Dominance-related seasonal song production is unrelated to circulating testosterone in a subtropical songbird

PublishedJournal ArticleCirculating testosterone (T) is widely considered to play a key role in the production of sexual displays by male vertebrates. While numerous studies support a role for circulating T in promoting the production of song in male birds, this understanding is based primarily on evidence from seasonally breeding northern temperate species, leaving it unclear whether this mechanism generalizes to other regions of the world. Here we investigate whether variation in circulating levels of T can explain the marked within- and among-individual variation in male song performance observed in a subtropical population of the year-round territorial white-browed sparrow weaver (Plocepasser mahali mahali). Our findings reveal that both circulating T and male song production peaked at a similar time point, halfway through the population-level breeding season. However, while dominant males were more likely to sing and sang for longer than subordinate males, within-group paired comparisons revealed no dominance-related differences in circulating T. Moreover, comparisons both among and within individual dominant males revealed that song duration, syllable rate and proportion of time spent singing were all unrelated to circulating T. Together, our findings suggest that natural variation in male song production, at least in this population of white-browed sparrow weavers, is achieved principally through mechanisms other than variation in circulating T concentration. More widely, our results are in line with the view that male song production is not exclusively regulated by gonadally synthesized steroids.J.E.Y. was supported by a University of Bristol Post-Graduate Scholarship, A.J.Y. and A.N.R. were supported by BBSRC David Phillips Research Fellowships (BB/H022716/1 and BB/C520555/1)

Surveillance of heat-related illness in small animals presenting to veterinary practices in the UK between 2013-2018

Background: Heat-related illness (HRI) can affect all companion animals and is likely to become more common as global temperatures rise. The misconception that HRI is primarily a result of dogs being trapped in hot cars, highlights a lack of awareness of HRI risk factors within the UK companion animal population. Aim: This project aimed to review all species of small animal presentations of HRI to UK veterinary practices participating in the Small Animal Veterinary Surveillance Network (SAVSNET), describe the inciting triggers and seasonality of HRI events, and review the clinical grade of canine patients presenting with HRI. Methods: Electronic consultation records were submitted by volunteer veterinary practices across Great Britain to SAVSNET. Cases were defined as animals presented for consultation with strong evidence of current, or recent heat induced illness during the study period (2013-2018). Results: The HRI cases included 146 dogs, 16 cats, eight guinea pigs, three rabbits and one ferret. Of the 118 HRI cases with a recorded trigger, exercise was the primary trigger for dogs presenting (73.5%); seven (6.9%) canine HRI events followed vehicular confinement. Environmental HRI was recorded as a trigger for the remaining dogs (19.6%), and for all cats, guinea pigs, rabbits and the ferret. Brachycephalic breeds comprised 21.2% of canine HRI cases, and all rabbits were brachycephalic breeds. Dogs presented for HRI between April and October, with 42.5% during July, typically the UK’s hottest month of the year. Cats with HRI were presented between May and September, with 75.0% during June and July. The smaller companion species - ferrets, rabbits and guinea pigs – were presented during the UK’s summer months June to August. Conclusion: This study highlights the risk of HRI to all pet animals during the UK’s warmer summer months (June to August). The findings support previous claims that exercise is the most common trigger of HRI in dogs, whilst environmental HRI (a hot ambient temperature) accounted for all HRI events in cats, rabbits, guinea pigs and ferrets. Both brachycephalic dogs and rabbits were overrepresented, adding further evidence that owners of these animals should be particularly vigilant for HRI during hot weather

The Role of SurA PPIase Domains in Preventing Aggregation of the Outer Membrane Proteins tOmpA and OmpT

SurA is a conserved ATP-independent periplasmic chaperone involved in the biogenesis of outer-membrane proteins (OMPs). Escherichia coli SurA has a core domain and two peptidylprolyl isomerase (PPIase) domains, the role(s) of which remain unresolved. Here we show that while SurA homologues in early proteobacteria typically contain one or no PPIase domains, the presence of two PPIase domains is common in SurA in later proteobacteria, implying an evolutionary advantage for this domain architecture. Bioinformatics analysis of > 350,000 OMP sequences showed that their length, hydrophobicity and aggregation propensity are similar across the proteobacterial classes, ruling out a simple correlation between SurA domain architecture and these properties of OMP sequences. To investigate the role of the PPIase domains in SurA activity, we deleted one or both PPIase domains from E. coli SurA and investigated the ability of the resulting proteins to bind and prevent the aggregation of tOmpA (19 kDa) and OmpT (33 kDa). The results show that wild-type SurA inhibits the aggregation of both OMPs, as do the cytoplasmic OMP chaperones trigger factor and SecB. However, while the ability of SurA to bind and prevent tOmpA aggregation does not depend on its PPIase domains, deletion of even a single PPIase domain ablates the ability of SurA to prevent OmpT aggregation. The results demonstrate that the core domain of SurA endows its generic chaperone ability, while the presence of PPIase domains enhances its chaperone activity for specific OMPs, suggesting one reason for the conservation of multiple PPIase domains in SurA in proteobacteria

Design and Synthesis of Cysteine-Specific Labels for Photo-Crosslinking Studies

Chemical cross-linking mass-spectrometry (XL-MS) represents a powerful methodology to map ligand/biomacromolecule interactions, particularly where conventional methods such as X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy or cryo-electron microscopy (EM) are not feasible. In this manuscript, we describe the design and synthesis of two new photo-crosslinking reagents that can be used to specifically label free thiols through either maleimido or methanethiosulfonate groups and facilitate PXL-MS workflows. Both crosslinkers are based on light sensitive diazirines – precursors of highly reactive carbenes which offer additional advantages over alternative crosslinking groups such as benzophenones and aryl nitrenes given the controlled rapid and more indiscriminate reactivity

Visualizing and trapping transient oligomers in amyloid assembly pathways

Oligomers which form during amyloid fibril assembly are considered to be key contributors towards amyloid disease. However, understanding how such intermediates form, their structure, and mechanisms of toxicity presents significant challenges due to their transient and heterogeneous nature. Here, we discuss two different strategies for addressing these challenges: use of (1) methods capable of detecting lowly-populated species within complex mixtures, such as NMR, single particle methods (including fluorescence and force spectroscopy), and mass spectrometry; and (2) chemical and biological tools to bias the amyloid energy landscape towards specific oligomeric states. While the former methods are well suited to following the kinetics of amyloid assembly and obtaining low-resolution structural information, the latter are capable of producing oligomer samples for high-resolution structural studies and inferring structure-toxicity relationships. Together, these different approaches should enable a clearer picture to be gained of the nature and role of oligomeric intermediates in amyloid formation and disease

Calling by Concluding Sentinels: Coordinating Cooperation or Revealing Risk?

Efficient cooperation requires effective coordination of individual contributions to the cooperative behaviour. Most social birds and mammals involved in cooperation produce a range of vocalisations, which may be important in regulating both individual contributions and the combined group effort. Here we investigate the role of a specific call in regulating cooperative sentinel behaviour in pied babblers (Turdoides bicolor). ‘Fast-rate chuck’ calls are often given by sentinels as they finish guard bouts and may potentially coordinate the rotation of individuals as sentinels, minimising time without a sentinel, or may signal the presence or absence of predators, regulating the onset of the subsequent sentinel bout. We ask (i) when fast-rate chuck calls are given and (ii) what effect they have on the interval between sentinel bouts. Contrary to expectation, we find little evidence that these calls are involved in regulating the pied babbler sentinel system: observations revealed that their utterance is influenced only marginally by wind conditions and not at all by habitat, while observations and experimental playback showed that the giving of these calls has no effect on inter-bout interval. We conclude that pied babblers do not seem to call at the end of a sentinel bout to maximise the efficiency of this cooperative act, but may use vocalisations at this stage to influence more individually driven behaviours

Modulation of Amyloidogenic Protein Self-Assembly Using Tethered Small Molecules

Protein–protein interactions (PPIs) are involved in many of life’s essential biological functions yet are also an underlying cause of several human diseases, including amyloidosis. The modulation of PPIs presents opportunities to gain mechanistic insights into amyloid assembly, particularly through the use of methods which can trap specific intermediates for detailed study. Such information can also provide a starting point for drug discovery. Here, we demonstrate that covalently tethered small molecule fragments can be used to stabilize specific oligomers during amyloid fibril formation, facilitating the structural characterization of these assembly intermediates. We exemplify the power of covalent tethering using the naturally occurring truncated variant (ΔN6) of the human protein β2-microglobulin (β2m), which assembles into amyloid fibrils associated with dialysis-related amyloidosis. Using this approach, we have trapped tetramers formed by ΔN6 under conditions which would normally lead to fibril formation and found that the degree of tetramer stabilization depends on the site of the covalent tether and the nature of the protein–fragment interaction. The covalent protein–ligand linkage enabled structural characterization of these trapped, off-pathway oligomers using X-ray crystallography and NMR, providing insight into why tetramer stabilization inhibits amyloid assembly. Our findings highlight the power of “post-translational chemical modification” as a tool to study biological molecular mechanisms

"Just old age" - a qualitative investigation of owner and veterinary professional experiences of and attitudes to ageing in dogs in the UK.

ObjectivesMany UK dogs live into old age, but owners may not recognise or report age-associated signs of disease which lead to negative welfare. This study investigated dog owner and veterinary professional experiences and attitudes towards ageing in dogs, how health care is offered, barriers to its delivery, and some best-practice solutions.Materials and methodsIn-depth semi-structured interviews were conducted with 15 owners of 21 dogs (aged 8 to 17 years mean: 13) and 11 veterinary professional (eight veterinary surgeons, two nurses and one physiotherapist). Open-text responses from 61 dog owner were collected using an online survey. Transcripts and survey responses were inductively coded into themes.ResultsFour themes were constructed: "just old age", barriers to care, trust in veterinary surgeons, and tools to improve health care. Age-related changes were mostly perceived as "just old age" by dog owner. Many dogs were no longer vaccinated and did not attend check-ups unless owners identified a problem. The greatest barriers to health care were finances (dog owner), owner awareness, willingness to act and consultation time (veterinary professional). Trust in veterinary professional was more likely when dog owner experienced continuity, prioritisation of care, clear communication and an accessible, knowledgeable and empathic veterinary professional. Participants suggested that senior health care and communication between dog owner and veterinary professional could be improved through questionnaires, and evidence-based online information.Clinical significanceOpportunities to educate owners on which clinical signs represent healthy or pathological ageing are being missed. Resources should be developed to guide on best-practice discussions in consultations, encourage more owners to recognise clinical signs and to seek and trust veterinary advice

Inter-domain dynamics in the chaperone SurA and multi-site binding to its outer membrane protein clients

The periplasmic chaperone SurA plays a key role in outer membrane protein (OMP) biogenesis. E. coli SurA comprises a core domain and two peptidylprolyl isomerase domains (P1 and P2), but its mechanisms of client binding and chaperone function have remained unclear. Here, we use chemical cross-linking, hydrogen-deuterium exchange mass spectrometry, single-molecule FRET and molecular dynamics simulations to map the client binding site(s) on SurA and interrogate the role of conformational dynamics in OMP recognition. We demonstrate that SurA samples an array of conformations in solution in which P2 primarily lies closer to the core/P1 domains than suggested in the SurA crystal structure. OMP binding sites are located primarily in the core domain, and OMP binding results in conformational changes between the core/P1 domains. Together, the results suggest that unfolded OMP substrates bind in a cradle formed between the SurA domains, with structural flexibility between domains assisting OMP recognition, binding and release