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J Castelli-Gair

Publications and source records attributed to J Castelli-Gair.

8 recordsLinked to original sources

Study of the posterior spiracles of Drosophila as a model to understand the genetic and cellular mechanisms controlling morphogenesis.

We have studied the posterior spiracles of Drosophila as a model to link patterning genes and morphogenesis. A genetic cascade of transcription factors downstream of the Hox gene Abdominal-B subdivides the primordia of the posterior spiracles into two cell populations that develop using two different morphogenetic mechanisms. The inner cells that give rise to the spiracular chamber invaginate by elongating into "bottle-shaped" cells. The surrounding cells give rise to a protruding stigmatophore by changing their relative positions in a process similar to convergent extension. The genetic cascades regulating spiracular chamber, stigmatophore, and trachea morphogenesis are different but coordinated to form a functional tracheal system. In the posterior spiracle, this coordination involves the control of the initiation of cell invagination that starts in the cells closer to the trachea primordium and spreads posteriorly. As a result, the opening of the tracheal system shifts back from the spiracular branch of the trachea into the posterior spiracle cells. We analyze the contribution of the ems gene to this coordination. In ems mutants, invagination of the spiracle cells adjacent to the trachea does not occur, but more posterior cells of the spiracle invaginate normally. This results in a spiracle without a lumen and with the tracheal opening located outside it.

Animals↗

The lines gene of Drosophila is required for specific functions of the Abdominal-B HOX protein.

The Hox genes encode homeobox transcription factors that control the formation of segment specific structures in the anterior-posterior axis. HOX proteins regulate the transcription of downstream targets acting both as repressors and as activators. Due to the similarity of their homeoboxes it is likely that much of the specificity of HOX proteins is determined by interaction with transcriptional cofactors, but few HOX cofactor proteins have yet been described. Here I present genetic evidence showing that lines, a segment polarity gene of Drosophila, is required for the function of the Abdominal-B protein. In lines mutant embryos Abdominal-B protein expression is normal but incapable of promoting its normal functions: formation of the posterior spiracles and specification of an eighth abdominal denticle belt. These defects arise because in lines mutant embryos the Abdominal-B protein cannot activate its direct target empty spiracles or other downstream genes while it can function as a repressor of Ultrabithorax and abdominal-A. The lines gene seems to be required exclusively for Abdominal-B but not for the function of other Hox genes.

Abdomen↗

Implications of the spatial and temporal regulation of Hox genes on development and evolution.

Nearly 20 years have passed since Ed Lewis revealed the importance of Hox genes in the specification of different segments in the anterior-posterior axis of the fly. Pioneering studies by several authors, among others Garcia-Bellido and his student Ginés Morata, helped to elaborate a theory of segmental specification that was strengthened with the arrival of molecular techniques to the field of Developmental Biology. The conservation of Hox genes in metazoans at the level of sequence, function and complex organization has resulted in the export of this Drosophila theory as a paradigm to interpret the development of axial specification in organisms less amenable to experimental study. There are two main ways to interpret how Hox genes work in Drosophila. One considering Hox genes as "segment identity" factors giving global properties to the segments in which they are active. Another considering Hox genes as encoding spatially restricted transcription factors required for a number decisions taken at the cellular level. Here I use published and unpublished experimental data to illustrate that early activation of the Hox genes does not establish a gene code that leads to "segment identity". I will stress the point that Hox expression patterns develop with the embryo, that there are many genes involved in this modulation, and that the changing pattern of expression is important to achieve the final shape of the animal. I will show that, by interpreting Hox gene function in this way, some apparently paradoxical results in the Hox field can be answered. Finally, I discuss the implications of dynamic Hox gene expression on the evolution of segment morphology.

Animals↗

How the Hox gene Ultrabithorax specifies two different segments: the significance of spatial and temporal regulation within metameres.

In Drosophila, the Hox gene Ultrabithorax (Ubx) specifies the development of two different metameres--parasegment 5, which is entirely thoracic, and parasegment 6, which includes most of the first abdominal segment. Here we investigate how a single Hox gene can specify two such different morphologies. We show that, in the early embryo, cells respond similarly to UBX protein in both parasegments. The differences between parasegments 5 and 6 can be explained by the different spatial and temporal pattern of UBX protein expression in these two metameres. We find no evidence for multiple threshold responses to different levels of UBX protein. We examine in particular the role of Ubx in limb development. We show that UBX protein will repress limb primordia before 7 hours, when Ubx is expressed in the abdomen, but not later, when UBX is first expressed in the T3 limb primordium. The regulation of one downstream target of UBX, the Distalless gene, provides a model for this transition at the molecular level.

Animals↗

Dissecting the temporal requirements for homeotic gene function.

Homeotic genes confer identity to the different segments of Drosophila. These genes are expressed in many cell types over long periods of time. To determine when the homeotic genes are required for specific developmental events we have expressed the Ultrabithorax, abdominal-A and Abdominal-Bm proteins at different times during development using the GAL4 targeting technique. We find that early transient homeotic gene expression has no lasting effects on the differentiation of the larval epidermis, but it switches the fate of other cell types irreversibly (e.g. the spiracle primordia). We describe one cell type in the peripheral nervous system that makes sequential, independent responses to homeotic gene expression. We also provide evidence that supports the hypothesis of in vivo competition between the bithorax complex proteins for the regulation of their down-stream targets.

Animals↗

The evolving role of Hox genes in arthropods.

Comparisons between Hox genes in different arthropods suggest that the diversity of Antennapedia-class homeotic genes present in modern insects had already arisen before the divergence of insects and crustaceans, probably during the Cambrian. Hox gene duplications are therefore unlikely to have occurred concomitantly with trunk segment diversification in the lineage leading to insects. Available data suggest that domains of homeotic gene expression are also generally conserved among insects, but changes in Hox gene regulation may have played a significant role in segment diversification. Differences that have been documented alter specific aspects of Hox gene regulation within segments and correlate with alterations in segment morphology rather than overt homeotic transformations. The Drosophila Hox cluster contains several homeobox genes that are not homeotic genes--bicoid, fushi-tarazu and zen. the role of these genes during early development has been studied in some detail. It appears to be without parallel among the vertebrate Hox genes. No well conserved homologues of these genes have been found in other taxa, suggesting that they are evolving faster than the homeotic genes. Relatively divergent Antp-class genes isolated from other insects are probably homologues of fushi-tarazu, but these are almost unrecognisable outside of their homeodomains, and have accumulated approximately 10 times as many changes in their homeodomains as have homeotic genes in the same comparisons. They show conserved patterns of expression in the nervous system, but not during early development.

Animals↗

Function of an Ultrabithorax minigene in imaginal cells.

An Ultrabithorax (Ubx) minigene constructed from three key Ubx control regions is capable of supporting development of Ubx null mutants throughout larval life and beyond to pharate flies, thereby rescuing the larval lethality due to the homeotic mutation. The cuticle of these flies shows that the minigene provides at least partial Ubx function in each of the four compartments whose morphogenetic pathways are determined by Ubx. We analyse beta-galactosidase patterns in imaginal discs conferred by each individual Ubx control region. From the comparison of these patterns with Ubx expression in Cbx mutants, we infer that long-range repressor elements in the chromosomal Ubx gene play an important role in the generation of Ubx expression patterns in imaginal discs. Expression and function of our Ubx minigenes indicate that Ubx control regions are capable of functioning properly out of context and detached from their normal chromosomal location within the homeotic gene complex.

Animals↗