In vitro assembly of gap junctions

Gap junction structures were assembled in vitro from octyl-β- -glucopyranoside-solubilized components of lens fiber cell membranes. Individual pore structures (connexons), short double-membrane structures, and other amorphous material were evident in the solubilized mixture. Following the removal of the detergent by dialysis, these connexons associated to form single- and double-layered, two-dimensional hexagonal arrays (unit cell size a = B = 8.5 nm). The formation of larger arrays was dependent on the lipid-to-protein ratio and the presence of Mg²⁺ ions. Crystallographic analysis of electron micrographs revealed that lens junctional connexons consisted of six subunits surrounding a stain-filled channel. Upon further detergent treatment, in vitro assembled gap junctions were insoluble and formed three-dimensional stacks while other components were solubilized. SDS-PAGE and mass data from scanning transmission electron microscopy strongly suggest that a 38-kDa polypeptide, which is a processed form of the lens specific gap junction protein MP70, is a major component of the arrays. The in vitro assembly of gap junctions opens new avenues for the structural analysis of gap junctions and for the study of the intermolecular interactions of connexons during junctional assembly.     

Quantitative trait loci associated with maximal exercise endurance in mice.

The role of genetics in the determination of maximal exercise endurance is unclear. Six- to nine-week-old F2 mice (n = 99; 60 female, 39 male), derived from an intercross of two inbred strains that had previously been phenotyped as having high maximal exercise endurance (Balb/cJ) and low maximal exercise endurance (DBA/2J), were treadmill tested to estimate exercise endurance. Selective genotyping of the F2 cohort (n = 12 high exercise endurance; n = 12 low exercise endurance) identified a significant quantitative trait locus (QTL) on chromosome X (53.7 cM, DXMit121) in the entire cohort and a suggestive QTL on chromosome 8 (36.1 cM, D8Mit359) in the female mice. Fine mapping with the entire F2 cohort and additional informative markers confirmed and narrowed the QTLs. The chromosome 8 QTL (EE8(F)) is homologous with two suggestive human QTLs and one significant rat QTL previously linked with exercise endurance. No effect of sex (P = 0.33) or body weight (P = 0.79) on exercise endurance was found in the F2 cohort. These data indicate that genetic factors in distinct chromosomal regions may affect maximal exercise endurance in the inbred mouse. Whereas multiple genes are located in the identified QTL that could functionally affect exercise endurance, this study serves as a foundation for further investigations delineating the identity of genetic factors influencing maximum exercise endurance.     

The influence of plant spacing on density-dependent versus frequency-dependent spore transmission of the anther smut Microbotryum violaceum

The anther smut fungus Microbotryum violaceum is a pollinator-transmitted plant disease. As for other vector-borne diseases, frequency-dependent transmission patterns are predicted, in contrast to the density-dependent transmission of passively spread diseases. Frequency dependence will, however, only arise if vectors compensate for varying plant spacings. To test this assumption, we set up experimental populations of the host plant, Silene latifolia, with varying disease density (number of diseased plants per plot) and frequency (proportion of plants diseased), and three different plant spacings. We measured spore deposition on healthy flowers in these plots on two dates. Spore deposition decreased considerably from the first to the second census, perhaps related to the concomitant decrease in inflorescence sizes of diseased plants. At our first census, spore deposition rates varied with disease frequency, and the effect of frequency depended on plant spacing. While spore deposition was positively frequency dependent at the 1.5-m inter-plant spacing, no effect of disease frequency was found at a spacings of 0.5 m or 3 m. Nor was there an effect of disease density on spore deposition at the first census. At the later census, on the other hand, spore deposition increased almost significantly with increasing disease density (P = 0.08). This difference in deposition pattern together with a significant decrease in spore receipt indicates changes in pollinator spectrum and/or activity. The correlation of spore numbers among flowers within plants, an indication for intra-plant moves by vectors, was significant at 0.5 m and 1.5 m but not at 3 m. Floral traits and sex of individual plants influenced the number of spores they received. On the first census date, spore deposition increased with increasing inflorescence size in female but not in male plants. On the second census date, neither sex nor number of open flowers had an effect on spore receipt. None of the experimental plants became infected, however, probably because of the unusually hot and dry weather. Received: 14 May 1998 / Accepted: 19 November 1998     

Genetic diversity in natural populations: a fundamental component of plant-microbe interactions

Genetic diversity for plant defense against microbial pathogens has been studied either by analyzing sequences of defense genes or by testing phenotypic responses to pathogens under experimental conditions. These two approaches give different but complementary information but, till date, only rare attempts at their integration have been made. Here we discuss the advances made, because of the two approaches, in understanding plant-pathogen coevolution and propose ways of integrating the two.     

Early inbreeding depression in the sexually polymorphic plant Dianthus sylvestris (Caryophyllaceae): Effects of selfing and biparental inbreeding among sex morphs

Predominantly outcrossing plant species are expected to accumulate recessive deleterious mutations, which can be purged when in a homozygous state following selfing. Individuals may vary in their genetic load because of different selfing histories, which could lead to differences in inbreeding depression among families. Lineage-dependent inbreeding depression can appear in gynodioecious species if obligatory outcrossed females are more likely to produce female offspring and if partially selfing hermaphrodites are more likely to produce hermaphrodites. We investigated inbreeding depression at the zygote, seed, and germination stages in the gynomonoecious-gynodioecious Dianthus sylvestris, including pure-sexed plants and a mixed morph. We performed hand-pollinations on 56 plants, belonging to the three morphs, each receiving 2-3 cross treatments (out-, sib- and self-pollination) on multiple flowers. Effects of cross treatments varied among stages and influenced seed provisioning, with sibling competition mainly occurring within outcrossed fruits. We found significant inbreeding depression for seed mass and germination and cumulative early inbreeding depression varied greatly among families. Among sex morphs, we found that females and hermaphrodites differed in biparental inbreeding depression, whereas uniparental was similar for all. Significant inbreeding depression levels may play a role in female maintenance in this species, and individual variation in association with sex-lineages proclivity is discussed.