The context and outcome of aggressive interactions between individuals has important fitness consequences. Displacements—an aggressive interaction wherein one individual is chased from a location by another—also have implications for social hierarchy formation and geographic distribution in songbirds. Morphological correlates, like body size, and social correlates, such as dominance rank, have been shown to mediate displacements in songbirds. However, the role of the physical environment, namely temperature, humidity, and time of day, which may influence an individual’s energy needs and thus displacement motivation, has remained understudied. We monitored songbird feeding and displacement behaviors using computerized automated feeders. We observed asymmetric differences across species in displacement involvement. To identify the conditions of the social and physical environment that are associated with the occurrence and outcome of songbird displacements at supplemental feeders, we use the machine learning approach, random forest, which is a novel method to the fields of ornithology and animal behavior. From our random forest models, we found that the attributes of the physical environment (i.e., humidity and the time of day) are associated with the occurrence of a displacement event, whereas the attributes of the social environment (i.e., species of the displacer and displaced individuals) are associated with which species are involved. These results provide context to develop further observational and experimental hypotheses to tease apart the inner workings of these multifactorial behaviors on a larger scale and provide a proof of concept for our analytical methods in the study of avian behavior.

Research across a range of biological subdisciplines and scales, ranging from molecular to ecosystemic, provides ample evidence that living systems generally exhibit both a degree of resistance to disruption and an ability to recover following disturbance. Not only do mechanisms of robustness and resilience exist across and between systems, but those mechanisms exhibit ubiquitous and scalable commonalities in pattern and function. Mechanisms such as redundancy, plasticity, interconnectivity, and coordination of subunits appear to be crucial internal players in the determination of stability. Similarly, factors external to the system such as the amplitude, frequency, and predictability of disruptors, or the prevalence of key limiting resources, may constrain pathways of response. In the face of a rapidly changing environment, there is a pressing need to develop a common framework for describing, assessing, and predicting robustness and resilience within and across living systems.

On the Qinghai-Tibet Plateau, extreme environmental conditions have imposed intense selective pressure on the evolution of phenotypic traits of wild animals. To date, limited information is available on behavioral and ecological traits concerning niche differentiation among sympatric animals on the Qinghai-Tibet Plateau, especially during winter when the environments are most severe. Here, we studied the seasonal variations in habitat occurrence, territorial behavior, and diet in two sympatric snow finches (the white-rumped snow finch, Onychostruthus taczanowskii, WRSF; and the rufous-necked snow finch, Pyrgilauda ruficollis, RNSF) to determine convergence and divergence of ecological traits in such severe climatic conditions. Our results showed that: (i) WRSF occupied rural areas as a dominant species throughout the annual cycle while RNSF occupied the rural areas in summer and then shifted to human-occupied areas in winter and spring; (ii) WRSFs exhibited robust aggressive behavior and territoriality during winter relative to RNSFs; (iii) the diets of both species varied with the season but did not vary between species except that WRSF ate significantly more seeds but RNSF consumed more starchy material derived from human food waste during winter. Therefore, the separations in the spatial niche and territoriality between WRSF and RNSF, especially in winter, may contribute to alleviating the pressure of interspecific competition, and promoting the coexistence of the two sympatric snow finches in the extreme environments on the Qinghai-Tibet Plateau.

The extreme climatic conditions (ECCs) of the Qinghai-Tibet Plateau impose strong selective pressures on the evolution of phenotypic traits in free-living animals. It is not well understood how animals on the Qinghai-Tibet Plateau modify their adrenocortical functions in response to both predictable and unpredictable events of ECCs, especially when the available resources are lowest during the wintering life-history stage. To uncover potential physiological mechanisms, we studied the life history stage dependent features of morphology, the plasma corticosterone response to acute stress and brain glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) mRNA expression in two sympatric snow finches: the white-rumped snow finch (Onychostruthus taczanowskii, WRSF); and the rufous-necked snow finch, Pyrgilauda ruficollis, RNSF) in Qinghai Province, China. Our results showed that (a) baseline corticosterone and stressor-induced corticosterone levels significantly varied with life history stage, but not between the species; (b) in WRSF, GR mRNA expression in the paraventricular nucleus was higher in the wintering stage compared to the pre-basic molt stage. There were no differences in hippocampus MR mRNA expression between stages in either species; (c) in the wintering stage, the suppression of corticosterone secretion in both species was an unexpected strategy in free-living animals. Both convergent and divergent phenotypic traits of adrenocortical responses to acute stress in two sympatric snow finches contribute to our understanding of the coping mechanisms of closely related species in the severe winter on the Qinghai-Tibet Plateau.

Authors Jason E Davis , Mary Beth Kolozsvary , Karolina M Pajerowska-Mukhtar, and Bo Zhang, 1 Radford University, Dept. of Biology, jdavis319@radford.edu 2 Siena College, Dept. of Environmental Studies and Sciences, mkolozsvary@siena.edu 3 University of Alabama at Birmingham, Dept. of Biology, kmukhtar@uab.edu Oklahoma State University, Department of Natural Ecology Resource and Management, bozhangophelia@gmail.com

Recurrent tumors are the predominant cause of breast cancer‐related deaths, therefore there is an urgent need for more effective therapeutic strategies. However, a critical barrier to their development is a lack of understanding of the molecular mechanisms involved in breast tumor recurrence. Recently, primary cilia, microtubule‐based organelles that project from the surface of vertebrate cells, have emerged into the spotlight for their link to numerous human diseases, including cancer. However, the relationship between primary cilia and breast cancer recurrence remains unknown. Here, using a genetically engineered mouse model of breast cancer, which closely recapitulates the clinical phenomenon of breast cancer recurrence, we have uncovered that primary cilia are more highly enriched in recurrent mammary tumors than in both primary mammary tumors and asymptomatic/contralateral mouse mammary tissues. Congruently, we found that the percentage of ciliated cells was substantially elevated in recurrent breast cancer cells compared to primary breast cancer cells. Moreover, RNA‐sequence analysis also revealed that GLI2 and GLI3, which are major downstream effectors of the morphogenic Sonic Hedgehog signaling pathway and are also well‐known to localize to primary cilia, were upregulated in recurrent breast cancer cells compared to primary breast cancer cells. Collectively, these data suggest that primary cilia may have a role in breast cancer recurrence. Our study also indicates that, at least within the recurrent stages of the disease, the primary cilium may serve as a specialized oncogenic signaling organelle.Support or Funding InformationTo Sarah C. Goetz: R00 HD076444 (from the National Institute of Child Health and Human Development).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Prostate cancer is one of the most common cancers in men and is the second leading cause of cancer‐related mortality in men. It is well known that enhanced activation of the MAPK pathway Raf‐MEK‐ERK1/2 directly correlates with the progression, androgen independence and poor prognosis of prostate cancer. Recent studies have also shown that the chemokine receptor CXCR4 is highly expressed in prostate cancer and contributes to prostate tumor growth and metastasis. However, the molecular mechanisms underlying the hyper‐activation of MAPK and the function of CXCR4 remain poorly elucidated. Here, we demonstrated that CXCL12‐induced CXCR4 activation markedly enhanced the activation of ADP‐ribosylation factor 1 (ARF1), a Ras‐related small G protein, in a time‐ and concentration‐dependent fashion, and siRNA‐mediated depletion of ARF1 dramatically inhibited MAPK activation by CXCR4 in prostate cancer cells. More interestingly, we found that ARF1 can mimic Ras to directly and activation‐dependently interact with Raf1, specifically its Ras‐binding domain. Mutations that disrupted ARF1 activation considerably attenuated ARF1‐Raf1 interaction and MAPK activation. These data suggest that, through its direct interaction with Raf1, ARF1 regulates the MAPK activation by CXCR4 in prostate cancer cells. We then investigated the role of ARF1 in prostate tumorigenesis. We found that ARF1 expression and activation were markedly elevated in human prostate cancer cells and tissues. Depletion of ARF1 significantly inhibited prostate cancer cell proliferation in vitro and xenograft tumor growth in vivo , whereas over‐expression of ARF1 produced opposing effects. These data have revealed an unappreciated function of ARF1 in prostate cancer. Altogether, our studies have identified a novel CXCR4‐ARF1‐MAPK signaling pathway which plays an important role in prostate tumorigenesis and may represent a key molecular target for therapeutic innovation (R01GM076167). Support or Funding Information R01GM076167

Molecular mechanisms governing the anterograde trafficking of nascent G protein-coupled receptors (GPCRs) are poorly understood. Here, we have studied the regulation of cell surface transport of α2-adrenergic receptors (α2-ARs) by GGA3 (Golgi-localized, γ-adaptin ear domain homology, ADP ribosylation factor-binding protein 3), a multidomain clathrin adaptor protein that sorts cargo proteins at the trans-Golgi network (TGN) to the endosome/lysosome pathway. By using an inducible system, we demonstrated that GGA3 knockdown significantly inhibited the cell surface expression of newly synthesized α2B-AR without altering overall receptor synthesis and internalization. The receptors were arrested in the TGN. Furthermore, GGA3 knockdown attenuated α2B-AR-mediated signaling, including extracellular signal-regulated kinase 1/2 (ERK1/2) activation and cyclic AMP (cAMP) inhibition. More interestingly, GGA3 physically interacted with α2B-AR, and the interaction sites were identified as the triple Arg motif in the third intracellular loop of the receptor and the acidic motif EDWE in the VHS domain of GGA3. In contrast, α2A-AR did not interact with GGA3 and its cell surface export and signaling were not affected by GGA3 knockdown. These data reveal a novel function of GGA3 in export trafficking of a GPCR that is mediated via a specific interaction with the receptor.

ADP-ribosylation factor 1 (ARF1) is a crucial regulator in vesicle-mediated membrane trafficking and involved in the activation of signaling molecules. However, virtually nothing is known about its function in prostate cancer. Here we have demonstrated that ARF1 expression is significantly elevated in prostate cancer cells and human tissues and that the expression levels of ARF1 correlate with the activation of mitogen-activated protein kinases (MAPK) ERK1/2. Furthermore, we have shown that overexpression and knockdown of ARF1 produce opposing effects on prostate cancer cell proliferation, anchorage-independent growth and tumor growth in mouse xenograft models and that ARF1-mediated cell proliferation can be abolished by the Raf1 inhibitor GW5074 and the MEK inhibitors U0126 and PD98059. Moreover, inhibition of ARF1 activation achieved by mutating Thr48 abolishes ARF1's abilities to activate the ERK1/2 and to promote cell proliferation. These data demonstrate that the aberrant MAPK signaling in prostate cancer is, at least in part, under the control of ARF1 and that, similar to Ras, ARF1 is a critical regulator in prostate cancer progression. These data also suggest that ARF1 may represent a key molecular target for prostate cancer therapeutics and diagnosis.

Intracellular trafficking and precise targeting to specific locations of G protein-coupled receptors (GPCRs) control the physiological functions of the receptors. Compared to the extensive efforts dedicated to understanding the events involved in the endocytic and recycling pathways, the molecular mechanisms underlying the transport of the GPCR superfamily from the endoplasmic reticulum (ER) through the Golgi to the plasma membrane are relatively less well defined. Over the past years, we have used α(2B)-adrenergic receptor (α(2B)-AR) as a model to define the factors that control GPCR export trafficking. In this chapter, we will review specific motifs identified to mediate the export of nascent α(2B)-AR from the ER and the Golgi and discuss the possible underlying mechanisms. As these motifs are highly conserved among GPCRs, they may provide common mechanisms for export trafficking of these receptors.

The amphipathic helix 8 in the membrane-proximal C-terminus is a structurally conserved feature of class A seven transmembrane-spanning G protein-coupled receptors (GPCRs). Mutations of this helical motif often cause receptor misfolding, defective cell surface transport and dysfunction. Surprisingly, we demonstrated here that a single point mutation at Lys308 in helix 8 markedly enhanced the steady-state surface density of the angiotensin II type 1a receptor (AT1aR). Consistent with the enhanced cell surface expression, Lys308 mutation significantly augmented AT1aR-mediated mitogen-activated protein kinase ERK1/2 activation, inositol phosphate production, and vascular smooth muscle cell migration. This mutation also increased the overall expression of AT1aR without altering receptor degradation. More interestingly, Lys308 mutation abolished AT1aR interaction with β-COP, a component of COPI transport vesicles, and impaired AT1aR responsiveness to the inhibition of Rab6 GTPase involved in the Golgi-to-ER retrograde pathway. Furthermore, these functions of Lys308 were largely dependent on its positively charged property. These data reveal previously unappreciated functions of helix 8 and novel mechanisms governing the cell surface transport and function of AT1aR.

Animals inhabiting urban areas must simultaneously cope with the unique challenges presented by this novel habitat type while exploiting the distinctive opportunities it offers. The costs and benefits of urban living are often assumed to be consistent across time, but may in fact vary depending on the habitat features influencing them. Here we examine the glucocorticoid levels and body condition of song sparrows (Melospiza melodia) resident at urban and rural sites over four consecutive years to determine whether these traits, which may be linked to the relative costs and benefits of these respective habitats, are consistent over time. Glucocorticoid levels and body condition varied by year in both habitat types. While habitat alone did not influence glucocorticoid levels, there was a significant interaction between year and habitat, indicating that glucocorticoids differ between habitats in some years but not others. There was no discernable effect of habitat alone on body condition. Overall, these data suggest that the costs and benefits of inhabiting urban versus rural habitats differ substantially from year to year.

Mitogen-activated protein kinases (MAPK) can be activated by a number of biochemical pathways through distinct signaling molecules. We have recently revealed a novel function for the Ras-like small GTPase ADP-ribosylation factor 1 (ARF1) in mediating the activation of Raf1–MEK–ERK1/2 pathway by G protein-coupled receptors [Dong C, Li C and Wu G (2011) J Biol Chem 286, 43,361–43,369]. Here, we have further defined the underlying mechanism and the possible function of ARF1-mediated MAPK pathway. We demonstrated that the blockage of ARF1 activation and the disruption of ARF1 localization to the Golgi by mutating Thr48, a highly conserved residue involved in the exchange of GDP for GTP, and the myristoylation site Gly2 abolished ARF1's ability to activate ERK1/2. In addition, treatment with Golgi structure disrupting agents markedly attenuated ARF1-mediated ERK1/2 activation. Furthermore, ARF1 significantly promoted cell proliferation. More interestingly, ARF1 activated 90kDa ribosomal S6 kinase 1 (RSK1) without influencing Elk-1 activation and ERK2 translocation to the nuclei. These data demonstrate that, once activated, ARF1 activates the MAPK pathway likely using the Golgi as a main platform, which in turn activates the cytoplasmic RSK1, leading to cell proliferation.

Organisms frequently need to adjust physiological mechanisms to successfully breed in novel habitats. To explore how some populations physiologically acclimate to novel environmental conditions while others do not, we examine three subspecies of the white-crowned sparrow, Zonotrichia leucophrys. Of these subspecies, Z. l. pugetensis has expanded its breeding range to high altitude over the last 60 years. We investigate physiological acclimation to high altitude conditions by comparing circulating levels of glucocorticoids among Z. l. gambelii, which only breeds at high altitude, Z. l. nuttalli, which only breeds at low altitude, a population of Z. l. pugetensis that breeds at low altitude, and a Z. l. pugetensis population that now breeds at high altitude. Glucocorticoids mediate physiological and behavioral responses to environmental conditions and are constitutively secreted, but can also be released facultatively. We hypothesized that elevation of the glucocorticoid corticosterone (CORT) may facilitate breeding in high altitude environments. We tested this hypothesis by comparing baseline and stress-induced CORT levels of subspecies breeding at low altitude, Z. l. pugetensis and Z. l. nuttalli, to subspecies breeding at high altitude, Z. l. pugetensis and Z. l. gambelii. We found that populations breeding at high altitude exhibit higher baseline and stress-induced levels of CORT. Additionally, we found that Z. l. pugetensis exhibit greater variation in the stress-induced CORT response. These results suggest an importance of modulation of hormonal mechanisms in facilitating breeding in high altitude environments, and that variation in these mechanisms may be associated with facilitating altitudinal range expansion.

Understanding the interactions of an organism and its environment is essential for us to integrate ultimate and proximate causation on a global scale. Organism–environment interaction includes all organisms including animals, plants, and non-eukaryotes, etc. because all of them are responsive to environmental change including those that are human-induced. A mechanistic approach is important for us to understand why some organisms can cope with change and others cannot. Here, we present three examples of environments ("the three poles") that are changing rapidly and how avian species typical of these ecosystems are responding. These examples include apparently adaptive responses to change in climate (i.e. the predictable environment) in one species in which a lengthened breeding season now allows multiple breeding attempts. Why other species are unable to respond in a similar way remains unclear. A second example describes how changing weather (i.e. the unpredictable) may have disastrous results for breeding success in a species adapted to an extreme cold environment. Implications for climate change in which weather extremes will become more common again suggest a mechanistic approach will be important to understand how organisms may respond. The third example outlines a scenario in which multiple human-induced rapid changes (a combination of predictable and unpredictable such as development, habitat change, introduction of invasive species and climate change) may influence indigenous species in different ways. Organism–environment interaction is a fundamental concept that may unify ultimate and proximate causation and point the way for future investigations striving to understand coping mechanisms in a world where both predictable and unpredictable components of the environment are changing.