Identification of the Molecular Basis of Morphological Variation in Avian Beaks

Vertebrates, particularly birds, show extremely variable species-specific morphology in craniofacial structures. Cranial neural crest cells give rise to all the cartilage and bone of the face, and transplantation experiments have shown that these cells contain species-specific patterning information. First, I employed custom cross-species microarrays to analyze the spectrum of developmental signaling pathway and transcription factor gene expression changes in neural crest cells of the developing beaks for the chicken, duck, and quail, both before and after morphological variation is evident. I found that neural crest cells have established a species-specific gene expression profile that predates morphological variation. In addition to expression changes in the Bmp and Calmodulin pathways, previously associated with morphological variation in Darwin\u27s finches, I observed dramatic changes in a number of Wnt signaling components in the broad-billed duck. Second, given that deletion of the microRNA processing gene DICER in neural crest cells results in loss of nearly all facial structures, I utilized high-throughput sequencing to describe the microRNAs that are expressed and/or differentially expressed among the same neural crest samples used for the microarray analysis. In remarkable contrast to relatively unchanged pattern of transcription factor gene expression, microRNA expression is highly dynamic during stages when avians acquire species-specific morphology. The microRNA expression profiles also suggest that the transition from multipotent, proliferative neural crest cells into cells differentiating to form the tissues of the face may be delayed in the duck relative to the chicken and quail. This prolonged period of proliferation in duck neural crest may contribute to the increase in beak size and width of the adult duck bill versus the chicken beak. Finally, I illustrate examples of how these genomic data sets can initiate new avenues of investigation and testable hypotheses. I found that the Wnt pathway acts upstream of the Bmp pathway and induces regional changes in growth of the developing beak. I also correlate changes in expression of miR-222 in the frontonasal prominence with alterations in protein: but not mRNA) levels of one of its target genes, the cell cycle regulator p27(KIP1). I then identified seven mature microRNAs that appear to be specific to the avian lineage. Using PCR, I confirmed that two of these, miR-2954 and miR-2954*, are conserved across the avian lineage, from ratites to songbirds

Growth of Sr1-xCaxRuO3 thin films by metalorganic aerosol deposition

We report the growth of thin films of Sr1-xCaxRuO3 on SrTiO3 and MgO substrates by metalorganic aerosol deposition. The structure and microstructure is characterized by X-ray diffraction and room-temperature scanning tunnelling microscopy (STM), respectively. STM indicates in-plane epitaxy and a small surface roughness for films on SrTiO3. The high-quality of the films is supported by large residual resistivity ratios up to 29.Comment: 4 Pages, 2 Figures, submitted to Proceedings of ICM 2009 (Karlsruhe

Constraint and diversification of developmental trajectories in cichlid facial morphologies

Background A major goal of evolutionary biology is to understand the origins of phenotypic diversity. Changes in development, for instance heterochrony, can be a potent source of phenotypic variation. On the other hand, development can also constrain the spectrum of phenotypes that can be produced. In order to understand these dual roles of development in evolution, we examined the developmental trajectory of a trait central to the extensive adaptive radiation of East African cichlid fishes: craniofacial adaptations that allow optimal exploitation of ecological niches. Specifically, we use geometric morphometric analysis to compare morphological ontogenies among six species of Lake Malawi cichlids (n \u3e 500 individuals) that span a major ecomorphological axis. We further evaluate how modulation of Wnt signaling impacts the long-term developmental trajectory of facial development. Results We find that, despite drastic differences in adult craniofacial morphologies, there are general similarities in the path of craniofacial ontogeny among species, suggesting that natural selection is working within a conserved developmental program. However, we also detect species-specific differences in the timing, direction, and/or duration of particular developmental trajectories, including evidence of heterochrony. Previous work in cichlids and other systems suggests that species-specific differences in adult morphology are due to changes in molecular signaling pathways that regulate early craniofacial development. In support of this, we demonstrate that modulation of Wnt signaling at early stages can shift a developmental trajectory into morphospace normally occupied by another species. However, without sustained modulation, craniofacial shape can recover by juvenile stages. This underscores the idea that craniofacial development is robust and that adult head shapes are the product of many molecular changes acting over extended periods of development. Conclusions Our results are consistent with the hypothesis that development acts to both constrain and promote morphological diversity. They also illustrate the modular nature of the craniofacial skeleton and hence the ability of selection to act upon distinct anatomical features in an independent manner. We propose that trophic diversity among cichlids has been achieved via shifts in both specific (e.g., stage-specific changes in gene expression) and global (e.g., heterochrony) ontogenetic processes acting within a conserved developmental program

Genetic analyses in Lake Malawi cichlids identify new roles for Fgf signaling in scale shape variation

Elasmoid scales are the most common epithelial appendage among vertebrates, however an understanding of the genetic mechanisms that underlie variation in scale shape is lacking. Using an F2 mapping cross between morphologically distinct cichlid species, we identified \u3e40 QTL for scale shape at different body positions. We show that while certain regions of the genome regulate variation in multiple scales, most are specific to scales at distinct positions. This suggests a degree of regional modularity in scale development. We also identified a single QTL for variation in scale shape disparity across the body. Finally, we screened a QTL hotspot for candidate loci, and identified the Fgf receptor fgfr1b as a prime target. Quantitative rtPCR and small molecule manipulation support a role for Fgf signaling in shaping cichlid scales. While Fgfs have previously been implicated in scale loss, these data reveal new roles for the pathway in scale shape variation

The Cyprinodon variegatus genome reveals gene expression changes underlying differences in skull morphology among closely related species

Genes in durophage intersection set at 15 dpf. This is a comma separated table of the genes in the 15 dpf durophage intersection set. Given are edgeR results for each pairwise comparison. Columns indicating whether a gene is included in the intersection set at a threshold of 1.5 or 2 fold are provided. (CSV 13 kb

An RNA interference-based screen of transcription factor genes identifies pathways necessary for sensory regeneration in the avian inner ear

Sensory hair cells of the inner ear are the mechano-electric transducers of sound and head motion. In mammals, damage to sensory hair cells leads to hearing or balance deficits. Non-mammalian vertebrates such as birds can regenerate hair cells after injury. In a previous study, we characterized transcription factor gene expression during chicken hair cell regeneration. In those studies, a laser micro-beam or ototoxic antibiotics were used to damage the sensory epithelia (SE). The current study focused on 27 genes that were up-regulated in regenerating SE compared to untreated SE in the previous study. Those genes were knocked down by siRNA, to determine their requirement for supporting cell proliferation and to measure resulting changes in the larger network of gene expression. We identified 11 genes necessary for proliferation and also identified novel interactive relationships between many of them. Defined components of the WNT, PAX and AP1 pathways were shown to be required for supporting cell proliferation. These pathways intersect on WNT4, which is also necessary for proliferation. Among the required genes, the CCAAT enhancer binding protein, CEBPG, acts downstream of Jun Kinase and JUND in the AP1 pathway. The WNT co-receptor LRP5 acts downstream of CEBPG as does the transcription factor BTAF1. Both of these genes are also necessary for supporting cell proliferation. This is the first large scale screen of its type and suggests an important intersection between the AP1 pathway, the PAX pathway and WNT signaling in the regulation of supporting cell proliferation during inner ear hair cell regeneration

Thin films of fluorinated 3d-metal phthalocyanines as chemical sensors of ammonia: an optical spectroscopy study

A comparative study of the sensor response toward gaseous ammonia of hexadecafluorinated 3d-metal phthalocyanine (MPcF16, MCu(II), Co(II), Zn(II), Ni(II)) thin films was performed using complementary experimental (viz., surface plasmon resonance, SPR, and IR absorption spectroscopy) along with theoretical (density functional theory calculations, DFT) techniques. SPR measurements revealed changes of both thickness and optical parameters (refraction indices and extinction coefficients) of the MPcF16 films caused by adsorption of NH3. The MPcF16 species studied exhibited the following order of sensor response: ZnPcF16>CoPcF16≥CuPcF16>NiPcF16. A good correlation was found between the DFT calculated (B3LYP/6-311++G(2df,p)) binding energies, experimentally measured shift of the selected IR bands, and the optical sensor response. Apart from this, we performed a detailed assignment of all intense..