The myosin filament: V. Intermediate voltage electron microscopy and optical diffraction studies of the substructure

Using a 200 kV electron microscope (JEM 200 A), thick (up to 0.4 μm) crosssections of the myosin filaments of vertebrate striated muscle were studied. It was found that: (a) with increasing section thickness the cross-sectional profiles of the shaft of the filament were increasingly more triangular and in sections 0.4 μm thick each apex of the triangle was clearly blunted. This unique cross-sectional profile is predicted by the model proposed by Pepe (1966,1967) in which 12 parallel structural units are packed to form a triangular profile with a structural unit missing at each apex of the triangle. (b) With increasing section thickness the substructure of the myosin filament was enhanced, with the best substructure visible in sections 0.2 μm to 0.3 μm thick. This strongly supports parallel alignment of structural units in the shaft of the filament as proposed by Pepe (1966,1967). (c) The substructure spacing, determined by optical diffraction from electron micrographs of cross-sections of individual myosin filaments or groups of filaments is about 4 nm. (d) The different optical diffraction patterns observed from individual myosin filaments can be explained if the projection of each structural unit in the plane of the section has an elongated profile. With a substructure spacing of 4 nm an elongated cross-sectional profile could be produced by having two myosin molecules per structural unit. Models drawn with two myosin molecules per structural unit in the model proposed by Pepe (1966,1967) gave optical diffraction patterns similar to those observed from individual filaments. (e) The different optical diffraction patterns observed from individual myosin filaments can be explained if the elongated profiles in each structural unit are similarly oriented but with the orientation changing along the length of the filament. The change in orientation per unit length of the filament must be small enough to maintain an elongated profile for the projection of the structural unit in the plane of the sections 0.3 μm thick. All of these observations and conclusions strongly support the model for the myosin filament proposed by Pepe (1966,1967).     

Food plant choices of two goldenrod beetles: Relation to plant quality

Summary The leaf beetles Trirhabda borealis and T. virgata are specialist herbivores of meadow goldenrods, Solidago spp., in central New York. Upon emergence, both larvae and adults must actively search for food plants, and must choose among the five goldenrod species (S. canadensis, S. gigantea, S. graminifolia, S. juncea and S. rugosa) that co-occur in old fields. The relationship between Trihabda foraging decisions and plant quality was examined by comparing food preferences in the field with the performance of beetles caged on each host. Trirhabda adults were highly selective in their use of food plants. Adults of T. borealis preferred a single host, S. canadensis, while T. virgata adults were most common on S. canadensis and S. gigantea. These preferences were not strictly related to variation in plant quality. In the laboratory, T. borealis performed equally well on four goldenrods (but completely failed to reproduce when fed S. graminifolia), and T. virgata performed equally well on all five hosts.Larval feeding preferences in each beetle species were broader, and were more in accord with subtle variation in plant quality. Newly-hatched T. borealis larvae readily colonized four hosts and rejected only S. graminifolia, which conferred the lowest survivorship and the slowest growth. Larvae of T. virgata accepted each host, even though growth rates were somewhat slower on S. graminifolia and S. juncea.Ontogenetic differences in host preference and host tolerance may have evolved because of the different host-finding abilities of each beetle stage. During host search, the sluggish, newly-emerged larvae may be more food-limited and accept even marginally inferior food plants. The relatively mobile adults are more discriminating and use a subset of suitable hosts. The intermingled dispersion of Solidago species in old fields results in frequent larval colonization of hosts seldom used by adults. In diverse plant communities, ultimate patterns of food plant choice can be a complex function of at least three factors: intrinisic plant quality, local plant dispersion, and the host-search abilities of the insect forager.     

The size of the extracellular space in the isolated midgut ofManduca sexta

The size of the extracellular space in the isolated midgut ofManduca sexta measured statically with labeled sucrose and with labeled inulin was 48 and 50 percent of tissue water respectively. Both labeled sulfate and labeled mannitol yielded loading spaces which approached the volume of the tissue water with increasing pulse time and are not valid markers of the extracellular space. A washout method yielded unreliable values for the sucrose and inulin spaces and its use cannot be justified using presently available midgut chamber designs. Values of the sucrose space measured statistically in this study and other studies using different Lepidopteran larvae and different perfusion chambers range from 42 to 48 percent of the tissue water. Agreement between published values of the sulfate space and these values of the sucrose space is fortuitous, the labeling pulses being short enough to yield approximately half saturation of the actual sulfate loading space.     

Genetics of Ia-like alloantigens in chickens and linkage with B major histocompatibility complex

Two sets of backcross matings were performed to test for linkage between genes coding for the Ia-like antigens ("Ia") and the B erythrocyte antigens (Ea-B) of the chicken. Evidence is presented which indicates that the "Ia" antigens are determined by a single codominant locus and that the Ea-B and "Ia" loci are on the same chromosome. Failure to detect a single recombinant between the Ea-B and "Ia" loci out of 208 progeny suggests close linkage of the two genes with a map distance of up to about 2 centimorgans. The "Ia" genes are thus included in the B major histocompatibility complex of the chicken.     

Technique for determining total bacterial virus counts in complex aqueous systems.

A direct method is described for measuring bacteriophage concentrations in complex aqueous systems. Conditions for sample clarification, phage recognition, and recovery were optimized. In contrast to the plaque assay, this procedure permits quantitation of total numbers of phages independent of bacterial host. Also, the modifications increase the sensitivity of the sedimentation assay, permitting detection of particles at a minimum concentration of 10(4) per ml. Maximal total phage concentrations in the aqueous phase of sewage and activated sludge mixed liquor were 1.3 x 10(6) and 4.3 x 10(7) per ml, respectively.     

The prosthetic group of methylamine dehydrogenase from Pseudomonas AM1: evidence for a quinone structure

The g-value and linewidth of ESR spectra of methylamine dehydrogenase (primary-amine:(acceptor) oxidoreductase (deaminating) EC 1.4.99.-) and methanol dehydrogenase (alcohol:(acceptor) oxidoreductase, EC 1.1.99.8) are very similar. This similarity is also reflected in electron-nuclear double resonance (ENDOR) results, the coupling constants of two protons in one enzyme equalling those in the other. The presence of a third proton in the ENDOR spectrum of methylamine dehydrogenase suggests a different structure or a different kind of interaction which can be related to the finding that the resolved ROSTHETIC GROUP IS PROTEIN-BOUND. The bound prosthetic group has a high redox-potential, supporting the conclusion from the ESR and ENDOR results that it is a quinone derivative.     

Early events in neuromuscular junction formation in vitro: induction of acetylcholine receptor clusters in the postsynaptic membrane and morphology of newly formed synapses

The development of clusters of acetylcholine (ACh) receptors at newly formed synapses between embryonic chick spinal cord and muscle cells grown in vitro has been studied by iontophoretic mapping with ACh. A semi-automated technique using on-line computer analysis of ACh responses and a photographic system to record the position of each ACh application permit the rapid construction of extensive and detailed maps of ACh sensitivity. Clusters of receptors, evident as peaks of ACh sensitivity, are present on many uninnervated myotubes. The distribution of ACh sensitivity closely parallels the distribution of 125I-alpha-bungarotoxin binding sites on the same muscle cell. In all cases where individual myotubes were adequately mapped before and after synapse formation, ingrowing axons induced new clusters of receptors rather than seeking out preexisting clusters. Synapses can form at active growth cones within 3 h of nerve-muscle contact. New receptor clusters can appear beneath neurites within a few hours. Many of the uninnervated clusters on innervated myotubes disappear with time. In contrast, receptor clusters on uninnervated myotubes remain in the same location for many hours. Synaptic clusters and clusters on uninervated myotubes are stable even though individual receptors are metabolized rapidly. The morphology of several identified sites of transmitter release was examined. At the scanning EM level, synapses appeared as small, rough-surfaced varicosities with filopodia that radiated outwards over the muscle surface. One synapse was studied by transmission EM. Acetylcholinesterase and a basement lamina were present within the synaptic cleft.     

Mutations affecting the formation of acetohydroxy acid synthase II in Escherichia coli K-12.

Summary Genetic mapping experiments have established that two recently isolated valine-resistant mutants of the K-12 strain of Escherichia coli have lesions lying between ilvE and rbs. These lesions allowed expression of the ilvG gene, specifying the valine-insensitive acetohydroxy acid synthase (synthase II) and an increased expression of the ilvEDA operon. In this respect, they resembled an earlier described ilvO lesion that was reported to lie between ilvA and ilvC. All three lesions were cis-dominant in cis-trans tests. Reexamination of the earlier studied ilvO lesion revealed that it, too, lies between ilvE and rbs. Valine-sensitive derivatives with lesions presumed to be in ilvG were selected from each of the valine-resistant strains. In two of the valine-resistant strains, the ilvG mutations were on the rbs side of ilvO, indicating a gene order rbs-ilvG-ilvO-ilvE-ilvD-ilvA-ilvC. In one of the recently isolated valine-resistant stocks, however, the apparent ilvG mutation was found to be between ilvE and the aline resistance marker. This finding suggests that either ilvO and ilvG mutations are interspersed or there is another locus, ilvR, that behaves phenotypically like ilvO and which lies between ilvG and rbs.     

Alkaloids of Zanthoxylum monophyllum and Z. punctatum

The known alkaloids N-methylcorydine and magnoflorine were major constituents of stems and branches of Zanthoxylum punctatum (Rutaceae). Berberine was the major alkaloid of Z. monophyllum, which also contained a new pyrano-2-quinolone alkaloid, zanthophylline, and its desmethyl analog. Zanthophylline has an —NCH₂OAc functional group which is readily hydrolyzed by dilute acid. Methyl vanillate and the furocoumarin columbianetin were also found in Z. monophyllum.