GALVANOTROPISM AND "REVERSAL OF INHIBITION" BY STRYCHNINE

The cathodically galvanotropic orientation of nemerteans, Lineus, and the anodic orientation of the gephyrean Echiurus, are reversed by the action of strychnine under conditions such that the typical "reversal of inhibition" induced by this substance is apparent. Nicotine does not give this result. Since it is necessary to assume that the strychnine effect is due to action upon the central ganglia, and since the galvanotropic effect depends upon action of the current on nerve cell bodies of the central ganglia, it must be assumed that the locus of reversal by strychnine is not perikaryal, but presumably synaptic.     

ON REVERSAL OF GEOTROPISM IN ASTERINA

A certain level of strychninization induces in Asterina reversal of geotropism from the normally geonegative movement to a persistent downward creeping. The effect of an attached float producing upward pull is to induce upward creeping, under these conditions, whereas normally it leads to downward movement. This reversal cannot be regarded as due to a mere intensification of the sensory effect of tension. It must be understood as representing a true reversal of inhibition. The temporary reversal of geotropism following mechanical disturbances (in the absence of strychnine) is interpreted in the same way.     

CYTOLOGICAL STUDIES ON THE ANTIMETABOLITE ACTION OF 2,6-DIAMINOPURINE IN VICIA FABA ROOTS

At a concentration of 9.6 x 10^–5 M, 2,6-diaminopurine (DAP) completely inhibited cell enlargement, cell division, and DNA synthesis (determined by microphotometric measurement of Feulgen dye) in Vicia faba roots. Inhibition of cell enlargement was partially reversed by adenine, guanine, xanthine, adenosine, and desoxyadenosine. Guanine and the nucleosides gave the greatest reversal, suggesting that one point of DAP action upon cell enlargement is a disruption of nucleoside or nucleotide metabolism, possibly during pentosenucleic acid synthesis. DAP inhibited cell division by preventing onset of prophase. At the concentrations used it had no significant effect on the rate or appearance of mitoses in progress. Inhibition of entrance into prophase was not directly due to inhibition of DNA synthesis since approximately half of the inhibited nuclei had the doubled (4C) amount of DNA. Adenine competitively reversed DAP inhibition of cell division, giving an inhibition index of about 0.5. Guanine gave a slight reversal while xanthine, hypoxanthine, adenosine, and desoxyadenosine were inactive. A basic need for free adenine for the onset of mitosis was suggested by this reversal pattern. Meristems treated with DAP contained almost no nuclei with intermediate amounts of DNA, indicating that DAP prevented the onset of DNA synthesis while allowing that underway to reach completion. The inhibition of DNA synthesis was reversed by adenine, adenosine, and desoxyadenosine although synthesis appeared to proceed at a slower rate in reversals than in controls. Inhibition of DNA synthesis by DAP is probably through nucleoside or nucleotide metabolism. A small general depression of DNA content of nuclei in the reversal treatments was observed. This deviation from DNA "constancy" cannot be adequately explained at present although it may be a result of direct incorporation of DAP into DNA. The possible purine precursor, 4-amino-5-imidazolecarboxamide gave no reversal of DAP inhibition of cell elongation and cell division and only a slight possible reversal of inhibition of DNA synthesis.     

Differential inhibition of the release of endogenous and newly synthesized acetylcholine from Torpedo synaptosomes by presynaptic muscarinic receptors

Activation of Torpedo presynaptic muscarinic acetylcholine (ACh) receptors with the agonist oxotremorine (20 μM) results in the inhibition of Ca²⁺-dependent release of endogenous ACh from Torpedo synaptosomes. This effect is reversed by the muscarinic antagonist atropine (1 μM) which, by itself, has no effect. In contrast, under the same conditions the amount of newly synthesized radiolabeled [³H]ACh released is not affected by muscarinic ligands. These findings suggest that presynaptic muscarinic inhibition in the Torpedo is due to interference with the mobilization of ACh from a storage pool.     

THE MECHANISMS OF TROPISTIC REACTIONS AND THE STRYCHNINE EFFECT IN DAPHNIA

1. Experiments with strychnine were performed to test two assumptions important in the development of a theory for the mechanisms involved in the tropisms exhibited by Daphnia. 2. After a brief interval in strychnine solution Daphnia exhibits a reversal of the primary sign (a) of phototropism, from negative to positive; and (b) of galvanotropism, from anodic to cathodic. In both cases the orientation of the body remains the same. 3. The mechanism responsible for the sign of phototropism and galvanotropism in Daphnia is therefore distinct from that underlying orientation. 4. Evidence is obtained indicating that changes in sign of tropism, produced by changes in illumination or by subjection to strychnine, involve the control of antagonistic muscles in the swimming appendages which are reciprocally innervated.     

Light-controlled Cycocel Reversal of Coumarin Inhibition of Lettuce Seed Germination and Root Growth

Lettuce seed germination or lettuce root elongation after germination in water was inhibited by coumarin and these inhibitions were reversed by Cycocel. 2.53 × 10³ M Cycocel reversed the inhibition of seed germination by 6.8 × 10⁴ M coumarin. and 6.32 × lO^?4 M Cycocel reversed the inhibition of root elongation by 3.4 × 10³ M coumarin. No other analogs of Cycocel reversed these coumarin induced inhibitions of growth. Cycocel reversal of coumarin inhibition of lettuce seed germination occurred in red light but not in far-red light or darkness. The red-far-red system was photoreversible. Cycocel and kinetin appear to act similarly in reversing coumarin inhibition of germination. Gibberellin A₃ and IAA were unable to reverse these coumarin induced inhibitions. A common mechanism is suggested for Cycocel reversal of coumarin and lAA inhibition of growth.     

Inhibition of the mitochondrial respiratory chain by alkylhydroxynaphthoquines: Reversal on discharge of the energized state

Inhibition of mitochondrial respiration by alkylhydroxynaphthoquinones may be reversed by addition of a variety of uncouplers including substituted phenols, carbonyl cyanide phenylhydrazones, divalent cations and univalent cations in the presence of ionophoretic antibiotics. A likely explanation for such reversibility is the requirement that the anionic inhibitor be transported to a site of action within the mitochondrion. Support for this view includes (1) failure to obtain reversal of inhibition with submitochondrial particles, (2) release of inhibition by a competing anion, succinate, (3) augmentation of inhibition when a divalent cation is taken up, (4) the chemical diversity of uncouplers that release inhibition and (5) inhibiton by uncoupling compounds of the uptake of labeled alkylhydroxynaphthoquinones. It is suggested that a similar explanation may apply to two other inhibitors of the cytochrome b–c region, antimycin and alkylhydroxyquinoline-N-oxides.     

Regeneration of photoreceptor organs in freshwater planarians at different levels of accumulation of natural methylmercury compounds

The effects of natural methylmercury compounds on regeneration of photoreceptor organs were studied in three freshwater planarians: Polycelis tenuis, Dugesia lugubris, and D. tigrina. Accumulation of methyl mercury in the planarian body suppressed regeneration of P. tenuis with numerous photoreceptor organs to a greater extent than in two other planarians that have only two eyes. High methyl mercury concentrations inhibited the restoration of photoreceptor organs in asexual and sexual D. tigrina races     

Effect of polyamines on palmitoyl-coenzyme A-caused inhibition of glucose-6-phosphate dehydrogenase from baker's yeast

Aliphatic polyamines reversed the inhibition of baker's yeast glucose-6-phosphate dehydrogenase by palmitoyl-CoA. The reversal effects of the polyamines on the inhibition were related not only to the number of amino (imino) groups but also to the carbon chain length of polyamine. The relieving constant (K_r) was calculated to determine the absolute concentration of each examined polyamine required to relieve the enzyme from the inhibition. A smaller K_r value indicates a stronger relieving activity. An ethylamine derivative with a large number of amino groups was more effective in reversing the palmitoyl-CoA inhibition. The K_r values of long-chain aliphatic diamines, triamines, and tetramines were smaller than those of short-chain ones. However, n-butylamine and ornithine, composed of four carbons like putrescine, had markedly larger K_r values than putrescine. Substitution of one or two amino groups by carboxyl or hydroxyl groups also appeared to relieve the enzyme from the inhibition. These results suggest that the reversal effect on palmitoyl-CoA inhibition is one of the important roles of polyamines. Furthermore, it is possible that the interaction of palmitoyl-CoA and polyamines physiologically regulates the pentose monophosphate cycle in baker's yeast.     

On the Reversal of the Inhibition of Photosynthesis Induced by Sodium Dimethyldithiocarbamate and Tetramethylthiuram disulphide

The reversal of 80–100 per cent inhibition of photosynthesis in the green alga Enteromorpha linza (L.) J. Ag., obtained with tetramethylthiuram disulphide (TMTD) and the corresponding sodium dithiocarhamate (Na-DMDT), was investigated. The inhibition obtained by the two compounds proves to be almost completely reversible by simply washing away the inhibitors. Copper(II) sulphate partially reversed the inhibition obtained with Na-DMDT while reduced glutathione (GSH) partially reversed inhibition obtained with both Na-DMDT and TMTD. The time to complete the reversal varied between 12 and 33 min. Almost no reversal of the TMTD induced inhibition was achieved with CuSO₄. Na-DMDT and CuSO₄ reacted instantaneously with each other. GSH most probably reduced TMTD stoichiometrically to DMDT. This reaction was complete- within 6 seconds. No reaction took place between GSH and Na-DMDT. The interpretation of The results eliminates the most conventional explanation of the mechanism of the inhibition of photosynthesis in E. linza with Na-DMDT, viz. complex formation with functional metals. The reaction between TMTD and functional thiol groups in the alga is the most plausible inhibition mechanism. It is inferred from the results that Na-DMDT is most probably oxidized by E. linza to TMTD and effects inhibition in this form. (PDF DAMAGE)     

Chloroplast Response to Low Leaf Water Potentials: II. Role of Osmotic Potential

Electron transport in chloroplasts isolated from desiccated sunflower (Helianthus annuus L. cv. Russian Mammoth) leaves was compared with electron transport in sunflower chloroplasts in sorbitol-containing media having various osmotic potentials. In media having low osmotic potentials and dichloroindophenol as electron acceptor, the activity for electron transport was inhibited, but the inhibition was much less than that due to comparable desiccation in vivo. The inhibition at low osmotic potentials was rapidly reversed by returning the chloroplasts to media having high osmotic potentials, but the activity of chloroplasts from desiccated tissue showed no reversal when the chloroplasts were placed in media having high osmotic potentials. Nevertheless, the inhibition of chloroplast activity due to desiccation in vivo was basically reversible, because chloroplasts recovered quickly when they were rehydrated in vivo. The large differences between desiccation in vivo and exposure to low osmotic potential in vivo indicate that osmotic solutions did not reproduce the effects of tissue desiccation. It is concluded that decreases in the Gibbs free energy of water due to decreased osmotic potentials probably have only a small effect on electron transport in chloroplasts from desiccated tissue and do not account for the major effects of leaf desiccation on electron transport.     

Spoltud-1 is a chromatoid body component required for planarian long-term stem cell self-renewal

Freshwater planarians exhibit a striking power of regeneration, based on a population of undifferentiated totipotent stem cells, called neoblasts. These somatic stem cells have several characteristics resembling those of germ line stem cells in other animals, such as the presence of perinuclear RNA granules (chromatoid bodies). We have isolated a Tudor domain-containing gene in the planarian species Schmidtea polychroa, Spoltud-1, and show that it is expressed in neoblast cells, germ line cells and central nervous system, and during embryonic development. Within the neoblasts, Spoltud-1 protein is enriched in chromatoid bodies. Spoltud-1 RNAi eliminates protein expression after 3 weeks, and abolishes the power of regeneration of planarians after 7 weeks. Neoblast cells are eliminated by the RNAi treatment, disappearing at the end rather than gradually during the process. Neoblasts with no detectable Spoltud-1 protein are able to proliferate and differentiate. These results suggest that Spoltud-1 is required for long term stem cell self renewal.     

The mechanism of ciliary movement. 3. Theory of suppression of reversal by electrical potential of cilia reversed by barium ions

It is well known that Paramecium (1) is cathodally galvanotactic in mixtures of sodium and calcium salts because of ciliary reversal at the cathodal end, and that (2) the cilia also may be reversed in solutions rich in monovalent cations or in barium or manganese ions, and (3) the organisms swim backward in these solutions. It also is well known (Kamada, '31) that Paramecium in solutions of barium salts (4) is anodally galvanotactic at low electric potentials, (5) is cathodally galvanotactic at higher potentials, and (6) again becomes anodally galvanotactic if the potential is lowered, but these results have never been explained. However, they can be explained if the membrane is assumed to act as an ion exchanger. Cilia are reversed by barium because barium replaces some of the calcium on the membrane. When a low cathodal potential is applied, the barium, because of its high electrophoretic velocity (Ba++> Ca++> Na+) is removed, thereby causing a suppression of the reversal. If the potential is increased, calcium is also removed, leaving mostly sodium on the membrane, and this causes a return of the reversal. Lowering the potential again causes a suppression of the reversal. Changes at the anodal end can be explained in a comparable manner.     

Expression pattern of the expanded noggin gene family in the planarian Schmidtea mediterranea

Noggin genes are mainly known as inhibitors of the Bone Morphogenetic Protein (BMP) signalling pathway. Noggin genes play an important role in various developmental processes such as axis formation and neural differentiation. In vertebrates, inhibition of the BMP pathway is usually carried out together with other inhibitory molecules: chordin and follistatin. Recently, it has been shown in planarians that the BMP pathway has a conserved function in the maintenance and re-establishment of the dorsoventral axis during homeostasis and regeneration. In an attempt to further characterize the BMP pathway in this model we have undertaken an in silico search of noggin genes in the genome of Schmidtea mediterranea. In contrast to other systems in which between one and four noggin genes have been reported, ten genes containing a noggin domain are present in S. mediterranea. These genes have been classified into two groups: noggin genes (two genes) and noggin-like genes (eight genes). Noggin-like genes are characterized by the presence of an insertion of 50–60 amino acids in the middle of the noggin domain. Here, we report the characterization of this expanded family of noggin genes in planarians as well as their expression patterns in both intact and regenerating animals. In situ hybridizations show that planarian noggin genes are expressed in a variety of cell types located in different regions of the planarian body.     

Some Properties of Partially Purified Pyruvate Kinase from Euglena gracilis Klebs var. bacillaris

A method of purification of pyruvate kinase (EC 2.7.1.40) from light-grown Euglena gracilis var. bacillaris was developed which yielded an enzyme preparation purified 115-fold over crude extracts. During organelle formation, levels of pyruvate kinase in extracts prepared from cells engaged in light-induced chloroplast development do not change significantly. The enzyme has a molecular weight of approximately 240,000 and a requirement for both K⁺ and Mg²⁺. Fructose 1,6-diphosphate activates the enzyme when the concentration of phosphoenol-pyruvate is limiting; it does not activate when the concentration of ADP is limiting. ATP, citrate, and Ca²⁺ are inhibitors of the enzyme and inhibit the fructose 1,6-diphosphate stimulation of the enzyme activity. ATP inhibition is only partially reversed by high concentrations of fructose 1,6-diphosphate. Further reversal of inhibition can be achieved by dialysis. Ca²⁺-dependent inhibition can be reversed by a chelating agent but not by increased concentrations of Mg²⁺.     

Pharmacological assessment of methamphetamine-induced behavioral hyperactivity mediated by dopaminergic transmission in planarian Dugesia japonica

The freshwater planarian Dugesia japonica has a simple central nervous system (CNS) and can regenerate complete organs, even a functional brain. Recent studies demonstrated that there is a great variety of neuronal-related genes, specifically expressed in several domains of the planarian brain. We identified a planarian dat gene, named it D. japonica dopamine transporter (Djdat), and analyzed its expression and function. Both in situ hybridization and immunofluorescence revealed that localization of Djdat mRNA and protein was the same as that of D. japonica tyrosine hydroxylase (DjTH). Although, dopamine (DA) content in Djdat(RNAi) planarians was not altered, Djdat(RNAi) planarians showed increased spontaneous locomotion. The hyperactivity in the Djdat(RNAi) planarians was significantly suppressed by SCH23390 or sulpiride pretreatment, which are D₁ or D₂ receptor antagonists, respectively. These results suggest that planarians have a Djdat ortholog and the ability to regulate dopaminergic neurotransmission and association with spontaneous locomotion.     

Epinephrine, Propranolol, and the Sucrose-Ammonium Inhibition of Flowering in Lemna paucicostata 6746

The sucrose-ammonium inhibition of flowering Lemna paucicostata 6746 in continuous blue light or in short days was partially overcome by epinephrine. This reversal was prevented by propranolol, an antagonist of epinephrine in animals. In ammonium-free medium, propranolol inhibited flowering, and this inhibition was completely overcome by epinephrine. Increased levels of Ca²⁺, Pi and nitrate partially reversed the inhibition by propranolol. Concentrations of cAMP, adenine, and adenosine that partially overcame the sucrose-ammonium inhibition did not affect flowering in cultures treated with propranolol. The possibility is discussed that the effects on flowering of sucrose-ammonium, propranolol, and epinephrine were due to altered intracellular levels of cAMP or of a cAMP-like compound.     

Red Light-inhibited Mesocotyl Elongation in Maize Seedlings: I. The Auxin Hypothesis

Red light-inhibited mesocotyl elongation, which occurs in intact Zea mays L. seedlings, was studied in excised segments which included the coleoptile (or parts therefrom) and apical centimeter of the mesocotyl. Experiments took into account, first, the ability of the segments to regenerate auxin supply sites, and, second, that auxin uptake can be greatly reduced if there is no cut surface, apical to the elongating cells, to act as a port of entry. In all cases, auxin completely reversed the inhibition of elongation by light. The results support the hypothesis that light regulates mesocotyl elongation by controlling auxin supply from the coleoptile. Sucrose concentration had no effect on auxin reversal of light-inhibited elongation, but relatively high concentrations of gibberellic acid (10 μm) could substitute for auxin in this system.     

REVERSAL OF REACTION BY MEANS OF STRYCHNINE IN PLANARIANS AND STARFISH

Two cases have been described, that of the marine planarian Bdelloura and that of the starfish Asterias forbesii, in which strychnine reverses reciprocal inhibition. These facts indicate that the nervous systems of these invertebrates function in a manner similar to those of the earthworm and vertebrates. Moreover, it would seem that strychnine acts upon some chemical component of the neuron which is always present in synaptic structures but which also occurs in the simpler neurons of lower forms. The fact that strychnine is without this characteristic effect on such forms as medusa and sea anemone, indicates that the nervous systems of the starfish and planarian have chemical affinities with the vertebrates which the cœlenterates do not possess.