Acetylcholine responses of identified neurons in Helix pomatia--I. Interactions between acetylcholine-induced and potential-dependent membrane conductances

The influence of potential-dependent membrane conductances on amplitude and time course of acetylcholine (ACh) responses was studied. The investigations were performed on the identified neurons B1 and B3 of the buccal ganglion of Helix pomatia. The neurons B1 and B3 were depolarized by ACh. The depolarization was accompanied by a decrease of membrane resistance. An inward rectification occurring negative to the resting membrane potential (RMP) reduced the amplitude of the ACh depolarizations. An outward rectification occurring positive to the RMP consisted of two parts and ceiled the ACh responses. The early outward current reduced the amplitude and modified the time course of ACh responses. Local responses or axonal action potentials increased the amplitude of the ACh depolarizations.     

Disomic and ditelosomic alien chromosome additions in beet (Beta vulgaris), carrying an extra chromosome of B. procumbens or telosome of B. patellaris.

Alien chromosome transmission through the female germ line as well as meiosis in pollen mother cells were studied in disomic and ditelosomic alien chromosome additions of beet. Beta vulgaris, carrying an extra pair of chromosomes or telosomes of B. procumbens or B. patellaris, respectively. The alien chromosomes carried genes for resistance to the beet cyst nematode, Heterodera schachtii, and screening for this resistance was used to select plants with or without the alien chromosomes. A great variation for alien chromosome transmission was recorded and plants carrying two extra alien chromosomes were recovered in the backcross progenies of the disomic or ditelosomic additions. However, in these progenies the average frequencies of plants without alien chromosomes (86%) did not clearly differ from that in similar progenies of the original monosomic or monotelosomic chromosome additions, indicating that doubling the number of the alien chromosome did not enlarge their transmission to the next generation. The alien chromosomes fully paired at pachytene and desynapsed again before diakinesis, indicating decreased chiasma formation. At second metaphase nearly 60% of the cells had one extra chromosome, and the remaining cells carried two or no extra chromosomes in about equal proportions. The tetrads looked fully normal. The expected relation between the average number of alien chromosomes in the germ cells and in the plants of the progenies did not show up, indicating a strong selection favouring the female gametes without alien chromosomes.     

Quantification of individual flagellate - bacteria interactions within semi-natural biofilms

Here we present a new approach to quantify food-web interactions within semi-natural biofilms by combining the establishment of biofilms from natural rivers in flow cells with video microscopy. In a first application of this approach, we focused on the surface-gliding heterotrophic flagellates (HF) Neobodo designis, Rhynchomonas nasuta and Planomonas sp. It was shown that the three HF generally ingested single biofilm-associated bacteria whereas bacteria within microcolonies were attacked but not ingested. However, grazing strategies differed considerably. While the kinetoplastids N. designis and R. nasuta displayed long search and short handling times, Planomonas sp. showed the opposite grazing characteristics. The latter behaviour resulted in a high relative predation success of 80% (precent of attacked prey ingested), whereas the relative predation success of the two kinetoplastids was only 20%. However, the two contrasting strategies resulted in similar ingestion rates for Planomonas sp. and N. designis of 0.5 to 0.6 ingestions flagellates(-1) minute(-1), respectively. Our results showed distinct differences in the feeding behaviour of three flagellates having similar life forms and provide direct evidence that microcolony formation in biofilms protects bacteria from grazing by HF in situ. The new approach provides individual-based insights into the complex food web interactions within biofilms.     

Role of the box C/D motif in localization of small nucleolar RNAs to coiled bodies and nucleoli.

Small nucleolar RNAs (snoRNAs) are a large family of eukaryotic RNAs that function within the nucleolus in the biogenesis of ribosomes. One major class of snoRNAs is the box C/D snoRNAs named for their conserved box C and box D sequence elements. We have investigated the involvement of cis-acting sequences and intranuclear structures in the localization of box C/D snoRNAs to the nucleolus by assaying the intranuclear distribution of fluorescently labeled U3, U8, and U14 snoRNAs injected into Xenopus oocyte nuclei. Analysis of an extensive panel of U3 RNA variants showed that the box C/D motif, comprised of box C', box D, and the 3' terminal stem of U3, is necessary and sufficient for the nucleolar localization of U3 snoRNA. Disruption of the elements of the box C/D motif of U8 and U14 snoRNAs also prevented nucleolar localization, indicating that all box C/D snoRNAs use a common nucleolar-targeting mechanism. Finally, we found that wild-type box C/D snoRNAs transiently associate with coiled bodies before they localize to nucleoli and that variant RNAs that lack an intact box C/D motif are detained within coiled bodies. These results suggest that coiled bodies play a role in the biogenesis and/or intranuclear transport of box C/D snoRNAs.     

In vitro induction of haploid plants from unpollinated ovules and ovaries of the sugarbeet (Beta vulgaris L.)

Summary Haploid plantlets from male fertile and male sterile sugarbeet plants could be induced at frequencies up to 2.2% using ovule culture. Ovary culture on media without charcoal resulted in a similar induction frequency. Plant development was inhibited by callus development originating from the mother tissue. When the callus parts were removed and the ovule transferred to a new medium without 2,4 D, callus formation could be inhibited by adding 0.5% charcoal to the medium. Up to 6.1% haploids were induced. Chromosome counts in leaf tips, chloroplast counts and isozyme patterns revealed that all plants were haploid and originated from the haploid cells of the embryo sac. Root tips showed spontaneous polyploidisation.     

The novel C-terminal KCNQ1 mutation M520R alters protein trafficking

The long QT-syndrome is characterized by a prolongation of the QT-interval and tachyarrhythmias causing syncopes and sudden death. We identified the missense mutation M520R in the calmodulin binding domain of the Kv7.1 channel from a German family with long QT-syndrome. Heterologous expression of the mutant did not reveal any whole-cell currents independent of the auxiliary subunit KCNE1. Co-expression of the wild-type Kv7.1 channels and the mutant showed that the mutant did not have a dominant negative effect. In immunocytochemical assays of transfected COS-1 cells wild-type Kv7.1 showed an immunopositive labeling of the plasma membrane. For M520R no plasma membrane staining was visible, instead a strong signal in the ER was observed. These results indicate that the LQT1 mutation M520R leads to ER-retention and dysfunctional trafficking of the mutant channel resulting in haploinsufficiency.     

Epigenetic effects of selenium and their implications for health

Alterations of epigenetic marks are linked to normal development and cellular differentiation as well as to the progression of common chronic diseases. The plasticity of these marks provides potential for disease therapies and prevention strategies. Macro- and micro-nutrients have been shown to modulate disease risk in part via effects on the epigenome. The essential micronutrient selenium affects human health outcomes, e.g., cancers, cardiovascular and autoimmune diseases, via selenoproteins and through a range of biologically active dietary selenocompounds and metabolism products thereof. This review provides an assessment of the current literature regarding epigenetic effects of dietary and synthetic selenocompounds, which include the modulation of marks and editors of epigenetic information and interference with one-carbon metabolism, which provides the methyl donor for DNA methylation. The relevance of a selenium-epigenome interaction for human health is discussed, and we also indicate where future studies will be helpful to gain a deeper understanding of epigenetic effects elicited by selenium.     

Diurnal actions of melatonin on epileptic activity in hippocampal slices of rats

Since melatonin receptors have been found in the hippocampus of mammals it has been suggested that melatonin can modulate neuronal functions of hippocampal cells. The effect of melatonin (10 nM/l and 1 microM/l) on frequency and amplitude of epileptiform field potentials (EFP) elicited by low Mg(2+) or by bicuculline was tested in the CA1 region of hippocampal slices of rats. In the low Mg(2+) model, melatonin, applied in a near physiological concentration of 10 nM/l, exerts no effect on EFP in slices prepared at night or during the day. In a concentration of 1 microM/l, however, melatonin enhances the frequency of EFP to approximately 140% in slices prepared during the day. This effect was suppressed through simultaneous administration of the melatonin receptor antagonist luzindole (10 microM/l). In contrast, melatonin did not affect epileptic activity in slices prepared at night. Epileptiform discharges elicited by blocking the GABAergic inhibition (bicuculline model) were not affected by melatonin, either during the day or at night. The results indicate that melatonin affects epileptic activity in a diurnal manner and that the action of melatonin is different in relation to the epilepsy model.     

Detection of electrophysiological indicators of neurotoxicity in human and rat brain slices by a three-dimensional microelectrode array

Electrophysiological techniques for the assessment of in vitro neurotoxicology have several advantages over other currently-used methods (for example, morphological techniques), including the ability to detect damage at a very early stage. Novel recording techniques based on microelectrode arrays are available, and could improve recording power. In this study, we investigated how a three-dimensional microelectrode array detects the electrophysiological endpoints of neurotoxicity. We conclude that electrophysiology sensitively reveals neurotoxic actions, and that three-dimensional microelectrode arrays could be proposed for use in neurotoxicology as recording tools that allow easy and sensitive multisite recording, from both rodent and human brain tissue.