Mouse mutagenesis: From gene to phenotype and back again

AbstractScreens for genetic mutations have been instrumental in identifying genes needed to execute particular biological processes. They have also helped to resolve the function of individual genes. Now the notion of large-scale mutagenesis screens in mouse, an experimental model for humans, is becoming a reality

A review of interactions between cetaceans and fisheries in the Azores

Author Posting. © John Wiley & Sons, 2010. This is the author's version of the work. It is posted here by permission of John Wiley & Sons for personal use, not for redistribution. The definitive version was published in Aquatic Conservation: Marine and Freshwater Ecosystems 21 (2011): 17-27, doi:10.1002/aqc.11581. Interactions between cetaceans and fishing activity in the Archipelago of the Azores were examined using information contained in grey literature and previously unpublished data collected by observer programmes and research projects from 1998 to 2006. Together with a brief description of the economics, gear, fishing effort, and past and ongoing monitoring projects, levels of cetacean bycatch and interference were reported for each major fishery. 2. Cetaceans were present in 7% (n=973) and interfered in 3% (n=452) of the fishing events monitored by observers aboard tuna-fishing vessels. Interference resulted in a significantly higher proportion of events with zero catches but it was also associated with higher tuna catches. 3. There was a decreasing trend in the proportion of tuna-fishing events with cetacean presence or interference throughout this study, as well as a reduction in the estimates of dolphins captured annually by the whole fleet. 4. Observers reported cetacean depredation in 16% of the sets for demersal species and in 2% of the sets for swordfish. Cetacean presence and depredation were associated with higher overall catches and higher catches per unit effort in demersal fisheries. Bottlenose dolphins (Tursiops truncatus) were responsible for most depredation events in demersal fisheries, whereas in the swordfish fishery, depredation was associated with the presence of killer whales (Orcinus orca). There were no reports of cetacean bycatch in these fisheries. There were also no reports of cetaceans interacting in the experimental deep-sea fisheries that were examined. 5. Available data suggests that levels of interaction between cetaceans and Azorean fisheries are generally low and that the economic impact of cetacean interference is probably small. However, for several traditional fisheries there are no accurate data to determine levels of cetacean interaction. We recommend that existing observer programmes be expanded to increase observer coverage of the demersal and swordfish fisheries and allow monitoring of other existing and emerging fisheries.M.A.S. was supported by an FCT postdoctoral (SFRH/BPD/29841/2006) grant, and R.P. was supported by an FCT doctoral grant (SFRH/BD/32520/2006)

Renal agenesis in mice homozygous for a gene trap mutation in the gene encoding heparan sulfate 2-sulfotransferase

Heparan sulfate proteoglycans have been implicated in the presentation of a number of secreted signaling molecules to their signal-transducing receptors. We have characterized a gene trap mutation in the gene encoding a heparan sulfate biosynthetic enzyme, heparan sulfate 2-sulfotransferase (HS2ST). Transgenic mice were generated from embryonic stem cells harboring this insertion. lacZ reporter gene activity in heterozygous embryos demonstrates that the gene is expressed differentially during embryogenesis, presumably directing dynamic changes in heparan sulfate structure. Moreover, mice homozygous for the Hs2st gene trap allele die in the neonatal period, exhibiting bilateral renal agenesis and defects of the eye and the skeleton. Analysis of kidney development in Hs2st mutants reveals that the gene is not required for two early events—ureteric bud outgrowth from the Wolffian duct and initial induction of Pax-2 expression in the metanephric mesenchyme. It is required, however, for mesenchymal condensation around the ureteric bud and initiation of branching morphogenesis. Because 2-O-sulfation has been shown to influence the functional interactions of ligands with heparan sulfate in vitro, we discuss the possibility that the Hs2st mutant phenotype is a consequence of compromised interactions between growth factors and their signal-transducing receptors. These data provide the first genetic evidence that the regulated synthesis of differentially glycosylated proteoglycans can affect morphogenesis during vertebrate development

Growth and Early Postimplantation Defects in Mice Deficient for the Bromodomain-Containing Protein Brd4†

In a gene trap screen we recovered a mouse mutant line in which an insertion generated a null allele of the Brd4 gene. Brd4 belongs to the Fsh/Brd family, a group of structurally related proteins characterized by the association of two bromodomains and one extraterminal domain. Members of this family include Brd2/Ring3/Fsrg1 in mammals, fs(1)h in Drosophila, and Bdf1 in Saccharomyces cerevisiae. Brd4 heterozygotes display pre- and postnatal growth defects associated with a reduced proliferation rate. These mice also exhibit a variety of anatomical abnormalities: head malformations, absence of subcutaneous fat, cataracts, and abnormal liver cells. In primary cell cultures, heterozygous cells also display reduced proliferation rates and moderate sensitivity to methyl methanesulfonate. Embryos nullizygous for Brd4 die shortly after implantation and are compromised in their ability to maintain an inner cell mass in vitro, suggesting a role in fundamental cellular processes. Finally, sequence comparisons suggest that Brd4 is likely to correspond to the Brd-like element of the mediator of transcriptional regulation isolated by Y. W. Jiang, P. Veschambre, H. Erdjument-Bromage, P. Tempst, J. W. Conaway, R. C. Conaway, and R. D. Kornberg (Proc. Natl. Acad. Sci. USA 95:8538-8543, 1998) and the Brd4 mutant phenotype is discussed in light of this result. Together, our results provide the first genetic evidence for an in vivo role in mammals for a member of the Fsh/Brd family

Targeted deletion of the novel cytoplasmic dynein mD2LIC disrupts the embryonic organiser, formation of the body axes and specification of ventral cell fates

Dyneins have been implicated in left-right axis determination during embryonic development and in a variety of human genetic syndromes. In this paper, we study the recently discovered mouse dynein 2 light intermediate chain (mD2LIC), which is believed to be involved in retrograde intraflagella transport and which, like left-right dynein, is expressed in the node of the mouse embryo. Cells of the ventral node of mouse embryos lacking mD2LIC have an altered morphology and lack monocilia, and expression of Foxa2 and Shh in this structure is reduced or completely absent. At later stages, consistent with the absence of nodal cilia, mD2LIC is required for the establishment of the left-right axis and for normal expression of Nodal, and the ventral neural tube fails to express Shh, Foxa2 and Ebaf. mD2LIC also functions indirectly in the survival of anterior definitive endoderm and in the maintenance of the anterior neural ridge, probably through maintenance of Foxa2 /Hnf3β expression. Together, our results indicate that mD2LIC is required to maintain or establish ventral cell fates and for correct signalling by the organiser and midline, and they identify the first embryonic function of a vertebrate cytoplasmic dynein

Targeted deletion of the novel cytoplasmic dynein mD2LIC disrupts the embryonic organiser, formation of the body axes and specification of ventral cell fates

Dyneins have been implicated in left-right axis determination during embryonic development and in a variety of human genetic syndromes. In this paper, we study the recently discovered mouse dynein 2 light intermediate chain (mD2LIC), which is believed to be involved in retrograde intraflagella transport and which, like left-right dynein, is expressed in the node of the mouse embryo. Cells of the ventral node of mouse embryos lacking mD2LIC have an altered morphology and lack monocilia, and expression of Foxa2 and Shh in this structure is reduced or completely absent. At later stages, consistent with the absence of nodal cilia, mD2LIC is required for the establishment of the left-right axis and for normal expression of Nodal, and the ventral neural tube fails to express Shh, Foxa2 and Ebaf. mD2LIC also functions indirectly in the survival of anterior definitive endoderm and in the maintenance of the anterior neural ridge, probably through maintenance of Foxa2 /Hnf3β expression. Together, our results indicate that mD2LIC is required to maintain or establish ventral cell fates and for correct signalling by the organiser and midline, and they identify the first embryonic function of a vertebrate cytoplasmic dynein