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[Free amino acids in the central nervous system of skates (Batomorpha)].

The distribution of free amino acids in strongly and weakly electric fishes (skates and rays) varies in relation to species, brain region and functional peculiarities. In comparison to the weakly electric fishes (Raja clavata and Myliobatis aquila) a number of brain regions of the strongly electric species (Torpedo marmorata and T. ocellata) show considerably higher concentrations of arginine, glycine and tyrosine, and lower concentrations of aspartic acid. The electric organ of the strongly electric fishes was characterized by the lack of tyrosine and glycine, by traces of GABA, and by considerably higher concentrations of alanine, serine, cysteine, and histidine than in the brain of the same species. The electromotor center Lobus electricus of the stronlgy electric fishes, in contrast to other brain regions, shows exceptionally high concentrations of tyrosine and alanine. In comparison to the mammalian brain, levels of brain alanine, glycine, cysteine and cystine are higher in the four fish species studied. The results are discussed in relation to the morphophysiological organization of the fish central nervous system and of the mechanisms of electrogenesis.

Amino Acids

A comparative study of the glycolipids of human, bird and fish testes and of human sperm.

The glycolipids of human testis and sperm have been compared. Both adult testis and the sperm exhibited remarkably complex, but generally similar, patterns of glycolipids. In particular, both contained appreciable amounts of the sulfogalactosylmonoalkylmonoacylglycerol, recently shown to be the principal glycolipid of the testis and sperm of a number of animals. In contrast, immature (prebuteral) human testis did not contain this compound. To extend knowledge on the possible distribution of sulfogalactosylmonoalkylmonoacylglycerol in the testes of other chordates, we have also analysed the glycolipids of the testes of a number of birds and fish. None of the testes from these species contained the above compound. Instead, sulfogalactosylceramide was found to be a major glycolipid of the testis of mature fowl, duck and skate-fish and sulfogalactosylglucosylceramide of the testis of mature salmon and trout. Immature duck testis contained only a trace of sulfogalactosylceramide. These studies reveal intriguing differences between the sulfatides of various chordates, lend support to the concept that sulfatides increase markedly in testis at a specific stage of spermatogenesis and suggest an important role for sulfatides in testicular and spermatozoal function.

Adult

[The effect of pentagastrin and ornithine-tetragastrin on motor activity of the stomach in skates and scorpion-fish].

Low doses of pentagastrin (50 mug/kg b. w.) have no physiological effect on motor activity of the stomach in the skate Dasyatis pastinaca. Average doses (100-200 mug/kg) stimulate the activity, whereas high ones (300 mug/kg) inhibit the frequency of stomach contractions, slightly increasing their amplitude. Ornithine tetragastrin in a dose 2000 mug/kg does not affect motor activity of the stomach in skates. In the scorpion-fish Scorpaena porcus, ornithine tetragastrin (1000 and 2000 mug/kg) inhibits motor activity of the stomach.

Animals

Patterns of purine nucleotides in fish erythrocytes.

1. The purine nucleotides were determined in the whole blood of 9 fresh water teleosts and 2 marine selachians. 2. GTP and ATP accounted for 88-99% of the total erythrocytes purines. 3. The ATP/ADP ratio ranged from 11 to 60 in the erythrocytes of the fish examined. 4. GTP is widely distributed in fish erythrocytes but its level ranged from 1 to 33 nmol/mg Hb (0.4 to 9 mumol/ml erythrocyte). 5. Lepomis and Esox exhibited a GTP/ATP ratio as elevated as in Anguilla; moreover the concentration of GTP per mol of Hb (physiologically most indicative) is higher in Lepomis, Esox, Ictalurus and Silurus than in Anguilla.

Animals

[The fatty acid composition of the gangliosides in different parts of the vertebrate brain].

Comparative studies on gangliosides from the brain of higher (mammals, birds) and lower (amphibians, cartilaginous and teleost fishes) vertebrates indicate that for each of the parts of the brain the same regularities in changes of fatty acid composition are typical as for the total brain in a series of vertebrates. All brain structures of the mammalian brain exhibit higher saturation of ganglioside fatty acids, higher content of stearic acid (C18:0) and lower relative content of long chain (C22-C24) fatty acids, especially that of C24:1, as compared to the corresponding brain structures of lower vertebrates. Fatty acid composition in birds is more close to mammalian one with respect to some of the features, while with respect to the other ones it occupies an intermediate position between lower vertebrates and mammals. In the series more phylogenetically ancient--more recent brain parts (medulla oblongata--midbrain--forebrain), the content of content of stearic acid and the degree of saturation ganglioside fatty acids increase, whereas the relative content of acids with a long chain (C22-C24) in them decreases (with the exception of amphibians). In all the brain structures of higher elasmobranch fishes, fatty acid composition of gangliosides exhibits more progressive features of chemical organization as compared to that in homologous structures of lower elasmobranch fishes.

Animals

Developmental analysis of the cone photoreceptor-less little skate retina reveals distinct Onecut1 isoforms.

The retinal development of elasmobranchs, the subclass comprising sharks, skates, and rays, remains poorly understood. This group is diverse in retinal phenotype, with many sharks and rays possessing rods together with one or more cone types. In contrast, the little skate (Leucoraja erinacea) has only a single rod photoreceptor type, which has been reported to exhibit some physiological and anatomical properties associated with cones. To investigate how this unusual photoreceptor system develops, we first identified an embryonic stage of early photoreceptor formation based on otx2 expression. We then developed a retinal electroporation approach to test whether a onecut1-dependent cone-associated reporter could be activated in the embryonic skate retina. Activation of this reporter was not detected, indicating that the corresponding enhancer is not robustly active under the conditions tested. To assess developmental changes in gene expression, we generated bulk RNA-seq datasets from embryonic, hatchling, and adult retinas. These analyses showed strong embryonic expression of onecut1, increasing expression of rod-associated genes through development, and pseudogenization or loss of multiple cone-enriched genes. We further identified a developmentally regulated onecut1 splice isoform containing an additional 48 amino acid sequence between the CUT and homeodomain DNA-binding domains. This spacer-containing isoform, termed LSOC1X2, was most abundant in the embryonic retina. To test whether LSOC1X2 retained regulatory activity, we assayed it in a mouse retinal reporter system. Both skate Onecut1 isoforms activated the ThrbCRM1 reporter in this heterologous context. Together, these findings identify a novel, developmentally regulated retinal onecut1 isoform in the little skate and establish it as a candidate regulator for future studies of photoreceptor development in this species and its elasmobranch relatives.

Animals

[The characteristics of the interaction of glycogenolytic enzymes with glycogen in endothermic and ectothermic animals].

It has been shown that the dependence of glycogenphosphorylase kinase activity on the quantity of glycogen added corresponds to the glycogenphosphorylase saturation curve by glycogen obtained during investigation of complex formation with the muscle glycogenphosphorylase of the animals: rat, frog Rana temporaria and fish Raja clavata under identical conditions. We suggest that glycogenolytic enzymes organize regular structure on glycogen particle. Glycogenphosphorylases of heat-loving animals have high affinity to glycogen that is accounted for necessity to keep the structure in natural conditions.

Adenosine Triphosphate

Differences in the dopaminergic innervation of the electroreceptive and mechanoreceptive medullary lateral line nuclei of the ray, Raja radiata.

An immunohistochemical study was made of the octavolateral centres in the medulla of a cartilaginous fish, using an antibody specific for dopamine. No immunopositive cell bodies were observed but dopaminergic fibres were present in parts of the region. The overlying cerebellar crest was devoid of dopamine; the dorsal (electroreceptive) nucleus was poorly innervated, but the medial (mechanoreceptive) nucleus received a rich dopaminergic innervation comprising very fine, varicose fibres.

Animals

Rod outer segments are designed for optimum photon detection.

The proposal that rod outer segment length is optimal with respect to photon absorption and noise control is extended and tested in a number of species. We find good agreement with our optimality criterion in duplex retinae where rods act as detectors of one or a few photons, but not in all rod retinae nor in those which are exposed to significant photic environmental noise.

Absorption

Rhodopsin regeneration in the normal and in the detached/replaced retina of the skate.

The bleaching and regeneration of rhodopsin in the skate retina was studied by means of fundus reflectometry, both in the normal eyecup preparation and after the retina had been detached and then replaced on the surface of the pigment epithelium (RPE). After bleaching virtually all the rhodopsin in the retinal test area of the normal eyecup, more than 90% of the photopigment was reformed after about 2 hr in darkness; over most of this time course, rhodopsin density rose linearly at a rate of 0.875% min-1 with a half-time of 55 min. Detaching the retina from its pigment epithelium resulted in a number of abnormalities, both structural and functional. Histological examination of the detached/replaced (D/R) retina showed striking alterations in the structural integrity of the RPE cells at their interface with the neural retina. The cells appeared vacuolated and misshapen, and the apical processes of the RPE, which normally ensheath the receptor outer segments, were shredded and free of their association with the visual cells. These morphological changes, as well as dilution of the IRBP content of the subretinal space caused by separation of the tissues, appear to be the main factors contributing to the functional abnormalities in rhodopsin kinetics. But despite these abnormalities and the persistent detachment, the rate of regeneration and the amount of rhodopsin reformed after bleaching were reduced by less than 50% of their normal values. The fact that a significant fraction of the bleached rhodopsin was regenerated under these conditions indicates that 11-cis retinal formed in the RPE was able to traverse a much greater than normal subretinal space to reach the opsin-bearing photoreceptor membranes.

Animals

Structure and function of the external gill filaments of embryonic skates (Raja erinacea).

We have investigated structure and function of the external gill filaments, which occur transiently in the embryonic little skate, Raja erinacea. Approximately 25-30 days after spawning (body mass 0.03-0.05 g) external gill filaments appear as an outgrowth from the caudal side of the gill arches. These filaments are thread-like, each containing one afferent and one efferent blood vessel, and by day 70-75 (body mass 0.4-0.5 g) they reach their maximum size at a length of about 1 cm and a blood vessel diameter of 70-80 microns. Subsequent resorption of the filaments is characterized by a decrease in both length and diameter of the blood vessel. By day 90-95 (body mass 0.9-1.0 g) the external gill filaments are completely resorbed and replaced by internal gills. Blood velocity, measured in these external filaments, increased with development from 0.1 mm.sec-1 to about 0.7 min.sec-1, and decreased again during resorption. Blood flow, calculated therefrom with blood vessel diameter, showed a similar maximum curve. A model analysis supports the hypothesis that in a full grown filament respiratory gas exchange is mainly perfusion-limited and can contribute significantly to the total oxygen uptake of the embryo. Analysis of the results indicates, however, that the gill filaments are not adequate as a gas exchange organ for later developmental stages.

Animals

Immunoreactivity of skate electrocytes towards monoclonal antibodies against human dystrophin and dystrophin-related (DMDL) protein.

Monoclonal antibodies against human dystrophin have been used to demonstrate the existence of a dystrophin-like protein in the electrocytes of skate electric organ. This protein is also present in skate muscle and resembles that found in Torpedo electric organ. Monoclonal antibodies against a human autosomal homologue of dystrophin (DMDL protein) did not detect a similar protein in skate or Torpedo. Immunocytochemical staining of the innervated and non-innervated faces of the electrocyte membrane was obtained using the anti-dystrophin antibodies only.

Animals

Identification of vitellogenin in the little skate (Raja erinacea).

1. Vitellogenin was isolated from mature female skates by selective precipitation with MgCl2/EDTA followed by chromatography on DEAE-cellulose columns. 2. A single monomer of approximately 205 kDa was identified on 6.0% SDS-PAGE gels. 3. In addition, isolation of yolk proteins with ammonium sulfate yielded proteins of 94 and 38 kDa (putative phosvitins) and putative lipovitellins of ca 105, 91 and 67 kDa. 4. In vivo phosphate incorporation in female and male skates implanted with estradiol indicated that vitellogenin was phosphorylated. 5. Total protein phosphate incorporation was significantly higher in females than male skates. 6. In male skates treated with estradiol, phosphate incorporation increased from 2 days after implantation to a maximum at approximately 11 days after implantation. 7. Determination of the rate of disappearance of 32P-labeled protein suggests a half-life of ca 200 hr in normal female skate plasma.

Animals

Identification of spinal electromotoneurons in the ray Raja clavata (Rajidae).

Retrograde transport of horseradish peroxidase injected ipsilaterally in several parts of the electric organ and light microscopy were employed to study motoneurons and electromotoneurons of the spinal cord in the weakly electric ray Raja clavata (Rajidae). The horseradish peroxidase-labelled electromotoneurons form a chain of cells located ventrolaterally in the gray matter of the spinal cord segments adjacent to the electric organ. The diameters of cells in the electromotor nucleus vary from 70 to 110 microns. When comparing other motoneurons of the spinal cord with electromotoneurons, the latter reveal no segmentation, whereas motoneurons yield accumulations which are close to metametric ones and are essentially smaller in size (averaging 33 microns in diameter).

Animals

Comparison of cadmium, mercury and calcium accumulations by isolated hepatocytes of the small skate (Raja erinacea) and rat.

1. Accumulation of calcium, cadmium and mercury by isolated hepatocytes of the small skate (Raja erinacea) and rat was examined at 14 and 37 degrees C, respectively. 2. Metal uptakes by both species were biphasic, with rat cells accumulating more metal than the skate cells. 3. Total accumulation after 30 min was in the order: mercury = cadmium much greater than calcium. 4. In both species calcium and cadmium accumulations were reduced at 4 degrees C, while mercury accumulation was not. 5. Cd accumulation was increased by Cu and Hg in both species. 6. Hg accumulation was inhibited by Cu in both species, and increased by Cd only in the rat cells.

Animals

Cadmium inhibits stimulus-response coupling in skate (Raja erinacea) electric organ.

1. The effects of cadmium on stimulus-response coupling in the skate (Raja erinacea) electric organ were examined. 2. Cadmium decreased the evoked electrical discharge and the evoked release of 3H-ACh in a concentration-related fashion. 3. Cadmium (100 microM) also blocked voltage-dependent 45Ca uptake. 4. Both d-tubocurarine and nifedipine blocked Ca uptake and evoked potential, but not 3H-ACh release, thus most of the 45Ca uptake measured was post-synaptic through L Ca channels. 5. Nickel, cadmium, and verapamil inhibited 3H-ACh release and evoked potential, indicating a block of pre-synaptic T Ca channels.

Acetylcholine

cDNA that encodes active agrin.

Agrin is thought to mediate the motor neuron-induced aggregation of AChRs and AChE on the surface of muscle fibers at neuromuscular junctions. We have isolated a cDNA from a chick brain library that, based on sequence homology and expression experiments, codes for active agrin. Examination of the sequence reveals considerable similarity to homologous cDNAs previously isolated from ray and rat libraries. A conspicuous difference is an insertion of 33 bp in chick agrin cDNA, which endows the encoded protein with AChR/AChE aggregating activity. Homologous transcripts having the 33 bp insertion were detected in the ray CNS, which indicates that an insertion of similar size is conserved in agrin in many, if not all, vertebrate species. Results of in situ hybridization studies and PCR experiments on mRNA isolated from motor neuron-enriched fractions of the spinal cord indicate that, consistent with the agrin hypothesis, motor neurons contain transcripts that code for active agrin.

Agrin

S-laminin expression in adult and developing retinae: a potential cue for photoreceptor morphogenesis.

The development of the neural retina follows a stereotyped time course that begins with an undifferentiated neuroepithelium populated by multipotential progenitor cells and ends with a highly differentiated tissue containing diverse cell types. The identities of the factors that guide this differentiation have remained elusive; a likely location for such factors, however, is the extracellular environment. Here, we show that the extracellular matrix component s-laminin is present in the neural retina, that s-laminin expression parallels the differentiation of rod photoreceptors, that photoreceptors interact with s-laminin in vitro, and that antibodies to s-laminin profoundly reduce the appearance of cells that express rhodopsin in vitro. These data suggest that s-laminin plays a role in the differentiation of the neural retina and provide evidence that the composition of the extracellular matrix may be an important determinant of retinal differentiation.

Animals