Inactivation of phosphorylase is a major component of the mechanism by which insulin stimulates hepatic glycogen synthesis.

Multiple signalling pathways are involved in the mechanism by which insulin stimulates hepatic glycogen synthesis. In this study we used selective inhibitors of glycogen synthase kinase-3 (GSK-3) and an allosteric inhibitor of phosphorylase (CP-91149) that causes dephosphorylation of phosphorylase a, to determine the relative contributions of inactivation of GSK-3 and dephosphorylation of phosphorylase a as alternative pathways in the stimulation of glycogen synthesis by insulin in hepatocytes. GSK-3 inhibitors (SB-216763 and Li+) caused a greater activation of glycogen synthase than insulin (90% vs. 40%) but a smaller stimulation of glycogen synthesis (30% vs. 150%). The contribution of GSK-3 inactivation to insulin stimulation of glycogen synthesis was estimated to be less than 20%. Dephosphorylation of phosphorylase a with CP-91149 caused activation of glycogen synthase and translocation of the protein from a soluble to a particulate fraction and mimicked the stimulation of glycogen synthesis by insulin. The stimulation of glycogen synthesis by phosphorylase inactivation cannot be explained by either inhibition of glycogen degradation or activation of glycogen synthase alone and suggests an additional role for translocation of synthase. Titrations with the phosphorylase inactivator showed that stimulation of glycogen synthesis by insulin can be largely accounted for by inactivation of phosphorylase over a wide range of activities of phosphorylase a. We conclude that a signalling pathway involving dephosphorylation of phosphorylase a leading to both activation and translocation of glycogen synthase is a critical component of the mechanism by which insulin stimulates hepatic glycogen synthesis. Selective inactivation of phosphorylase can mimic insulin stimulation of hepatic glycogen synthesis.     

The importance of carotenoids in signaling during aggressive interactions between male firemouth cichlids (Cichlasoma meeki)

Male firemouth cichlids, Cichlasoma meeki, have red pigmentationcovering large areas of their ventral surface, which is displayedduring aggressive interactions. We manipulated the levels ofred pigmentation by assigning the fish to one of two diets,which were as similar as possible except that one was high incarotenoids while the other was low in carotenoids. During diadictrials under white light, fish kept on the high carotenoid dietwon a higher proportion of contests than fish kept on the lowcarotenoid diet Under green light, where differences in rednesscannot be discriminated, there was no effect of diet on theoutcome of contests. These experiments demonstrate that it isthe effect of the diet on red pigmentation that is importantrather than some confounding variable such as differential growthrates. The weight of the two fish was also important; therewas a tendency for the heavier fish to win more contests. Themass effect was subordinate to color under white light but wasthe dominant factor under green light The nature of the contestsunder the different light conditions also varied; the displayin which the red pigmentation is most obvious was not used undergreen light, but was common under white light This suggeststhat the display strategies are flexible and can be alteredaccording to which displays are most effective in a given environmentPrevious studies of other species of fish and birds have shownthat the degree of redness influences mate choice and is affectedby parasite infestations. We propose that carotenoid pigmentationis likely to reflect a general quality, influenced by severalfactors, rather than a context-specific quality such as fightingability.     

An experimental analysis of mate choice in the wren: a monomorphic, polygynous passerine

Male wrens (Troglodytes troglodytes) construct nests that areused in their display to females. Previous work has suggestedthat the number of vacant nests may be used as a mate choicecue. Correlational data from 1992 confirmed that females appearedto be assessing die number of vacant nests on a male's territoryand preferentially mating with males with more nests. Male taillengdi was also correlated widi mating success. In 1993 thenumbers of nests on territories was experimentally manipulated,the female setdement patterns confirmed that die number of vacantnests did mediate mate choice. Male tail length failed to explainadditional variance in mating success when die variance explainedby the experimental manipulation was removed, suggesting diatdie original correlation arose because both tail length andmating success were correlated widi a confounding variable.The structure of the vegetation in a male's territory influencedmating success. This appeared to be due to nests surviving betterin territories widi dense vegetation. Males on territories inwhich nests survive well had longer tails. Male-male competitionfor good territories may explain die observed effects of malemorphology on mating success. Furdier analysis of die nest choicedata showed diat all nests had an equal chance of being usedby a female. The fact diat all nests had an equal probabilityof being chosen by a female means diat each additional nestbuilt by a male wren results in die same increase in matingsuccess. This suggests diat die benefits to males of nest buildingincrease linearly. The number of nests on a territory will beaffected by various factors such as predation pressure, nestbuilding rate, and vegetation structure. The information diatfemales are getting by assessing such a signal is discussed.     

Altered SOS induction associated with mutations in recF, recO and recR

The SOS system of Escherichia coli aids survival following damage to DNA by promoting DNA repair while cell division is delayed. Induction of the SOS response is dependent on RecA and also on the product of recF. We show that normal induction also requires the products of recO and recR. SOS induction was monitored using a sfiA-lacZ fusion strain. Induction was delayed to a similar degree by mutation in recF, recO or recR. A similar effect was observed following overexpression of RecR from a recombinant recR ⁺plasmid. We show that the overexpression of RecR also reduces the UV resistance of a recBC sbcBC strain and of a sfiA strain, but not of a rec ⁺ sfiA ⁺strain. The implications of these data for the kinetics of DNA repair are discussed.     

Gene inactivation in the plant pathogen Glomerella cingulata: three strategies for the disruption of the pectin lyase gene pnIA

The feasibility of performing routine transformation-mediated mutagenesis in Glomerella cingulata was analysed by adopting three one-step gene disruption strategies targeted at the pectin lyase gene pnIA. The efficiencies of disruption following transformation with gene replacement- or gene truncation-disruption vectors were compared. To effect replacement-disruption, G. cingulata was transformed with a vector carrying DNA from the pnlA locus in which the majority of the coding sequence had been replaced by the gene for hygromycin B resistance. Two of the five transformants investigated contained an inactivated pnlA gene (pnlA ^– );both also contained ectopically integrated vector sequences. The efficacy of gene disruption by transformation with two gene truncation-disruption vectors was also assessed. Both vectors carried a 5and 3truncated copy of the pnlA coding sequence, adjacent to the gene for hygromycin B resistance. The promoter sequences controlling the selectable marker differed in the two vectors. In one vector the homologous G. cingulata gpdA promoter controlled hygromycin B phosphotransferase expression (homologous truncation vector), whereas in the second vector promoter elements were from the Aspergillus nidulans gpdA gene (heterologous truncation vector). Following transformation with the homologous truncation vector, nine transformants were analysed by Southern hybridisation; no transformants contained a disrupted pnlA gene. Of nineteen heterologous truncation vector transformants, three contained a disrupted pnlA gene; Southern analysis revealed single integrations of vector sequence at pnlA in two of these transformants. pnlA mRNA was not detected by Northern hybridisation in pnlA-transformants. pnlA-transformants failed to produce a PNLA protein with a pI identical to one normally detected in wild-type isolates by silver and activity staining of isoelectric focussing gels. Pathogenesis on Capsicum and apple was unaffected by disruption of the pnlA gene, indicating that the corresponding gene product, PNLA, is not essential for pathogenicity. Gene disruption is a feasible method for selectively mutating defined loci in G. cingulata for functional analysis of the corresponding gene products.     

Genetical and physical assignments of equine microsatellites—first integration of anchored markers in horse genome mapping

Twenty equine microsatellites were isolated from a genomic phage library, and their genetical and physical localization was sought by linkage mapping and fluorescent in situ hybridization (FISH). Nineteen of the markers were found to be polymorphic with, in most cases, heterozygosities exceeding 50%. The markers were mapped in a Swedish reference family for gene mapping, comprising eight half-sib families from Standardbred and Icelandic horse sires. Segregation was analyzed against a set of 35 other markers typed in the pedigree. Thirteen of the microsatellites showed linkage to at least one other marker, with a total of 21 markers being involved in these linkages. In parallel, 18 of the microsatellites could be assigned to their chromosomal region by FISH. These assignments involved eight equine autosomes: ECA1, 2, 4, 6, 9, 10, 15, and 16. The genetical and physical mappings revealed by this study represent a significant extension of the current knowledge of the equine genome map. Received: 24 September 1996 / Accepted: 1 December 1996