A screen for round egg mutants in Drosophila identifies tricornered, furry, and misshapen as regulators of egg chamber elongation.

The elongation of tissues and organs during embryonic development results from the coordinate polarization of cell behaviors with respect to the elongation axis. Within the Drosophila melanogaster ovary, initially spherical egg chambers lengthen dramatically as they develop to create the elliptical shape of the mature egg. This morphogenesis depends on an unusual form of planar polarity within the egg chamber's outer epithelial cell layer known as the follicle cells. Disruption of follicle cell planar polarity leads to the production of round rather than elongated eggs; however, the molecular mechanisms that control this tissue organization are poorly understood. Starting from a broadly based forward genetic screen, we have isolated 12 new round egg complementation groups, and have identified four of the mutated genes. In mapping the largest complementation group to the fat2 locus, we unexpectedly discovered a high incidence of cryptic fat2 mutations in the backgrounds of publicly available stocks. Three other complementation groups correspond to the genes encoding the cytoplasmic signaling proteins Tricornered (Trc), Furry (Fry), and Misshapen (Msn). Trc and Fry are known members of an NDR kinase signaling pathway, and as a Ste20-like kinase, Msn may function upstream of Trc. We show that all three proteins are required for follicle cell planar polarity at early stages of egg chamber elongation and that Trc shows a planar polarized distribution at the basal follicle cell surface. These results indicate that this new mutant collection is likely to provide novel insight into the molecular mechanisms controlling follicle cell planar polarity and egg chamber elongation

Polarity determination in breast tissue: desmosomal adhesion, myoepithelial cells, and laminin 1

In all epithelial organs, apicobasal polarity determines functional integrity and contributes to the maintenance of tissue and organ specificity. In the breast, the functional unit is a polar double-layered tube consisting of luminal epithelial cells surrounded by myoepithelial cells and a basement membrane. It is far from clear how this double-layered structure is established and how polarity is maintained. Two recent papers have shed some light onto this intriguing problem in mammary gland biology. The results point to desmosomes and laminin 1 as having crucial roles. However, some questions remain

Dynein Regulates Epithelial Polarity and the Apical Localization of stardust A mRNA

Intense investigation has identified an elaborate protein network controlling epithelial polarity. Although precise subcellular targeting of apical and basolateral determinants is required for epithelial architecture, little is known about how the individual determinant proteins become localized within the cell. Through a genetic screen for epithelial defects in the Drosophila follicle cells, we have found that the cytoplasmic Dynein motor is an essential regulator of apico–basal polarity. Our data suggest that Dynein acts through the cytoplasmic scaffolding protein Stardust (Sdt) to localize the transmembrane protein Crumbs, in part through the apical targeting of specific sdt mRNA isoforms. We have mapped the sdt mRNA localization signal to an alternatively spliced coding exon. Intriguingly, the presence or absence of this exon corresponds to a developmental switch in sdt mRNA localization in which apical transcripts are only found during early stages of epithelial development, while unlocalized transcripts predominate in mature epithelia. This work represents the first demonstration that Dynein is required for epithelial polarity and suggests that mRNA localization may have a functional role in the regulation of apico–basal organization. Moreover, we introduce a unique mechanism in which alternative splicing of a coding exon is used to control mRNA localization during development

Domains controlling cell polarity and proliferation in the Drosophila tumor suppressor Scribble

Cell polarity and cell proliferation can be coupled in animal tissues, but how they are coupled is not understood. In Drosophila imaginal discs, loss of the neoplastic tumor suppressor gene scribble (scrib), which encodes a multidomain scaffolding protein, disrupts epithelial organization and also causes unchecked proliferation. Using an allelic series of mutations along with rescuing transgenes, we have identified domain requirements for polarity, proliferation control, and other Scrib functions. The leucine-rich repeats (LRR) tether Scrib to the plasma membrane, are both necessary and sufficient to organize a polarized epithelial monolayer, and are required for all proliferation control. The PDZ domains, which recruit the LRR to the junctional complex, are dispensable for overall epithelial organization. PDZ domain absence leads to mild polarity defects accompanied by moderate overproliferation, but the PDZ domains alone are insufficient to provide any Scrib function in mutant discs. We suggest a model in which Scrib, via the activity of the LRR, governs proliferation primarily by regulating apicobasal polarity

Endosomal entry regulates Notch receptor activation in Drosophila melanogaster

Signaling through the transmembrane receptor Notch is widely used throughout animal development and is a major regulator of cell proliferation and differentiation. During canonical Notch signaling, internalization and recycling of Notch ligands controls signaling activity, but the involvement of endocytosis in activation of Notch itself is not well understood. To address this question, we systematically assessed Notch localization, processing, and signaling in a comprehensive set of Drosophila melanogaster mutants that block access of cargo to different endocytic compartments. We find that γ-secretase cleavage and signaling of endogenous Notch is reduced in mutants that impair entry into the early endosome but is enhanced in mutants that increase endosomal retention. In mutants that block endosomal entry, we also uncover an alternative, low-efficiency Notch trafficking route that can contribute to signaling. Our data show that endosomal access of the Notch receptor is critical to achieve physiological levels of signaling and further suggest that altered residence in distinct endocytic compartments could underlie pathologies involving aberrant Notch pathway activation

Book Reviews and Libel Proceedings

The American journal of International Law has been informed of the initiation in France of penal proceedings against the editor in chief of the European journal of International Law (EJIL), by virtue of a complaint filed by an author of a book reviewed on a Web site affiliated with the Ejll.1 We share the concerns of other professional societies regarding the potential of such litigation for chilling academic discourse. 2 We also take this opportunity to explain the practice of the AJIL concerning communications from authors who object to book reviews published in our pages, and to state our position on the important questions of academic freedom involved

The Functions of Auxilin and Rab11 in Drosophila Suggest That the Fundamental Role of Ligand Endocytosis in Notch Signaling Cells Is Not Recycling

Notch signaling requires ligand internalization by the signal sending cells. Two endocytic proteins, epsin and auxilin, are essential for ligand internalization and signaling. Epsin promotes clathrin-coated vesicle formation, and auxilin uncoats clathrin from newly internalized vesicles. Two hypotheses have been advanced to explain the requirement for ligand endocytosis. One idea is that after ligand/receptor binding, ligand endocytosis leads to receptor activation by pulling on the receptor, which either exposes a cleavage site on the extracellular domain, or dissociates two receptor subunits. Alternatively, ligand internalization prior to receptor binding, followed by trafficking through an endosomal pathway and recycling to the plasma membrane may enable ligand activation. Activation could mean ligand modification or ligand transcytosis to a membrane environment conducive to signaling. A key piece of evidence supporting the recycling model is the requirement in signaling cells for Rab11, which encodes a GTPase critical for endosomal recycling. Here, we use Drosophila Rab11 and auxilin mutants to test the ligand recycling hypothesis. First, we find that Rab11 is dispensable for several Notch signaling events in the eye disc. Second, we find that Drosophila female germline cells, the one cell type known to signal without clathrin, also do not require auxilin to signal. Third, we find that much of the requirement for auxilin in Notch signaling was bypassed by overexpression of both clathrin heavy chain and epsin. Thus, the main role of auxilin in Notch signaling is not to produce uncoated ligand-containing vesicles, but to maintain the pool of free clathrin. Taken together, these results argue strongly that at least in some cell types, the primary function of Notch ligand endocytosis is not for ligand recycling