Genetic analysis of lung function in inbred mice suggests vitamin D receptor as a candidate gene

Vitamin D receptor (VDR) polymorphisms are associated with an increased asthma incidence in human populations; however, observations in Vdr knockout mice are unclear. The aim of our study was to determine the influence of the genetic variation in Vdr among inbred strains on lung resistance (i.e., dynamic and airway resistance). In an intercross between the strains C57BL/6J (B6) and KK/HlJ (KK), we identified that a significant QTL for dynamic resistance on Chr X was interacting with a QTL on Chr 15. The Chr 15 QTL peak was located in close proximity to the Vdr locus. We further examined if phenotypes of several inbred strains with varying Vdr genotypes differed. Strains with a B6-like genotype on the Vdr locus had significantly lower airway resistance than strains with a KK-like genotype. Vdr knockout mice were examined for dynamic resistance and showed significantly higher resistance than mice with one (i.e., heterozygous) or both copies (i.e., wild-type) of the Vdr. In comparison to B6, the strain A/J is more resistant but carries the same genotype at the Vdr locus. Dietary vitamin D manipulation in the strain A/J did not rescue the high airway resistance phenotype. Finally, we observed that serum vitamin D does not correlate significantly with lung resistance parameters in a survey of 18 strains. Conclusively, Vdr contributes to the phenotypic variation of lung resistance in inbred mice but other molecules in the Vdr pathway and extended network [i.e., Chr X gene(s)] may contribute as well.     

Tissue specific reactions of sorghum roots to the mycoherbicide Fusarium oxysporum f. sp. strigae versus the pathogenic F. proliferatum

Fusarium oxysporum f. sp. strigae (Foxy 2) is a mycoherbicide against Striga hermonthica. To ensure the safe use of this biocontrol agent, and as part of the risk assessment, this study was aimed at providing cytological evidence that Foxy 2 does not possess pathogenic behavior towards the non-target host sorghum. Therefore, we compared the infection processes and sorghum root tissue reactions towards the pathogenic F. proliferatum to that of Foxy 2 using light- and transmission electron microscopy. Given that during the growth process, hyphae could get into the central cylinder, tissue specific reactions of sorghum to Foxy 2 were also investigated by wounding the roots (exposing the vascular system), and testing for proliferation of hyphae within the vessels. Results showed that 2 weeks after sowing, F. proliferatum had invaded and destroyed all cell types including the central cylinder while Foxy 2 hyphae were located around the outer endodermal layer and were not able to penetrate the latter. There was an increase in blue autoflourescence in the central cylinder and especially the endodermis, probably due to increased phenolics in Foxy 2 infected roots which was not the case for F. proliferatum. This might contribute to the inability of Foxy 2 to penetrate the endodermis. Transmission electron microscopy showed extensive degradation of endodermis and vessel walls into thin translucent layers by F. proliferatum but not by Foxy 2. In the mechanically wounded and infected roots, Foxy 2 could invade the central cylinder close to the wound but was not identified a few millimeters away from the wound. This implies that it was not able to grow within or destroy the central cylinder even when already present within it; probably due to the observed increased phenolics. Thus, exposure of the vascular system did not serve as a route for the invasion of Foxy 2 which therefore could not further cause tracheomycosis. Therefore, Foxy 2 could be seen as a fungus well suited for biocontrol.     

Host-parasite interaction during subepidermal sporulation and pustule opening in rust fungi (Pucciniales)

Protoplasma - In late infection stages, rust fungi sporulate by building blister-like structures (pustules) in the mesophyll under the epidermal layer of their hosts. I investigated the...     

A survey of airway responsiveness in 36 inbred mouse strains facilitates gene mapping studies and identification of quantitative trait loci

Airway hyper-responsiveness (AHR) is a critical phenotype of human asthma and animal models of asthma. Other studies have measured AHR in nine mouse strains, but only six strains have been used to identify genetic loci underlying AHR. Our goals were to increase the genetic diversity of available strains by surveying 27 additional strains, to apply haplotype association mapping to the 36-strain survey, and to identify new genetic determinants for AHR. We derived AHR from the increase in airway resistance in females subjected to increasing levels of methacholine concentrations. We used haplotype association mapping to identify associations between AHR and haplotypes on chromosomes 3, 5, 8, 12, 13, and 14. And we used bioinformatics techniques to narrow the identified region on chromosome 13, reducing the region to 29 candidate genes, with 11 of considerable interest. Our combined use of haplotype association mapping with bioinformatics tools is the first study of its kind for AHR on these 36 strains of mice. Our analyses have narrowed the possible QTL genes and will facilitate the discovery of novel genes that regulate AHR in mice.     

Mouse genome-wide association study identifies polymorphisms on chromosomes 4, 11, and 15 for age-related cardiac fibrosis

Dystrophic cardiac calcinosis (DCC), also called epicardial and myocardial fibrosis and mineralization, has been detected in mice of a number of laboratory inbred strains, most commonly C3H/HeJ and DBA/2J. In previous mouse breeding studies between these DCC susceptible and the DCC-resistant strain C57BL/6J, 4 genetic loci harboring genes involved in DCC inheritance were identified and subsequently termed Dyscalc loci 1 through 4. Here, we report susceptibility to cardiac fibrosis, a sub-phenotype of DCC, at 12 and 20 months of age and close to natural death in a survey of 28 inbred mouse strains. Eight strains showed cardiac fibrosis with highest frequency and severity in the moribund mice. Using genotype and phenotype information of the 28 investigated strains, we performed genome-wide association studies (GWAS) and identified the most significant associations on chromosome (Chr) 15 at 72 million base pairs (Mb) (P < 10^?13) and Chr 4 at 122 Mb (P < 10^?11) and 134 Mb (P < 10^?7). At the Chr 15 locus, Col22a1 and Kcnk9 were identified. Both have been reported to be morphologically and functionally important in the heart muscle. The strongest Chr 4 associations were located approximately 6 Mb away from the Dyscalc 2 quantitative trait locus peak within the boundaries of the Extl1 gene and in close proximity to the Trim63 and Cap1 genes. In addition, a single-nucleotide polymorphism association was found on chromosome 11. This study provides evidence for more than the previously reported 4 genetic loci determining cardiac fibrosis and DCC. The study also highlights the power of GWAS in the mouse for dissecting complex genetic traits.     

Oxalic Acid Has an Additional,Detoxifying Function in Sclerotinia sclerotiorum Pathogenesis

The mechanism of the diseases caused by the necrotroph plant pathogen Sclerotinia sclerotiorum is not well understood. To investigate the role of oxalic acid during infection high resolution, light-, scanning-, transmission electron microscopy and various histochemical staining methods were used. Our inoculation method allowed us to follow degradation of host plant tissue around single hyphae and to observe the reaction of host cells in direct contact with single invading hyphae. After penetration the outer epidermal cell wall matrix appeared degraded around subcuticular hyphae (12-24 hpi). Calcium oxalate crystals were detected in advanced (36-48 hpi) and late (72 hpi) infection stages, but not in early stages. In early infection stages, surprisingly, no toxic effect of oxalic acid eventually secreted by S. sclerotiorum was observed. As oxalic acid is a common metabolite in plants, we propose that attacked host cells are able to metabolize oxalic acid in the early infection stage and translocate it to their vacuoles where it is stored as calcium oxalate. The effects, observed on healthy tissue upon external application of oxalic acid to non-infected, living tissue and cell wall degradation of dead host cells starting at the inner side of the walls support this idea. The results indicate that oxalic acid concentrations in the early stage of infection stay below the toxic level. In plant and fungi oxalic acid/calcium oxalate plays an important role in calcium regulation. Oxalic acid likely could quench calcium ions released during cell wall breakdown to protect growing hyphae from toxic calcium concentrations in the infection area. As calcium antimonate-precipitates were found in vesicles of young hyphae, we propose that calcium is translocated to the older parts of hyphae and detoxified by building non-toxic, stable oxalate crystals. We propose an infection model where oxalic acid plays a detoxifying role in late infection stages.     

Synchronized drumming enhances activity in the caudate and facilitates prosocial commitment--if the rhythm comes easily

Why does chanting, drumming or dancing together make people feel united? Here we investigate the neural mechanisms underlying interpersonal synchrony and its subsequent effects on prosocial behavior among synchronized individuals. We hypothesized that areas of the brain associated with the processing of reward would be active when individuals experience synchrony during drumming, and that these reward signals would increase prosocial behavior toward this synchronous drum partner. 18 female non-musicians were scanned with functional magnetic resonance imaging while they drummed a rhythm, in alternating blocks, with two different experimenters: one drumming in-synchrony and the other out-of-synchrony relative to the participant. In the last scanning part, which served as the experimental manipulation for the following prosocial behavioral test, one of the experimenters drummed with one half of the participants in-synchrony and with the other out-of-synchrony. After scanning, this experimenter “accidentally” dropped eight pencils, and the number of pencils collected by the participants was used as a measure of prosocial commitment. Results revealed that participants who mastered the novel rhythm easily before scanning showed increased activity in the caudate during synchronous drumming. The same area also responded to monetary reward in a localizer task with the same participants. The activity in the caudate during experiencing synchronous drumming also predicted the number of pencils the participants later collected to help the synchronous experimenter of the manipulation run. In addition, participants collected more pencils to help the experimenter when she had drummed in-synchrony than out-of-synchrony during the manipulation run. By showing an overlap in activated areas during synchronized drumming and monetary reward, our findings suggest that interpersonal synchrony is related to the brain''s reward system.     

Untersuchungen über die Endospermschleime der Samen von Cercis siliquastrum L. und Ononis natrix L

Die Samen der Caesalpiniacee Cercis siliquastrum sowie die der Papilionacee Ononis natrix enthalten im unterschiedlich stark entwickelten Endosperm quellbare Zellwandschleime. Die Samen von Ononis natrix besitzen ein vergleichsweise gering entwickeltes Endosperm, jedoch sind die einzelnen Zellen bis zur völligen Reduktion des Lumens schleimerfüllt. Bei Wasserzugabe quellen diese Zellwandschleime beider Arten unterschiedlich stark auf. Die Anfärbbarkeit der Endospermschleime wird anhand der Samen von Cercis siliquastrum für verschiedene Farbstoffe beschrieben. Zur näheren Charakterisierung der Quellbarkeit der Endospermschleime werden Quellungsindex sowie Viskositätsveränderung wäßriger Lösungen untersucht. Besonderes Interesse gilt der Untersuchung der Endospermschleime während der Reifung und Keimung der Samen. Dazu werden aus zerkleinerten unreifen, reifen und gekeimten Samen die Schleimstoffe extrahiert, sauer hydrolysiert und chromatographisch analysiert. Anhand chromatogrammspek-tralphotometrischer Untersuchungen ließ sich für die unterschiedlichen Entwicklungsstadien der Samen das Verhältnis von Galaktose/Mannose ermitteln. Dabei wurden zwischen reifenden, reifen und gekeimten Samen von Ononis natrix nur geringe Schwankungen in der Zusammensetzung des Galaktomannans nachgewiesen. Demgegenüber nimmt bei heranreifenden Samen von Cereis siliquastrum der Galaktosegehalt des Galaktomannans bis zur Reife zu und während der Keimung deutlich ab.     

A major X-linked locus affects kidney function in mice

Chronic kidney disease is a common disease with increasing prevalence in the western population. One common reason for chronic kidney failure is diabetic nephropathy. Diabetic nephropathy and hyperglycemia are characteristics of the mouse inbred strain KK/HlJ, which is predominantly used as a model for metabolic syndrome due to its inherited glucose intolerance and insulin resistance. We used KK/HlJ, an albuminuria-sensitive strain, and C57BL/6J, an albuminuria-resistant strain, to perform a quantitative trait locus (QTL) cross to identify the genetic basis for chronic kidney failure. Albumin–creatinine ratio (ACR) was measured in 130 F2 male offspring. One significant QTL was identified on chromosome (Chr) X and four suggestive QTL were found on Chrs 6, 7, 12, and 13. Narrowing of the QTL region was focused on the X-linked QTL and performed by incorporating genotype and expression analyses for genes located in the region. From the 485 genes identified in the X-linked QTL region, a few candidate genes were identified using a combination of bioinformatic evidence based on genomic comparison of the parental strains and known function in urine homeostasis. Finally, this study demonstrates the significance of the X chromosome in the genetic determination of albuminuria.