The chemical basis of morphogenesis

It is suggested that a system of chemical substances, called morphogens, reacting together and diffusing through a tissue, is adequate to account for the main phenomena of morphogenesis. Such a system, although it may originally be quite homogeneous, may later develop a pattern or structure due to an instability of the homogeneous equilibrium, which is triggered off by random disturbances. Such reaction-diffusion systems are considered in some detail in the case of an isolated ring of cells, a mathematically convenient, though biologically unusual system. The investigation is chiefly concerned with the onset of instability. It is found that there are six essentially different forms which this may take. In the most interesting form stationary waves appear on the ring. It is suggested that this might account, for instance, for the tentacle patterns onHydra and for whorled leaves. A system of reactions and diffusion on a sphere is also considered. Such a system appears to account for gastrulation. Another reaction system in two dimensions gives rise to patterns reminiscent of dappling. It is also suggested that stationary waves in two dimensions could account for the phenomena of phyllotaxis. The purpose of this paper is to discuss a possible mechanism by which the genes of a zygote may determine the anatomical structure of the resulting organism. The theory does not make any new hypotheses; it merely suggests that certain well-known physical laws are sufficient to account for many of the facts. The full understanding of the paper requires a good knowledge of mathematics, some biology, and some elementary chemistry. Since readers cannot be expected to be experts in all of these subjects, a number of elementary facts are explained, which can be found in text-books, but whose omission would make the paper difficult reading. Reprinted from thePhilosophical Transactions of the Royal Society (part B), Vol. 237, pp. 37–72 (1953) with the permission of the Royal Society, London.     

The biological and chemical basis for tissue-selective amyloid disease

Factors controlling the onset and progression of extracellular amyloid diseases remain largely unknown. Central to disease etiology is the efficiency of the endoplasmic reticulum (ER) machinery that targets destabilized mutant proteins for degradation and the enhanced tendency of these variants to aggregate if secreted. We demonstrate that mammalian cells secrete numerous transthyretin (TTR) disease-associated variants with wild-type efficiency in spite of compromised folding energetics. Only the most highly destabilized TTR variants are subjected to ER-associated degradation (ERAD) and then only in certain tissues, providing insight into tissue selective amyloidosis. Rather than a "quality control" standard based on wild-type stability, we find that ER-assisted folding (ERAF), based on global protein energetics, determines the extent of export. We propose that ERAF (influenced by the energetics of the protein fold, chaperone enzyme distributions, and metabolite chaperones) in competition with ERAD defines the unique secretory aptitude of each tissue.     

The genetic basis for indole-diterpene chemical diversity in filamentous fungi

Indole-diterpenes are a structurally diverse group of secondary metabolites with a common cyclic diterpene backbone derived from geranylgeranyl diphosphate and an indole group derived from indole-3-glycerol phosphate. Different types and patterns of ring substitutions and ring stereochemistry generate this structural diversity. This group of compounds is best known for their neurotoxic effects in mammals, causing syndromes such as ‘ryegrass staggers’ in sheep and cattle. Because many of the fungi that synthesise these compounds form symbiotic relationships with plants, insects, and other fungi, the synthesis of these compounds may confer an ecological advantage to these associations. Considerable recent progress has been made on understanding indole-diterpene biosynthesis in filamentous fungi, principally through the cloning and characterisation of the genes and gene products for paxilline biosynthesis in Penicillium paxilli. Important insights into how the indole-diterpene backbone is synthesised and decorated have been obtained using P. paxilli mutants in this pathway. This review provides an overview of these recent developments.     

The mechanical triggering of bioluminescence in marine dinoflagellates: chemical basis

In both photosynthetic (Pyrodinium bahamense, Gonyaulax polyedra, Pyrocystis Iunula, P. noctiluca, P. fusiformis) and nonphotosynthetic (Noctiluca miliaris) bioluminescent dinoflagellates chemical stimulation can by-pass mechanical stimulation. The effective ions are Ca⁺⁺, K⁺, NH₄⁺ and H⁺. Other chemicals found effective are those implicated in Ca⁺⁺ transport or binding. There are interspecies differences in degrees of mechanical and chemical stimulability. Photoinhibition of mechanical stimulability is the result of two effects, the first a reduction in total bioluminescence potential and the second a decrease in mechanical stimulability resulting experimentally in a decreased rate of light emission. This latter effect can be reversed with Ca⁺⁺ ions. Chemicals which bind Ca⁺⁺ or displace Ca⁺⁺ can mimic the effects of photoinhibition. The chemical inhibition of mechanical stimulability is also reversed by Ca⁺⁺ ions. A scheme is proposed which is consistent for all species examined.     

The chemical basis for sex recognition in Drosophila melanogaster

Chemical cues were recognized to play a predominant role in initiating male courtship behaviour in Drosophila melanogaster as measured by the number and duration of wing-vibration responses elicited in test males. The effect was associated with compounds specific to the female cuticle, for which we describe a simple extraction procedure. Female active extracts were compared with behaviourally non-active extracts from males, using gas-liquid and thin-layer chromatography. Using these preparative methods, long-chain hydrocarbons were isolated and activity was found only among unsaturated molecules. One, heptacosadiene, inducing the highest level of courtship, appears to be the main aphrodisiac pheromone of the female D. melanogaster. This compound is specific to females of the species and is the most abundant of their cuticular hydrocarbons.     

The chemical basis of the argentaffin reaction applied to glutaraldehyde fixed tissue

Summary A series of in vitro chemical tests was performed to elucidate the histochemical basis of the argentaffin technique in localizing 5-hydroxytryptamine within tissues fixed by glutaraldehyde. When glutaraldehyde is mixed with the amine a precipitate is formed which is trapped in the tissue at the site of 5-hydroxytryptamine storage. Chemically the precipitate is not a homogeneous compound. One aldehyde group of glutaraldehyde engages the amine group of 5-hydroxytryptamine in a condensation reaction to form a tetrahydro--carboline. The other aldehyde group is available in crypto form as a carbinolamine or its dehydration product and is readily released under the subsequent reaction conditions when ammoniacal silver hydroxide is added. Thus, metallic silver is precipitated at the site of 5-hydroxytryptamine storage in the tissues.This work was supported by grants from the Medical Research Council of Canada (MA 2182) and the Ontario Heart Association.     

The chemical basis of resistance of Sorghum bicolor to attack by Locusta migratoria

The resistance of young Sorghum bicolor to attack by Locusta migratoria (L.) has been investigated from a chemical standpoint. Absence of phagostimulants is not important. The release on biting of HCN from the non-deterrent cyanogenic glucoside, dhurrin, has been shown quantitatively to play an important part in the unpalatability. Correlation of feeding data with release rates of HCN showed that the increase in palatability of Sorghum did not closely correspond to the rapid fall in release of HCN as the plants aged, and when different concentrations of HCN were directly introduced into the mouthparts of Locusta using a cannula, the concentration of HCN released after the first growth stage was insufficient to account for the continued unpalatability. The effects on Locusta of other chemicals in Sorghum were monitored using a series of crude plant extracts. A lipid-soluble extract from plants at growth stage 1 and an extract of phenolic acids naturally released from the plants at stage 2 both showed deterrent effects. The individual phenolic acids were identified and all were found to occur naturally as esters with no deterrent properties. Estimates were made of the concentrations of free acids released by disruption of the plant tissues in several ways: maximum concentration occurs at growth stage 2. Bioassays showed that the deterrent effects of HCN and the phenolic acid mixture were additive and the change in palatability of the plant with increasing maturity is thus completely accounted for.

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Résumé On montre que Sorghum bicolor, au stade de plantule, est très peu apprécié par Locusta migratoria, mais qu'il devient de plus en plus appétissant au cours de sa croissance et de sa maturation. Ce changement n'est pas dû uniquement aux quantites croissantes de phagostimulants nutritifs.On a quantifié les changements du nivéau de la cyanogénèse chez Sorghum au fur et à mesure de sa croissance et on a mesuré le taux de dégagement de HCN par hydrolyse enzymatique après lésion des tissus. Par une technique nouvelle de biotitrage utilisant une canule implantée en permanence à travers le labium, on a montré que les concentrations de HCN dégagées naturellement par les jeunes plantes ont un effet répulsif sur Locusta. On a montré que la cyanogénése est la cause du caractère inappétissant des très jeunes plantes mais son niveau n'est pas suffisamment élevé pour expliquer le maintien d'une appétence réduite.Au second stade de croissance étudié, les acides phénoliques produits par la plante accroissent la répulsion causée par la cyanogénèse. On a trouvé que ces acides étaient présents dans la plante sous forme d'esters et qu'ils étaient dégagés enzymatiquement lors de lésions causées aux tissus par l'insecte s'alimentant, de la même façon que HCN. Aucun des acides phénoliques n'est dégagé à une concentration suffisante pour être répulsif, mais le mélange est répulsif à cause de l'effet cumulé des différentes substances chimiques.Le changement du caractère appétissant de Sorghum bicolor (var. 65D) pour Locusta migratoria est attribuable aux effects combinés de la cyanogénése et du dégagement d'acide phénolique.

     

The pharmacology of neurosteroidogenesis

In adrenal cortex and other steroidogenic tissues including glial cells, the conversion of cholesterol into pregnenolone is catalyzed by the cytochrome P450_scc located in the inner mitochondrial membrane. A complex mechanism operative in regulating cholesterol access to P450_scc limits the rate of pregnenolone biosynthesis. Participating in this mechanism are DBI (diazepam binding inhibitor), an endogenous peptide that is highly expressed in steroidogenic cells and some of the DBI processing products including DBI 17–50 (TTN). DBI and TTN activate steroidogenesis by binding to a specific receptor located in the outer mitochondrial membrane, termed mitochondrial DBI receptor complex (MDRC). MDRC is a hetero-oligomeric protein: only the subunit that includes the DBI and benzodiazepine (BZD) recognition sites has been cloned. Several 2-aryl-3-indoleacetamide derivatives (FGIN-1-X) with highly selective affinity (nM) for MDRC were synthesized which can stimulate steroidogenesis in mitochondrial preparations. These compounds stimulate adrenal cortex steroidogenesis in hypophysectomized rats but not in intact animals. Moreover, this steroidogenesis is inhibited by the isoquinoline carboxamide derivative PK 11195, a specific high affinity ligand for MDRC with a low intrinsic steroidogenic activity. Some of the FGIN-1-X derivatives stimulate brain pregnenolone accumulation in adrenalectomized-castrated rats. The FGIN-1-X derivatives that increase brain pregnenolone content, elicit antineophobic activity and antagonize punished behavior in the Vogel conflict test in rats. These actions of FGIN-1-X are resistant to inhibition by flumazenil, a specific inhibitor of BZD action in GABA_A receptors but are antagonized by PK 11195, a specific blocker of the steroidogenesis activation via MDRC stimulation. It is postulated that the pharmacological action of FGIN-1-X depends on a positive modulation of the GABA action on GABA_A receptors mediated by the stimulation of brain neurosteroid production.