Hormonally mediated maternal effects in birds

The main aim of this thesis was to investigate the effects of environmental conditions, particularly unpredictable or potentially negative ones, on the maternal transmission of the primary avian stress hormone, corticosterone, to developing embryos. We currently lack information on the extent to which conditions in the maternal environment are transmitted to the offspring in birds via egg compositional changes. It is possible that maternally derived hormonal signals communicate information about the external environment to developing embryos and directly influence the fitness of their offspring in a negative or positive way. I found, using captive zebra finches, that the experimental stressor of unpredictable food availability (as these birds are used to ad libitum food) experienced by mothers can elevate yolk CORT concentrations, but only when combined with the additional demand of laying a replacement clutch (potentially a buffering system to prevent mild stressors impacting on CORT transmission to the embryo). I then looked at yolk CORT concentrations in two populations of gulls (herring and lesser black-backed gulls) in which the population trajectories differed depending on environmental conditions (potentially a reflection of different exposures to stressful stimuli). The results however did not support this hypothesis, as there were no differences according to habitat type or between species (where they coexist). This would suggest that the different environmental circumstances (harsher for the herring gull) experienced by these two species are not reflected in differences in their eggs (at least in terms of CORT). This could be the result of the eggs being buffered from the maternal CORT environment or it may be that the difficult environmental conditions are not occurring during the breeding season. We also identified that experimental human disturbance during the laying period does not appear to elevate yolk CORT concentrations, although there was a trend for concentrations to be higher following the loss of the first clutch in the herring gull (as seen in the zebra finches). I also measured yolk CORT concentrations in Common Eider eggs and looked for differences according to the degree of nest shelter. I found no relationship between shelter and yolk CORT, but birds that laid in more sheltered sites had, on average, smaller eggs. This may indicate lesser quality birds are nesting in the sheltered sites and that yolk CORT is not affected by maternal condition. Finally, I looked at another mechanism through which information relating to the maternal environment could be transferred to the embryo. I investigated whether there were any links between maternally derived immunity and CORT by comparing the anti-microbial lysozyme and CORT concentrations in the albumen. I found no correlation between CORT and lysozyme, suggesting that CORT may not affect lysozyme production. It may be that other factors such as colony density and ‘cleanliness’ are more important in determining the concentrations of lysozyme deposited in the egg or that lysozyme production is not sufficiently costly to be influenced by the maternal stress state. The overall theme of my findings is that CORT concentrations in eggs do not appear to vary much with maternal environments. I will discuss these findings in their broader ecological and evolutionary context and discuss if stress hormones are indeed being used as adaptive signals for preparing the embryo for its postnatal environment

A Review of Thick-Billed Murre Banding in the Canadian Arctic, 1950–2010

Banding of Thick-billed Murres Uria lomvia in the Canadian Arctic was initiated by L.M. Tuck in the 1950s, when he visited three of the largest breeding colonies in Canada. Up to 2010, banding had been carried out at eight of the 10 major breeding colonies, with totals of more than 1000 birds banded at Coburg Island and Cape Hay, Bylot Island, in the High Arctic and at Digges Sound and Coats Island in northern Hudson Bay. Because murres are long-lived birds, large-scale banding can continue to provide useful results for decades. A total of about 89 000 bandings in Arctic Canada resulted in 1757 usable recoveries up to 2010, the vast majority of which were birds killed by hunters in West Greenland or Newfoundland and Labrador. There was no apparent change in the large-scale geographical pattern of recoveries over the period reviewed, but the proportion of bands recovered has fallen. Several periods of higher- or lower-than-expected recoveries can be attributed to particular events: anomalous ice conditions, intensive gill-net fisheries, and oiling at sea. Thus, banding provided a useful tool not only for identifying migration and wintering areas, but also for identifying transient sources of increased mortality.Les débuts du baguage des guillemots de Brünnich Uria lomvia dans l’Arctique canadien remontent aux années 1950, sous l’égide de L.M. Tuck à l’occasion de sa visite de trois des plus grandes colonies de nidification au Canada. Jusqu’en 2010, le baguage a été effectué à huit des dix grandes colonies de nidification, pour un total de plus de 1 000 oiseaux bagués à l’île Coburg, à cap Hay et à l’île Bylot, dans l’Extrême-Arctique, ainsi qu’au détroit de Digges et à l’île Coats, dans le nord de la baie d’Hudson. Puisque les guillemots de Brünnich sont des oiseaux qui vivent longtemps, le baguage à grande échelle peut donner des résultats pendant des années. Jusqu’en 2010, les quelque 89 000 baguages effectués dans l’Arctique canadien se sont traduits par 1 757 récupérations d’oiseaux utilisables, la vaste majorité d’entre eux ayant été tuée par des chasseurs de l’ouest du Groenland ou de Terre-Neuve-Labrador. Il n’y avait aucun changement apparent dans le modèle géographique à grande échelle des oiseaux récupérés au cours de la période examinée, mais la proportion de bagues récupérées a diminué. Plusieurs périodes de récupérations plus grandes ou moins grandes que prévu peuvent être attribuées à des événements particuliers : un état anormal des glaces, la pêche intensive au filet maillant et le mazoutage en mer. Par conséquent, le baguage a représenté un outil utile non seulement pour déterminer les lieux de migration et les aires d’hivernage, mais aussi pour repérer les sources transitoires de mortalité accrue.Mots clés : L.M. Tuck, guillemot de Brünnich, Uria lomvia, chasse, baguage, population

Mapping the cellular electrophysiology of rat sympathetic preganglionic neurones to their roles in cardiorespiratory reflex integration:A whole cell recording study in situ

Sympathetic preganglionic neurones (SPNs) convey sympathetic activity flowing from the CNS to the periphery to reach the target organs. Although previous in vivo and in vitro cell recording studies have explored their electrophysiological characteristics, it has not been possible to relate these characteristics to their roles in cardiorespiratory reflex integration. We used the working heart–brainstem preparation to make whole cell patch clamp recordings from T3–4 SPNs (n = 98). These SPNs were classified by their distinct responses to activation of the peripheral chemoreflex, diving response and arterial baroreflex, allowing the discrimination of muscle vasoconstrictor-like (MVC(like), 39%) from cutaneous vasoconstrictor-like (CVC(like), 28%) SPNs. The MVC(like) SPNs have higher baseline firing frequencies (2.52 ± 0.33 Hz vs. CVC(like) 1.34 ± 0.17 Hz, P = 0.007). The CVC(like) have longer after-hyperpolarisations (314 ± 36 ms vs. MVC(like) 191 ± 13 ms, P < 0.001) and lower input resistance (346 ± 49  MΩ vs. MVC(like) 496 ± 41 MΩ, P < 0.05). MVC(like) firing was respiratory-modulated with peak discharge in the late inspiratory/early expiratory phase and this activity was generated by both a tonic and respiratory-modulated barrage of synaptic events that were blocked by intrathecal kynurenate. In contrast, the activity of CVC(like) SPNs was underpinned by rhythmical membrane potential oscillations suggestive of gap junctional coupling. Thus, we have related the intrinsic electrophysiological properties of two classes of SPNs in situ to their roles in cardiorespiratory reflex integration and have shown that they deploy different cellular mechanisms that are likely to influence how they integrate and shape the distinctive sympathetic outputs

Clustering of eastern North Pacific tropical cyclone tracks: ENSO and MJO effects

A probabilistic clustering technique is used to describe tropical cyclone tracks in the eastern North Pacific, on the basis of their shape and location. The best track data set is decomposed in terms of three clusters; these clusters are analyzed in terms of genesis location, trajectory, landfall, intensity, seasonality, and their relationships with the El Niño–Southern Oscillation (ENSO) and Madden-Julian Oscillation (MJO). Longitudinal track location plays a strong discriminating role in the regression mixture model's solution, with the average track orientation becoming more zonal toward the west. This progression encapsulates well the relationship between tropical cyclones over the eastern tropical Pacific and the MJO or ENSO. Two of the clusters describe tropical cyclones (TCs) with tracks that lie near the coast of Mexico and Central America. The most frequent cluster contains tracks that trend west-northwestward, while the second most frequent one has genesis locations that lie slightly to the southeast of those in the most frequent cluster and tracks that run typically parallel to the Central American coast. This second cluster is shown to be significantly associated with the westerly phase of the MJO. The third, least frequent cluster contains TCs with westward trajectories over the central and eastern equatorial Pacific; some of these TCs have an impact on Hawaii and other islands, as far as the central and western North Pacific regions. The least frequent cluster is strongly related to ENSO, while the others are not; it occurs significantly more frequently during El Niño conditions. Examination of the large-scale patterns of atmospheric circulation and sea surface temperature associated with each of our three clusters are consistent with previous studies. Anomalous low-level westerly zonal winds from the monsoon trough and MJO meet anomalous easterlies near the region of genesis in each cluster

The evolution of Runx genes II. The C-terminal Groucho recruitment motif is present in both eumetazoans and homoscleromorphs but absent in a haplosclerid demosponge

Background. The Runt DNA binding domain (Runx) defines a metazoan family of sequence-specific transcription factors with essential roles in animal ontogeny and stem cell based development. Depending on cis-regulatory context, Runx proteins mediate either transcriptional activation or repression. In many contexts Runx-mediated repression is carried out by Groucho/TLE, recruited to the transcriptional complex via a C-terminal WRPY sequence motif that is found encoded in all heretofore known Runx genes. Findings. Full-length Runx genes were identified in the recently sequenced genomes of phylogenetically diverse metazoans, including placozoans and sponges, the most basally branching members of that clade. No sequences with significant similarity to the Runt domain were found in the genome of the choanoflagellate Monosiga brevicollis, confirming that Runx is a metazoan apomorphy. A contig assembled from genomic sequences of the haplosclerid demosponge Amphimedon queenslandica was used to construct a model of the single Runx gene from that species, AmqRunx, the veracity of which was confirmed by expressed sequences. The encoded sequence of the Runx protein OscRunx from the homoscleromorph sponge Oscarella carmella was also obtained from assembled ESTs. Remarkably, a syntenic linkage between Runx and Supt3h, previously reported in vertebrates, is conserved in A. queenslandica. Whereas OscRunx encodes a C-terminal Groucho-recruitment motif, AmqRunx does not, although a Groucho homologue is found in the A. queenslandica genome. Conclusion. Our results are consistent with the hypothesis that sponges are paraphyletic, and suggest that Runx-WRPY mediated recruitment of Groucho to cis-regulatory sequences originated in the ancestors of eumetazoans following their divergence from demosponges

The evolution of Runx genes I. A comparative study of sequences from phylogenetically diverse model organisms

BACKGROUND: Runx genes encode proteins defined by the highly conserved Runt DNA-binding domain. Studies of Runx genes and proteins in model organisms indicate that they are key transcriptional regulators of animal development. However, little is known about Runx gene evolution. RESULTS: A phylogenetically broad sampling of publicly available Runx gene sequences was collected. In addition to the published sequences from mouse, sea urchin, Drosophila melanogaster and Caenorhabditis elegans, we collected several previously uncharacterised Runx sequences from public genome sequence databases. Among deuterostomes, mouse and pufferfish each contain three Runx genes, while the tunicate Ciona intestinalis and the sea urchin Strongylocentrotus purpuratus were each found to have only one Runx gene. Among protostomes, C. elegans has a single Runx gene, while Anopheles gambiae has three and D. melanogaster has four, including two genes that have not been previously described. Comparative sequence analysis reveals two highly conserved introns, one within and one just downstream of the Runt domain. All vertebrate Runx genes utilize two alternative promoters. CONCLUSIONS: In the current public sequence database, the Runt domain is found only in bilaterians, suggesting that it may be a metazoan invention. Bilaterians appear to ancestrally contain a single Runx gene, suggesting that the multiple Runx genes in vertebrates and insects arose by independent duplication events within those respective lineages. At least two introns were present in the primordial bilaterian Runx gene. Alternative promoter usage arose prior to the duplication events that gave rise to three Runx genes in vertebrates

Assessment of Canopy Chlorophyll Content Retrieval in Maize and Soybean: Implications of Hysteresis on the Development of Generic Algorithms

Canopy chlorophyll content (Chl) closely relates to plant photosynthetic capacity, nitrogen status and productivity. The goal of this study is to develop remote sensing techniques for accurate estimation of canopy Chl during the entire growing season without re-parameterization of algorithms for two contrasting crop species, maize and soybean. These two crops represent different biochemical mechanisms of photosynthesis, leaf structure and canopy architecture. The relationships between canopy Chl and reflectance, collected at close range and resampled to bands of the Multi Spectral Instrument (MSI) aboard Sentinel-2, were analyzed in samples taken across the entirety of the growing seasons in three irrigated and rainfed sites located in eastern Nebraska between 2001 and 2005. Crop phenology was a factor strongly influencing the reflectance of both maize and soybean. Substantial hysteresis of the reflectance vs. canopy Chl relationship existed between the vegetative and reproductive stages. The effect of the hysteresis on vegetation indices (VI), applied for canopy Chl estimation, depended on the bands used and their formulation. The hysteresis greatly affected the accuracy of canopy Chl estimation by widely-used VIs with near infrared (NIR) and red reflectance (e.g., normalized difference vegetation index (NDVI), enhanced vegetation index (EVI) and simple ratio (SR)). VIs that use red edge and NIR bands (e.g., red edge chlorophyll index (CIred edge), red edge NDVI and the MERIS terrestrial chlorophyll index (MTCI)) were minimally affected by crop phenology (i.e., they exhibited little hysteresis) and were able to accurately estimate canopy Chl in two crops without algorithm re-parameterization and, thus, were found to be the best candidates for generic algorithms to estimate crop Chl using the surface reflectance products of MSI Sentinel-2

Evaluation of developmental phenotypes produced by morpholino antisense targeting of a sea urchin Runx gene

BACKGROUND: Runx transcription factors are important regulators of metazoan development. The sea urchin Runx gene SpRunt was previously identified as a trans-activator of the CyIIIa actin gene, a differentiation marker of larval aboral ectoderm. Here we extend the functional analysis of SpRunt, using morpholino antisense oligonucleotides (morpholinos) to interfere with SpRunt expression in the embryo. RESULTS: The developmental effects of four different SpRunt-specific morpholinos were evaluated. The two morpholinos most effective at knocking down SpRunt produce an identical mitotic catastrophe phenotype at late cleavage stage that is an artifact of coincidental mis-targeting to histone mRNA, providing a cautionary example of the insufficiency of two different morpholinos as a control for specificity. The other two morpholinos produce gastrula stage proliferation and differentiation defects that are rescued by exogenous SpRunt mRNA. The expression of 22 genes involved in cell proliferation and differentiation was analyzed in the latter embryos by quantitative polymerase chain reaction. Knockdown of SpRunt was found to perturb the expression of differentiation markers in all of the major tissue territories as well as the expression of cell cycle control genes, including cyclin B and cyclin D. CONCLUSIONS: SpRunt is essential for embryonic development, and is required globally to coordinate cell proliferation and differentiation

Runx-dependent expression of PKC is critical for cell survival in the sea urchin embryo

BACKGROUND: Runx transcription factors play critical roles in the developmental control of cell fate and contribute variously as oncoproteins and tumor suppressors to leukemia and other cancers. To discover fundamental Runx functions in the cell biology of animal development, we have employed morpholino antisense-mediated knockdown of the sea urchin Runx protein SpRunt-1. Previously we showed that embryos depleted of SpRunt-1 arrest development at early gastrula stage and underexpress the conventional protein kinase C SpPKC1. RESULTS: We report here that SpRunt-1 deficiency leads to ectopic cell proliferation and extensive apoptosis. Suppression of the apoptosis by pharmacological inhibition of caspase-3 prevents the ectopic proliferation and rescues gastrulation, indicating that many of the overt defects obtained by knockdown of SpRunt-1 are secondary to the apoptosis. Inhibition or knockdown of SpPKC1 also causes apoptosis, while cell survival is rescued in SpRunt-1 morphant embryos coinjected with SpPKC1 mRNA, suggesting that the apoptosis associated with SpRunt-1 deficiency is caused by the deficit in SpPKC1 expression. Chromatin immunoprecipitation indicates that SpRunt-1 interacts physically with SpPKC1 in vivo, and cis-regulatory analysis shows that this interaction activates SpPKC1 transcription. CONCLUSIONS: Our results show that Runx-dependent activation of SpPKC1 is essential for maintaining protein kinase C activity at levels conducive to cell survival during embryogenesis

Toward Generic Models for Green LAI Estimation in Maize and Soybean: Satellite Observations

Informative spectral bands for green leaf area index (LAI) estimation in two crops were identified and generic models for soybean and maize were developed and validated using spectral data taken at close range. The objective of this paper was to test developed models using Aqua and Terra MODIS, Landsat TM and ETM+, ENVISAT MERIS surface reflectance products, and simulated data of the recently-launched Sentinel 2 MSI and Sentinel 3 OLCI. Special emphasis was placed on testing generic models which require no re-parameterization for these species. Four techniques were investigated: support vector machines (SVM), neural network (NN), multiple linear regression (MLR), and vegetation indices (VI). For each technique two types of models were tested based on (a) reflectance data, taken at close range and resampled to simulate spectral bands of satellite sensors; and (b) surface reflectance satellite products. Both types of models were validated using MODIS, TM/ETM+, and MERIS data. MERIS was used as a prototype of OLCI Sentinel-3 data which allowed for assessment of the anticipated accuracy of OLCI. All models tested provided a robust and consistent selection of spectral bands related to green LAI in crops representing a wide range of biochemical and structural traits. The MERIS observations had the lowest errors (around 11%) compared to the remaining satellites with observational data. Sentinel 2 MSI and OLCI Sentinel 3 estimates, based on simulated data, had errors below 8%. However the accuracy of these models with actual MSI and OLCI surface reflectance products remains to be determined