Generation of an Fsp1 (fibroblast‐specific protein 1)‐Flpo transgenic mouse strain

Recombination systems represent a major breakthrough in the field of genetic model engineering. The Flp recombinases (Flp, Flpe, and Flpo) bind and cleave DNA Frt sites. We created a transgenic mouse strain ([Fsp1‐Flpo]) expressing the Flpo recombinase in fibroblasts. This strain was obtained by random insertion inside mouse zygotes after pronuclear injection. Flpo expression was placed under the control of the promoter of Fsp1 (fibroblast‐specific protein 1) gene, whose expression starts after gastrulation at Day 8.5 in cells of mesenchymal origin. We verified the correct expression and function of the Flpo enzyme by several ex vivo and in vivo approaches. The [Fsp1‐Flpo] strain represents a genuine tool to further target the recombination of transgenes with Frt sites specifically in cells of mesenchymal origin or with a fibroblastic phenotype.     

Length-weight relationship of twelve mesopelagic fishes from the western Tropical Atlantic

Length-weight relationship parameters were calculated for twelve mesopelagic fish species from the western Tropical Atlantic: Diretmus argenteus, Melamphaes polylepis, Bolinichthys distofax, Diaphus lucidus, Diaphus splendidus, Electrona risso, Hygophum taaningi, Taaningichthys bathyphilus, Melanolagus bericoides, Winteria telescopa, Diplophos taenia, Astronesthes similus. Data was collected off northeastern Brazil from April 9th to May 6th, 2017. Hauls were conducted during day and night at 47 stations by using a micronekton trawl (body mesh: 40 mm, cod-end mesh: 10 mm) from 10 to 1,113 m depth. The material was fixed in a 4% formalin solution for 1 month and then preserved in a 70% alcohol solution for proximally 6 months before processing for length (nearest 0.1 cm of standard length) and weight (nearest 0.01 g of total weight). A new maximum standard length for Winteria telescopa is also provided.     

Crucial role for LKB1 to AMPKα2 axis in the regulation of CD36-mediated long-chain fatty acid uptake into cardiomyocytes

Enhanced contractile activity increases cardiac long-chain fatty acid (LCFA) uptake via translocation of CD36 to the sarcolemma, similarly to increase in glucose uptake via GLUT4 translocation. AMP-activated protein kinase (AMPK) is assumed to mediate contraction-induced LCFA utilization. However, which catalytic isoform (AMPKα1 versus AMPKα2) is involved, is unknown. Furthermore, no studies have been performed on the role of LKB1, a kinase with AMPKK activity, on the regulation of cardiac LCFA utilization. Using different mouse models (AMPKα2-kinase-dead, AMPKα2-knockout and LKB1-knockout mice), we tested whether LKB1 and/or AMPK are required for stimulation of LCFA and glucose utilization upon treatment of cardiomyocytes with compounds (oligomycin/AICAR/dipyridamole) which induce CD36 translocation similar to that seen upon contraction. In AMPKα2- kinase-dead cardiomyocytes, the stimulating effects of oligomycin and AICAR on palmitate and deoxyglucose uptake and palmitate oxidation were almost completely lost. Moreover, in AMPKα2- and LKB1-knockout cardiomyocytes, oligomycin-induced LCFA and deoxyglucose uptake were completely abolished. However, the stimulatory effect of dipyridamole on palmitate uptake and oxidation was preserved in AMPKα2-kinase-dead cardiomyocytes. In conclusion, in the heart there is a signaling axis consisting of LKB1 and AMPKα2 which activation results in enhanced LCFA utilization, similarly to enhanced glucose uptake. In addition, an unknown dipyridamole-activated pathway can stimulate cardiac LCFA utilization by activating signaling components downstream of AMPK.     

Rab11b Regulates the Apical Recycling of the Cystic Fibrosis Transmembrane Conductance Regulator in Polarized Intestinal Epithelial Cells

The cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP/PKA-activated anion channel, undergoes efficient apical recycling in polarized epithelia. The regulatory mechanisms underlying CFTR recycling are understood poorly, yet this process is required for proper channel copy number at the apical membrane, and it is defective in the common CFTR mutant, ΔF508. Herein, we investigated the function of Rab11 isoforms in regulating CFTR trafficking in T84 cells, a colonic epithelial line that expresses CFTR endogenously. Western blotting of immunoisolated Rab11a or Rab11b vesicles revealed localization of endogenous CFTR within both compartments. CFTR function assays performed on T84 cells expressing the Rab11a or Rab11b GDP-locked S25N mutants demonstrated that only the Rab11b mutant inhibited 80% of the cAMP-activated halide efflux and that only the constitutively active Rab11b-Q70L increased the rate constant for stimulated halide efflux. Similarly, RNAi knockdown of Rab11b, but not Rab11a, reduced by 50% the CFTR-mediated anion conductance response. In polarized T84 monolayers, adenoviral expression of Rab11b-S25N resulted in a 70% inhibition of forskolin-stimulated transepithelial anion secretion and a 50% decrease in apical membrane CFTR as assessed by cell surface biotinylation. Biotin protection assays revealed a robust inhibition of CFTR recycling in polarized T84 cells expressing Rab11b-S25N, demonstrating the selective requirement for the Rab11b isoform. This is the first report detailing apical CFTR recycling in a native expression system and to demonstrate that Rab11b regulates apical recycling in polarized epithelial cells.     

Effects of Tissue Collection Methods on Morphometrics and Survival of Captive Neonatal Northern Bobwhite

ABSTRACT We assessed effects of tissue collection methods (i.e., patagial microbiopsy and down feathers) and chick age at sampling on morphometrics and 21-day survival of 600 captive neonatal northern bobwhite (Colinus virginianus). We observed minimal effects on morphometrics and no difference in survival among patagial microbiopsy (x̄ = 0.96 ± 0.03), down feathers (x̄ = 0.92 ± 0.04), and control (x̄ = 0.86 ± 0.05) methods. DNA analysis from patagial microbiopsy, down feather, and egg tooth samples showed greater concentrations of DNA from patagial microbiopsy (x̄ = 10.28 ± 1.74 μg/ml) than either down feather (x̄ = 4.10 ± 1.74 μg/ml) or egg teeth (x̄ = 2.35 ± 1.74 μg/ml).     

Estimating Duration of Short-Term Acute Effects of Capture Handling and Radiomarking

ABSTRACT Radiotelemetry is used extensively in zoographic studies of wildlife species, including northern bobwhite (Colinus virginianus). These studies assume that radiomarking does not affect survival of marked individuals. However, most researchers implicitly acknowledge that capture, handling, and radiomarking may have short-term deleterious effects on individuals and, therefore, include in analyses only animals that survive an adjustment period of arbitrary length (often 7 days) following capture and marking. Length of adjustment period is rarely empirically based and may potentially bias survival estimates. We outline an analytical approach to determine an appropriate adjustment period and illustrate this approach by examining effects of time-since-marking on survival of 410 northern bobwhite captured during winter from 1997 to 2001, in Mississippi, USA. We modeled daily survival rates using time-since-marking as a covariate in the nest-survival model of Program MARK. Although survival varied among and within years, we found no evidence to suggest that standard adjustment periods of 7–14 days were appropriate for our sample. If adjustment periods are used in radiotelemetry studies, those that are empirically based may be more appropriate than arbitrarily set periods.     

Population Growth and Demography of Common Loons in the Northern United States

ABSTRACT We used recent developments in theoretical population ecology to construct basic models of common loon (Gavia immer) demography and population dynamics. We parameterized these models using existing survival estimates and data from long-term monitoring of loon productivity and abundance. Our models include deterministic, 2-stage, density-independent matrix models, yielding population growth-rate estimates (λ) of 0.99 and 1.01 for intensively studied populations in our Wisconsin, USA, and New Hampshire, USA, study areas, respectively. Perturbation analysis of these models indicated that estimated growth rate is extremely sensitive to adult survival, as expected for this long-lived species. Also, we examined 20 years of count data for the 2 areas and evaluated support for a set of count-based models of population growth. We detected no temporal trend in Wisconsin, which would be consistent with fluctuation around an average equilibrium state but could also result from data limitations. For New Hampshire, the model set included varying formulations of density dependence and partitioning of stochasticity that were enabled by the annual sampling resolution. The best model for New Hampshire included density regulation of population growth and, along with the demographic analyses for both areas, provided insight into the possible importance of breeding habitat availability and the abundance of nonbreeding adults. Based on these results, we recommend that conservation organizations include nonbreeder abundance in common loon monitoring efforts and that additional emphasis be placed on identifying and managing human influences on adult loon survival.     

Factors Influencing Reporting and Harvest Probabilities in North American Geese

ABSTRACT We assessed variation in reporting probabilities of standard bands among species, populations, harvest locations, and size classes of North American geese to enable estimation of unbiased harvest probabilities. We included reward (US$10, $20, $30, $50, or $100) and control ($0) banded geese from 16 recognized goose populations of 4 species: Canada (Branta canadensis), cackling (B. hutchinsii), Ross's (Chen rossii), and snow geese (C. caerulescens). We incorporated spatially explicit direct recoveries and live recaptures into a multinomial model to estimate reporting, harvest, and band-retention probabilities. We compared various models for estimating harvest probabilities at country (United States vs. Canada), flyway (5 administrative regions), and harvest area (i.e., flyways divided into northern and southern sections) scales. Mean reporting probability of standard bands was 0.73 (95% CI = 0.69–0.77). Point estimates of reporting probabilities for goose populations or spatial units varied from 0.52 to 0.93, but confidence intervals for individual estimates overlapped and model selection indicated that models with species, population, or spatial effects were less parsimonious than those without these effects. Our estimates were similar to recently reported estimates for mallards (Anas platyrhynchos). We provide current harvest probability estimates for these populations using our direct measures of reporting probability, improving the accuracy of previous estimates obtained from recovery probabilities alone. Goose managers and researchers throughout North America can use our reporting probabilities to correct recovery probabilities estimated from standard banding operations for deriving spatially explicit harvest probabilities.