Esterase polymorphisms in muscle and brain of four fresh water fishes belonging to the family Cyprinidae

Esterase polymorphism in muscle and brain tissues of four species of freshwater fishes belonging to the family cyprinidae are examined using polyacrylamide gel electrophoresis. The enzymes are classified according to their substrate specificity and inhibitor sensitivity. Distribution of six classes of esterases; viz. carboxyl, acetyl, aryl, choline, Esdp and ER-esterases in the two tissues of the four species is reported.     

Identical reactivity of brain and erythrocyte cholinesterases of some mammals

The paper deals with a comparative study of various aspects of reactivity (substrate and inhibitor specificity, sensitivity to action of hydrophobic organophosphorus inhibitors, capability for reactivation) of cholinesterase preparations from brain, erythrocytes, and serum of a group of mammals (human, rabbit, rat, cattle, dog, and cat). It has turned out that at preservation of the species and tissue specificity, catalytic properties of the brain and erythrocyte cholinesterases of the same animal species are very close to each other.     

Cholinesterase activity in tissues of some crab species from the Japan Sea

The comparative study of the cholinesterase activity in some crab species was carried out for the first time with use of a set of thiocholine substrates. The substrate specificity was studied in stellar nerve, heart, and hemolymph of three crab species. The crab hemolymph was shown to be characterized by the highest enzyme activity. The enzyme from various crab organs has different structure o substrate specificity. Properties of crab enzymes were compared with acetylcholinesterase (AChE) of human blood erythrocytes, butyrylcholinesterase (BuChE) of horse blood serum, enzyme of squids and bivalve molluscs. The obtained data allow the conclusion to be made on differences in properties of enzymes both at the interspecies and at the tissue levels.     

Comparative Study of Substrate–Inhibitor Specificity of Brain Cholinesterase Activity of Pacific Salmon Fish of the Salmonidae Family

Salmon fish of the Salmonidae family, representatives of the Pacific salmon Oncorhynchus genus, the humpback salmon O. gorbuscha Wielb., dog salmon O. keta, coho salmon O. kisutch, blueback salmon O. nerka, and chinook salmon O. tschawytscha, and also of noble salmons genus, Salmo—black salmon S. salar, caught in the northwest water area of the Pacific ocean, are studied in the work. Iodides of acetyl-, propionyl-, and butyrylthiocholine were used as substrates. Carbamate proserine and 5 organophosphorus inhibitors were studied as inhibitors. Testing of homogeneity of cholinesterase activity in the brain tissue has revealed only one enzyme in the coho salmon, while several enzymes, in the dog salmon and humpback salmon, which can be an enzymologic argument for the favor of hypothesis about the presence of interspecies groups among Pacific salmons of the Oncorhynchus genus. Interspecies differences in substrate specificity of the brain tissue in the studied salmonid species are found. The fish enzymes of the fishes have shown a high sensitivity to proserine. Only in the case of diisopropylfluorophosphate, both interspecies and intergenus differences are revealed.     

Species-specific differences in the substrate-inhibitory specificity of cholinesterases from optical ganglia of squids of the Gonatidae family

A comparative study was carried out of the substrate and inhibitory specificity of cholinesterase preparations from squids, representatives of 3 genes and 5 species of the Gonatidae family:Berryteuthis (B. magister andB. anonichos),Gonatus (G. kamtschaticus andG. tinro), andGonatipsis (G. borealis), that have overlapping habitation areals in the Bering Sea. As substrates, there were used bromides of acetylthiocholine, propionylthiocholine, and butyrylthiocholine, as organophosphorus inhibitors, diisopropylfluorophosphate, a cation-containing inhibitor, and 4 hydrophobic compounds. The homogeneity of the cholinesterase activity in these preparations has been shown, the intergenus and interspecies differences in the enzyme properties are revealed, and also the peculiarity of properties of enzymes from Gonatidae squids is emphasized in comparison with cholinesterase from the Pacific squidTodarodes paciflcus and “standard” mammalian enzymes (from human erythrocytes and horse blood serum). The revealed interspecies differences are discussed in terms of evolutionary development of the Gonatidae family.     

Neuroecology of cartilaginous fishes: the functional implications of brain scaling

It is a widely accepted view that neural development can reflect morphological adaptations and sensory specializations. The aim of this review is to give a broad overview of the current status of brain data available for cartilaginous fishes and examine how perspectives on allometric scaling of brain size across this group of fishes has changed within the last 50 years with the addition of new data and more rigorous statistical analyses. The current knowledge of neuroanatomy in cartilaginous fishes is reviewed and data on brain size (encephalization, n = 151) and interspecific variation in brain organization (n = 84) has been explored to ascertain scaling relationships across this clade. It is determined whether similar patterns of brain organization, termed cerebrotypes, exist in species that share certain lifestyle characteristics. Clear patterns of brain organization exist across cartilaginous fishes, irrespective of phylogenetic grouping and, although this study was not a functional analysis, it provides further evidence that chondrichthyan brain structures might have developed in conjunction with specific behaviours or enhanced cognitive capabilities. Larger brains, with well-developed telencephala and large, highly foliated cerebella are reported in species that occupy complex reef or oceanic habitats, potentially identifying a reef-associated cerebrotype. In contrast, benthic and benthopelagic demersal species comprise the group with the smallest brains, with a relatively reduced telencephalon and a smooth cerebellar corpus. There is also evidence herein of a bathyal cerebrotype; deep-sea benthopelagic sharks possess relatively small brains and show a clear relative hypertrophy of the medulla oblongata. Despite the patterns observed and documented, significant gaps in the literature have been highlighted. Brain mass data are only currently available on c. 16% of all chondrichthyan species, and only 8% of species have data available on their brain organization, with far less on subsections of major brain areas that receive distinct sensory input. The interspecific variability in brain organization further stresses the importance of performing functional studies on a greater range of species. Only an expansive data set, comprised of species that span a variety of habitats and taxonomic groups, with widely disparate behavioural repertoires, combined with further functional analyses, will help shed light on the extent to which chondrichthyan brains have evolved as a consequence of behaviour, habitat and lifestyle in addition to phylogeny.     

Histochemical characterization of cholinesterase activity in the frog brain with special reference to its localization on the wall of blood vessels

Synopsis The nature of the cholinesterase activity present on the wall of blood vessels in amphibian brain has been studied histochemically. It is concluded that the enzyme involved is acetylcholinesterase rather than butyrylcholinesterase, which more frequently occurs in the blood vessels of the brain of other vertebrates. The histochemical results are in agreement with most biochemical data concerning substrate specificity and inhibitor response for both acetylcholinesterase and butyrylcholinesterase. The two main hypotheses put forward by others in order to explain cholinesterase activity in brain vessels are discussed briefly.     

Cholinesterases from plant tissues. VI. Preliminary characterization of enzymes from Solanum melongena L. and Zea mays L.

Enzymes capable of hydrolyzing esters of thiocholine have been assayed in extracts of Solanum melongena L. (eggplant) and Zea Mays L. (corn). The enzymes from both species are inhibited by the anti-cholinesterases neostigmine, physostigmine, and 284c51 and by AMO-1618, a plant growth retardant and they both have pH optima near pH 8.0. The enzyme from eggplant is maximally active at a substrate concentration of 0.15 mM acetylthiocholine and is inhibited at higher substrate concentrations. On the basis of this last property, the magnitude of inhibition by the various inhibitors, and the substrate specificity, we conclude that the enzyme from eggplant, but not that from corn, is a cholinesterase.     

Substrate Specificity of Cholinesterases in Various Representatives of the Animal Kingdom

This review summarizes the literature data as well as experimental results obtained at our Institute over a period of 50 years on the substrate specificity of cholinesterases–acetylcholine acetylhydrolases (EC 3.1.1.7) and acylcholine acylhydrolases (EC 3.1.1.8). The parameters of enzymatic hydrolysis of oxo- and thiocholine and β-methylcholine esters in different organs and tissues were analyzed in 66 animal species including 22 chordate, 20 insect, 1 mite, 17 mollusk, 4 nematode, and 2 flatworm species. Our substrate specificity studies and extensive data on the inhibitory specificity obtained using irreversible organophosphorous inhibitors and reversible effectors unequivocally indicate that the cholinesterase family is characterized by a clear-cut species and tissue specificity.     

Comparative-enzymological study of cholinesterase of the Pacific squid <Emphasis Type="Italic">Todarodes pacificus</Emphasis>

Summarized are results of the 40-year studies of the Russian biochemists on the comparative-enzymological characteristics of cholinesterase of optic ganglia of the Pacific squid Todarodes pacificus. The review includes comparative evaluation of the cholinesterase activity of various hydrobiont tissues, the proof of enzymatic homogeneity of tissues of the Pacific squid optic ganglia, data on substrate specificity with study of 18 ester substrates as well as detailed study of inhibitory specificity (61 irreversible inhibitors and 49 reversible onium inhibitors). Peculiarity of properties of this enzyme as compared with vertebrate and invertebrate cholinesterases is shown.     

Species diversity and conservation status of cartilaginous fishes (Chondrichthyes) of Russian waters

The paper considers the composition of the fauna of cartilaginous fishes (Chondrichthyes) in the waters adjacent to the coast of the Russian Federation. Its features are analyzed in the seas of basins of different oceans. The conservation status of cartilaginous fishes in the waters of Russia is assessed according to the criteria of the International Union for Conservation of Nature. The current state of individual threatened species is briefly characterized.     

Cholinesterases from flounder muscle. Purification and characterization of glycosyl-phosphatidylinositol-anchored and collagen-tailed forms differing in substrate specificity

Flounder (Platichthys flesus) muscle contains two types of cholinesterases, that differ in molecular form and in substrate specificity. Both enzymes were purified by affinity chromatography. About 8% of cholinesterase activity could be attributed to collagen-tailed asymmetric acetylcholinesterase sedimenting at 17S, 13S and 9S, which showed catalytic properties of a true acetylcholinesterase. 92% of cholinesterase activity corresponded to an amphiphilic dimeric enzyme sedimenting at 6S in the presence of Triton X-100. Treatment with phospholipase C yielded a hydrophilic form and uncovered an epitope called the cross-reacting determinant, which is found in the hydrophilic form of a number of glycosyl-phosphatidylinositol-anchored proteins. This enzyme showed catalytic properties intermediate to those of acetylcholinesterase and butyrylcholinesterase. It hydrolyzed acetylthiocholine, propionylthiocholine, butyrylthiocholine and benzoylthiocholine. The Km and the maximal velocity decreased with the length and hydrophobicity of the acyl chain. At high substrate concentrations the enzyme was inhibited. The p(IC50) values for BW284C51 and ethopropazine were between those found for acetylcholinesterase and butylcholinesterase. For purified detergent-soluble cholinesterase a specific activity of 8000 IU/mg protein, a turnover number of 2.8 x 10(7) h-1, and 1 active site/subunit were determined.     

Substrate–Inhibitor Analysis of Cholinesterase from Hemolymph of the Pacific Gastropod Mollusc Neptunea eulimata

Based on our own and literature data, a detailed substrate–inhibitor analysis of catalytic properties of cholinesterase of hemolymph of the Pacific gastropod mollusc Neptunea eulimata has been carried out. Using specific substrates and organophosphorus inhibitors, homogeneity of the enzyme preparation has been shown. The study of the substrate specificity, using 8 substrates—choline, fluorogenic and chromogenic esters—has revealed some features of similarity with mammalian erythrocyte cholinesterase. At the same time, testing of a large group (33 compounds) of organophosphorus inhibitors with various structure, including hydrophobic inhibitors studied for the first time, has established both quantitative and qualitative differences from the mammalian blood enzymes. It is concluded that cholinesterase from the neptunea hemolymph cannot be ascribed unanimously to the type of acetylhydrolases of acetylcholine (EC 3.1.1.7).     

Systematic position of fish species and ganglioside composition and content

The ganglioside content in brain of cartilaginous and bony fishes studied varies from 110 to 581 and from 104 to 595 micrograms sialic acid per g of wet weight respectively. A high degree of alkali lability and the predominance of C18-sphingosine and N-acetylneuraminic acid are typical of fish brain gangliosides. A high content of oligosialogangliosides with four and more residues of sialic acid and the predominance of gangliosides with gangliotetraosyl carbohydrate chain are characteristic for teleost brain. No pronounced difference was revealed in ganglioside composition and content of clupeomorphs and percomorphs. Gangliosides with short (lactosyl and gangliotriaosyl) carbohydrate chain predominate in brain of all cartilaginous fishes studied. A statistically significant difference was found in ganglioside content, relative oligosialoganglioside content and ganglioside fatty acid composition of squalomorphs and rajiformes, on one hand, and dasyatiformes and galeomorphs, on the other hand.     

Head size constrains forebrain development and evolution in ray-finned fishes

In ray-finned fishes, which comprise nearly half of all vertebrate species, the telencephalon does not evaginate, as it does in other vertebrates, but instead everts. No detailed explanation for this species difference has ever been offered. Here we propose that telencephalic eversion evolved because ray-finned fish embryos are so small that their telencephalon cannot evaginate but must, instead, squeeze into the space just dorsal to the developing nasal epithelia and rostral to the eyes-morphogenetic movements that amount to eversion. Evidence for this hypothesis derives from cladistic analyses, which show that early ray-finned fishes reduced their adult body size and adopted a novel reproductive strategy, based on the production of myriad minute young. Because body size tends to be inversely proportional to brain:body ratio, this phylogenetic reduction in body size implies that embryonic ray-finned fishes should have proportionately larger brains than embryos of species whose telencephalons evaginate. This prediction was confirmed by comparing serially sectioned heads of representative ray-finned and cartilaginous fish embryos at several stages of development. The brain, excluding its ventricles, occupies 36-46% of the cranial cavity in embryonic ray-finned fishes, but less than 20% in embryonic sharks. Moreover, three-dimensional reconstructions show that in embryonic ray-finned fishes the telencephalon has no room for a full-fledged evagination; instead, it spreads into the spaces just dorsal and caudal to the developing nasal epithelia. These morphogenetic movements, in conjunction with a thinning of the forebrain roof, generate telencephalic eversion.     

Redistribution of membrane 5''-nucleotidase in rabbit peritoneal polymorphonuclear leucocytes during phagocytosis

Calmodulin-dependent glycogen synthase kinase isolated from skeletal muscle and synapsin I kinase II isolated from brain have several properties that are very similar. These properties include: substrate and site-specificities, immunological cross-reactivity, and phosphopeptide maps following limited proteolysis. Both enzymes phosphorylate a wide variety of substrate proteins. The two kinases may represent different isozymes of a multifunctional calmodulin-dependent protein kinase that mediates many of the actions of Ca2+ in various tissues. Therefore, we propose the name 'calmodulin-dependent multi-protein kinase' for this broad specificity enzyme.     

Brachyurins--serine collagenolytic enzymes from crabs

The properties of brachyurins, proteolytic enzymes belonging to a new subfamily of chymotrypsin-like proteases, are considered. These enzymes, found in various species of crustacean, exhibit mixed substrate specificity and a marked collagenolytic activity. The enzymatic and physicochemical properties of brachyurins I and their primary and spatial structures are discussed in detail. A separate chapter is devoted to the preparations of collagenases from the hepatopancreas of king crab: their action on the damaged skin and use in medicine.     

Brachyurins, Serine Collagenolytic Enzymes from Crabs

The properties of brachyurins, proteolytic enzymes belonging to a new subfamily of chymotrypsin-like proteases, are considered. These enzymes, found in various species of crustacean, exhibit mixed substrate specificity and a marked collagenolytic activity. The enzymatic and physicochemical properties of brachyurins I and their primary and spatial structures are discussed in detail. A separate chapter is devoted to the preparations of collagenases from the hepatopancreas of king crab: their action on the damaged skin and use in medicine.     

Pseudomonas aeruginosa cholinesterase and phosphorylcholine phosphatase: two enzymes contributing to corneal infection

Choline, acetylcholine and betaine used as the sole carbon, nitrogen or carbon and nitrogen source increase cholinesterase activity in addition to phosphorylcholine phosphatase and phospholipase C activities in Pseudomonas aeruginosa. The cholinesterase activity catalyses the hydrolysis of acetylthiocholine (Km approx. 0.13 mM) and propionylthiocholine (Km approx. 0.26 mM), but not butyrylthiocholine, which is a pure competitive inhibitor (Ki 0.05 mM). Increasing choline concentrations in the assay mixture decreased the affinity of cholinesterase for acetylthiocholine, but in all cases prevented inhibition raised by high substrate concentrations. Considering the properties of these enzymes, and the fact that in the corneal epithelium there exists a high acetylcholine concentration and that Pseudomonas aeruginosa produces corneal infection, it is proposed that these enzymes acting coordinately might contribute to the breakdown of the corneal epithelial membrane.     

Extracellular Fluid Proteins of Goldfish Brain: Evidence for the Presence of Proteases and Esterases

Preparations of enriched fractions of extracellular fluid (ECF) proteins from goldfish brain were found to contain protease(s) and esterase(s). The N-substituted furanacryloyl (FA) peptides FA-Phe-Gly-Gly and FA-Phe-OMe were used as model substrates for determining protease and esterase activity, respectively, in a spectrophotometric assay. Studies of the profile of substrate specificity and identification of the types of compounds that were effective as inhibitors showed that these ECF enzymes have some distinctive properties. GSH, but not GSSG, and EDTA inhibited the protease(s) without influencing the esterase(s), whereas L-1-tosylamide-2-phenylethylchloromethyl ketone blocked both protease and esterase activities of ECF. Most of the protease and esterase properties of ECF could be bound to concanavalin A-Sepharose affinity chromatographic columns in association with ependymin--a brain extracellular protein. These observations indicate that ECF may contain a metalloprotease(s) and raise the possibility that the ependymins might be a substrate for these ECF enzymes.