Clark R. Coffman

Expression of an extracellular deletion of Xotch diverts cell fate in Xenopus embryos

Xotch is a Xenopus homolog of Notch, a receptor involved in cell fate decisions in Drosophila. Using an extracellular deletion construct, Xotch delta E, we show that Xotch has a similar role in Xenopus embryos. Broad expression causes the loss of dorsal structures and the expansion and disorganization of the brain. Single blastomere injections of Xotch delta E induce autonomous neural and mesodermal hypertrophy, even in the absence of cell division. Xotch delta E inhibits the early expression of epidermal and neural crest markers yet enhances and extends the response of animal caps to mesodermal and neural induction. Our data suggest a mechanism for the function of Notch homologs in which they delay differentiation and leave undetermined cells competent to respond to later inductive signals.

Programmed cell death of primordial germ cells in Drosophila is regulated by p53 and the Outsiders monocarboxylate transporter.

Primordial germ cell development uses programmed cell death to remove abnormal, misplaced or excess cells. Precise control of this process is essential to maintain the continuity and integrity of the germline, and to prevent germ cells from colonizing locations other than the gonads. Through careful analyses of primordial germ cell distribution in developing Drosophila melanogaster embryos, we show that normal germ cell development involves extensive programmed cell death during stages 10-12 of embryogenesis. This germ cell death is mediated by Drosophila p53 (p53). Mutations in p53 result in excess primordial germ cells that are ectopic to the gonads. Initial movements of the germ cells appear normal, and wild-type numbers of germ cells populate the gonads, indicating that p53 is required for germ cell death, but not migration. To our knowledge, this is the first report of a loss-of-function phenotype for Drosophila p53 in a non-sensitized background. The p53 phenotype is remarkably similar to that of outsiders (out) mutants. Here, we show that the out gene encodes a putative monocarboxylate transporter. Mutations in p53 and out show nonallelic noncomplementation. Interestingly, overexpression of p53 in primordial germ cells of out mutant embryos partially suppresses the out germ cell death phenotype, suggesting that p53 functions in germ cells either downstream of out or in a closely linked pathway. These findings inform models in which signaling between p53 and cellular metabolism are integrated to regulate programmed cell death decisions.

Growth cone interactions with a glial cell line from embryonic Xenopus retina

We have isolated a nonneuronal cell line from Xenopus retinal neuroepithelium (XR1 cell line). On the basis of immunocytochemical characterization using monoclonal antibodies generated in our laboratory as well as several other glial-specific antibodies, we have established that the XR1 cells are derived from embryonic astroglia. A monolayer of XR1 cells serves as an excellent substrate upon which embryonic retinal explants attach and elaborate neurites. This neurite outgrowth promoting activity appears not to be secreted into the medium, as medium conditioned by XR1 cells is ineffective in promoting outgrowth. Cell-free substrates were prepared to examine whether outgrowth promoting activity is also associated with the XR1 extracellular matrix (ECM). Substrates derived from XR1 cells grown on collagen are still capable of promoting outgrowth following osmotic shock and chemical extraction. This activity does not appear to be associated with laminin or fibronectin. Scanning electron microscopy was used to examine growth cones of retinal axons on XR1 cells and other substrates that supported neurite outgrowth. Growth cones and neurites growing on a monolayer of XR1 cells, or on collagen conditioned by XR1 cells, closely resemble the growth cones of retinal ganglion cells in vivo. A polyclonal antiserum (NOB1) generated against XR1 cells effectively and specifically inhibits neurite outgrowth on XR1-conditioned collagen. We therefore propose that neurite outgrowth promoting factors produced by these cells are associated with the extracellular matrix and may be glial specific.

Xenopus fibrillin is expressed in the organizer and is the earliest component of matrix at the developing notochord‐somite boundary

We identify a Xenopus fibrillin homolog (XF), and show that its earliest developmental expression is in presumptive dorsal mesoderm at gastrulation, and that XF expression is regulated by mesoderm‐inducing factors in animal cap assays. XF protein is also first detected in presumptive mesoderm, but is concentrated specifically into extracellular‐matrix structures that begin to develop de novo by mid‐gastrulation at both of the bilateral presumptive notochord‐somite boundaries. Later in embryogenesis, XF protein is localized to the extracellular matrix at tissue boundaries, where it is found surrounding the notochord, the somites, and the neural tube, as well as under the epidermis. This pattern of protein deposition combines to give the appearance of an “embryonic skeleton,” suggesting that one role for XF is to serve as a mechanical element in the embryo prior to bone deposition. Developmental Dynamics 235:1974–1983, 2006.

Cell migration and programmed cell death of Drosophila germ cells.

Abstract: Cell migration and programmed cell death are essential components of animal development and homeostasis, and the germ cells of Drosophila provide a simple genetic system to study the molecular mechanisms that govern these important cellular processes. Detailed descriptions of germ cell migration in Drosophila were accomplished long ago, but most genetic and molecular analyses of the process have occurred within the past 10 years. A few of the genes required for germ cell migration have been identified, and a very interesting picture is emerging. However, a process as complex as cell migration must involve the functions of many more molecules. In addition, cell migration and cell death mechanisms are often linked, as it is important to eliminate cells that are misplaced and could present a danger to the organism. In Drosophila, genes involved in germ cell migration can also affect programmed cell death. Currently, very little is known about how germ cells ectopic to the gonads are eliminated. To date, only four genes have been reported with roles in germ cell death, and three of these have additional functions in germ cell pathfinding. The nature of the cell death program has not been elucidated. Here, I provide a brief review of Drosophila germ cell migration and programmed cell death at both the descriptive and molecular levels. Many questions remain to be answered, but advances made in recent years are providing useful insights into these critical biological phenomena.

An evolutionarily conserved arginine is essential for Tre1 G protein-coupled receptor function during germ cell migration in Drosophila melanogaster.

Background G protein-coupled receptors (GPCRs) play central roles in mediating cellular responses to environmental signals leading to changes in cell physiology and behaviors, including cell migration. Numerous clinical pathologies including metastasis, an invasive form of cell migration, have been linked to abnormal GPCR signaling. While the structures of some GPCRs have been defined, the in vivo roles of conserved amino acid residues and their relationships to receptor function are not fully understood. Trapped in endoderm 1 (Tre1) is an orphan receptor of the rhodopsin class that is necessary for primordial germ cell migration in Drosophila melanogaster embryos. In this study, we employ molecular genetic approaches to identify residues in Tre1 that are critical to its functions in germ cell migration. Methodology/Principal Findings First, we show that the previously reported scattershot mutation is an allele of tre1. The scattershot allele results in an in-frame deletion of 8 amino acids at the junction of the third transmembrane domain and the second intracellular loop of Tre1 that dramatically impairs the function of this GPCR in germ cell migration. To further refine the molecular basis for this phenotype, we assayed the effects of single amino acid substitutions in transgenic animals and determined that the arginine within the evolutionarily conserved E/N/DRY motif is critical for receptor function in mediating germ cell migration within an intact developing embryo. Conclusions/Significance These structure-function studies of GPCR signaling in native contexts will inform future studies into the basic biology of this large and clinically important family of receptors.

DNA Damage‐Induced Programmed Cell Death: Potential Roles in Germ Cell Development

Abstract: The detection of DNA damage is necessary to protect against proliferation of potentially harmful cells and often results in cell cycle arrest and programmed cell death. Key components of DNA damage signaling networks include ATM, CHK2, p53, and Bax. Mutations in these damage signaling systems are linked to tumorigenesis and developmental abnormalities. Expression of some of these genes in primordial germ cells (PGCs) argues that PGCs may utilize DNA damage‐induced signaling mechanisms to select against germ cells that are genetically defective, thus maintaining the integrity of the germline. This paper summarizes the roles of these DNA damage signaling molecules and addresses their potential involvement in germ cell development.

Improved Student Learning through a Faculty Learning Community: How Faculty Collaboration Transformed a Large-Enrollment Course from Lecture to Student Centered

The authors describe how a faculty learning community was leveraged to implement active-learning strategies and improve student learning in a large-enrollment introductory course.

G protein-coupled receptor roles in cell migration and cell death decisions.

Abstract: Recognition of external conditions and the elicitation of appropriate responses are critical to a cells ability to adjust to various developmental and environmental cues. G protein‐coupled receptors (GPCRs) are a large class of receptors that act to relay external information into the cell by initiating signaling pathways that allow the cell to adapt to its present conditions. There are numerous ligands that activate GPCRs to initiate a multitude of intracellular signaling cascades involved in critical decisions including cell growth, differentiation, proliferation, migration, survival, and death. This article focuses on the signaling pathways involved in cell migration, survival, and death decisions with an emphasis on germ cells from various organisms.

Study strategies and beliefs about learning as a function of academic achievement and achievement goals.

ABSTRACT Prior research by Hartwig and Dunlosky [(2012). Study strategies of college students: Are self-testing and scheduling related to achievement? Psychonomic Bulletin & Review, 19(1), 126–134] has demonstrated that beliefs about learning and study strategies endorsed by students are related to academic achievement: higher performing students tend to choose more effective study strategies and are more aware of the benefits of self-testing. We examined whether students’ achievement goals, independent of academic achievement, predicted beliefs about learning and endorsement of study strategies. We administered Hartwig and Dunlosky’s survey, along with the Achievement Goals Questionnaire [Elliot, A. J., & McGregor, H. A. (2001). A 2 × 2 achievement goal framework. Journal of Personality & Social Psychology, 80, 501–519] to a large undergraduate biology course. Similar to results by Hartwig and Dunlosky, we found that high-performing students (relative to low-performing students) were more likely to endorse self-testing, less likely to cram, and more likely to plan a study schedule ahead of time. Independent of achievement, however, achievement goals were stronger predictors of certain study behaviours. In particular, avoidance goals (e.g., fear of failure) coincided with increased use of cramming and the tendency to be driven by impending deadlines. Results suggest that individual differences in student achievement, as well as the underlying reasons for achievement, are important predictors of students’ approaches to studying.