Transforming growth factor-[beta]1 regulates steady-state PTH/PTHrP receptor mRNA levels and PTHrP binding in ROS 17/2.8 osteosarcoma cells

The effect of transforming growth factor [beta]1 (TGF-[beta]1) on the expression of mRNA for the parathyroid hormone receptor and binding of iodinated parathyroid hormone-related protein in ROS 17/2.8 osteosarcoma cells was evaluated. TGF-[beta]1 stimulated a 2-7-fold increase in steady state mRNA levels for the parathyroid hormone receptor at a maximal dose of 5 ng/ml, with increased levels of expression at 6 h of TGF-[beta]1-incubation, and peak levels at 8-24 h. Receptor binding studies revealed a significant increase in PTHrP-specific binding with TGF-[beta]1 doses as low as 0.5 ng/ml and a 55% increase in numbers of receptors with no alteration in binding affinity with 5.0 ng/ml TGF-[beta]1. Time course studies indicated that receptor binding was increased at 24 h with peak levels reached at 48 h of treatment. PTH-stimulated cAMP levels were significantly increased in ROS 17/2.8 cells treated with TGF-[beta]1 (0.5 ng/ml) for 48 h. These data indicate that TGF-[beta]1 upregulates steady-state mRNA, ligand binding and PTH/PTHrP receptor signaling in rat osteosarcoma cells. The effects of TGF-[beta]1 on bone may be attributed in part to regulation of the PTH/PTHrP receptor at the molecular level.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31617/1/0000549.pd

Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression

Multiple, complex molecular events characterize cancer development and progression(1,2). Deciphering the molecular networks that distinguish organ- confined disease from metastatic disease may lead to the identification of critical biomarkers for cancer invasion and disease aggressiveness. Although gene and protein expression have been extensively profiled in human tumours, little is known about the global metabolomic alterations that characterize neoplastic progression. Using a combination of high- throughput liquid- and- gas- chromatography- based mass spectrometry, we profiled more than 1,126 metabolites across 262 clinical samples related to prostate cancer ( 42 tissues and 110 each of urine and plasma). These unbiased metabolomic profiles were able to distinguish benign prostate, clinically localized prostate cancer and metastatic disease. Sarcosine, an N- methyl derivative of the amino acid glycine, was identified as a differential metabolite that was highly increased during prostate cancer progression to metastasis and can be detected non- invasively in urine. Sarcosine levels were also increased in invasive prostate cancer cell lines relative to benign prostate epithelial cells. Knockdown of glycine- N- methyl transferase, the enzyme that generates sarcosine from glycine, attenuated prostate cancer invasion. Addition of exogenous sarcosine or knockdown of the enzyme that leads to sarcosine degradation, sarcosine dehydrogenase, induced an invasive phenotype in benign prostate epithelial cells. Androgen receptor and the ERG gene fusion product coordinately regulate components of the sarcosine pathway. Here, by profiling the metabolomic alterations of prostate cancer progression, we reveal sarcosine as a potentially important metabolic intermediary of cancer cell invasion and aggressivity.Early Detection Research Network ; National Institutes of Health ; MTTC ; Clinical Translational Science Award ; Fund for Discovery of the University of Michigan Comprehensive Cancer Center ; University of Michigan Cancer Biostatistics Training Grant ; Doris Duke Charitable FoundationWe thank J. Granger for help in manuscript preparation, J. Siddiqui and R. Varambally for help with the clinical database, and A. Vellaichamy and S. Pullela for technical assistance. We thank K. Pienta for access to metastatic prostate cancer samples from the University of Michigan Prostate SPORE rapid autopsy programme. This work is supported in part by the Early Detection Research Network (A.M.C., J.T.W.), National Institutes of Health (A.S., S.P., J.B., T.M.R., D.G., G.S.O. and A.M.C.) and an MTTC grant (G.S.O. and A.S.). A.M.C. is supported by a Clinical Translational Science Award from the Burroughs Welcome Foundation. A. S. is supported by a grant from the Fund for Discovery of the University of Michigan Comprehensive Cancer Center. L. M. P. is supported by the University of Michigan Cancer Biostatistics Training Grant. A. M. C and S. P. are supported by the Doris Duke Charitable Foundation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/62661/1/nature07762.pd

Determining crystal structures through crowdsourcing and coursework

We show here that computer game players can build high-quality crystal structures. Introduction of a new feature into the computer game Foldit allows players to build and real-space refine structures into electron density maps. To assess the usefulness of this feature, we held a crystallographic model-building competition between trained crystallographers, undergraduate students, Foldit players and automatic model-building algorithms. After removal of disordered residues, a team of Foldit players achieved the most accurate structure. Analysing the target protein of the competition, YPL067C, uncovered a new family of histidine triad proteins apparently involved in the prevention of amyloid toxicity. From this study, we conclude that crystallographers can utilize crowdsourcing to interpret electron density information and to produce structure solutions of the highest quality

Spatial movements and social networks in juvenile male song sparrows

The time between fledging and breeding is a critical period in songbird ontogeny, but the behavior of young songbirds in the wild is relatively unstudied. The types of social relationships juveniles form with other individuals can provide insight into the process through which they learn complex behaviors crucial for survival, territory establishment, and mate attraction. We used radio telemetry to observe social associations of young male song sparrows (Melospiza melodia) from May to November. Juvenile song sparrows were frequently observed in social flocks and generally associated with more birds in the summer than in the autumn months. Most juvenile subjects formed stable social relationships with other birds and were seen with the same individual on up to 60% of the days observed. The strongest associations occurred with other juvenile males, and these individuals were often seen <1 m from the subject, even when the subject moved large distances between tracking observations. Associations also had long-term behavioral consequences as subjects were more likely to establish territories near their associates and learn shared song types. Our results indicate that male song sparrows spend a large percentage of the juvenile life stage forming social relationships and suggest that these associations may be important for the ecology of young birds and the ontogeny of their behaviors

Indirect reciprocity:song sparrows distrust aggressive neighbours based on eavesdropping

The evolution of cooperation between unrelated individuals has been a central issue in evolutionary biology. The main problem in most theories of cooperation is how a cooperative player selects individuals to 'trust' so that he does not get exploited by noncooperators. While early models emphasized the role of direct experience with individuals in deciding who to trust, more recent work has shown that individuals can eavesdrop on interactions between other individuals to identify cooperators and noncooperators. This second route to cooperation is called indirect reciprocity. In spatially structured populations with repeated interactions between players, both sources of information (direct experience and observed reputation) are readily available. Most models and empirical studies to date, however, have considered indirect reciprocity only in one-shot interactions when direct experience is not available. We examined the role of indirect reciprocity in the maintenance of mutual restraint in aggression (Dear Enemy cooperation) between territorial male song sparrows, Melospiza melodia. We found that territory owners eavesdropped on simulated defections by a neighbour (intrusions onto a third bird's territory) and subsequently retaliated against these defecting neighbours. Taken together with our previous results, these results suggest that both direct and indirect reciprocity can be at work in repeated-interaction scenarios, and together lead to emergence of cooperation. (C) 2010 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.</p

Good neighbour, bad neighbour: song sparrows retaliate against aggressive rivals

Many territorial animals, despite being in direct competition for resources such as space, food and mates, show reduced aggression towards their neighbours. This situation is called the Dear Enemy effect. One explanation of the Dear Enemy effect is that it is due to a conditional strategy like Tit for Tat where territory holders cooperate by reducing aggression towards neighbours that also show reduced aggression, but retaliate against aggressive neighbours. Previous research found evidence for such a conditional strategy in migratory species but not in species with long-term association between neighbours, suggesting that long-term neighbours might be engaged in more ‘forgiving’ strategies. We tested this hypothesis in male song sparrows, Melodia melospiza, which are resident year-round in our population (leading to long-term associations between neighbouring birds) and display the Dear Enemy effect. We found that following a simulated intrusion by a neighbour, song sparrow males responded more strongly to playback of this neighbour than to playback of a neutral neighbour from their respective boundaries, consistent with a conditional retaliation strategy. We suggest that the primary effect of an intrusion by a neighbour might be to increase the perceived risk of cuckoldry by the intruding male, and increased aggression and vigilance towards this neighbour might be a strategy to prevent cuckoldry

Immediate and long-term effects of testosterone on song plasticity and learning in juvenile song sparrows

Steroid sex hormones play critical roles in the development of brain regions used for vocal learning. It has been suggested that puberty-induced increases in circulating testosterone (T) levels crystallize a bird's repertoire and inhibit future song learning. Previous studies show that early administration of T crystallizes song repertoires but have not addressed whether new songs can be learned after this premature crystallization. We brought 8 juvenile song sparrows (Melospiza melodia) into the laboratory in the late summer and implanted half of them with subcutaneous T pellets for a two week period in October. Birds treated with T tripled their singing rates and crystallized normal songs in 2 weeks. After T removal, subjects were tutored by 4 new adults. Birds previously treated with T tended toward learning fewer new songs post T, consistent with the hypothesis that T helps to close the song learning phase. However, one T-treated bird proceeded to learn several new songs in the spring, despite singing perfectly crystallized songs in the fall. His small crystallized fall repertoire and initial lag behind other subjects in song development suggest that this individual may have had limited early song learning experience. We conclude that an exposure to testosterone sufficient for crystallization of a normal song repertoire does not necessarily prevent future song learning and suggest that early social experiences might override the effects of hormones in closing song learning. (c) 2012 Elsevier B.V. All rights reserved.</p

Who initiates extrapair mating in song sparrows?

Extrapair paternity (EPP) is a common feature of many mating systems. Although molecular methods have made it possible to document the rate of EPP across numerous taxa, we still lack an understanding of how and why EPP happens. Behavioral data on mating interactions are needed to answer this question. We employed radiotelemetry to follow the movement patterns of tagged song sparrows (Melospiza melodia) during their nesting cycle. We found that females and males commonly forayed onto neighboring territories in the prefertile period, frequently together. In the fertile period, however, the foray rate dropped significantly, and females largely stayed on their own territories. Concurrently in the fertile period, both the time mates spent in proximity and intrusion rates on their territories increased compared with the prefertile period. After the female started laying eggs, proximity of the mates to one another declined again, and some males started to foray onto neighboring territories, in most cases into territories where there was a fertile female. Thus, females do not seem to seek out particular males for extrapair copulations (EPCs) in their fertile period, but rather it is the males who actively seek extrapair mating. Males therefore face a trade-off between ensuring paternity at home and pursuing copulations elsewhere. Agonistic interactions between extrapair males and females were almost entirely absent, suggesting that males do not force females into copulations. Therefore, in song sparrows, males have to initiate EPCs by seeking out fertile females, suggesting that males probably drive the pattern of EPP in our population.</p