Sexual differentiation of brain and behavior in the zebra finch: Critical periods for effects of early estrogen treatment

In order to determine the critical period(s) during which estrogen alters sexually dimorphic behavior and neuroanatomy in zebra finches (Poephila guttata), nestlings were injected daily 20 μg estradiol benzoate (EB) during posthatching week 1, week 2, week 3, or weeks 1, 2, and 3. At 7 months of age, birds were implanted with testosterone propionate and tested with female partners for singing, dancing, and copulatory mounting. Brains were subsequently processed for morphometry, and the volumes of the song system nuclei HVC, area X, and RA and the soma sizes and densities of neurons in RA were determined. Males given EB during week 1 failed to mount. Females given EB during week 1 were fully masculinized with respect to dancing and RA neuron soma size and density, and were partially masculinized with respect to song nuclei volumes and singing. Treatment beginning after week 1 was ineffective or less effective for all measures. Only for RA neuron measures was treatment for all three weeks more effective than week 1 treatment. Thus the first post-hatching week is the most influential period of those tested for effects of exogenous estrogen on sexual differentiation in this species, and is a period during which both masculinization of females and demasculinization of males is possible. 1994 John Wiley & Sons, Inc.     

Tissue distribution and subcellular localization of the family of Kidney Ankyrin Repeat Domain (KANK) proteins

Kidney Ankyrin Repeat-containing Proteins (KANKs) comprise a family of four evolutionary conserved proteins (KANK1 to 4) that localize to the belt of mature focal adhesions (FAs) where they regulate integrin-mediated adhesion, actomyosin contractility, and link FAs to the cortical microtubule stabilization complex (CMSC). The human KANK proteins were first identified in kidney and have been associated with kidney cancer and nephrotic syndrome. Here, we report the distributions and subcellular localizations of the four Kank mRNAs and proteins in mouse tissues. We found that the KANK family members display distinct and rarely overlapping expression patterns. Whereas KANK1 is expressed at the basal side of epithelial cells of all tissues tested, KANK2 expression is mainly observed at the plasma membrane and/or cytoplasm of mesenchymal cells and KANK3 exclusively in vascular and lymphatic endothelial cells. KANK4 shows the least widespread expression pattern and when present, overlaps with KANK2 in contractile cells, such as smooth muscle cells and pericytes. Our findings show that KANKs are widely expressed in a cell type-specific manner, which suggests that they have cell- and tissue-specific functions.     

Measuring Information-Transfer Delays

In complex networks such as gene networks, traffic systems or brain circuits it is important to understand how long it takes for the different parts of the network to effectively influence one another. In the brain, for example, axonal delays between brain areas can amount to several tens of milliseconds, adding an intrinsic component to any timing-based processing of information. Inferring neural interaction delays is thus needed to interpret the information transfer revealed by any analysis of directed interactions across brain structures. However, a robust estimation of interaction delays from neural activity faces several challenges if modeling assumptions on interaction mechanisms are wrong or cannot be made. Here, we propose a robust estimator for neuronal interaction delays rooted in an information-theoretic framework, which allows a model-free exploration of interactions. In particular, we extend transfer entropy to account for delayed source-target interactions, while crucially retaining the conditioning on the embedded target state at the immediately previous time step. We prove that this particular extension is indeed guaranteed to identify interaction delays between two coupled systems and is the only relevant option in keeping with Wiener’s principle of causality. We demonstrate the performance of our approach in detecting interaction delays on finite data by numerical simulations of stochastic and deterministic processes, as well as on local field potential recordings. We also show the ability of the extended transfer entropy to detect the presence of multiple delays, as well as feedback loops. While evaluated on neuroscience data, we expect the estimator to be useful in other fields dealing with network dynamics.     

Uneven Rates of Landscape Change as a Source of Bias in Roadside Wildlife Surveys

ABSTRACT Roadside survey data have been used frequently to assess species occurrence and population trends and to establish conservation priorities. However, most studies using such data assume that samples are representative of either the amount of habitat or its rate of change at larger spatial scales. We tested both of these assumptions for the Breeding Bird Survey (BBS) from 1974 to 2001 in New Brunswick, Canada. Our study focused on mature forest—a cover type that we predicted would be characterized by rapid change due to human activities and that is of high ecological importance. We also sought to determine whether land cover changes adjacent to BBS routes were related to bird population trends detected in BBS data. Within all 3 time periods examined (1970s, 1980s, and 1990s), the amount of mature forest adjacent to BBS routes was significantly lower than in surrounding 1° blocks of latitude and longitude. This could be problematic for studies that use roadside data to compare the relative abundance of species. On average, mature forest declined at a rate of-1.5% per year over the 28-year study period. We detected no significant difference in the rate of change between degree blocks and BBS routes over this time span. However, in the 1970s and 1980s, mature forest declined more rapidly in degree blocks (-2.7%/yr) than adjacent to BBS routes (-0.5/yr). We also found that the BBS trend for a mature forest-associated species, blackburnian warbler (Dendroica fusca), was correlated with the trend in mature forest along BBS routes. This, combined with slower rates of mature forest change along routes in the 1970s and 1980s, suggests that BBS data may have underestimated population declines during this period. It is important that research be conducted to test for potential biases in roadside surveys caused by uneven rates of landscape change, particularly in regions characterized by rapid habitat alteration.     

Glucagon signalling in the dorsal vagal complex is sufficient and necessary for high‐protein feeding to regulate glucose homeostasis in vivo

High‐protein feeding acutely lowers postprandial glucose concentration compared to low‐protein feeding, despite a dichotomous rise of circulating glucagon levels. The physiological role of this glucagon rise has been largely overlooked. We here first report that glucagon signalling in the dorsal vagal complex (DVC) of the brain is sufficient to lower glucose production by activating a Gcgr–PKA–ERK–K_ATP channel signalling cascade in the DVC of rats in vivo. We further demonstrate that direct blockade of DVC Gcgr signalling negates the acute ability of high‐ vs. low‐protein feeding to reduce plasma glucose concentration, indicating that the elevated circulating glucagon during high‐protein feeding acts in the brain to lower plasma glucose levels. These data revise the physiological role of glucagon and argue that brain glucagon signalling contributes to glucose homeostasis during dietary protein intake.     

MET and PI3K/mTOR as a Potential Combinatorial Therapeutic Target in Malignant Pleural Mesothelioma

Malignant pleural mesothelioma (MPM) is an aggressive disease with a poor prognosis. Studies have shown that both MET and its key downstream intracellular signaling partners, PI3K and mTOR, are overexpressed in MPM. Here we determined the combinatorial therapeutic efficacy of a new generation small molecule inhibitor of MET, ARQ 197, and dual PI3K/mTOR inhibitors NVP-BEZ235 and GDC-0980 in mesothelioma cell and mouse xenograft models. Cell viability results show that mesothelioma cell lines were sensitive to ARQ 197, NVP-BEZ235 and GDC-0980 inhibitors. The combined use of ARQ 197 with either NVP-BEZ235 or GDC-0980, was synergistic (CI<1). Significant delay in wound healing was observed with ARQ 197 (p<0.001) with no added advantage of combining it with either NVP-BEZ235 or GDC-0980. ARQ 197 alone mainly induced apoptosis (20±2.36%) that was preceded by suppression of MAPK activity, while all the three suppressed cell cycle progression. Both GDC-0980 and NVP-BEZ235 strongly inhibited activities of PI3K and mTOR as evidenced from the phosphorylation status of AKT and S6 kinase. The above observation was further substantiated by the finding that a majority of the MPM archival samples tested revealed highly active AKT. While the single use of ARQ 197 and GDC-0980 inhibited significantly the growth of MPM xenografts (p<0.05, p<0.001 respectively) in mice, the combination of the above two drugs was highly synergistic (p<0.001). Our results suggest that the combined use of ARQ 197/NVP-BEZ235 and ARQ 197/GDC-0980 is far more effective than the use of the drugs singly in suppressing MPM tumor growth and motility and therefore merit further translational studies.     

Effect of fire on benthic algal assemblage structure and recolonization in intermittent streams

Abstract: Dry biofilm on rocks and other substrata forms an important drought refuge for benthic algae in intermittent streams following the cessation of flow. This dry biofilm is potentially susceptible to disturbance from bushfires, including direct burning and/or scorching and damage from radiant heat, particularly when streams are dry. Therefore, damage to dry biofilms by fire has the potential to influence algal recolonization and assemblage structure in intermittent streams following commencement of flow. The influence of fire on benthic algal assemblages and recolonization was examined in intermittent streams of the Grampians National Park, Victoria, Australia, using a field survey and manipulative field experiment. The field survey compared assemblages in two intermittent streams within a recently burnt area (within 5 months of the fire) with two intermittent streams within an unburnt area. The two burnt streams were still flowing during the fire so most biofilms were not likely to be directly exposed to flames. Considerable site‐to‐site and stream‐to‐stream variation was detected during the field survey, which may have obscured potential differences attributable to indirect effects of the fire. The manipulative field experiment occurred in two intermittent streams and consisted of five treatments chosen to replicate various characteristics of bushfires that may influence dry biofilms: dry biofilm exposed directly to fire; dry biofilm exposed to radiant heat; dry biofilm exposed to ash; and two procedural controls. After exposure to the different treatments, rocks were replaced in the streams and algae were sampled 7 days after flow commenced. Differences occurred across treatments, but treatment differences were inconsistent across the two streams. For example, direct exposure to fire reduced the abundance of recolonizing algae and altered assemblage structure in both streams, while radiant heat had an effect on assemblage structure in one stream only. The manipulative field experiment is likely to have represented the intensity of a small bushfire only. Nonetheless, significant differences across treatments were detected, so these experimental results suggest that fire can damage dry biofilms, and hence, influence algal recolonization and assemblage structure in intermittent streams.