The bell-shaped Ca2+ dependence of the inositol 1,4, 5-trisphosphate-induced Ca2+ release is modulated by Ca2+/calmodulin.

Calmodulin inhibits inositol 1,4,5-trisphosphate (IP3) binding to the IP3 receptor in both a Ca2+-dependent and a Ca2+-independent way. Because there are no functional data on the modulation of the IP3-induced Ca2+ release by calmodulin at various Ca2+ concentrations, we have studied how cytosolic Ca2+ and Sr2+ interfere with the effects of calmodulin on the IP3-induced Ca2+ release in permeabilized A7r5 cells. We now report that calmodulin inhibited Ca2+ release through the IP3 receptor with an IC50 of 4.6 microM if the cytosolic Ca2+ concentration was 0.3 microM or higher. This inhibition was particularly pronounced at low IP3 concentrations. In contrast, calmodulin did not affect IP3-induced Ca2+ release if the cytosolic Ca2+ concentration was below 0.3 microM. Calmodulin also inhibited Ca2+ release through the IP3 receptor in the presence of at least 10 microM Sr2+. We conclude that cytosolic Ca2+ or Sr2+ are absolutely required for the calmodulin-induced inhibition of the IP3-induced Ca2+ release and that this dependence represents the formation of the Ca2+/calmodulin or Sr2+/calmodulin complex.     

Calcium transport systems in the LLC-PK1 renal epithelial established cell line

ATP-dependent calcium uptake was measured in membrane vesicles prepared from the renal epithelial LLC-PK1 established cell line. The relative contribution of the nonmitochondrial versus the mitochondrial calcium uptake is larger in LLC-PK1 cell homogenates than in homogenates from renal cortex. Two types of calcium pump, characterized by the formation of calcium-dependent phosphointermediates of 135 kDa and 115 kDa, were found in membrane fractions from LLC-PK1 cells. The 135 kDa calcium pump was also detected by 125I-labelled calmodulin overlay. Although the subcellular localization in LLC-PK1 cell membranes could not be unambiguously determined, it is conceivable that the 135 kDa and the 115 kDa molecules represent the plasma membrane calcium pump and the endoplasmic reticulum calcium pump respectively, in agreement with what was found for renal cortex preparations. Extravesicular sodium partially inhibits ATP-driven calcium uptake in a plasma-membrane-enriched fraction of the LLC-PK1 cells. The effect is potentiated by a vesicle inside-negative membrane potential. Although the effect is less pronounced than in renal cortex basal-lateral membranes, this observation suggests that an Na+-Ca2+ exchange mechanism is also present in LLC-PK1 cells. ATP-dependent calcium uptake in nonmitochondrial intracellular stores was investigated, using saponin-permeabilized cells. Permeabilized LLC-PK1 cells lowered the free calcium concentration in the medium to less than 0.4 microM. More than 60% of the accumulated calcium can be released by addition of inositol 1,4,5-trisphosphate. Our data indicate that the LLC-PK1 cell line can be successfully used as model system for the study of renal calcium handling.     

Diversity concept in ecology

Hierarchy of systems organization is used as a framework in advancing methodological guidelines for posing correct questions related to ecological diversity.Diversity if defined in general terms as a property of a set of elements dependent on and determines: by the epistemological perspective. Ontological diversity, because it is indefinite, is regarded as unmeasurable.Ecological diversity — be it species, spatial, reproductive or trophic, is a particular case of diversity of matter and must be precisely defined on the studied level of system organization.Diversity measurements combining more than one level of organization are information-void, for data become irretrievable as a result of such a treatment.Hypotheses explaining diversity sources are briefly surveyed. An integrated model interrelating them is constructed as a result of some basic views on the structure of matter coupled with the empirical knowledge of involved factors. The model reflects hierarchies of systems, their dynamics, and the complexities of factors affecting diversity. The model's properties suggest that it is conceptually related to an imaginary model of organization of ecological systems. Parameters recognized as invariant components of this complexity are: habitat differentiation, ecological specialization of species, ambiental changes, and integration of this system.Finally, it is shown that the peculiar shape of the species abundances curve is a necessary consequence of stratification of the system structure, which is an intrinsic attribute of hierarchical organization.     

Satelliten-Telemetrie beim Wei?storch (Ciconia ciconia) auf dem Wegzug — eine Pilotstudie

Zusammenfassung (1) 1991 konnten erstmals 4 mit Kleinsendern ausgerüstete Weißstörche mit Hilfe der Satelliten-Telemetrie auf Teilstrecken ihres Wegzugs bis zu 46 Tage lang verfolgt werden. Die japanischen Sender betrugen nur etwa 2 % des Körpergewichts der Vögel; die Ortung erfolgte durch das ARGOS-System. Die Versuchsvögel zeigten völlig normales Zugverhalten. — (2) Drei der in Brandenburg und Sachsen-Anhalt markierten Vögel waren Ostzieher und konnten über Strecken von etwa 640–4700 km verfolgt werden, 1 Storch bis zur ägyptisch-sudanesischen Grenze. Ein Westzieher konnte rund 1400 km bis zu den Pyrenäen geortet werden. — (3) Die Vögel wanderten individuell recht verschieden. 2 zogen weitgehend kontinuierlich bis in den Sudan bzw. zu den Pyrenäen, die anderen legten längere Pausen ein. Die ermittelten Zugstrecken verliefen recht geradlinig; Richtungsänderungen erfolgten vor allem an der Donau, den Karpaten, am Mittelmeer und auf der Sinai-Halbinsel. Tagesetappen betrugen mindestens bis zu 370 km, in einem Fall in 21 Tagen durchschnittlich 224 km/Tag. Die Zuggeschwindigkeit lag in der Größenordnung von 30–90 km/h. — (4) Verbesserte Sender mit längerer Lebensdauer und mehreren Ortungen pro Tag dürften es bald ermöglichen, individuelle Wanderrouten von Weißstörchen und anderen Großvögeln praktisch lückenlos zu ermitteln. Begleitmannschaften werden zudem die Zug- und Rastökologie mit Sendern ausgerüsteter Vögel mit erfassen können. Damit dürfte der Vogelschutz auf dem Zug eine neue Dimension gewinnen.

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Satellite tracking of White Storks during the autumn migratory period — a pilot study

Summary (1) In 1991 parts of the routes of White Storks migrating in autumn could be recorded for the first time by satellite tracking. Four individuals could be followed for up to 46 days. Transmitter weight accounted for only about 2 % of body mass. Locations were obtained by the ARGOS system. Migratory behaviour of the experimental birds appeared to be absolutely normal. — (2) The birds were equipped with transmitters in eastern Germany. Three of them followed the eastern migration route and could be tracked from 640 up to 4700 km, the latter reaching the borders of Egypt and Sudan. A western migrant could be followed over a distance of about 1400 km towards the Pyrenees. — (3) Migration showed considerable individual variation. Whereas in two birds migration was largely continuous towards the Sudan and the Pyrenees, respectively, the other birds rested for longer periods. The tracked migration routes were fairly straight. Marked directional shifts occurred towards the Danube valley, at the Carpathian mountains, the Mediterranean and on the Sinai. Capacity per day was at least 370 km. One bird covered 224 km/day on average during a period of 21 days. Migration speed ranged in the magnitude of 30–90 km/h. — (4) Improved transmitters with increased lifetime giving several locations per day will presumably allow to record migration routes of White Storks and other large birds more completely in the near future. Escorts should then be able to closely analyse the ecology of migration and staging of their test birds. These possibilities may give a new dimension to bird conservation measures during migration.

     

Isolation, characterization, and localization of the inositol 1,4,5-trisphosphate receptor protein in Xenopus laevis oocytes.

Inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) induces Ca2+ oscillations and waves in Xenopus laevis oocytes. Microsomes from oocytes exhibit high-affinity binding for Ins(1,4,5)P3, and demonstrate Ins(1,4,5)P3-induced Ca2+ release. The Ins(1,4,5)P3 receptor (InsP3R) was purified from oocyte microsomes as a large tetrameric complex and shown to have a monomer molecular mass of 256 kDa, compared with 273 kDa for the brain InsP3R. Binding to the oocyte receptor is highly specific for Ins(1,4,5)P3 and is inhibited by heparin (IC50, 2 micrograms/ml). Immunoblot analysis revealed that an antibody against the C-terminal sequence of the brain receptor recognized the oocyte receptor. These results, in addition to the difference in pattern obtained after limited proteolysis, suggest that the oocyte InsP3R is a new shorter isoform of the mammalian brain type I InsP3R. Immunofluorescence experiments indicated the presence of the InsP3R in the cortical layer and the perinuclear endoplasmic reticulum of the oocyte. However, immunological and biochemical experiments did not reveal the presence of the ryanodine receptor. The presence of an InsP3R and the absence of a ryanodine receptor support the importance of Ins(1,4,5)P3 in Ca2+ handling by oocytes and particularly in the induction of Ca2+ oscillations and waves.     

Alpha-Helical Destabilization of the Bcl-2-BH4-Domain Peptide Abolishes Its Ability to Inhibit the IP3 Receptor

The anti-apoptotic Bcl-2 protein is the founding member and namesake of the Bcl-2-protein family. It has recently been demonstrated that Bcl-2, apart from its anti-apoptotic role at mitochondrial membranes, can also directly interact with the inositol 1,4,5-trisphosphate receptor (IP₃R), the primary Ca²⁺-release channel in the endoplasmic reticulum (ER). Bcl-2 can thereby reduce pro-apoptotic IP₃R-mediated Ca²⁺ release from the ER. Moreover, the Bcl-2 homology domain 4 (Bcl-2-BH4) has been identified as essential and sufficient for this IP₃R-mediated anti-apoptotic activity. In the present study, we investigated whether the reported inhibitory effect of a Bcl-2-BH4 peptide on the IP ₃R1 was related to the distinctive α-helical conformation of the BH4 domain peptide. We therefore designed a peptide with two glycine “hinges” replacing residues I14 and V15, of the wild-type Bcl-2-BH4 domain (Bcl-2-BH4-IV/GG). By comparing the structural and functional properties of the Bcl-2-BH4-IV/GG peptide with its native counterpart, we found that the variant contained reduced α-helicity, neither bound nor inhibited the IP ₃R1 channel, and in turn lost its anti-apoptotic effect. Similar results were obtained with other substitutions in Bcl-2-BH4 that destabilized the α-helix with concomitant loss of IP₃R inhibition. These results provide new insights for the further development of Bcl-2-BH4-derived peptides as specific inhibitors of the IP₃R with significant pharmacological implications.