Effect of sterol structure on ordered membrane domain (raft) stability in symmetric and asymmetric vesicles

Sterol structure influences liquid ordered domains in membranes, and the dependence of biological functions on sterol structure can help identify processes dependent on ordered domains. In this study we compared the effect of sterol structure on ordered domain formation in symmetric vesicles composed of mixtures of sphingomyelin, 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol, and in asymmetric vesicles in which sphingomyelin was introduced into the outer leaflet of vesicles composed of DOPC and cholesterol. In most cases, sterol behavior was similar in symmetric and asymmetric vesicles, with ordered domains most strongly stabilized by 7-dehydrocholesterol (7DHC) and cholesterol, stabilized to a moderate degree by lanosterol, epicholesterol and desmosterol, and very little if at all by 4-cholesten-3-one. However, in asymmetric vesicles desmosterol stabilized ordered domain almost as well as cholesterol, and to a much greater degree than epicholesterol, so that the ability to support ordered domains decreased in the order 7-DHC > cholesterol > desmosterol > lanosterol > epicholesterol > 4-cholesten-3-one. This contrasts with values for intermediate stabilizing sterols in symmetric vesicles in which the ranking was cholesterol > lanosterol ~ desmosterol ~ epicholesterol or prior studies in which the ranking was cholesterol ~ epicholesterol > lanosterol ~ desmosterol. The reasons for these differences are discussed. Based on these results, we re-evaluated our prior studies in cells and conclude that endocytosis levels and bacterial uptake are even more closely correlated with the ability of sterols to form ordered domains than previously thought, and do not necessarily require that a sterol have a 3β-OH group.     

Biologie, ökologie und brutpflege von coelotes terrestris (wider) (araneae,agelenidae) teil I. biologie und ökologie

Zusammenfassung Coelotes terrestris (Wider) hat seinen Verbreitungsschwerpunkt in den geschlossenen Laub- und Nadelwäldern besserer Bonität auf lehmigen Böden und kommt in erster Linie in Berggegenden vor. Von dort strahlt sie aber weit auf die Sandböden der Ebene aus und kompensiert dabei die Feuchtigkeitsdifferenz zwischen Lehm- und Sandboden durch ihren Aufenthalt in Gebieten mit hohem Grundwasserstand. Den wechselnden Feuchtigkeitsverhältnissen entsprechend ist ihr Vorkommen in Verbreitungsrandgebieten inselartig. Wenn sie stellenweise auch in Grenzgebieten möglicher Besiedlung, wie sie scheinbar trockene, dürftige Föhrenbestände darstellen, eine überraschend hohe Populationsdichte erreicht, ist dies nicht auf ein Optimum an Faktorenkonstellation schlechthin, sondern auf die Unmöglichkeit einer Ausbreitung in angrenzende Trockengebiete zurückzuführen. Coelotes ist eine Bodenspinne. Auf lehmigen Böden lebt sie in Laubstreu, in Bodenspalten unter den flach ausgebreiteten Wurzeln von Fichten, auf Sandboden in hochwüchsigem Moos (Hypneen, nicht Sphagnum), vor allem aber in der Rohhumus- und Streuschicht, die sockelartig den Fuß von Föhrenstämmen umgibt, und überall gerne unter großen, tiefliegenden Steinen. Bewohner von Kiefernstämmen legen ihre Netze mit Vorliebe zwischen Borke und umgebender Rohhumus-bzw. Streuschicht an. Um einen Kiefernstamm von nur 16 cm Durchmesser wurden bis zu fünf bewohnte Netze gefunden. Eine Prävalenz von Kiefernborke war im Wahlversuch nachweisbar, desgleichen eine solche von Rohhumus vor ebenso feuchtem Lehm- oder Sandboden verschiedener Körnigkeit. Dank seiner beträchtlichen Wasserkapazität und guten Wärme-Isolation bietet der Rohhumus diesen Spinnen auch in Trockengebieten eine Existenzmöglichkeit. Resistenzversuche ergaben, daß Coelotes zwar auf die Dauer eine weitgehende Wasserdampfsättigung der Luft benötigt, daß aber seine Widerstandsfähigkeit gegenüber einer relativen Luftfeuchtigkeit von 30–75% im Vergleich zu anderen feuchtigkeitssuchenden Bodentieren recht groß ist. Die Resistenz aller Jugendstadien ist höher als die der Erwachsenen. Während sich aber die Lebensdauer der noch im mütterlichen Netz verweilenden Kleinspinnen mit abnehmender Luftfeuchtigkeit merklich verkürzt, erweist sich die Widerstandsfähigkeit der halbwüchsigen Individuen, die überwintern, in allen Feuchtigkeitsbereichen als annähernd gleich hoch. Selbst in 75% relativer Luftfeuchtigkeit blieben adulte nur durchschnittlich 131 Std am Leben, bei 30% dagegen immerhin noch 40 Std. Jungspinnen hingegen halten in 30% relativer Luftfeuchtigkeit 2–3mal so lange aus.Bei Stammbewohnern ließ sich eine Bevorzugung der Ostseite (nicht Osthälfte!) und noch deutlicher eine Meidung der West- und Nordwestseite der Stämme nachweisen, wodurch sie sich - ihrer Hemihygrophilie entsprechend - dem unmittelbaren Abfluß des Regenwassers von den Stämmen entziehen. Nächst der genauen Ostrichtung werden aber auch die Nord- und Südseiten der Stämme stark frequentiert, die Zwischenhimmelsrichtungen dagegen auffallend benachteiligt. Als Erklärung dafür wird angenommen, daß die Spinnen durch vermutliche polarisationsoptische Orientierung primär der Ostseite zustreben. Ist diese schon besetzt, rücken sie zur Wahrung eines angemessenen Sicherheitsabstandes vom Nachbarindividuum um rund 90° des Kreisbogens ab und treffen somit auf die Nord- oder Südseite. - Die Überwinterung der und halbwüchsigen Tiere erfolgt 10–15 cm von den Kiefernstämmen entfernt im Rohhumus und zwar nur etwa 5 cm unter der Bodenoberfläche. Dabei bevorzugen sie die Ostseite und relativ trockene Stellen. Seltener überwintern sie in ihren alten Wohnröhren, die sie dann oben verschließen.Habilitationsschrift     

Green fluorescent protein (GFP) as a marker during pollen development

The transient expression of three mutant forms of green fluorescent protein (GFP) genes, GFP4, GFP5ER, and GFP4S65C, under several constitutive and pollenspecific promoters throughout pollen development in Nicotianatabacum, thaliana and Antirrhinummajus is described. Immature pollen of tobacco, Arabidopsis and snapdragon, isolated at different developmental stages, were bombarded with plasmids containing the GFP and cultured in vitro for several days until maturity. The expression of GFP was monitored every day during in vitro maturation, germination and pollination, as well as after in situ pollination. The expression pattern of each construct was compared in parallel experiments to that of ßglucuronidase (GUS) constructs expressed by the same promoters. The results show that the expression level of all three GFP mutant forms was dependent on the strength of the promoter used. The strongest promoter was the DC3 promoter, and no notable differences in the intensity and brightness of all three versions of GFP were observed. GFPexpressing pollen from tobacco and snapdragon developed in vitro for several days until maturity and germinated in vitro as well as on the surface of stigmata, strongly suggesting that all three GFPs are not toxic for the development of functional pollen. Furthermore, stably transformed tobacco plants expressing GFP under the control of the strong pollenexpressed DC3 and LAT52 promoters were not impaired in reproductive function, confirming that GFP can be used as a nondestructive marker for plant reproductive biology and development.     

Cloning, genomic structure, and expression profiles of TULIP1 (GARNL1), a brain-expressed candidate gene for 14q13-linked neurological phenotypes, and its murine homologue

Previously, we have described the clinical and molecular characterization of a de novo 14q13.1-q21.1 microdeletion, less than 3.5 Mb in size, in a patient with severe microcephaly, psychomotor retardation, and other clinical anomalies. Here we report the characterization of the genomic structure of the human tuberin-like protein gene 1 (TULIP1; approved gene symbol GARNL1), a CpGisland-associated, brain-expressed candidate gene for the neurological findings in our patient, and its murine homologue. The human TULIP1 gene was mapped to chromosome band 14q13.2 by fluorescence in situ hybridization of BAC clone RP11-355C3 (GenBank Accession No. AL160231), containing the 3' region of the gene. TULIP1 spans about 271 kb of human genomic DNA and is divided into 41 exons. An untranscribed, processed pseudogene of TULIP1 was found on human chromosome band 9q31.1. The active locus TULIP1, encoding a predicted protein of 2036 amino acids, is expressed ubiquitously in pre- and postnatal human tissues. The murine homologue Tulip1 spans about 220 kb of mouse genomic DNA and is also divided into 41 exons, encoding a predicted protein of 2035 amino acids. No pseudogene could be found in the available mouse sequence data. Several splicing variants were found. Considering the location, expression profile, and predicted function, TULIP1 is a strong candidate for several neurological features seen in 14q deletion patients. Additionally we searched for mutations in the coding region of TULIP1 in subjects from a family with idiopathic basal ganglia calcification (IBGC; Fahr disease), previously linked to chromosome 14q. We identified two novel SNPs in the intron-exon boundaries; however, they did not segregate only with affected subjects in the predicted model of an autosomal dominant disease such as IBGC.     

Molecular microbial ecology of the gastrointestinal tract: from phylogeny to function

During the past decade it became evident that anaerobic cultivation-based approaches provides an incomplete picture of the microbial diversity in the GI tract, since at present only a minority of microbes can be obtained in culture. The application of molecular, mainly 16S ribosomal RNA (rRNA)-based approaches enables researchers to bypass the cultivation step and has proven its usefulness in studying the microbial composition in a variety of ecosystems, including the gastrointestinal (GI) tract. This critical review summarizes the impact of these culture-independent approaches on our knowledge of the ecology of the GI tract and provides directions for future studies which should emphasize function of specific strains, species and groups of microbes.     

MKK7 couples stress signalling to G2/M cell-cycle progression and cellular senescence

During the development of multicellular organisms, concerted actions of molecular signalling networks determine whether cells undergo proliferation, differentiation, death or ageing. Here we show that genetic inactivation of the stress signalling kinase, MKK7, a direct activator of JNKs in mice, results in embryonic lethality and impaired proliferation of hepatocytes. Beginning at passage 4-5, mkk7(-/-) mouse embryonic fibroblasts (MEFs) display impaired proliferation, premature senescence and G2/M cell cycle arrest. Similarly, loss of c-Jun or expression of a c-JunAA mutant in which the JNK phosphorylation sites were replaced with alanine results in a G2/M cell-cycle block. The G2/M cell-cycle kinase CDC2 was identified as a target for the MKK7-JNK-c-Jun pathway. These data show that the MKK7-JNK-c-Jun signalling pathway couples developmental and environmental cues to CDC2 expression, G2/M cell cycle progression and cellular senescence in fibroblasts.     

Pattern and process in Amazon tree turnover, 1976-2001

Previous work has shown that tree turnover, tree biomass and large liana densities have increased in mature tropical forest plots in the late twentieth century. These results point to a concerted shift in forest ecological processes that may already be having significant impacts on terrestrial carbon stocks, fluxes and biodiversity. However, the findings have proved controversial, partly because a rather limited number of permanent plots have been monitored for rather short periods. The aim of this paper is to characterize regional-scale patterns of 'tree turnover' (the rate with which trees die and recruit into a population) by using improved datasets now available for Amazonia that span the past 25 years. Specifically, we assess whether concerted changes in turnover are occurring, and if so whether they are general throughout the Amazon or restricted to one region or environmental zone. In addition, we ask whether they are driven by changes in recruitment, mortality or both. We find that: (i) trees 10 cm or more in diameter recruit and die twice as fast on the richer soils of southern and western Amazonia than on the poorer soils of eastern and central Amazonia; (ii) turnover rates have increased throughout Amazonia over the past two decades; (iii) mortality and recruitment rates have both increased significantly in every region and environmental zone, with the exception of mortality in eastern Amazonia; (iv) recruitment rates have consistently exceeded mortality rates; (v) absolute increases in recruitment and mortality rates are greatest in western Amazonian sites; and (vi) mortality appears to be lagging recruitment at regional scales. These spatial patterns and temporal trends are not caused by obvious artefacts in the data or the analyses. The trends cannot be directly driven by a mortality driver (such as increased drought or fragmentation-related death) because the biomass in these forests has simultaneously increased. Our findings therefore indicate that long-acting and widespread environmental changes are stimulating the growth and productivity of Amazon forests.     

Intraspecific sexual selection on a speciation trait, male coloration, in the Lake Victoria cichlid Pundamilia nyererei

The haplochromine cichlids of Lake Victoria constitute a classical example of explosive speciation. Extensive intra- and interspecific variation in male nuptial coloration and female mating preferences, in the absence of postzygotic isolation between species, has inspired the hypothesis that sexual selection has been a driving force in the origin of this species flock. This hypothesis rests on the premise that the phenotypic traits that underlie behavioural reproductive isolation between sister species diverged under sexual selection within a species. We test this premise in a Lake Victoria cichlid, by using laboratory experiments and field observations. We report that a male colour trait, which has previously been shown to be important for behavioural reproductive isolation between this species and a close relative, is under directional sexual selection by female mate choice within this species. This is consistent with the hypothesis that female choice has driven the divergence in male coloration between the two species. We also find that male territoriality is vital for male reproductive success and that multiple mating by females is common.