The effects of diet and digestive cycle on the gastrointestinal tract pH values in the goldfish, Carassius auratus L., Mozambique tilapia, Oreochromis mossambicus (Peters), and channel catfish, Ictalurus punctatus (Rafinesque)

The digestive tract pH values of the teleost fish Oreochromis mossambicus, Carassius auratus and Ictalurus punctatus were determined in fish that were (1) starved, (2) fed herbivorous, omnivorous and carnivorous diets and (3) fed altered pH diets. The digestive tract pH profile was determined throughout an 8 h cycle. The herbivorous or carnivorous nature of the diet did not affect the pH values in any of the species. Significant differences in gut pH values were observed when the fish ingested diets with altered pHs. There was a difference between the starved and baseline fed values in Carassius auratus which was not observed in the other species. The pH values in the digestive tracts of the three species varied during the digestive cycle.     

Molecular Characterization and Functional Commonality of Nucleophosmin/Nucleoplasmin in Two Cyprinid Fish

Nucleophosmin/nucleoplasmin has been studied mostly in mammals and amphibians. To clarify the characteristics and function of nucleophosmin/nucleoplasmin in teleost fish, we cloned a full-length cDNA sequence from two cyprinid fish, Carassius auratus gibelio and Carassius auratus. Molecular characterization and multiple sequence alignments suggested that they are the homologs of nucleophosmin. RT-PCR and Western blot detected a specific expression in gonads, and immunofluorescence localization revealed their distribution in oogenic and spermatogenic cells. Furthermore, a sperm decondensation function was demonstrated by immunodepletion and in vitro sperm decondensation experiments. The data suggest that the cloned nucleophosmin should share expressional and functional characterization with nucleoplasmin and therefore provide novel evidence for a functional commonality of nucleophosmin and nucleoplasmin in fish.     

An immunohistochemical study of Carassius RF amide in the stomach, intestine, and pancreas of the Japanese butterfly ray, Gymnura japonica

Carassius RFamide (C-RFa) is a brain-gut peptide, isolated originally from the brain of a teleost fish, Japanese crucian carp (Carassius auratus langsdorfii). It is a novel FMRFamide-related peptide sharing homologies with mammalian prolactin-related peptide. We observed the localization of C-RFa-like immunoreactivity (C-RFa-IR) in the stomach, intestine, and pancreas of Japanese butterfly ray using an immunohistochemical technique. In the whole gastrointestinal tract, C-RFa-IR was observed in smooth muscle cells, and in the Meissner’s and Auerbach’s plexus ganglia. Furthermore, in the stomach C-RFa-IR was also observed at the bottom of the fundic glands in both the body and pyloric region. In the intestine C-RFa-IR was observed in epithelial cells at the tip of mucosal folds. In the pancreas, C-RFa-IR was observed in islet cells. Thus, we suggest that ray C-RFa may have functional roles in both peripheral neurotransmission and endocrine function of the gastroenteropancreatic system.     

Effect of copper ions on hematological and cytogenetic parameters of freshwater fishes <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

The effect of copper ions on the characteristics of white blood cells in Carassius auratus gibelio was studied. Changes in characteristics of cytogenetic instability in erythrocytes and in gill epithelial cells as a response to the effect of copper solutions were evaluated. The frequency of appearance of gill epithelial apoptotic cells was analyzed. The possibility of complex use of hematological indices and characteristics of cytogenetic instability to evaluate the effect of xenobiotics on fish organisms was shown.     

Genetic identification of larvae and juveniles reveals the difference in the spawning site among Cyprininae fish species/subspecies in Lake Biwa

To understand the differences in the spawning sites among Cyprininae fishes in Lake Biwa, we conducted periodic sampling of larvae and juveniles at three sites (irrigation ditch, St. 1; river, St. 2; and satellite lake, St. 3). On the basis of species/subspecies identification by using RAPD analysis, we examined the species composition of the larvae and juveniles at these three sampling sites. The number of specimens was 616, 68, and 117 at St. 1, St. 2, and St. 3, respectively. Based on morphological and genetic identification, the specimens were found to include nine fish species/subspecies, namely, Carassius auratus grandoculis, Carassius cuvieri, Carassius auratus langsdorfii, Cyprinus carpio, Sarcocheilichthys sp., Silurus asotus, Oryzias latipes, Odontobutis obscura obscura, and Rhinogobius sp. The species composition at the three sites also differed. Among the Cyprininae fishes, C. auratus grandoculis, C. auratus langsdorfii, and Cyprinus carpio were found in abundance at St. 1; C. cuvieri was not collected from St. 1 but was found at the other two sites, particularly St. 3. Among the other fishes, Rhinogobius sp. was collected at St. 1 and St. 3, whereas the other four occurred only at St. 1. These results suggest that the selection of spawning sites by C. cuvieri differs to a certain extent from that of the other Cyprininae fishes, and the irrigation ditch in the lake is an important habitat for the larvae and juveniles of native fish species.     

Surface folds in superficial epidermal cells of three species of teleost fish

Scanning electron microscope studies of fish epidermis demonstrate whorled arrangements of microfolds on the surface of superficial epidermal cells in the following species of teleost fish: the guppyfish Poecilia reticulata (Peters), the goldfish Carassius auratus auratus (L.), and the clown anemone fish Amphiprion percula (Lacépède). The pattern is most pronounced in epidermal cells covering the scales, but is also seen in the epidermal cells of the tail and body fins. The whorled pattern was not present on the gill epithelium or the surface of the eyes. These observations differ from transmission electron microscope studies which have described the skin surface as being composed of microvilli.     

Chromosome synapsis and recombination during meiotic division in gynogenetic triploid ginbuna,Carassius auratus langsdorfii

Electron microscope examinations of primary oocytes of gynogenetic triploid ginbuna (Carassius auratus langsdorfii) and diploid gengoroubuna (Carassius auratus cuvieri) were carried out. Typical synaptonemal complexes were observed in both subspecies. In addition, clear differences in PGM-A electrophoretic patterns between a female parent ginbuna and her offspring were detected. It was concluded that synapsis and recombination occur between at least some homologous chromosomes in triploid ginbuna.     

Genomic organization and evolution of the 5S ribosomal DNA in Tilapiini fishes

Summary 5S rDNA sequences present an intense dynamism and have proved to be valuable as genetic markers to distinguish closed related species and also in the understanding of the evolutionary dynamic of repetitive sequences in the genomes. In order to identify patterns of 5S rDNA organization and their evolution in the genome of fish species, such genomic segment was investigated in the tilapias Oreochromis niloticus and Tilapia rendalli, and in the hybrid O. urolepis hornorum × O. mossambicus. A dual 5S rDNA system was identified in the three analyzed tilapia samples. Although each 5S rDNA class was conserved among the three samples, a distinct 5S rDNA genome organization pattern could be evidenced for each sample. The presence of a dual 5S rDNA system seems to be a general trait among non-related teleost fish orders, suggesting that evolutionary events of duplication have occurred before the divergence of the main groups of teleost fishes.     

NORs and counterstain-enhanced fluorescence studies in Cyprinidae of different ploidy level

The ribosomal RNA gene expression in the genomes of evolutionary diploid (Scardinius erythrophthalmus, Leucaspius delineatus, Tinca tinca) and polyploid species (Cyprinus carpio, Carassius carassius, Carassius auratus gibelio, Carassius auratus auratus) of Cyprinidae has been investigated by means of a silver nitrate technique. The diploid species investigated exhibited only one pair of chromosomes with nucleolus organizers (NOR). Higher numbers of rRNA-expressing chromosomal sites in several evolutionary polyploid species (Carassins) gave evidence against a complete functional diploidization, at least with regard to the NOR bearing chromosomes in these species. The NORs displayed a heterochromatic brilliant chromomycin A₃ fluorescence. No distamycin-A/DAPI-bright heterochromatic blocks were detected in the genomes of the Cyprinidae.     

Structural characterization of the melano-macrophage centres (MMC) of goldfish Carassius auratus

In the present work we have studied the organization of melano-macrophage centres (MMCs) in the peripheral lymphoid organs, including spleen, pro- and mesonephros, of the goldfish, Carassius auratus, in an attempt to clarify their cellular composition, origins and functional relationships. Histological analysis demonstrated a similar organization in the three organs on the basis of closely packed phagocytic cells containing abundant pigment. The MMCs of Carassius auratus are found throughout the parenchyma of spleen and kidney and show a close association with the vascular system, i.e. splenic ellipsoids, sinusoids of red pulp and renal blood sinuses. They exhibit distinct degree of development from small groups of actively phagocytic macrophages to large, totally or partially encapsulated centres, where effete phagocytic cells are filled by cell debris. Ultrastructural and histochemical data suggest that the main inclusion observed in the MMCs of Carassius auratus is lipofuscin. Haemosiderin occurs in lesser amounts and melanin is almost restricted to kidney MMCs,--mainly mesonephros--. Our results suggest various non-specific physiological roles for the teleost MMCs, including tissue breakdown and erythrocyte catabolism.     

Novel Immune-type Receptor Genes and the Origins of Adaptive and Innate Immune Recognition

The prototypic forms of teleost novel immune-type receptors(NITRs) consist of a variable (V) region, a unique V-like C2(V/C2) domain, a transmembrane region and a cytoplasmic tailcontaining immunoreceptor tyrosine-based inhibition motifs (ITIMs).NITRs encode diversified V regions in large multigene familiesbut do not undergo somatic rearrangement. Studies in four differentbony fish model systems have identified a number of differentorganizational forms of NITRs. Specifically, NITR genes encodeN-terminal ectodomains of the V-type but otherwise vary in the:total number of extracellular immunoglobulin domains, numberand location of joining (J) region-like motifs, presence oftransmembrane regions, presence of charged residues within transmembraneregions, presence of cytoplasmic tails, and/or distributionof ITIM(s) within the cytoplasmic tails. V region-containingNITRs constitute a far more complex family than recognized originallyand currently include individual members that potentially functionthrough inhibitory as well as activating mechanisms. The genomicorganization of the NITR gene cluster as well as the structuraldiversity and overall architecture of the NITR proteins is reminiscentof genes encoded at the mammalian leukocyte receptor cluster(LRC); however, there presently is no functional evidence tosupport an orthologous relationship between NITR and LRC geneproducts. Comparisons of the predicted structures of the NITRshave identified several short regions of sequence identity anda novel cloning strategy has been devised that selects for secretoryand transmembrane proteins that encode these short motifs. Usingthis approach, related genes termed immune-type receptors (ITRs)have been identified in cartilaginous fish. Taken together,these studies indicate that leukocyte regulatory receptors,including those that mediate natural killer function, mighthave emerged early in vertebrate evolution and that the NITR/ITRgenes represent a new and potentially highly significant linkbetween innate and adaptive immune responses.     

Possible Reason for the Expansion of Carassius auratus (Linnaeus, 1758) (Teleostei,Cyprinidae) in the Danube River Basin

The author found quite a close negative relationship between commercial catches of predatory fishes and of German carp (Carassius auratus) in the lower Danube. He supposes that the main reason for the population explosion and subsequent expansion of German carp in the Danube River basin seems to be a shift in the balance of the fish community due to a reduction in the density of predatory species.     

Cloning,expression, and characterization of fugu CD4, the first ectothermic animal CD4

We have cloned and sequenced the first ectothermic animal CD4 gene from fugu, Takifugu rubripes, using a public database of the third draft sequence of the fugu genome. The fugu CD4 gene encodes a predicted protein of 463 amino acids containing four extracellular immunoglobulin (Ig)-like domains, a transmembrane region, and a cytoplasmic tail. Fugu CD4 shares low identity of about 15–20% with avian and mammalian CD4 proteins. Unlike avian and mammalian CD4, fugu CD4 lacks the Cys pair of the first Ig-like domain, but has a unique possible disulfide bond in the third domain. These differences suggest that fugu CD4 may have a different structure that could affect binding of major histocompatibility complex class II molecules and subsequent T-cell activation. In the putative fugu cytoplasmic region, the protein tyrosine kinase p56^lck binding motif is conserved. The predicted fugu CD4 gene is composed of 12 exons, differing from other CD4 genes, but showing conserved synteny and many conserved sequence motifs in the promoter region. RT-PCR analysis demonstrated that the fugu CD4 gene is expressed predominantly in lymphoid tissues. We also show that fugu CD4 can be expressed on the surface of cells via transfection. Molecular characterization of CD4 in fish provides insights into the evolution of both the CD4 molecule and the immune system.     

Interspecific differences of respiratory functions of some freshwater fish species

Hemoglobin affinity to oxygen, Bohr effect, sodium, potassium, and magnesium ion concentrations of erythrocytes in the three fish species from Rybinsk Reservoir (golden carp Carassius auratus gibelio, bream Abramis brama and pike Esox lucius) immediately after catching and during acclimation to laboratory conditions were studied. Our own and literature data on some other fish species were analyzed. The relationship between the functional features of the hemoglobin and ionic environment was found, and the changes taking place during acclimation and interspecific differences of these indices were shown.     

G6f-Like Is an ITAM-Containing Collagen Receptor in Thrombocytes

Collagen activates mammalian platelets through a complex of the immunoglobulin (Ig) receptor GPVI and the Fc receptor γ-chain, which has an immunoreceptor tyrosine-based activation motif (ITAM). Cross-linking of GPVI mediates activation through the sequential activation of Src and Syk family kinases and activation of PLCγ2. Nucleated thrombocytes in fish are activated by collagen but lack an ortholog of GPVI. In this study we show that collagen activates trout thrombocytes in whole blood and under flow conditions through a Src kinase driven pathway. We identify the Ig receptor G6f-like as a collagen receptor and demonstrate in a cell line assay that it signals through its cytoplasmic ITAM. Using a morpholino for in vivo knock-down of G6f-like levels in zebrafish, we observed a marked delay or absence of occlusion of the venous and arterial systems in response to laser injury. Thus, G6f-like is a physiologically relevant collagen receptor in fish thrombocytes which signals through the same ITAM-based signalling pathway as mammalian GPVI, providing a novel example of convergent evolution.     

Isolation and characterization of microsatellite markers from Fangzheng silver crucian carp,Carassius auratus gibelio (Bloch), and cross‐amplification in the closely related species crucian carp,Carassius auratus auratus (Linnaeus)

We report the development of 20 microsatellite markers for Fangzheng silver crucian carp, Carassius auratus gibelio (Bloch). Nineteen out of 20 showed polymorphism with alleles ranging from two to 14. These loci were screened to amplify the closely related species crucian carp, Carassius auratus auratus (Linnaeus) and all of them can amplify DNA with the size similar to the former. The origin of silver crucian carp is in issue and the population genetic structure is still unclear. Microsatellite markers isolated from the silver crucian carp and their utility in the crucian carp will be useful for these researches.     

Ultrastructure of Elasmobranch and Teleost Thrombocytes

Zapata, A., Carrato, A. 1980. Ultrastructure of elasmobranch and teleost thrombocytes. (Departamento de Morfología Microscópica, Facultad de Biología, Universidad Complutense, Madrid, Spain.) — Acta zool. (Stockh.) 61 (3): 179–182. The ultrastructure of circulating thrombocytes of Raja clavata and Torpedo marmorata (Elasmobranchs) and Rutilus rutilus and Gobio gobio (Teleosts) was described. In elasmobranchs, only one thrombocytic cell type was found, while teleosts presented two types. These cells contained microtubules and cytoplasmic granules, whose functional significance related to clotting processes is analyzed.     

表面活性剂对鱼体内铅的蓄积、分布和清除的影响

研究了重金属铅在鲫鱼体内蓄积、分布和清除规律以及阴离子表面活性剂十二烷基苯磺酸钠对鱼体内铅的蓄积、分布和清除的影响。结果表明,铅进入鲫鱼体内后,在不同部位中的分布有差异,铅的含量从高到低依次为内脏,头部,腹肌,背肌。鱼体内铅的含量随着暴露时间的延长而增大;表面活性剂十二烷基苯磺酸钠能延缓铅在鱼体内的蓄积,加快鱼体内铅的清除。鱼体内铅的清除速度比蓄积慢。