LH-β and FSH-β mRNA expression in nesting and post-breeding three-spined stickleback, Gasterosteus aculeatus, and effects of castration on expression of LH-β, FSH-β and spiggin mRNA

The mRNA expression of the LH- and FSH- subunits were measured in nesting and post-breeding male three-spined sticklebacks, Gasterosteus aculetaus, as well as in castrated and sham-operated nesting males. Furthermore, expression of an androgen induced kidney protein, spiggin, and 11-ketotestosterone (11KT) levels, were measured in the castrated and sham-operated males. Nesting males had significantly higher levels of both LH- and FSH- mRNA expression compared to post-breeding males. Furthermore, sham-operated males had significantly higher levels of LH- mRNA and spiggin mRNA expression than the castrated fish. Expression of FSH-, on the other hand, did not differ between castrated and sham-operated males. There were strong positive individual correlations between circulating levels of 11KT on the one hand and expressions of LH- and spiggin mRNA, whereas the correlation between 11KT levels and FSH- mRNA was weak. The negative effect of castration on -LH mRNA indicates that gonadal hormones stimulate this expression, whereas this was not the case for -FSH. The observed decline in -LH expression after the end of the breeding season may be the result of cessation of the gonadal stimulation of the pituitary. On the other hand, it is not likely that this can explain the decline in FSH- expression.     

RAPD analysis of interspecific relationships in presumably apomictic Cotoneaster species

A few of the approximately 300 Cotoneaster species described are diploid but the majority appear to be polyploid. The occurrence of apomixis inpolyploid Cotoneaster species has been reported but never proven with genetic markers. We have used 76 polymorphic RAPD markers to investigate the breeding system and phenetic grouping of some critical taxa, including a total of 19 plant accessions representing 13 mostly European species in the series Cotoneaster. Three to four individual plants, raised from seed from the same original plant, were analyzed for each of three accessions to investigate the possible occurrence of apomictic seed set. Absolutely congruent RAPD profiles were encountered among seedlings from one accession, whereas we found one or two marker differences among seedlings from the other two accessions. Genetic similarities among the different accessions were analyzed with a UPGMA-derived dendrogram. The most deviant taxon was the Chinese C. albokermesinus. A group withC. soczavianus and C. tomentosus was rather isolated from the remainder, as was also C. kullensis. Among the remaining taxa, two well supported clusters were found: (1) C. antoninae and C. uralensis, and (2) C. integerrimus and C. raboutensis, whereas the other five species (C. canescens, C. niger, C. scandinavicus, C. juranus, C. cambricus) formed a poorly supported cluster with no clear substructuring. A principal coordinate analysis yielded results that were in good correspondence with the dendrogram. Again C. albokermesinus appears to be totally isolated from the other species. In addition, two well-defined and rather isolated groups were found: (1) C. tomentosus and C. soczavianus, and (2) C. antoninae and C. uralensis, with the remainder comprising a loosely defined group. This revised version was published online in July 2006 with corrections to the Cover Date.     

Global allocation rules for patterns of biomass partitioning in seed plants

A general allometric model has been derived to predict intraspecific and interspecific scaling relationships among seed plant leaf, stem, and root biomass. Analysis of a large compendium of standing organ biomass sampled across a broad sampling of taxa inhabiting diverse ecological habitats supports the relations predicted by the model and defines the boundary conditions for above- and below-ground biomass partitioning. These canonical biomass relations are insensitive to phyletic affiliation (conifers versus angiosperms) and variation in averaged local environmental conditions. The model thus identifies and defines the limits that have guided the diversification of seed plant biomass allocation strategies.     

Nutrient and persistent pollutant deposition on the Bothnian Bay ice and snow fields

A study of atmospheric deposited nutrients and some persistent pollutants has been carried out on the ice and snow fields of the subarctic Bothnian Bay, the northernmost basin of the Baltic Sea. Total amounts of 600, 500 and 400 tons NO₃-N, NH₄-N and N_org-N, respectively are deposited in the snow while the corresponding amounts for P_tot-P is 40 tons. The corresponding amount for PCB and lindane are 1.0 and 0.2 kg, respectively. The measurements were carried out on the snow-covered ice four to six weeks old. A part of the deposited snow has been incorporated -into the snow-ice and an attempt to estimate its mean thickness and its amount of nutrients has been made. The total amounts now reach 1700, 1300 and 1100 tons of NO₃-N and NH₄-N and N_org-N, respectively in the snow and ice together. The results obtained support the use of land-based stations in estimates of seasonal atmospheric nutrient deposition to the Bothnian Bay. The observed concentrations of chloroorganic compounds correspond to those land-based observations reported from the same latitude in the northern hemisphere and reported in literature.     

Rapid chemically induced changes of PtdIns(4,5)P2 gate KCNQ ion channels

To resolve the controversy about messengers regulating KCNQ ion channels during phospholipase C-mediated suppression of current, we designed translocatable enzymes that quickly alter the phosphoinositide composition of the plasma membrane after application of a chemical cue. The KCNQ current falls rapidly to zero when phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2 or PI(4,5)P2] is depleted without changing Ca2+, diacylglycerol, or inositol 1,4,5-trisphosphate. Current rises by 30% when PI(4,5)P2 is overproduced and does not change when phosphatidylinositol 3,4,5-trisphosphate is raised. Hence, the depletion of PI(4,5)P2 suffices to suppress current fully, and other second messengers are not needed. Our approach is ideally suited to study biological signaling networks involving membrane phosphoinositides.     

Modulation of postendocytic sorting of G protein-coupled receptors

Recycling of the mu opioid receptor to the plasma membrane after endocytosis promotes rapid resensitization of signal transduction, whereas targeting of the delta opioid receptor (DOR) to lysosomes causes proteolytic down-regulation. We identified a protein that binds preferentially to the cytoplasmic tail of the DOR as a candidate heterotrimeric GTP-binding protein (G protein)-coupled receptor-associated sorting protein (GASP). Disruption of the DOR-GASP interaction through receptor mutation or overexpression of a dominant negative fragment of GASP inhibited receptor trafficking to lysosomes and promoted recycling. The GASP family of proteins may modulate lysosomal sorting and functional down-regulation of a variety of G protein-coupled receptors.     

Directional spread of an alpha-herpesvirus in the nervous system

Pseudorabies virus (PRV), an alpha-herpesvirus, is capable of spreading between synaptically connected neurons in diverse hosts. In this report, two lines of experimentation are summarized that provide insight into the mechanism of virus spread in neurons. First, techniques were developed to measure the transport dynamics of capsids in infected neurons. Individual viral capsids labeled with green fluorescent protein (GFP) were visualized and tracked as they moved in axons away from infected neuronal cell bodies in culture during egress. Second, the effects of three viral membrane proteins (gE, gI and Us9) on the localization of envelope, tegument, and capsid proteins in infected, cultured sympathetic neurons were determined. These three proteins are necessary for spread of infection from pre-synaptic neurons to post-synaptic neurons in vivo (anterograde spread). Us9 mutants apparently are defective in anterograde spread in neural circuits because essential viral membrane proteins such as gB are not transported to axon terminals to facilitate spread to the connected neuron. By contrast, gE and gI mutants manifest their phenotype because these proteins most likely function at the axon terminal of the infected neuron to promote spread. These two sets of experiments are consistent with a model for herpesvirus spread in neurons first suggested by Cunningham and colleagues where capsids and envelope proteins, but not whole virions, are transported separately into the axon.