Telomere shortening impairs regeneration of the olfactory epithelium in response to injury but not under homeostatic conditions

Atrophy of the olfactory epithelium (OE) associated with impaired olfaction and dry nose represents one of the most common phenotypes of human aging. Impairment in regeneration of a functional olfactory epithelium can also occur in response to injury due to infection or nasal surgery. These complications occur more frequently in aged patients. Although age is the most unifying risk factor for atrophic changes and functional decline of the olfactory epithelium, little is known about molecular mechanisms that could influence maintenance and repair of the olfactory epithelium. Here, we analyzed the influence of telomere shortening (a basic mechanism of cellular aging) on homeostasis and regenerative reserve in response to chemical induced injury of the OE in late generation telomere knockout mice (G3 mTerc^−/−) with short telomeres compared to wild type mice (mTerc^+/+) with long telomeres. The study revealed no significant influence of telomere shortening on homeostatic maintenance of the OE during mouse aging. In contrast, the regenerative response to chemical induced injury of the OE was significantly impaired in G3 mTerc^−/− mice compared to mTerc^+/+ mice. Seven days after chemical induced damage, G3 mTerc^−/− mice exhibited significantly enlarged areas of persisting atrophy compared to mTerc^+/+ mice (p = 0.031). Telomere dysfunction was associated with impairments in cell proliferation in the regenerating epithelium. Deletion of the cell cycle inhibitor, Cdkn1a (p21) rescued defects in OE regeneration in telomere dysfunctional mice. Together, these data indicate that telomere shortening impairs the regenerative capacity of the OE by impairing cell cycle progression in a p21-dependent manner. These findings could be relevant for the impairment in OE function in elderly people.     

Invasion of dwarf bamboo into alpine snow-meadows in northern Japan: pattern of expansion and impact on species diversity

Recently, a dwarf bamboo species,Sasa kurilensis; Poaceae, has invaded into alpine snow-meadows in the wilderness area of the Taisetsu Mountains, northern Japan. This dwarf bamboo species has a wide distribution range from lowland to alpine sites of snowy regions. Because of the formation of dense evergreen culms and an extensive rhizome system, other plants are excluded following invasion by this dwarf bamboo, resulting in low species diversity. Dwarf bamboo originally inhabited the leeward slopes of alpine dwarf pine (Pinus pumila) clumps in alpine regions. During the last 32 years, however, dwarf bamboo has expanded its distribution area by up to 47% toward snow-meadows, especially on southeastern facing slopes. This rapid change may be related to the decrease in soil moisture and expansion of the annual growing period caused by the recent acceleration of snowmelt time. A multiyear census revealed that the density of bamboo culms increased 30-150% during 2 years, and the annual expansion of bamboo rhizomes was 39 cm on average. In addition to the expansion of bamboo clumps by vegetative growth, the possibility of migration by seed dispersal was also suggested by a genet analysis. With the increase in culm density, the species richness of snow-meadow vegetation decreased to less than one-quarter of the original level due to intense shading by dwarf bamboo. The rapid vegetation change in these almost pristine alpine environments isolated from the human activity implies that global climate change already influences the alpine ecosystem.     

Resilient Plant–Bird Interactions in a Volcanic Island Ecosystem: Pollination of Japanese Camellia Mediated by the Japanese White-Eye

Observations of interspecies interactions during volcanic activity provide important opportunities to study how organisms respond to environmental devastation. Japanese camellia (Camellia japonica L.) and its main avian pollinator, the Japanese White-eye (Zosterops japonica), offer an excellent example of such an interaction as key members of the biotic community on Miyake-jima, which erupted in 2000 and continues to emit volcanic gases. Both species exhibit higher resistance to volcanic damage than other species. We examined the effects of volcanic activity on this plant–pollinator system by estimating pollen flow and the genetic diversity of the next generation. The results showed that despite a decrease in Camellia flowers, the partitioning of allelic richness among mother-tree pollen pools and seeds decreased while the migration rate of pollen from outside the study plot and the pollen donor diversity within a fruit increased as the index of volcanic damage increased. In areas with low food (flower) density due to volcanic damage, Z. japonica ranged over larger areas to satisfy its energy needs rather than moving to areas with higher food density. Consequently, the genetic diversity of the seeds (the next plant generation) increased with the index of volcanic damage. The results were consistent with previously published data on the movement of Z. japonica based on radio tracking and the genetic diversity of Camellia pollen adhering to pollinators. Overall, our results indicated that compensation mechanisms ensured better pollination after volcanic disturbance.     

An Immunohistochemical Analysis of Tissue Thrombin Expression in the Human Atria

Objective

Thrombin, the final coagulation product of the coagulation cascade, has been demonstrated to have many physiological effects, including pro-fibrotic actions via protease-activated receptor (PAR)-1. Recent investigations have demonstrated that activation of the cardiac local coagulation system was associated with atrial fibrillation. However, the distribution of thrombin in the heart, especially difference between the atria and the ventricle, remains to be clarified. We herein investigated the expression of thrombin and other related proteins, as well as tissue fibrosis, in the human left atria and left ventricle.

Methods

We examined the expression of thrombin and other related molecules in the autopsied hearts of patients with and without atrial fibrillation. An immunohistochemical analysis was performed in the left atria and the left ventricle.

Results

The thrombin was immunohistologically detected in both the left atria and the left ventricles. Other than in the myocardium, the expression of thrombin was observed in the endocardium and the subendocardium of the left atrium. Thrombin was more highly expressed in the left atrium compared to the left ventricle, which was concomitant with more tissue fibrosis and inflammation, as detected by CD68 expression, in the left atrium. We also confirmed the expression of prothrombin in the left atrium. The expression of PAR-1 was observed in the endocardium, subendocardium and myocardium in the left atrium. In patients with atrial fibrillation, strong thrombin expression was observed in the left atrium.

Conclusions

The strong expression levels of thrombin, prothrombin and PAR-1 were demonstrated in the atrial tissues of human autopsied hearts.     

Domestication Process of the Goat Revealed by an Analysis of the Nearly Complete Mitochondrial Protein-Encoding Genes

Goats (Capra hircus) are one of the oldest domesticated species, and they are kept all over the world as an essential resource for meat, milk, and fiber. Although recent archeological and molecular biological studies suggested that they originated in West Asia, their domestication processes such as the timing of population expansion and the dynamics of their selection pressures are little known. With the aim of addressing these issues, the nearly complete mitochondrial protein-encoding genes were determined from East, Southeast, and South Asian populations. Our coalescent time estimations suggest that the timing of their major population expansions was in the Late Pleistocene and significantly predates the beginning of their domestication in the Neolithic era (≈10,000 years ago). The ω (ratio of non-synonymous rate/synonymous substitution rate) for each lineage was also estimated. We found that the ω of the globally distributed haplogroup A which is inherited by more than 90% of goats examined, turned out to be extremely low, suggesting that they are under severe selection pressure probably due to their large population size. Conversely, the ω of the Asian-specific haplogroup B inherited by about 5% of goats was relatively high. Although recent molecular studies suggest that domestication of animals may tend to relax selective constraints, the opposite pattern observed in our goat mitochondrial genome data indicates the process of domestication is more complex than may be presently appreciated and cannot be explained only by a simple relaxation model.     

Targeted Gene Deletion of miRNAs in Mice by TALEN System

Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.     

A Critical Role of Fatty Acid Binding Protein 4 and 5 (FABP4/5) in the Systemic Response to Fasting

During prolonged fasting, fatty acid (FA) released from adipose tissue is a major energy source for peripheral tissues, including the heart, skeletal muscle and liver. We recently showed that FA binding protein 4 (FABP4) and FABP5, which are abundantly expressed in adipocytes and macrophages, are prominently expressed in capillary endothelial cells in the heart and skeletal muscle. In addition, mice deficient for both FABP4 and FABP5 (FABP4/5 DKO mice) exhibited defective uptake of FA with compensatory up-regulation of glucose consumption in these tissues during fasting. Here we showed that deletion of FABP4/5 resulted in a marked perturbation of metabolism in response to prolonged fasting, including hyperketotic hypoglycemia and hepatic steatosis. Blood glucose levels were reduced, whereas the levels of non-esterified FA (NEFA) and ketone bodies were markedly increased during fasting. In addition, the uptake of the ¹²⁵I-BMIPP FA analogue in the DKO livers was markedly increased after fasting. Consistent with an increased influx of NEFA into the liver, DKO mice showed marked hepatic steatosis after a 48-hr fast. Although gluconeogenesis was observed shortly after fasting, the substrates for gluconeogenesis were reduced during prolonged fasting, resulting in insufficient gluconeogenesis and enhanced hypoglycemia. These metabolic responses to prolonged fasting in DKO mice were readily reversed by re-feeding. Taken together, these data strongly suggested that a maladaptive response to fasting in DKO mice occurred as a result of an increased influx of NEFA into the liver and pronounced hypoglycemia. Together with our previous study, the metabolic consequence found in the present study is likely to be attributed to an impairment of FA uptake in the heart and skeletal muscle. Thus, our data provided evidence that peripheral uptake of FA via capillary endothelial FABP4/5 is crucial for systemic metabolism and may establish FABP4/5 as potentially novel targets for the modulation of energy homeostasis.     

Plasma ADMA concentrations associate with aerobic fitness in postmenopausal women

Aims

Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of nitric oxide (NO) synthase, an enzyme responsible for the generation of NO. Plasma concentrations of ADMA increase in the elderly and in postmenopausal women. In fact, an elevated ADMA level is a risk factor of cardiovascular disease. Aerobic exercise has a beneficial effect on cardiovascular disease. However, the relationship between ADMA and aerobic fitness is unknown. The aim of this study was to determine whether plasma ADMA concentrations correlate with aerobic fitness levels in postmenopausal women.

Main methods

Thirty healthy postmenopausal women aged 50–76 years participated in this study. We measured plasma concentrations of ADMA and oxygen consumption at the ventilatory threshold (VO_2VT) as an index of aerobic fitness. Subjects were divided into the low aerobic fitness (Low fitness) and high aerobic fitness (High fitness) groups, and the dividing line was set at the median VO_2VT value.

Key findings

VO_2VT was significantly higher in the High fitness group than in the Low fitness group (P < 0.01). The plasma ADMA concentrations in the High fitness group were significantly lower than those in the Low fitness group (P < 0.05). There was a negative correlation between plasma ADMA concentrations and VO_2VT (r = − 0.532, P < 0.01).

Significance

We found that plasma ADMA concentrations were associated with aerobic fitness in postmenopausal women. The results of this study suggest that habitual aerobic exercise may decrease plasma ADMA concentrations.     

Frequency-dependent herbivory by a leaf beetle,Phaedon brassicae,on hairy and glabrous plants of Arabidopsis halleri subsp. gemmifera

Frequency-dependent prey choice by natural enemies may influence the coexistence of multiple prey types, but little is known about whether frequency-dependent foraging choice occurs in herbivory on plants showing resistance polymorphism within a single population. Here we examined frequency-dependent foraging by a crucifer-feeding leaf beetle, Phaedon brassicae, on trichome-producing (hairy) and trichomeless (glabrous) plants coexisting within a natural population of the perennial herb Arabidopsis halleri subsp. gemmifera. Larvae of P. brassicae fed on hairy leaves showed slower growth than those fed on glabrous leaves. Although adult beetles consumed similar amounts of leaves when they were fed either hairy or glabrous leaves in no-choice conditions, our choice experiment showed that adult beetles fed at less than the proportionally expected level on hairy leaves compared to glabrous leaves when the hairy leaves were less or equally abundant. Both types of leaves were consumed at the proportionally expected levels when the hairy leaves were more abundant than the glabrous leaves. In a natural population, the leaf damage on the hairy plants was negatively correlated with the local proportion of the glabrous plants in a 1-m diameter patch across 2 years, while correlations between the leaf damage on the glabrous plants and their proportion differed between the 2 years. Additionally, we found five glucosinolates in leaves of A. halleri, but their accumulation did not differ between hairy and glabrous plants. Our experimental results indicate that hairy plants incur less herbivory by P. brassicae when glabrous plants are abundant. The field pattern provides evidence suggestive of frequency-dependent herbivory acting on hairy plants. The present study highlights one of the putative mechanisms of maintaining plant resistance polymorphism.