the Hylobates phylogeny in contrast to the absence of basal species suggested in the previous results (Figure 2). The divergence times were newly estimated with a Bayesian relaxed clock approach using the protein sequences of mtgenomes from 22 individuals (indicated in the revised Figure 2), but in which the three tree priors of the previously observed monophylies (H. lar-H. pileatus, H. klossii-H. moloch and H. agilis-H. muelleri) were no longer set. We obtained somewhat younger estimates with the narrower intervals of 95% highest poster density (HPD) for dating the splitting events compared to the older estimates resulting from the previous analysis (Table 1). Namely, the results of corrected Bayesian estimation suggested that the timings of most splitting events in the Hylobatidae family occurred from the Late Miocene to the Early Pleistocene (11.61-0.78 million years ago) and the Hylobates radiation began some 3.32 million years ago. Therefore, accordingly, we would like to replace the Table 4 of partition characteristics with a new revised table, and replace the Figure 2 of phylogenetic tree with a new revised figure as well. Moreover, a new table of corrected Bayesian estimates is also provided to replace the previous Table 1." The correct Figure 2 can be seen here: The correct legend for Figure 2 is:Figure 2. Phylogenetic tree of gibbons and outgroup primates based on the mtDNA concatenated dataset. The phylogenetic relationships among gibbons and six outgroup primates were inferred from the mtDNA concatenated dataset, including three partitioned sets: ribosomal RNA, transfer RNA and protein-coding gene. The maximum likelihood (ML) and Bayesian methods were used to reconstruct phylogenetic trees. Both analyses produced the same topology and their support values are indicated by circles on the nodes of the Bayesian tree shown here. Individuals used in the estimation of divergence times are marked with an asterisk. The corrected Table 1 can be seen here: The corrected Table 4 can be seen here

Fossils and molecular data are two independent sources of information that should in principle provide consistent inferences of when evolutionary lineages diverged. Here we use an alternative approach to genetic inference of species split times in recent human and ape evolution that is independent of the fossil record. We first use genetic parentage information on a large number of wild chimpanzees and mountain gorillas to directly infer their average generation times. We then compare these generation time estimates with those of humans and apply recent estimates of the human mutation rate per generation to derive estimates of split times of great apes and humans that are independent of fossil calibration. We date the human–chimpanzee split to at least 7–8 million years and the population split between Neanderthals and modern humans to 400,000–800,000 y ago. This suggests that molecular divergence dates may not be in conflict with the attribution of 6- to 7-million-y-old fossils to the human lineage and 400,000-y-old fossils to the Neanderthal lineage.

The neuronal network is tightly regulated by a large variety of locally connected GABAergic neurons. Neuregulin1 (Nrg1) and its receptor ErbB4 are master regulators in the morphological and functional development of excitatory synapses in GABAergic neurons. We previously showed that the immunoglobulin-like cell adhesion molecule, nectin-like molecule-2 (Necl-2)/CADM1, interacts with the ErbB3 and ErbB4 receptors, and that the interaction of Necl-2 with ErbB3 inhibits the Nrg1-induced ErbB3/ErbB2 signaling in epithelial cells. Here, we examined the role of the interaction of Necl-2 with ErbB4 in GABAergic neurons. Necl-2 was co-expressed with ErbB4 in parvalbumin-positive GABAergic neurons in the mouse hippocampus and co-localized with ErbB4 at excitatory synapses. Necl-2 knockdown enhanced the Nrg1-induced phosphorylation of ErbB4. Moreover, overexpression of PTPN13, which is a tyrosine phosphatase bound to the cytoplasmic tail of Necl-2, suppressed the Nrg1-induced development of excitatory synapses in GABAergic neurons through the inhibition of ErbB4 activity. These results indicate that Necl-2 interacts with ErbB4 and regulates the development of excitatory synapses via the regulation of ErbB4 activity in GABAergic neurons.