David Matthes

The semaphorin genes encode a family of transmembrane and secreted growth cone guidance molecules

In addition to its expression on subsets of axons, grasshopper Semaphorin I (Sema I, previously called Fasciclin [Fas] IV) is expressed on an epithelial stripe in the limb bud, where it functions in the guidance of two sensory growth cones as they abruptly turn upon encountering this sema I boundary. We report here on the cloning and characterization of two sema genes in Drosophila, one in human, and the identification of two related viral sequences, all of which encode proteins with conserved Semaphorin domains. Drosophila sema (D-Sema) I is a transmembrane protein, while D-Sema II and human Sema III are putative secreted proteins that are similar to the recently reported chick collapsin. D-Sema I and D-Sema II are expressed by subsets of neurons and muscles. Genetic analysis in Drosophila reveals that semall is an essential gene that is required for both proper adult behavior and survival.

Fasciclin IV: Sequence, expression, and function during growth cone guidance in the grasshopper embryo

Monoclonal antibody 6F8 was used to characterize and clone fasciclin IV, a new axonal glycoprotein in the grasshopper, and to study its function during growth cone guidance. Fasciclin IV is dynamically expressed on a subset of axon pathways in the developing CNS and on circumferential bands of epithelial cells in developing limb buds. One of these bands corresponds to the location where the growth cones of the Ti1 pioneer neurons make a characteristic turn while extending toward the CNS. Embryos cultured in the 6F8 antibody or Fab exhibit aberrant formation of this axon pathway. cDNA sequence analysis suggests that fasciclin IV has a signal sequence; long extracellular, transmembrane, and short cytoplasmic domains; and shows no homology with any protein in the available data bases. Thus, fasciclin IV appears to be a novel integral membrane protein that functions in growth cone guidance.

Semaphorin II can function as a selective inhibitor of specific synaptic arborizations

Previous studies showed that grasshopper semaphorin I, a transmembrane semaphorin, functions in vivo to steer a pair of growth cones, prevent defasciculation, and inhibit branching; and that chick collapsin, a secreted semaphorin, can function in vitro to cause growth cone collapse. Semaphorin II, a secreted semaphorin in Drosophila, is transiently expressed by a single large muscle during motoneuron outgrowth and synapse formation. To test the in vivo function of semaphorin II, we created transgenic Drosophila that generate ectopic semaphorin II expression by muscles that normally do not express it. The results show that semaphorin II can function in vivo as a selective target-derived signal that inhibits the formation of specific synaptic terminal arbors.

Semaphorin 4A is dynamically regulated during thymocyte development in mice

Semaphorins are important regulators of peripheral T and B-cell mediated immune responses in mice and humans. Modulatory roles of semaphorins in T cell development are also being characterized. We carefully analyzed the gene expression and protein levels of semaphorins 4A, 4D, and 7A at various developmental stages of T cell maturation in the thymus of C57BL/6 mice. Sema7a was expressed at very low levels, while Sema4d was abundant at all developmental stages of mouse thymocytes. We found the most interesting pattern of gene regulation and protein localization for semaphorin 4A. Both semaphorin 4A mRNA and protein were clearly detected on the earliest progenitors and were downregulated through thymic development. SEMA4A protein also showed a distinct cortico-medullary pattern of localization. Our findings contribute to an understanding of the complex roles played by semaphorins in the network of spatially and temporally regulated cues underpinning T cell development in the thymus.

Students Propose Genetic Solutions to Societal Problems

The Genetic Engineering Proposal Project, an IBI prize-winning module, teaches biology students to devise innovative bioproducts or solutions to environmental or health problems. In the Foundations of Biology sequence for entering biological sciences majors at the University of Minnesota, inquiry-based learning is woven throughout the classroom and laboratory. During the first semester lecture and discussion, students work in teams on a Genetic Engineering Proposal in which they propose a gene-based solution to a societal problem of their own choosing. Instructors coach the teams throughout the semester on experimental design and resources, as well as on data analysis, presentation strategies, team work, and research ethics. On the basis of outcomes from the nearly 3000 students who have taken the course over the past 6 years, the project has succeeded in engaging students in the intellectual work of biologists and the experience of science as creative inquiry.