Isolation of Allium pollen protoplasts

Studies on protoplasts isolation were carried out with mature pollen grains of 29 samples of species of Allium aflatunense, A. cepa, A. fistulosum, A. karataviense, A. longicuspis, A. nutans, A. odorum, A. sativum and A. schoenoprasum. Surface sterilized pollen grains drifted from crushed anthers were incubated in an enzyme solution containing 1% (w/v) cellulase Onozuka R-10, 1% (w/v) Macerozyme R-10, 0,5 mol l^-1 sucrose and the basal salts of Nitsch medium. Protoplasts were released within 3 to 120 min, either from the pollen grain, through a slightly disturbed germination pore (narrow aperture), or through a wider aperture, when the exine surrounding the germination pore was disturbed. For the first time, protoplasts were obtained from 13 genotypes of 6 Allium species, at a rate of 1 to 30% of the digested intact pollen grains, depending on the genotype.     

Glutamatergic plasticity by synaptic delivery of GluR-B(long)-containing AMPA receptors

Activity-driven delivery of AMPA receptors is proposed to mediate glutamatergic synaptic plasticity, both during development and learning. In hippocampal CA1 principal neurons, such trafficking is primarily mediated by the abundant GluR-A subunit. We now report a study of GluR-B(long), a C-terminal splice variant of the GluR-B subunit. GluR-B(long) synaptic delivery is regulated by two forms of activity. Spontaneous synaptic activity-driven GluR-B(long) transport maintains one-third of the steady-state AMPA receptor-mediated responses, while GluR-B(long) delivery following the induction of LTP is responsible for approximately 50% of the resulting potentiation at the hippocampal CA3 to CA1 synapses at the time of GluR-B(long) peak expression-the second postnatal week. Trafficking of GluR-B(long)-containing receptors thus mediates a GluR-A-independent form of glutamatergic synaptic plasticity in the juvenile hippocampus.     

NO participate in the communication between the first‐ and second‐order neurons in the trigeminal sensory tract in the dog

Histochemical, immunocytochemical and radioassay study was performed to detect the occurrence of NOS‐immunoreactive primary trigeminal sensory somata in the trigeminal ganglion, including their fiber components. Spinal trigeminal tract and sensory trigeminal nuclei were studied using the same methods. It was found that more than 30% of all somata in the trigeminal ganglion are NOS immunoreactive. Corresponding fibers were detected in the spinal trigeminal tract. NOS immunoreactive fibers of three different categories could be followed to terminate in the sensory trigeminal nuclei. Data presented here confirm that trigeminal sensory system is richly endowed with NOS and that NO is used to communicate between the first and second‐order trigeminal sensory neurons. Acknowledgements: Supported by VEGA Grant no. 2/3217/23PS9, STAA Grant no. 51‐013002 and by NIH grants NS 32794 and NS 40386 to M.M.     

EXPLORING THE NATURAL ROLE OF MICROCYSTINS—A REVIEW OF EFFECTS ON PHOTOAUTOTROPHIC ORGANISMS1

Cyanobacterial blooms and the production of cyanotoxins represent a serious global problem. Although the effects of a group of important cyanotoxins, microcystins (MCs), have been studied intensively in various organisms, little is known about the natural functions of these cyclic heptapeptides. MCs may have allelopathic effects. This paper summarizes the information from the studies that have investigated the effects of MCs on photoautotrophs in vitro and in vivo. Interactions with terrestrial plants, macrophytes, macroalgae, and planktonic microalgae are reported in detail with respect to the ecological relevancy of experimental conditions related to allelopathy. Our review shows that only a limited number of studies described harmful effects of MCs at concentrations that are typical for the environment. Consequently, the ability of MCs to act as general allelopathic compounds against photoautotrophs seems unlikely. However, further research is needed for definitive confirmation or rejection of the allelopathic hypothesis as well as, an explanation of the crucial question of MC function in the context of new information from evolutionary and molecular biology.     

Ca2+/recoverin dependent regulation of phosphorylation of the rhodopsin mutant R135L associated with retinitis pigmentosa

No single molecular mechanism accounts for the effect of mutations in rhodopsin associated with retinitis pigmentosa. Here we report on the specific effect of a Ca2+/recoverin upon phosphorylation of the autosomal dominant retinitis pigmentosa R135L rhodopsin mutant. This mutant shows specific features like impaired G-protein signaling but enhanced phosphorylation in the shut-off process. We now report that R135L hyperphosphorylation by rhodopsin kinase is less efficiently inhibited by Ca2+/recoverin than wild-type rhodopsin. This suggests an involvement of Ca2+/recoverin into the molecular pathogenic effect of the mutation in retinitis pigmentosa which is the cause of rod photoreceptor cell degeneration. This new proposed role of Ca2+/recoverin may be one of the specific features of the proposed new Type III class or rhodopsin mutations associated with retinitis pigmentosa.     

Separation and quantification of the cellular thiol pool of pea plants treated with heat, salt and atrazine

A novel procedure for the separation of the cellular thiol pool according to the molecular weight and localization of compounds with sulphydryl groups is presented. This simple and rapid method allows the differentiation of thiols into three major fractions-low molecular weight (LMT, primarily glutathione and free cysteine), protein-bound (TPT) and pellet-bound (PBT, associated with cell walls and broken organelles). Moreover, determination of the ratio between surface (readily reactive) thiols (ATG) and those that are more or less buried in the protein structure (BTG) can be achieved. In intact pea leaves, the amounts of the total thiols (LMT+PBT+TPT) varies from 2.5 to 4.8 micromol/g of fresh material. The data for LMT, PBT and TPT were related to each other in the approximate ratio 1:2:7. Treatments of pea plants with high temperature, salinity and low amounts of atrazine affect these sulphydryl types differently. For a greater understanding of the applicability of this method to physiological research, the main mechanisms leading to alterations in the cellular thiol pool are discussed. Furthermore, it is suggested that the proportion of available to buried thiols (ATG/BTG) in proteins could be used as a convenient marker for stress impacts.     

At a crossroads: human DNA tumor viruses and the host DNA damage response

Human DNA tumor viruses induce host cell proliferation in order to establish the necessary cellular milieu to replicate viral DNA. The consequence of such viral-programmed induction of proliferation coupled with the introduction of foreign replicating DNA structures makes these viruses particularly sensitive to the host DNA damage response machinery. In fact, sensors of DNA damage are often activated and modulated by DNA tumor viruses in both latent and lytic infection. This article focuses on the role of the DNA damage response during the life cycle of human DNA tumor viruses, with a particular emphasis on recent advances in our understanding of the role of the DNA damage response in EBV, Kaposi's sarcoma-associated herpesvirus and human papillomavirus infection.