The mechanism of the calcium signal and correlation with histamine release in 2H3 cells

Rat basophil leukemic (2H3) cells ( Siraganian , R.P., McGivney , A., Barsumian , E. L., Crews, F. T., Hirata , F., and Axelrod , J. (1982) Fed. Proc. 41, 30-34) loaded with fluorescent Ca2+ indicator quin 2 ( Tsien , R. Y. (1980) Biochemistry 19, 2396-2404) showed a rapid increase in free cytosol calcium concentration [( Ca]i) when histamine release was induced. Intracellular quin 2 concentrations up to 7 mM did not affect release of histamine in response to antigen (aggregated ovalbumin) or concanavalin A with cells primed with antigen-specific monoclonal IgE, or in response to Ca2+ ionophores. The [Ca]i increased from approximately 105 nM to a maximum of approximately 1200 nM within 2 to 3 min after antigenic stimulation and then declined slowly over 30 min toward the level in unstimulated cells. Histamine release was most rapid as [Ca]i reached the maximum value and then decreased continuously with [Ca]i over the subsequent 30 min. Neither the Ca signal nor histamine release was observed when the Ca2+ concentration in the medium [( Ca]o) was less than 50 microM, but both responses were restored on readdition of Ca2+ to 1 mM. The maximal Ca signal was obtained when [Ca]o was approximately greater than 1 mM and was half-maximal at [Ca]o congruent to 0.4 mM. In marked contrast [Ca]i in unstimulated cells varied very little with [Ca]o from 0.1 to 1 mM. Maintenance of the Ca signal required the continuous presence of stimulating ligand, external Ca2+, and the maintenance of cellular ATP; metabolic inhibitors blocked or reversed the Ca signal. La+ ions also caused a rapid and reversible block of the Ca signal and histamine release. The data are interpreted in a model in which the Ca signal is generated by a La3+-sensitive signal influx pathway that is functionally independent of the normal Ca2+ influx pathway in unstimulated cells, and that allows a 10-fold or greater increase in rate of Ca2+ entry. The Ca signal is maintained dynamically by the balance between the increased Ca2+ influx and active Ca2+ efflux across the plasma membrane.     

General method for the derivation and numerical solution of epithelial transport models

Summary A general method is presented for the formulation and numerical evaluation of mathematical models describing epithelial transport. The method is based on the principles of conservation of mass, and maintenance of electroneutrality within the cells and bathing solutions. It is therefore independent of the specific membrane transport mechanisms, and can be used to evaluate different models describing arbitrary transport processes (including passive, active and cotransport processes). Detailed numerical methods are presented that allow computation of steady-state and transient responses under open-circuit, current-clamp and voltage-clamp conditions, using a general-purpose laboratory minicomputer. To evaluate the utility of this approach, a specific model is presented that is consistent with the Koefoed-Johnson and Ussing hypothesis of sodium transport in tight epithelia (Acta Physiol. Scand. 42:298–308, 1958). This model considers passive transport of an arbitrary number of permeant solutes, active transport of sodium and potassium, and osmotically induced water transport across the apical and basolateral membranes. Results of the model are compared to published experimental measurements in rabbit urinary bladder epithelium.     

Corticotropin-releasing factor (CRF) stimulates locomotor activity in intact and hypophysectomized newts (Amphibia)

Locomotor activity of rough-skinned newts (Taricha granulosa) was significantly higher in intact and hypophysectomized males injected intracranially with 100 ng CRF (ovine corticotropin-releasing factor) than in those injected with 10 ng CRF or saline. In addition, an injection of corticosterone or dexamethasone failed to stimulate newt locomotor activity. These results provide evidence that CRF can act independently of pituitary hormones to stimulate locomotor activity in a nonmammalian vertebrate.     

Abdominal positioning interneurons in crayfish: projections to and synaptic activation by higher CNS centers

Electron microscopic observations suggest that venom from isolated nematocysts of the stinging tentacles of the Portuguese man-of-war, Physalia physalis, causes histamine release via a rapid, short-duration exocytosis of granules and a slower, long-duration lysis of mast cells. Fine structural changes in mast cells are concurrent with histamine release and are independent of the presence of leukocytes. Vesiculation of the plasma membrane and release of granules nearest the cell surface occur within 10 sec after exposure to 100 micrograms venom/10(5) cells. Released granules and granules retained in plasma membrane invaginations are fibrous and less electron opaque than more centrally located granules. Complex channels to the external medium continue to form, and within 1 min, characteristics of both degranulation and cytolysis are well advanced. Mitochondria are swollen or disrupted. Microridges are absent. Intracellular granules are significantly fewer in venom-treated mast cells, but are more widely separated than in controls. This suggests that degranulation occurs at early stages but is halted as cytolysis proceeds.     

The calcium signal and phosphatidylinositol breakdown in 2H3 cells

Phosphatidylinositol (PI) and its phosphorylated derivatives are rapidly broken down in 2H3 cells stimulated with antigen, with a time course which coincides with the generation of the Ca signal. Stimulated PI breakdown is absolutely dependent on Ca2+ in the medium with a concentration dependence similar to that of the Ca signal and histamine release described in the preceding paper. However, PI breakdown does not depend on the rise in free cytoplasmic Ca2+ concentration in stimulated cells over the range 100 nM to 1 microM. Thus, stimulation by the ionophore A23187 causes only a small increase in PI breakdown and the Ca signal stimulated by antigen can be selectively blocked with appropriate concentrations of Zn2+ (100 microM) or La3+ (10-100 microM) which have small or negligible effects on stimulated PI breakdown. Both PI breakdown and the Ca signal appear to depend on a common external Ca2+ site (or sites) with Km approximately equal to 0.4 mM, and the data are consistent with either independent activation of PI phosphodiesterase and the Ca signal after antigenic stimulation, or with PI breakdown as a component of the mechanism by which the Ca signal is generated.     

Gastrointestinal helminths of the northern bobwhite in Florida: 1968 and 1983

We collected 153 northern bobwhites (Colinus virginianus) over a 10-mo period from Tall Timbers Research Station near Tallahassee, Florida, USA. Five species of gastrointestinal helminths were encountered commonly (greater than 30% prevalence): Cyrnea colini, Raillietina cesticillus, R. colinia, Heterakis isolonche, and Trichostrongylus tenuis. Other helminths included Brachylaima sp., Rhabdometra odiosa, Mediorhynchus papillosis, Cheilospirura spinosa, Dispharynx nasuta, Gongylonema ingluvicola, and Tetrameres pattersoni. During the intervening 15 yr since the last year-round study of this population of birds, C. spinosa and T. pattersoni have declined markedly, and Strongyloides sp. probably has become extinct locally. Prevalence and intensity seem more likely to fluctuate in parasite species that have complex life cycles.     

Chromosomal localization of group-specific component by in situ hybridization

Group-specific component (GC), an alpha 2-globulin plasma protein synthesized primarily in the liver, is the major vitamin D-binding protein in plasma. It has two common phenotypes, GC1 and GC2, which appear in all human populations. Using the cDNA insert containing the entire coding sequence of GC2, the GC gene was mapped to human chromosomal bands 4q13----q21.1 by in situ hybridization.     

Cloning and expression of a cDNA encoding a maize glutathione-S-transferase in E. coli.

The isolation and characterization of a family of maize glutathione-S-transferases (GST's) has been described previously. These enzymes are designated GSTs I, II and III based on size, substrate specificity and responsiveness to safeners. GST III has been shown to act on the herbicide alachlor as well as the commonly used substrate 1-chloro-2,4-dinitrobenzene (CDNB). Clones were isolated from a maize cDNA library in lambda gt10. Three clones contained the entire coding region for GST III. The sequences of these clones were consistent with the known amino terminal GST III protein sequence. Moreover, expression of one of these clones in E. coli resulted in a GST activity as measured with both CDNB and alachlor, proving that at least one of the clones encodes an active GST III species. With the enzyme expressed in E. coli it will become possible to study enzyme structure-function relationships ex planta. While a number of different GST proteins are present in maize tissue the GST III gene is present in single or low copy in the genome.