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Multicolor fluorescent in situ mRNA hybridization (FISH) on whole mounts and sections
No full textIn situ hybridization involves the hybridization of an antisense RNA probe to an mRNA transcript and it is a powerful method for the characterization of gene expression in tissues, organs, or whole organisms. Performed as a whole mount (WISH), it allows the detection of mRNA transcripts in three dimensions, while combined with sectioning, either before or after hybridization, it provides gene expression information with cellular resolution. FISH relies on the fluorescence detection of probes and is the method of choice for the simultaneous detection of transcripts with similar or overlapping expression patterns, as each can be clearly distinguished by the selection of fluorophore. Here, we describe a protocol for performing multicolor FISH in Xenopus embryos in whole mounts and sections that can be further combined with antibody staining
Characterization of the aglomerular kidney in the bay pipefish syngnathus leptorhynchus
No full textBibliography: p. 94-101Some pages are in colour
Coral spawning synchronicity: the role of ciradian genes in circalunar rhythms
No full textBibliography: p. 198-242Many pages are in colour
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Role of p53 family members in the development of Xenopus laevis
No full texttextp53 family members play an essential role in tumor suppression and
development. In this report, the developing Xenopus embryos have been used to
analyze their role in development by introducing wild-type or dominant negative
proteins into early embryos and studying the phenotype with a variety of
techniques. Mutant p53 was functionally mapped in order to dissect the possible
mechanism underlying its tumorigenic property, which, surprisingly, is
independent of the oligomerization domain and does not target its family
members. The function of Xenopus p63 was characterized by overexpression of
the wild-type protein. Even though it lacks a typical transcriptional activation
domain, Xp63 possesses some p53-like activities in controlling the cell cycle and
apoptosis. The major sites of Xp63 expression are the epidermis and neural crest,
suggesting that it might play a role in the formation of these structures. This is
supported by the observation that neural crest formation is affected by
overexpression of dominant-negative Xp63. These results suggest that, while
sharing many similarities, the p53 family members have distinct functions during
early development. Together, these findings have offered a starting point for
future study of functional conservation and divergence of these molecules as to
their role in cancer and development and have established the Xenopus embryo as
an efficacious system for such studies.Biological Sciences, School o
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Characterization and evolution of peridinin-chlorophyll a binding protein gene families in symbiotic dinoflagellates
Get PDFThis dissertation consists of three integrated chapters. Chapter
One presents the first genomic characterization of the PCP gene family from a
symbiotic dinoflagellate, Symbiodinium sp. from Hippopus hippopus
(Symbiodinium 203). Symbiodinium 203 has long PCP genes organized like those
of Amphidinium carterae and Lingulodinium polyedra, but with a putative
promoter that is different from L. polyedra. There are at least 14 distinct coding
regions out of 36 ± 12 PCP genes in this family. Diversity of Symbiodinium 203's
PCP gene family appears to be consequence of low levels of concerted evolution
and acts as a primary source of variability in PCP isoforms. Predicted amino acid
substitutions in Symbiodinium 203's PCP apoproteins result in shifts of isoelectric
points, and protein modeling suggests that polymorphic sites may influence light harvesting of holoproteins. In Chapter Two, the first PCP coding sequences from
S. pilosum, Symbiodinium sp. from Dichocoenia stokesii, S. pulchrorum and S.
kawagutii were presented. Diverse PCP gene families occur in all major clades of
Symbiodinium and in both size classes of the gene. As with Symbiodinium 203 in
Chapter One, these PCP gene families do not appear to have been homogenized
through mechanisms leading to concerted evolution. The predicted PCP
apoproteins from S. pilosum and S. kawagutii have calculated isoelectric focusing
points that generally match values previously measured for these species, which
supports the hypothesis that genetic polymorphism is the primary source
generating differences in PCP isoforms. Protein modeling produced a putative
tertiary structure for S. pilosum apoproteins and was used to identify polymorphic
sites in S. pilosum and S. kawagutii PCPs that could affect spectral tuning of
peridinins. And Chapter Three contains the first phylogenetic analyses of the
evolution of dinoflagellate PCP gene families. The objective of this section is to
estimate the selective pressure at the codon level within PCP genes. PCP
polymorphism is ancient, however, the polymorphism is not maintained by
positive selection. Codon sites within PCP genes are evolving under purifying
selection and are subjected to net reduced levels of concerted evolution. Isoform
diversity is probably selected for within a functional range.Biological Sciences, School o
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Temperature Modulation of Biological Clocks in a Reef-Building Coral
No full textReef-building coral of the Great Barrier Reef demonstrate remarkably synchronized and precise timing of reproduction. They achieve this precision through sophisticated biological clock systems entrained by their local environment. This thesis explores how temperature and lunar light modulates the biological clocks of Acropora millepora by measuring transcriptomic changes with RNA-sequencing. Coral colonies were collected and maintained under artificial lunar light at the Heron Island Research Station under cool and warm temperature treatments corresponding to winter and summer averages at this locale. Individuals were sampled over a 24-hour period during a full lunar month to capture long-term daily profiles of gene expression. It was found that numerous biological clock genes were impacted by temperature and lunar light. Furthermore, temperature and lunar phase altered the expression of interesting hormonal pathways involved in reproductive behaviours. These data will help elucidate the mechanisms underlying the precise timing of reproduction in reef-building coral and the effect that different temperatures have on this process
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