The calcium-sensing receptor and its interacting proteins

Seven membrane-spanning, or G protein-coupled receptors were originally thought to act through het-erotrimeric G proteins that in turn activate intracellular enzymes or ion channels, creating relatively simple, linear signalling pathways. Although this basic model remains true in that this family does act via a relatively small number of G proteins, these signalling systems are considerably more complex because the receptors interact with or are located near additional proteins that are often unique to a receptor or subset of receptors. These additional proteins give receptors their unique signalling personalities. The extracellular Ca-sensing receptor (CaR) signals via Galpha(i), Galpha(q) and Galpha(12/13), but its effects in vivo demonstrate that the signalling pathways controlled by these subunits are not sufficient to explain all its biologic effects. Additional structural or signalling proteins that interact with the CaR may explain its behaviour more fully. Although the CaR is less well studied in this respect than other receptors, several CaR-interacting proteins such as filamin, a potential scaffolding protein, receptor activity modifying proteins (RAMPs) and potassium channels may contribute to the unique characteristics of the CaR. The CaR also appears to interact with additional proteins common to other G protein-coupled receptors such as arrestins, G protein receptor kinases, protein kinase C, caveolin and proteins in the ubiquitination pathway. These proteins probably represent a few initial members of CaR-based signalling complex. These and other proteins may not all be associated with the CaR in all tissues, but they form the basis for understanding the complete nature of CaR signalling.     

Picomolar inhibitors as transition-state probes of 5'-methylthioadenosine nucleosidases.

Transition states can be predicted from an enzyme's affinity to related transition-state analogues. 5'-Methylthioadenosine nucleosidases (MTANs) are involved in bacterial quorum sensing pathways and thus are targets for antibacterial drug design. The transition-state characteristics of six MTANs are compared by analyzing dissociation constants (K(d)) with a small array of representative transition-state analogues. These inhibitors mimic early or late dissociative transition states with K(d) values in the picomolar range. Our results indicate that the K(d) ratio for mimics of early and late transition states are useful in distinguishing between these states. By this criterion, the transition states of Neisseria meningitides and Helicobacter pylori MTANs are early dissociative, whereas Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae, and Klebsiella pneumoniae MTANs have late dissociative characters. This conclusion is confirmed independently by the characteristic [1'- (3)H] and [1'- (14)C] kinetic isotope effects (KIEs) of these enzymes. Large [1'- (3)H] and unity [1'- (14)C] KIEs are observed for late dissociative transition states, whereas early dissociative states showed close-to-unity [1'- (3)H] and significant [1'- (14)C] KIEs. K d values of various MTANs for individual transition-state analogues provide tentative information about transition-state structures due to varying catalytic efficiencies of enzymes. Comparing K d ratios for mimics of early and late transition states removes limitations inherent to the enzyme and provides a better predictive tool in discriminating between possible transition-state structures.     

Domain redistribution within ergosterol-containing model membranes in the presence of the antimicrobial compound fengycin

The cyclic lipopeptide fengycin, produced by Bacillus subtilis, exhibits its antimicrobial capabilities by altering the integrity of the cell membrane of plant pathogens. Previous work has correlated fengycin activity with membrane characteristics, such as sterol content. This work focused on the influence of fengycin on supported lipid bilayers containing varying levels of ergosterol. Total internal reflection fluorescence (TIRF) microscopy was used to visualize and distinguish ordered (L_β/L_o) and disordered (L_α/L_d) domains in the model membranes following exposure to low (50 μg) and high (500 μg) fengycin doses. Application of an initial low dose of fengycin to 0% and 3% ergosterol-containing bilayers resulted in redistribution of L_α/L_β and L_o/L_d domains, respectively, which the bilayers compensated and corrected for over time. These membranes were unable to tolerate a second 50 μg dose or a single high fengycin dose. The 6% ergosterol bilayers were able to tolerate sequential low doses of fengycin. Exposure of these bilayers to the high fengycin dose caused a decrease in the number of L_o domains, albeit less than that seen in the 0% and 3% ergosterol bilayers. Bilayers containing 12% ergosterol, exhibited the least amount of change after fengycin exposure. These were the only bilayer to exhibit an increase in area taken up by ordered domains. These results suggest fengycin may preferentially act on the L_β or L_o phase, the area in which ergosterol resides. Bilayers containing low levels of ergosterol appear to be more sensitive to the lipopeptide, suggesting ergosterol plays a role in buffering perturbations caused by fengycin.     

LAP2 Proteins Chaperone GLI1 Movement between the Lamina and Chromatin to Regulate Transcription

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Status and distribution of jaguarundi in Texas and Northeastern México: Making the case for extirpation and initiation of recovery in the United States

The jaguarundi (Puma yagouaroundi) is a small felid with a historical range from central Argentina through southern Texas. Information on the current distribution of this reclusive species is needed to inform recovery strategies in the United States where its last record was in 1986 in Texas. From 2003 to 2021, we conducted camera‐trap surveys across southern Texas and northern Tamaulipas, México to survey for medium‐sized wild cats (i.e., ocelots [Leopardus pardalis], bobcats [Lynx rufus], and jaguarundi). After 350,366 trap nights at 685 camera sites, we did not detect jaguarundis at 16 properties or along 2 highways (1050 km²) in Texas. However, we recorded 126 jaguarundi photographic detections in 15,784 trap nights on 2 properties (125.3 km²) in the northern Sierra of Tamaulipas, Tamaulipas, México. On these properties, latency to detection was 72 trap nights, with a 0.05 probability of detection per day and 0.73 photographic event rate every 100 trap nights. Due to a lack of confirmed class I sightings (e.g., specimen, photograph) in the 18 years of this study, and no other class I observations since 1986 in the United States, we conclude that the jaguarundi is likely extirpated from the United States. Based on survey effort and results from México, we would have expected to detect jaguarundis over the course of the study if still extant in Texas. We recommend that state and federal agencies consider jaguarundis as extirpated from the United States and initiate recovery actions as mandated in the federal jaguarundi recovery plan. These recovery actions include identification of suitable habitat in Texas, identification of robust populations in México, and re‐introduction of the jaguarundi to Texas.     

Quantification of long-term doxorubicin response dynamics in breast cancer cell lines to direct treatment schedules

While acquired chemoresistance is recognized as a key challenge to treating many types of cancer, the dynamics with which drug sensitivity changes after exposure are poorly characterized. Most chemotherapeutic regimens call for repeated dosing at regular intervals, and if drug sensitivity changes on a similar time scale then the treatment interval could be optimized to improve treatment performance. Theoretical work suggests that such optimal schedules exist, but experimental confirmation has been obstructed by the difficulty of deconvolving the simultaneous processes of death, adaptation, and regrowth taking place in cancer cell populations. Here we present a method of optimizing drug schedules in vitro through iterative application of experimentally calibrated models, and demonstrate its ability to characterize dynamic changes in sensitivity to the chemotherapeutic doxorubicin in three breast cancer cell lines subjected to treatment schedules varying in concentration, interval between pulse treatments, and number of sequential pulse treatments. Cell populations are monitored longitudinally through automated imaging for 600–800 hours, and this data is used to calibrate a family of cancer growth models, each consisting of a system of ordinary differential equations, derived from the bi-exponential model which characterizes resistant and sensitive subpopulations. We identify a model incorporating both a period of growth arrest in surviving cells and a delay in the death of chemosensitive cells which outperforms the original bi-exponential growth model in Akaike Information Criterion based model selection, and use the calibrated model to quantify the performance of each drug schedule. We find that the inter-treatment interval is a key variable in determining the performance of sequential dosing schedules and identify an optimal retreatment time for each cell line which extends regrowth time by 40%-239%, demonstrating that the time scale of changes in chemosensitivity following doxorubicin exposure allows optimization of drug scheduling by varying this inter-treatment interval.     

Investigating bee dietary preferences along a gradient of floral resources:how does resource use align with resource availability?

Bee dietary preferences,or the floral resources that they consistently collect,likely impact where a species can persist.For this reason it is likely that bee dietary preferences are dependent upon the composition of the plant community.In this study,we evaluated floral visits and pollen loads of the mining bee,Andrena angustitarsata Viereck,across a 630 km north-south range to understand dietary preferences along a floral resource gradient.Previous research,in a more geographically limited area,suggested this species was an eclectic oligolege on predominantly Apiaceae and in part Rosaceae.In the present study we found the species predominately visited and collected pollen from Apiaceae and Rosaceae,but visited 12 flower families and collected pollen from 32,distinguishing them as generalist foragers.The frequency of Apiaceae pollen on the bees and the species-level specialization index(a measure of visit specialization)were higher in regions with higher Apiaceae abundance.In addition Apiaceae and Rosaceae were the only plant families significantly preferred for pollen collection,regardless of floral abundance.We conclude that across our study region A.angustitarsata has a generalist dietary breadth,but also has dietary preference for Apiaceae and Rosaceae.Our study indicates that while bees may overall make generalist foraging decisions they may still prefer and likely benefit from selecting fewer flower taxa.     

Extracting spatial networks from capture–recapture data reveals individual site fidelity patterns within a marine mammal’s spatial range

1. Estimating the impacts of anthropogenic disturbances requires an understanding of the habitat‐use patterns of individuals within a population. This is especially the case when disturbances are localized within a population''s spatial range, as variation in habitat use within a population can drastically alter the distribution of impacts.
2. Here, we illustrate the potential for multilevel binomial models to generate spatial networks from capture–recapture data, a common data source used in wildlife studies to monitor population dynamics and habitat use. These spatial networks capture which regions of a population''s spatial distribution share similar/dissimilar individual usage patterns, and can be especially useful for detecting structured habitat use within the population''s spatial range.
3. Using simulations and 18 years of capture–recapture data from St. Lawrence Estuary (SLE) beluga, we show that this approach can successfully estimate the magnitude of similarities/dissimilarities in individual usage patterns across sectors, and identify sectors that share similar individual usage patterns that differ from other sectors, that is, structured habitat use. In the case of SLE beluga, this method identified multiple clusters of individuals, each preferentially using restricted areas within their summer range of the SLE.
4. Multilevel binomial models can be effective at estimating spatial structure in habitat use within wildlife populations sampled by capture–recapture of individuals, and can be especially useful when sampling effort is not evenly distributed. Our finding of a structured habitat use within the SLE beluga summer range has direct implications for estimating individual exposures to localized stressors, such as underwater noise from shipping or other activities.

     

No sex differences in evoked contractile properties after fatiguing isometric and isotonic exercise for the plantar flexors

Objectives:Females tend to fatigue less than males after isometric exercise, but less is clear for isotonic exercise. Further, there have been relatively few sex comparisons for fatigability of the plantar flexors (PFs). We sought to investigate potential sex differences in contractile properties after a sustained maximal voluntary isometric contraction (MVIC) and isotonic contractions.Methods:Twenty-seven physically active males (n=14; 22±2 yrs) and females (n=13; 21±2 yrs) randomly performed a 2 min MVIC and 120 concentric isotonic (30% MVIC) contractions for the PFs on separate visits. Before and after each fatiguing task, muscle activation was obtained from brief MVICs, which was followed (~2 sec) by tibial nerve stimulation at rest. Contractile properties including peak twitch, absolute and normalized time to peak twitch, and half relaxation time were calculated.Results:No sex differences existed for fatigue-induced changes in muscle activation (p=0.09-0.41; d=0.33-0.69) or contractile properties (p=0.19-0.96; d=0.06-0.94).Conclusions:Peripheral fatigue, as indicated by contractile parameters, did not differ between sexes after isometric or isotonic exercise. The PFs similar fiber type proportions between sexes or greater fiber type heterogeneity may explain why sex differences in fatigability, though common in other muscle groups (e.g., knee extensors), were not expressed in this muscle group.