Basolateral plasma membrane localization of ouabain-sensitive sodium transport sites in the secretory epithelium of the avian salt gland

The distribution of Na+ pump sites (Na+-K+-ATPase) in the secretory epithelium of the avian salt gland was demonstrated by freeze-dry autoradiographic analysis of [(3)H] ouabain binding sites. Kinetic studies indicated that near saturation of tissue binding sites occurred when slices of salt glands from salt-stressed ducks were exposed to 2.2 μM ouabain (containing 5 μCi/ml [(3)H]ouabain) for 90 min. Washing with label-free Ringer's solution for 90 min extracted only 10% of the inhibitor, an amount which corresponded to ouabain present in the tissue spaces labeled by [(14)C]insulin. Increasing the KCl concentration of the incubation medium reduced the rate of ouabain binding but not the maximal amount bound. In contrast to the low level of ouabain binding to salt glands of ducks maintained on a freshwater regimen, exposure to a salt water diet led to a more than threefold increase in binding within 9-11 days. This increase paralleled the similar increment in Na+-K+-ATPase activity described previously. [(3)H]ouabain binding sites were localized autoradiographically to the folded basolateral plasma membrane of the principal secretory cells. The luminal surfaces of these cells were unlabeled. Mitotically active peripheral cells were also unlabeled. The cell-specific pattern of [(3)H]ouabain binding to principal secretory cells and the membrane-specific localization of binding sites to the nonluminal surfaces of these cells were identical to the distribution of Na+-K+-ATPase as reflected by the cytochemical localization of ouabain-sensitive and K+-dependent nitrophenyl phosphatase activity. The relationship between the nonluminal localization of Na+-K+-ATPase and the possible role of the enzyme n NaCl secretion is considered in the light of physiological data on electrolyte transport in salt glands and other secretory epithelia.     

Les Cases de la Valenciana,a new early Miocene small-mammal locality from the Vallès-Penedès Basin (Catalonia,Spain)

Abstract

The Valles-Penedes Basin (Catalonia, Spain) is classical area for the study of Miocene land mammal faunas. Nevertheless, the early Miocene part of the record has deserved little attention as compared to younger intervals. Most notably, the small mammals of this age have not been described in detail, consequently hampering the correlation of the Valles-Penedes record with other chronological schemes. In this work we describe the rich and diverse small mammal fauna from Les Cases de la Valenciana site (Gelida, Alt Penedès) which includes marsupials, eulipotyphlans, lagomorphs and rodents. On the basis of the presence of the cricetids Megacricetodon and Democricetodon this site is correlated with European Neogene zone MN4, yielding an age of 17–16 Ma. However, the rodent assemblage is comparable to that of chronologically close localities of the Calatayud-Montalbán Basin (Aragon, Spain), indicating that the same biochronological scheme can be applied to both areas. In this way, the coexistence of the eomyids Ligerimys ellipticus and Ligerimys florancei coupled with the presence of Megacricetodon primitivus indicates a correlation with local biozone Ca of the Calatayud-Montalban Basin, ranging from 16.3 to 16 Ma. Finally, the correlation of other early Miocene sites of the Valles-Penedes Basin is discussed and refined.     

Electron crystallographic study of photosystem II of the cyanobacterium Synechococcus elongatus

The determination of the structure of PSII at high resolution is required in order to fully understand its reaction mechanisms. Two-dimensional crystals of purified highly active Synechococcus elongatus PSII dimers were obtained by in vitro reconstitution. Images of these crystals were recorded by electron cryo-microscopy, and their analysis revealed they belong to the two-sided plane group p22(1)2(1), with unit cell parameters a = 121 A, b = 333 A, and alpha = 90 degrees. From these crystals, a projection map was calculated to a resolution of approximately 16 A. The reliability of this projection map is confirmed by its close agreement with the recently presented three-dimensional model of the same complex obtained by X-ray crystallography. Comparison of the projection map of the Synechococcus elongatus PSII complex with data obtained by electron crystallography of the spinach PSII core dimer reveals a similar organization of the main transmembrane subunits. However, some differences in density distribution between the cyanobacterial and higher plant PSII complexes exist, especially in the outer region of the complex between CP43 and cytochrome b(559) and adjacent to the B-helix of the D1 protein. These differences are discussed in terms of the number and organization of some of the PSII low molecular weight subunits.     

Beta-adrenoceptor-blocking drugs and blood sugar control in diabetes mellitus.

The effects on diabetic control of the relative cardioselective beta-blocker metoprolol and the non-selective drug propranolol were compared in 20 hypertensive diabetic patients receiving diet alone or diet and oral hypoglycaemic agents. Each drug was given for one month in a double-blind, cross-over study. Fasting, noon, and mid-afternoon blood sugar concentrations rose by 1.0-1.5 mmol/l (18-27 mg/100 ml). The rise with propranolol was not significantly greater than with metoprolol. In a few patients the rise was clinically important. The small overall change observed in diabetic control should not deter the use of beta-blockers in non-insulin-dependent diabetics, provided control is carefully monitored at the onset of treatment.     

D-galactose uptake by snail intestine: competitive inhibition by phlorizin and sodium dependence

The net entry of galactose into the tissue of snail everted intestinal rings with 2 or 15 minute long incubation periods has been measured. With 10(-4) M phlorizin, the mediated transport is completely blocked while only the passive entry of sugar is produced. Lower concentrations of the glycoside partially inhibit transport according to competitive inhibition kinetics (K1 = 10(-7) M). The transport of galactose is Na+ dependent. In the absence of Na+, transport ceases and the sugar entry can be explained through simple diffusion. With 15 mM Na+ (control 71,4 mM) transport diminishes and a marked increase in the apparent Km with no changes in the Vmax is observed. One mM harmaline completely blocks galactose (0.5 mM) transport. One mM ouabain also makes transport null, but only after tissue preincubation with the inhibitor on the serosal side.     

Interactions among amino acid transport systems in snail Helix aspersa intestine

Interactions among the transport of diverse amino acids in everted intestine of snail Helix aspersa have been studied. The uptake of 0.5 mM methionine is clearly inhibited by high concentrations (40 mM) of leucine, and not by proline or lysine, whereas the last two amino acids inhibit cycloleucine uptake. Methionine strongly inhibits proline and lysine uptake, which is significantly inhibited by their analogs hydroxiproline and arginine, respectively. Results suggest that in Helix intestine the transport systems for basic amino acids and iminoacids are shared with high affinity by methionine whereas the neutral amino acids transport systems do not seem to be shared, or are so very weakly, by the basic ones or by the imino acids.     

Scanning electron microscopic studies of cilia

Summary A simple method for the preparation of ciliated epithelia for study with the scanning electron microscope is described. Ciliary groups are well preserved and it is possible to discern individual cilia and work out their numbers and orientation. Following scanning electron microscopical study some of the material was prepared for transmission electron microscopy and the ultrastructure of the tissue was found to be surprisingly well preserved. The tracheal epithelium of the rabbit, the olfactory epithelia of the goldfish and the rabbit, and the sensory epithelia in the statocyst of a cephalopod mollusc were examined with the scanning electron microscope to demonstrate the possibilities of the method. Acknowledgements. We would like to thank Professor J. Z. Young for his continued interest and support. The scanning electron microscope was purchased with a grant provided by the Science Research Council to Dr. Boyde, Mr. R. Willis helped in the initial stages of the study, Mr. G. Savage provided help with the goldfish material, Mr. S. Waterman provided much photographic assistance, and Mrs. N. Finney the secretarial assistance.