A new method for the reconstitution of the anion transport system of the human erythrocyte membrane

Summary The anion transport protein of the human erythrocyte membrane, band 3, was solubilized and purified in solutions of the non-ionic detergent Triton X-100. It was incorporated into spherical lipid bilayers by the following procedure: (1) Dry phosphatidylcholine was suspended in the protein solution. Octylglucopyranoside was added until the milky suspension became clear. (2) The sample was dialyzed overnight against detergentfree buffer. (3) Residual Triton X-100 was removed from the opalescent vesicle suspension by sucrose density gradient centrifugation and subsequent dialysis. Sulfate efflux from the vesicles was studied, under exchange conditions, using a filtration method. Three vesicle subpopulations could be distinguished by analyzing the time course of the efflux. One was nearly impermeable to sulfate, and efflux from another was due to leaks. The largest subpopulation, however, showed transport characteristics very similar to those of the anion transport system of the intact erythrocyte membrane: transport numbers (at 30°C) close to 20 sulfate molecules per band 3 and min, an activation energy of approx. 140 kJ/mol, a pH maximum at pH 6.2, saturation of the sulfate flux at sulfate concentrations around 100mm, inhibition of the flux by H₂DIDS and flufenamate (approx.K _l-values at 30°C: 0.1 and 0.7 m, respectively), and right-side-out orientation of the transport protein (as judged from the inhibition of sulfate efflux by up to 98% by externally added H₂DIDS). Thus, the system represents, for the first time, a reconstitution of all the major properties of the sulfate transport across the erythrocyte membrane.     

Characterization of the ugp region containing the genes for the phoB dependent sn-glycerol-3-phosphate transport system of Escherichia coli

Summary The ugp structural genes, coding for the pho regulon dependent sn-glycerol-3-phosphate transport system, were cloned in pBR322 and characterized. The expression of the cloned ugp system was phoB dependent. Cells containing the ugp plasmid overproduced the G3P binding protein upon phosphate starvation. Tn5 mutagenesis of the cloned DNA revealed that the ugp genes are organized in two separate operons which comprise at least four genes: ugpB and ugpD constitute one operon, ugpA and ugpC constitute the other. The structural gene for the G3P binding protein (G3PBP) is ugpB.The ugpC gene product was also synthesized in minicells as a polypeptide, with an apparent molecular weight of 40,000. No gene products could be assigned to the ugpA and ugpD genes. Hybridization experiments allowed the physical characterization of 20 kb of DNA adjacent to the ugp genes on the E. coli chromosome including the liv genes.     

Light-temperature interactions in the control of photosynthesis in Antarctic phytoplankton

Summary During October/November 1983 photosynthetic responses of natural phytoplankton from the Scotia Sea and Bransfield strait to light and temperature were examined in incubators. Both assimilation numbers at saturating light levels and the slopes of the light-limited portions of the photosynthesis versus irradiance curves were smaller than in algae from lower latitudes. However, both parameters increased significantly with rising temperatures. Light-saturated photosynthesis on the average exhibited a Q₁₀-value of ca. 4.2 between-1.5°C and +2°C. Light-limited photosynthesis between-1.5°C and +5°C rose at a rate corresponding to a Q₁₀-value of roughly 2.6. Above +5°C, temperature enhancement of both light-saturated and light-limited photosynthetic rates was minimal or absent. Our results suggest that under extremely low temperatures light-limited photosynthetic rates become temperature-dependent due to changes in maximum quantum yields.     

Ultrastructural differentiation of the epithelium and mucous glands of the esophagus in the chick embryo

Esophagi of White Leghron chick embryos, six days to hatching, were fixed in 5% glutaraldehyde in a cacodylate-sucrose buffer, postfixed in 1% OsO₄, dehydrated and embedded in Araldite. Tissues were sectioned with glass knives and viewed with a Philips EM 100C. The epithelium in early stages of development is characterized by small intercellular spaces, few cell processes and few desmosomes. In contrast the differentiated epithelium contains numerous cell processes, large intercellular spaces and numerous desmosomes. Mucin appeared in the mucous glands at 17 days.     

The genes of the lysosomal cysteine proteinases cathepsin B, H, L, and S map to different mouse chromosomes

The mouse genes for the lysosomal cysteine proteinases cathepsin B, H, L, and S were mapped to Chromosomes (Chrs) 14, 9, 13, and 3, respectively. Two of the DNA probes used in this study detected an additional, independently segregating locus. The cathepsin B-specific probe hybridized to a locus on Chr 2, and the cathepsin H probe to a locus on the X Chr. These loci either correspond to pseudogenes or to cathepsin B- and cathepsin H-related genes. The four cysteine proteinases mapped in this study lie within known regions of conserved synteny between mouse and human chromosomes, when compared with the corresponding positions of their human homologs. Assuming that the genes of the cysteine proteinase gene family arose from a common ancestral gene, our results suggest that these four cysteine proteinases had been dispersed over different chromosomes before separation of mouse and human in evolution. Received: 22 August 1996 / Accepted: 20 November 1996     

Distribution, morphology and projections of nitrergic and non-nitrergic submucosal neurons in the pig small intestine

 Sequential nitric oxide synthase immunohistochemistry and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry in pig small intestinal wholemounts revealed a complete colocalisation of the two nitrergic markers in submucous neurons. The external submucous plexus (ESP) contained nitrergic neurons throughout. In the internal submucous plexus (ISP) we found a moderate number of nitrergic neurons in the duodenum, while they were rare in the jejunum and nearly absent in the ileum. Combined NADPHd histochemistry and silver impregnation showed morphological ESP type III and VI neurons to be NADPHd positive whereas ESP type II, IV and V neurons were NADPHd negative. Axons of ESP type III, IV and VI neurons were often observed to enter interconnecting strands directed abluminally. ESP type II neurons projected mainly to the ISP. In special silver-impregnated wholemounts containing both external muscle layers and the abluminal part of the submucous layer, i.e. the myenteric plexus and the ESP, the great majority of impregnated axons within the interconnecting strands were observed to run between both plexuses and did not enter the circular muscle layer. We conclude that ESP type III and VI neurons are nitrergic while ESP type II, IV and V neurons are non-nitrergic. Furthermore, we assume that ESP type III, IV and VI neurons may represent a submucosal input to the myenteric plexus. Accepted: 26 August 1997     

Structure of the female reproductive tract of the apple snail. II. Scanning electron microscopy

The albumen gland of Pomacea paludosa, a prosobranch gastropod, contains two main ducts. The albumen gland duct consists of a single layer of secretory and non-secretory cells. The surface of the non-secretory cells is covered with cilia. Microvilli are associated with the luminal edges of the secretory cells. Globules of secretory products appear at the cell surfaces. The capsule gland duct coils through the albumen gland and is composed of two opposing faces each of two layers of cells. The upper layer consists of ciliated non-secretory cells and the microvilli covered necks of the goblet-shaped secretory cells. The bases of the secretory cells comprises the lower layer of cells. Differences in the arrangement of cellular processes and number exists between the duct epithelia.     

A field study on the dependence of the fecundity of Daphnia spec. on food concentration

Summary The dependency of the fecundity of Daphnia spec. on the food concentration was studied in Lake Constance. To eliminate the influence of temperature the number of eggs per adult female was plotted against the concentration of particulate carbon (<50 m) at the estimated time of the production of the eggs. The curve obtained is similar to the results of laboratory investigations. The minimum food concentration for egg production is slightly above 0.2 mg C/1; at 0.7 mg C/1 the curve reaches a plateau. The absolute values of the threshold concentration and the incipient limiting level are compared to laboratory results. It appears that the shift of the field curve to higher concentrations is caused by the imcomplete availability of the 50 m particulate carbon fraction to the daphnids. Daphnia in Lake Constance seem to be food limited during most of the year.