The horizontal cells in the lobula plate of the blowfly,Phaenicia sericata

SummarySummary Combining intracellular recording and dye injection techniques, the horizontal cells of the blowfly,Phaenicia (= Lucilia) sericata, were studied.Anatomy In each lobula plate, one finds a set of three cells, termed NH-, EH- and SH-cell. EH occurs in two distinct anatomical forms, EH1 and EH2, differing in their respective branching patterns of the axon at the frontal surface of the lobula plate. Each cell's dendrite covers approximately a third of the surface of the lobula plate corresponding to a third of the visual field of the ipsilateral eye. These dendrites possess postsynaptic spines. The axons of all three cells pass along the frontal surface of the lobula plate within the inner chiasma; they cross the optic peduncle and enter the central protocerebrum where they form a second arborization, the axonal arborization consisting of dorsally extending collaterals. The axons terminate in the posterior slope of the ventrolateral protocerebrum. The axonal arborization as well as the axonal terminals possess telodendritic knobs. Ultrastructural investigations show that the lobula plate-dendrite possesses exclusively postsynaptic chemical synapses, and that the axonal arborisation and the axonal terminals possess pre- as well as postsynaptic chemical synapses. The very endings of the axons are exclusively presynaptic.Physiology The horizontal cells respond to stimulation within the ipsi- and/or contralateral receptive field. Regressive motion within the contralateral receptive field induces EPSPs and action potentials of small amplitude (10–35 mV); progressive motion is ineffective. Within the ipsilateral receptive field, regressive motion hyperpolarizes the cell membrane whereas progressive motion induces a strong depolarizing membrane potential-shift with superimposed fast potential changes of noisy appearance. Thus, the horizontal cells respond to rotational movement of the surround around the high axis of the animal: clockwise rotation excites the horizontal cells of the right lobula plate and counterclockwise motion those of the left lobula plate, respectively. However, this compound potential behaviour can only be recorded in the lobula plate-axon and main dendrites, whereas the horizontal cells respond tocontralateral regressive motion with action potentialsonly in their axonal terminals in the posterior slope; no graded potentials can be recorded in this cell region if stimulation occurs within theipsilateral receptive field. It is discussed that the previously described graded potentials for the axonal terminals (Hausen 1976b) can only be measured if the cells are already damaged. The probable cause of this change in response behaviour from action potentials to a compound potential behaviour (consisting of graded potentials and action potentials though of small amplitude) is discussed.This research was supported by the Deutsche Forschungsgemeinschaft through grants Ec56/1a + b and a Heisenberg stipend EC 56/3, funds from the SFB 114, and a grant from the National Science Foundation (NSF BMS 74-21712) awarded to the author and L.G. Bishop. I am indebted to Dr. A. Whittle and particularly to Prof. K. Meller for their invaluable help in ultrathin sectioning and to Mrs. B. Decker who introduced me to the technique of cutting serial semithin sections. Prof. K. Hamdorf helped with many stimulating discussions. I am most grateful to Dr. W. Broughton for kindly correcting the English style.     

Role of cytoplasmic proteins in cold shock injury of Amoeba

Strains of Amoeba have been used to study the mechanisms of cellular injury induced by rapid cooling (cold shock). Cell viability was found to depend on the time and temperature of cold exposure, on the rate of cooling and on the morphology of the cells prior to chilling. All strains underwent a granuloplasmic contraction following undercooling to ?10 °C, although its extent varied; strains most damaged by cold shock exhibited the most violent cytoplasmic contractions. Cryomicroscopy demonstrated that the cellular contraction occurred upon rewarming, not during cooling. Cells damaged by cold shock were osmotically responsive, demonstrating that irreversible damage to the plasmalemma does not account for the phenomenon.Several compounds protected Amoeba against cold shock injury, glycerol and glucose being the most effective. With glycerol an optimum rate of cooling was observed upon cooling to ?10 °C, at both faster and slower cooling rates damage increased.The state of cellular actin in control cells and following cold shock was monitored by the DNase 1 inhibition assay and by electron microscopy. A comparatively “cold shock resistant” strain of A. proteus was found to contain less total actin per unit cellular protein than the more “sensitive” Amoeba sp. strain Bor. In the Bor strain a cold-induced aggregation of cytoplasmic filaments was evident in electron micrographs, presumably a crosslinking of preexisting F-actin.     

Discussion on the state of water in the myofilament lattice and other biological systems, based on the fact that the usual concepts of colloid stability can not explain the stability of the myofilament lattice

Application of the usual concepts of colloid stability shows that the in vivo spacings between the myofilaments, making up the contractile part of the muscle myofibrils, correspond to energies of 10(-4) to 10(-1) kT. Refinements in the calculations of the electrostatic and Van der Waals-London energies do not significantly modify these values. Therefore, theory does not predict the observed stability of the myofilament lattice. It is shown that the interfilament water very likely plays an active role in the myofilament lattice. More generally, the structure of water in living cells is probably different from that of bulk water.     

Biosynthesis of cyclopiazonic acids in Penicillium cyclopium: The isolation of dimethylallylpyrophosphate: Cyclo-acetoacetyltryptophanyl dimethylallyltransferase

During the production of β-cyclopiazonic acid (βCA) by Penicillium cyclopium, the following points are noted: (a) Dimethylallylpyrophosphate (DMAPP), which is incorporated into α-cyclopiazonic acid (αCA) in vivo, stimulates overall CA synthesis, whereas tryptophan, although incorporated into αCA, inhibits overall CA synthesis. (b) A previously suggested substrate, γ,γ-dimethylallyltryptophan, is not a precursor of αCA. (c) The accumulation of cyclo-acetoacetyl-l-tryptophanyl (cAATrp) is described in both the culture medium and mycelium with increasing growth. (d) A cell-free extract of mycelium will catalyze the conversion of exogenous cAATrp and exogenous DMAPP into βCA in 1:1 stoichiometry, the βCA being bound to a protein.     

The structure of steady-state enzyme kinetic equations: a graph-theoretical algorithm for obtaining conditions for reduction in degree by common-factor cancellation

Many enzyme kinetic steady-state equations are so complicated that analysis of their predictions, preferably by graph-theoretical methods, without deriving the equations is desirable. An example of such a method is given here. It is a graph-theoretical algorithm for determining non-trivial relations between the rate constants of a mechanism that cause the numerator and denominator of its rate expression have a common factor so permitting the degree of the expression to be reduced. An algorithm for writing the several different forms of the reduced rate equation is also given. The algorithm is applied to some standard simple enzymic mechanisms that give relatively complicated rate equations.In the case of a 2:2 equation, the sign of curvature in a double reciprocal plot depends on the sign of the expression in the common-factor condition.     

Fiber optic mapping of the Xenopus visual system: shift in the retinotectal projection during development

Two new techniques for assaying the retina to tectum connections in the lower vertebrate visual system are presented. These techniques allow defined regions of the retina to be stimulated, thus circumventing some of the difficulties of the more conventional retinotectal mapping techniques. Applying these techniques to the Xenopus visual system demonstrates that the retina-to-tectum projection shifts during development. The central part of the retinotectal projection moves medially and caudally about 150 microns (10% of the size of the tectum) in two weeks. The presence of such plasticity in a normal developing animal indicates that the plasticity previously observed in experimentally altered animals probably reflects a normal developmental process.     

The murine sublingual and submandibular mucins their isolation and characterization

From the mouse sublingual and submandibular glands high-molecular weight glycoproteins (mucins) were isolated. These mucins appeared to be homogeneous in polyacrylamide gel electrophoresis and in the analytical ultracentrifuge. S_20,w values of 10.9 and 5.5 were found for the sublingual and submandibular mucin respectively. With sodium dodecyl sulfate or N-acetylcysteine no subunits could be detected.Both mucins consisted for about 1/3 of protein and 2/3 of carbohydrate. Their mucin character was also denoted by the high content of serine plus threonine. Respectively 42 mol% and 34 mol% of the protein core of the sublingual and submandibular mucins consisted of these amino acids. The main sugars in these mucins were sialic acid, galactosamine, galactose, glucosamine and mannose. The molar ratio for the sublingual and submandibular mucin being 1.00 : 1.03 : 1.08 : 0.26 : 0.23 and 1.00 : 0.71 : 1.10 : 0.65 : 0.53, respectively.The sialic acid content of both mucins was about 25%. Fucose and sulfate, on the other hand, were less than 1%. The presence of sulfate was also indicated by preliminary studies in vivo on the incorporation of [³⁵SO₄] sulfate.     

The surface potential of and double-layer interaction force between surfaces characterized by multiple ionizable groups.

At the beginning of cleavage, a cell develops the form of a short central cylinder capped at the ends with ellipsoids. The strain pattern produced by this shape, with the application of constricting forces, guides the cell either to divide into two spherical cells or to take the shape of a long, thin cell which may remain a single cell or divide into two thin cells.     

Biomass accumulation and shading effects of epiphytes on leaves of the seagrass, Heterozostera tasmanica, in Victoria,Australia

A method is described for estimating the rate of accumulation of epiphyte biomass on leaves of the seagrass, Heterozostera tasmanica (Martens ex Aschers.) den Hartog and for estimating the effect of epiphyte biomass on photosynthesis of the seagrass. Epiphyte biomass was determined by comparison of the weight per unit area of epiphyte-covered and epiphyte-free leaf blades. Epiphyte weight increased as age of the seagrass leaves increased. Linear regression on epiphyte biomass vs. leaf age estimated the rate of biomass accumulation. Rates varied from 5.7 to 104 μg epiphyte dry weight per cm² of leaf surface per day at three sites in Western Port and Port Phillip Bay, Victoria. Rates of accumulation of epiphyte biomass were generally higher during December through March (summer) than in May (autumn), August (winter) or October (Spring). Light attenuation by epiphytes increase linearly with biomass. The rate of biomass accumulation of epiphytes was compared with leaf growth rate, ambient photon flux density in H. tasmanica beds and the photosynthesis—photon flux density curve of H. tasmanica. This comparison demonstrated that epiphyte biomass can accumulate fast enough to shade H. tasmanica leaves and significantly reduce the time (to less than one half of the leaf life span) in which positive net photosynthesis of the leaf blade is possible.     

On Zeeman's equations for the nerve impulse

A model for the nerve impulse due to Zeeman (1972) and based on catastrophe theory is compared with alternative models and criticisms of Zeeman's model by Sussmann and Zahler (1977, 1978) are assessed. The criticisms of Zeeman's motivation for his model are found to carry some weight. Sussmann and Zahler (1977, 1978) list numerous features of Zeeman's model which, they state, are not in agreement with experiment. These statements as they stand are largely erroneous, and the model still remains to be tested by a critical series of experiments. However, a detailed analysis reveals defects in Zeeman's model, not among those claimed by Sussmann and Zahler, showing that the explicit equations of the model cannot be correct. The possibility of a modified approach along similar lines and its ultimate adoption remains open.     

Comparison of the ultrastructure of stimulated and unstimulated mechanoreceptors in the taste hairs of the blowfly Phaenicia serricata

The structure of mechanoreceptors at the base of labeilar taste hairs of the blowfly Phaenicia serricata were examined in stimulated and unstimulated conditions (i.e. with the hair bent or unbent). Physiological recordings from the mechanoreceptor showed that the receptors responded when the hair is bent dorsally or ventrally and when the hair is bent at extreme angles. These conditions are the same as those placed on hairs in the anatomical studies. Bending the hair toward the ventral labellar surface caused the hair base to compress and indent the tubular body and its surrounding membrane and sheath at the distal end of the mechanoreceptor dendrite. In compressed tubular bodies, microtubules oriented longitudinally were bent and separated a greater distance from each other. Separation as much as 70 nm was observed in compressed tubular bodies as compared with a maximum of 26 nm between microtubules in tubular bodies of unbent hairs. The dense amorphous material between microtubules of compressed tubular bodies formed prominent bridges 18 nm thick connecting the microtubules at intervals of 48–74 nm. Thin 10 nm filaments were also evident in the spaces between microtubules. When the hair was bent toward the proximal end of the proboscis, the tip of the tubular body was bent about 15 °. The tubular body appears to function as a firm but resilient structure over which the dendritic membrane can be stretched during mechanostimulation. Comparison of morphology of bent and unbent hairs suggests a means by which mechanical force from the movement of the hair is transferred to the receptors by structures in the hair socket region. No differences were found in ciliary structures of stimulated and unstimulated receptors.     

Microtubule arrays in the cortex and near the germinal vesicle of immature starfish oocytes

An extensive array of long, crisscrossing microtubules has been discovered in the cortex of oocytes of the starfish Pisaster ochraceus. The microtubules were visualized in cortex preparations by indirect immunofluorescence microscopy using antibodies to tubulin. The cortical array of microtubules is present in all oocytes before and for about 30 min after the application of 1-methyladenine, the hormone that induces oocyte maturation. The presence of microtubules was confirmed by electron microscopy. The microtubules in this array are depolymerized when oocytes are treated with colchicine or nocodozole and are augmented when oocytes are treated with taxol. Dihydrocytochalasin B treatment of the oocytes causes the microtubules to aggregate, presumably by altering a microfilament network also found in the cortex. The distribution of microtubules was also explored in whole oocytes stained with antitubulin. One or two aster-like structures were observed adjacent to the germinal vesicle of each oocyte.     

Local energized proton hypotheses of mitochondrial oxidative phosphorylation

Two hypotheses are compared each interpreting mitochondrial energy transduction in terms of a localized form of proton activity. Their differences are seen to be profound and far-reaching. It is concluded that the "chemiosmotic/local energized proton dialogue" as conducted hitherto has offered a very incomplete and restricted analysis of the problems of mitochondrial oxidative phosphorylation.     

Flash photolysis-electron spin resonance study of the effect of o-phenanthroline and temperature on the decay time of the ESR signal B1 in reaction-center preparations and chromatophores of mutant and wild strains of Rhodopseudomonas spheroides and Rhodospirillum rubrum

We have studied the effect of o-phenanthroline and temperature on the decay rate of Signal B1 in reaction-center preparations and in chromatophores from Rhodopseudomonas spheroides and Rhodospirillum rubrum. We have shown that o-phenanthroline binds specifically to the reaction center protein (the binding center is probably at the iron) and when so bound inhibits the transfer of electrons from primary to secondary acceptors. We have also shown that the direct return decay time (A^? → P865⁺) increases with increasing temperature above approx. 150 K. This phenomenon has been interpreted within a quantum mechanical tunnelling model in which the distance of closest approach between P865⁺ and A^? increases about 2 Å between approx. 150 and 300 K.     

Assembly-disassembly of actin bundles in starfish oocytes: an analysis of actin-associated proteins in the isolated cortex

One rapid response of starfish oocytes to the maturation-inducing hormone, 1-methyladenine (1-MA), is the formation of transient actin-filled spikes on the cell surface. The presence and distribution of G- and F-actin and several actin-associated proteins were examined in cortices isolated from oocytes before, during, and after spike formation by using antibodies and the F-actin-specific stain, NBD-phallacidin. Before 1-MA addition, staining with antiactin and NBD-phallacidin indicates that most of the actin in the cortex is either G-actin or oligomeric actin, but rather little is F-actin. Application of the hormone results in the conversion and redistribution of this cortical actin into large bundles of F-actin which form the cores of spikes. When the spikes recede, F-actin disappears, and the amount of all forms of actin bound in the cortex appears to decrease. Antibodies to sea urchin egg myosin, fascin and a 220-kDa protein were used to examine these actin-associated proteins during the times that the organization of actin changes. Myosin and the 220-kDa protein are bound to the cortex and uniformly distributed before 1-MA application while fascin appears to be unbound. When spikes appear after 1-MA addition, fascin and the 220-kDa protein are localized coincidently with the spikes, whereas myosin remains uniformly distributed throughout the cortex and is excluded from the spikes. After spike resorption, fascin and the 220-kDa protein appear to lose their cortical binding while myosin retains its localization unchanged. These results indicate that actin, fascin and the 220-kDa protein undergo major organizational changes in the cortex in response to 1-MA.     

Seasonal variation in standing crop,density and leaf growth rate of the seagrass, Heterozostera tasmanica, in western Port and Port Phillip Bay,Victoria, Australia

Standing crop, density and leaf growth rate of Heterozostera tasmanica (Martens ex Aschers.) den Hartog along with light, temperature, nutrient and sediment characteristics were determined monthly for fifteen months at three study sites in Western Port and one site in Port Phillip Bay, Victoria, Australia. Erect vegetative stems of H. tasmanica were frequently branched, were present throughout the year and accounted for 25–60% of the above-sediment biomass, with the stem proportion higher during winter than summer. At three of the four sites there was a unimodal seasonal pattern in which minimum leaf standing crop (27–61 g dry wt. m^?2), density (600–2000 leaf cluster m^?2) and leaf productivity (0.34–0.77 g dry wt. m^?2 day^?1) generally occurred during winter (June–August) and maximum leaf standing crop (105–173 g dry wt. m^?2), density (2700–5000 leaf cluster m^?2) and leaf productivity (2.6–4.2 g dry wt. m^?2 day^?1) occurred during summer (December–February). A bimodal seasonal pattern with minimum standing crop and density during midsummer occurred at one site. This anomalous seasonal pattern may be due to exposure and desiccation stress during spring low tides. At the site receiving the lowest irradiance, standing crop, density and annual leaf production also were lowest, but length and width of leaves, shoot height and leaf growth rate per leaf cluster were the highest of the four study sites. On average, each leaf cluster at any one of the study sites produced 30–31 leaves per year with mean leaf turnover rates of 1.3–1.7% day^?1. Annual leaf production of H. tasmanica ranged from 410 to 640 g dry wt.m^?2 at the four sites.