Functional knock down of VCAM1 in mice mediated by endoplasmatic reticulum retained intrabodies

Functional knockdowns mediated by endoplasmatic reticulum-retained antibodies (ER intrabodies) are a promising tool for research because they allow functional interference on the protein level. We demonstrate for the first time that ER intrabodies can induce a knock-down phenotype in mice. Surface VCAM1 was suppressed in bone marrow of heterozygous and homozygous ER intrabody mice (iER-VCAM1 mice). iER-VCAM1 mice did not have a lethal phenotype, in contrast to the constitutive knockout of VCAM1, but adult mice exhibited physiological effects in the form of aberrant distribution of immature B-cells in blood and bone marrow. The capability to regulate knock-down strength may spark a new approach for the functional study of membrane and plasma proteins, which may especially be valuable for generating mouse models that more closely resemble disease states than classic knockouts do.     

Duration of strike-induced chemosensory searching in long-term captive rattlesnakes at national zoo,audubon zoo,and San Diego Zoo

Rattlesnakes typically strike and release adult rodent prey. Striking is followed by a sustained, high rate of tongue flicking that guides the snake to the envenomated, dead prey. Wild-caught rattlesnakes exhibited this chemosensory searching for about 2.5 h, and the present study demonstrated that long-term captive rattlesnakes (Crotalus atrox, C durissus, C horridus, C vegrandis, C unicolor) at three zoos did the same. Because these zoo-raised snakes had always been offered dead rodents and because the snakes had become accustomed to ingesting them without striking, the present snakes had rarely exercised their innate predatory repertoires. The duration of chemosensory searching in these snakes indicates that this important aspect of the predatory repertoire had not been degraded as a consequence of long-term captive husbandry.     

Compensatory substitutions and the evolution of the mitochondrial 12S rRNA gene in mammals

12S ribosomal RNA (rRNA) gene sequences from a suite of mammalian taxa (13 placentals, 4 marsupials, 1 monotreme), for which phylogenetic relationships are well established based on independent criteria, were employed to study the evolution of this gene. Phylogenetic analysis of 12S sequences produces a phylogeny that agrees with expectations. Base composition provides evidence for directional symmetrical substitution pressure in loops; in stems, base composition is much more even. Rates of nucleotide substitution are lower in stems than loops. Patterns of nucleotide substitution show an overall preference for transitions over transversions, with this difference more profound in stems than loops. Among different transversion pathways, there is a wide range of transformation frequencies. An analysis of compensatory substitutions shows that there is strong evidence for their occurrence and that a weighting factor of 0.61 should be applied in phylogenetic analyses to account for the dependence of mutations at stem positions relative to positions where changes are independent. Among stem variables (i.e., stem length, interaction distance, substitution rates, G+C content, and the percentage of bases that are paired), several significant correlations were discovered, but stem length and interaction distance are uncorrelated with other variables.      

Post-strike orientation of the prairie rattlesnake facilitates location of envenomated prey

Prairie rattlesnakes (Crotalus viridis) typically release adult rodent prey after envenomation. The post-strike head orientation of the snake may facilitate location of the trail left by the rodent. To examine this possibility, mice were presented using a pair of tongs so that no chemical cues were deposited on any surfaces. Snakes exhibited a change in head orientation after predatory strikes, bringing them closer to the departure bearing of the prey. In addition, when trail searching began snakes contacted the departure bearing of the prey first rather than the entry bearing. When rodent trails are available, we expect this initial contact to bias the snake towards selecting the post-envenomation trail.     

Population decline in California spotted owls near their southern range boundary

Species worldwide have begun to shift their range boundaries in response to climate change and other anthropogenic causes, with population declines at the trailing edge of a species' range often foreshadowing future changes in core parts of the range. Therefore, we analyzed a 30-year (1991–2019) data set for the California spotted owl (Strix occidentalis occidentalis) near its southern range boundary in southern California, USA, that included the largest regional population (San Bernardino Mountains) to estimate trends in territory occupancy and reproduction. We then assessed how these demographic rates were affected by habitat, wildfire, fuel treatments, and climate. Mean occupancy declined from 0.82 to 0.39 during our study, whereas reproductive output showed no temporal trends ( young/occupied territory). Territory extinction (extirpation) rates were relatively low in territories with more large trees (≥50 cm dbh), and colonization increased strongly with large tree density for low-elevation territories within the shrub-woodland ecotype but not for higher-elevation territories within mixed-conifer forest. High-severity wildfire had an adverse effect on occupancy: territory extinction rates steadily increased with the amount of high-severity fire within an owl territory during the previous 10 years, while colonization declined to nearly zero when ≥40% of a territory burned at high-severity during the previous 10 years. The effects of high-severity fire were unlikely to be confounded with post-fire fuel treatments, which primarily consisted of the removal, burning, or scattering of brush and small trees and snags (<40.6 cm dbh) and affected much smaller areas than high-severity fire. Of the 40 territories that received fuel treatments within 10 years of a fire, only 3 of them had post-fire fuel treatments that affected >5% of the territory, whereas average area burned at high severity for all 40 territories was 17%. Fuel treatments intended to modify fire behavior and reduce the likelihood of large, high-severity fires led to increases in territory extinction and colonization such that their net effect on occupancy was minimal. Our simulations of occupancy dynamics indicated that high-severity fire accounted for 9.6% of the observed decline in occupancy, whereas fuel treatments effectively accounted for none of the decline. Spotted owl reproductive output was lower at territories where fuel treatments occurred, but low- to moderate-severity fire resulted in much larger, population-level reductions in reproductive output (141 fewer young) from 2006–2019 than treatments (19 fewer young). Thus, the benefits of fuel treatments that reduce fire occurrence and severity appear to outweigh potential short-term costs to spotted owls and their habitat. Because high-severity fire only explained a modest amount of the long-term occupancy decline and much of the decline occurred in the 1990s before large fires occurred, additional factors are likely adversely affecting the owl population and merit further study. Nevertheless, the large observed population decline, limited evidence of owl dispersal among mountain ranges in the southern California metapopulation, and negative effects of increasingly large and severe fire suggest that California spotted owls at their southern range boundary are vulnerable to extirpation. In an era of climate change, owls in the core part of the range will likely become increasingly susceptible to warmer temperatures and increased severe fire activity in the future. Thus, the restoration of historical, low-severity fire regimes through fuels management while maintaining large trees is important to improving owl persistence.     

Contribution of Glucosinolate Transport to Arabidopsis Defense Responses

Accumulation of glucosinolates, a class of defense-related secondary metabolites found almost exclusively in the Capparales, is induced in response to a variety of biological stresses. It is often assumed that elevated glucosinolate levels result from de novo biosynthesis, but glucosinolate transport from other parts of the plant to the site of herbivory or pathogen infection can also contribute to the defense response. Several studies with Arabidopsis and other crucifers have demonstrated that glucosinolates from vegetative tissue are transported to developing seeds. Here we discuss evidence that long-chain aliphatic glucosinolates are transported to the site of herbivory in response to Myzus persicae (green peach aphid) feeding on Arabidopsis.Key Words: glucosinolate, transport, graft, Arabidopsis, Myzus persicae, aphid     

Chronic social defeat up‐regulates expression of the serotonin transporter in rat dorsal raphe nucleus and projection regions in a glucocorticoid‐dependent manner

Chronic stress and dysfunction of the serotonergic system in the brain have been considered two of the major risks for development of depression. In this study, adult Fischer 344 rats were subjected to a regimen of chronic social defeat (CSD). To mimic stressful conditions, some rats were not exposed to CSD, but instead treated with corticosterone (CORT) in oral solution while maintained in their home cage. Protein levels of the serotonin transporter (SERT) in the dorsal raphe nucleus (DRN), hippocampus, frontal cortex, and amygdala were examined by Western blotting or immunofluorescence staining. The results showed that CSD up‐regulated SERT protein levels in the DRN, hippocampus, frontal cortex, and amygdala regions. This up‐regulation was abolished or prevented by adrenalectomy, or treatment with antagonists of corticosteroid receptors mifepristone and spironolactone, alone or in combination. Similarly, up‐regulated SERT protein levels in these brain regions were also observed in rats treated with oral CORT ingestion, which was analogously prevented by treatment with mifepristone and spironolactone. Furthermore, both CSD‐ and CORT‐induced up‐regulation of SERT protein levels in the DRN and three brain regions were attenuated by simultaneous treatment with fluoxetine, an antidepressant that specifically inhibits serotonin reuptake. The results indicate that up‐regulation in SERT protein levels in the DRN and forebrain limbic structures caused by CSD regimen was mainly motivated by CORT through corticosteroid receptors. The present findings demonstrate that chronic stress is closely correlated with the serotonergic system by acting on the regulation of the SERT expression in the DRN and its projection regions, which may contribute to the development of depression.     

Effects of the number of markers per haplotype and clustering of haplotypes on the accuracy of QTL mapping and prediction of genomic breeding values

The aim of this paper was to compare the effect of haplotype definition on the precision of QTL-mapping and on the accuracy of predicted genomic breeding values. In a multiple QTL model using identity-by-descent (IBD) probabilities between haplotypes, various haplotype definitions were tested i.e. including 2, 6, 12 or 20 marker alleles and clustering base haplotypes related with an IBD probability of > 0.55, 0.75 or 0.95. Simulated data contained 1100 animals with known genotypes and phenotypes and 1000 animals with known genotypes and unknown phenotypes. Genomes comprising 3 Morgan were simulated and contained 74 polymorphic QTL and 383 polymorphic SNP markers with an average r² value of 0.14 between adjacent markers. The total number of haplotypes decreased up to 50% when the window size was increased from two to 20 markers and decreased by at least 50% when haplotypes related with an IBD probability of > 0.55 instead of > 0.95 were clustered. An intermediate window size led to more precise QTL mapping. Window size and clustering had a limited effect on the accuracy of predicted total breeding values, ranging from 0.79 to 0.81. Our conclusion is that different optimal window sizes should be used in QTL-mapping versus genome-wide breeding value prediction.