Functional study of the Caenorhabditis elegans secretory-excretory system using laser microsurgery

Individual cells of the Caenorhabditis elegans secretory-excretory system were ablated by laser microbeam in various larval stages. Effects on growth, molting, osmoregulation, fertility, longevity, and dauer larva formation were tested. Single-cell ablations did not prevent subsequent molting, but ablation of the pore cell or the duct cell resulted in the absence of the normal cuticular lining of the excretory duct following a molt. When the pore cell, duct cell, or excretory cell was ablated, the animals filled with fluid within 12-24 hr and died within a few days, producing very few progeny. Ablation of the excretory gland cell, on the other hand, had no obvious developmental or behavioral effects. Excretory activity was monitored in dauer larvae by observing pulsation of the excretory duct in conditions of differing osmolarity. The rate of pulsation was quite variable over time in conditions of low osmotic strength, but average five- to six-fold higher than that observed in buffered saline. These observations, combined with the effects of laser ablation, lead to the conclusion that one function of the excretory system is osmoregulation.     

A Ca2+-activated ATPase specifically released by Ca2+ shock from Paramecium tetraurelia

Deciliation of Paramecium tetraurelia by a Ca2+ shock procedure releases a discrete set of proteins which represent about 1% of the total cell protein. Marker enzymes for cytoplasm (hexokinase), endoplasmic reticulum (glucose-6-phosphatase), peroxisomes (catalase), and lysosomes (acid phosphatase) were not released by this treatment. Among the proteins selectively released is a Ca2+-dependent ATPase. This enzyme has a broad substrate specificity which includes GTP, ATP, and UTP, and it can be activated by Ca2+, Sr2+, or Ba2+, but not by Mg2+ or by monovalent cations. The crude enzyme has a specific activity of 2-3 mumol/min per mg; the optimal pH for activity is 7.5. ATPase, GTPase, and UTPase all reside in the same protein, which is inhibited by ruthenium red, is irreversibly denatured at 50 degrees C, and which has a sedimentation coefficient of 8-10 S. This enzyme is compared with other surface-derived ATPases of ciliated protozoans, and its possible roles are discussed.     

Plant and Soil Nematodes: Societal Impact and Focus for the Future.

Plant and soil nematodes significandy impact our lives. Therefore, we must understand and manage these complex organisms so that we may continue to develop and sustain our food production systems, our natural resources, our environment, and our quality of life. This publication looks specifically at soil and plant nematology. First, the societal impact of nematodes and benefits of nematology research are briefly presented. Next, the opportunities facing nematology in the next decade are outlined, as well as the resources needed to address these priorities. The safety and sustainability of U.S. food and fiber production depends on public and administrative understanding of the importance of nematodes, the drastic effects of nematodes on many agricultural and horticultural crops, and the current research priorities of nematology.     

Mutations Affecting the Chemosensory Neurons of Caenorhabditis Elegans

We have identified and characterized 95 mutations that reduce or abolish dye filling of amphid and phasmid neurons and that have little effect on viability, fertility or movement. Twenty-seven mutations occurred spontaneously in strains with a high frequency of transposon insertion. Sixty-eight were isolated after treatment with EMS. All of the mutations result in defects in one or more chemosensory responses, such as chemotaxis to ammonium chloride or formation of dauer larvae under conditions of starvation and overcrowding. Seventy-five of the mutations are alleles of 12 previously defined genes, mutations which were previously shown to lead to defects in amphid ultrastructure. We have assigned 20 mutations to 13 new genes, called dyf-1 through dyf-13. We expect that the genes represented by dye-filling defective mutants are important for the differentiation of amphid and phasmid chemosensilla.     

Modeling flux of free and protein-bound radioisotopes into the pulmonary interstitium

Several groups of investigators are using external detection of radiolabeled protein to study the flux of protein from plasma into the pulmonary interstitium. A basic assumption for these studies has been that the unbound (free) tracer concentration is small and insignificant. The purpose of this study is to evaluate how free tracer influences the determination of normalized slope index. A five-compartment model for the lung was used with transport equations for both unbound and bound nuclide flux. Parameters of the unbound and bound transport equations were varied to evaluate the sensitivity of normalized slope index to each parameter. The model was also compared with published protein flux data to investigate the validity of the transport model. Application of the model to external scan data provides a sensitive method for evaluating the flux of bound and unbound tracers into the pulmonary interstitium. We conclude that because the distribution volume for unbound tracer is large with respect to protein distribution volume, even a small amount of unbound tracer (2-5%) can create large errors in the determination of normalized slope index.