Studies with Cells From Marine Fish in Tissue Culture.

Clem, L. William; Moewus, Liselotte; Sigel, M. Michael

doi:10.3181/00379727-108-27059

Cited by 95 publications

(41 citation statements)

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“…The study of chromosomes in these animals has been expanding significantly thanks to the development of refined techniques of cell and tissue culture originally developed for mammals, but later adapted to the fish physiology (Clem et al, 1961;Booke, 1968;Wolf & Quimby, 1969;Denton, 1973) and the development of less expensive in vivo direct methods (Osuf-Costaz & Foresti, 1992). Because of the basal position that ray-finned fishes occupy in the phylogeny of vertebrates, studies on chromosomes of this group have provided valuable information for the understanding of different aspects such as mechanisms of sex determination and evolution of sex chromosomes, distribution of the nucleolus organizers regions (NOR), existence of supernumerary chromosomes and the role of polyploidy in evolution (Pisano et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Chromosome evolution in fishes: a new challenging proposal from Neotropical species

et al. 2014

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We present a database containing cytogenetic data of Neotropical actinopterygian fishes from Venezuela obtained in a single laboratory for the first time. The results of this study include 103 species belonging to 74 genera assigned to 45 families and 17 out of the 40 teleost orders. In the group of marine fishes, the modal diploid number was 2n=48 represented in 60% of the studied species, while in the freshwater fish group the modal diploid complement was 2n=54, represented in 21.21 % of the studied species. The average number of chromosomes and the mean FN were statistically higher in freshwater fish than in marine fish. The degree of diversification and karyotype variation was also higher in freshwater fish in contrast to a more conserved cytogenetic pattern in marine fish. In contrast to the assumption according to which 48 acrocentric chromosomes was basal chromosome number in fish, data here presented show that there is an obvious trend towards the reduction of the diploid number of chromosomes from values near 2n=60 with high number of biarmed chromosomes in more basal species to 2n=48 acrocentric elements in more derived Actinopterygii.Se presenta una base de datos que contiene los datos citogenéticos de peces Actinopterigios Neotropicales de Venezuela obtenidos por primera vez en un solo laboratorio. Los resultados de este estudio incluyen 103 especies pertenecientes a 74 géneros de 45 familias contenidas en 17 de los 40 órdenes de teleósteos. En el grupo de peces marinos, el número diploide modal fue 2n=48 representado en 60% de las especies estudiadas, mientras que en el grupo de peces de agua dulce el complemento diploide modal fue 2n=54, representado en el 21,21% de las especies estudiadas. El número de cromosomas y FN promedio fueron estadísticamente superiores en peces dulceacuícolas. El grado de diversificación y variación en el cariotipo también fue mayor en peces de agua dulce en contraste con un patrón citogenético más conservado en peces marinos. En contraposición a la suposición según la cual 48 cromosomas acrocentricos era el número cromosómico basal en los peces, los datos aquí presentados muestran que existe una evidente tendencia hacia la reducción del número de cromosomas desde valores cercanos a 2n=60 con alto número de cromosomas birrámeos en las especies más basales a 2n=48 elementos acrocéntricos en los actinopterigios más derivados.

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Section: Introductionmentioning

confidence: 99%

Chromosome evolution in fishes: a new challenging proposal from Neotropical species

et al. 2014

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“…All tissue culture reagents were obtained from Gibco. Epithelioma papillosum of carp (Cyprinus carpio) (EPC) (Wolf & Mann, 1980) and grunt-fin (Haemulon sciurus) tissue (GF) cells (Clem et al, 1961) were grown in minimal essential medium (MEM) with Hanks' balanced salts solution (Sigma), 10 mM HEPES (pH 7*3), 2 mM glutamine, 0.23 '70 NaHCO, and 10 '/o (v/v) heat-inactivated foetal bovine serum at 25 "C in a 5 % (v/v) CO, atmosphere. For culturing GF, MEM was supplemented with 58 mM NaCl for optimal growth.…”

Section: Methodsmentioning

confidence: 99%

Internalization and cytotoxicity are important virulence mechanisms in vibrio-f ishepithelial cell interactions

Wang

Oon

et al. 1998

Microbiology

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Vibrio anguillarum and Vibrio damselee are Gram-negative bacteria that cause systemic infections called vibriosis in fish. They can enter fish cells and survive as intracellular parasites. The host-pathogen interactions between these Vibrio species and the fish epithelial cell lines epithelioma papillosum of carp (EPC) and grunt-f in tissue (GF) cells, were examined using phase-contrast, scanning electron and confocal microscopy. In addition, potential signal transduction pathways that precede bacterial internalization were studied by using signal transduction inhibitors. Some Vibrio species induced morphological changes in fish cells and this allowed classification into a cytopathic group and a noncytopathic group. The cytopathic group could be subdivided into two invasive groups (I and II) and a cytotoxic group. Of the invasive strains V. anguillarum 811218-5W (group I) and GNirus/5(3) (group II), genistein, a tyrosine kinase inhibitor, only inhibited internalization of V. anguillarum GNirus/5(3) into EPC cells, whereas staurosporine, a protein kinase C inhibitor, accelerated internalization of both strains. Cytochalasin D, an inhibitor of microfilament polymerization, prevented internalization of both strains, whilst vincristin, a microtubule inhibitor, only inhibited internalization of V. anguillarum GNirus/5(3). For the cytotoxic strain V. damselae ATCC 33539, extracellular products (ECP) alone caused morphological changes in EPC and GF. Bacterial internalization may not be important in the pathogenesis of this group. The non-cytopathic strain V. anguillarum SU5/93(2) did not enter cells or induce any changes in EPC and GF monolayen. This study has identified some major differences between Vibrio species in their interactions with fish cells in vitm and will thus facilitate future studies of the molecular basis of pathogenesis of vibriosis.

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“…After wiping of the body surface with 70% ethanol cotton, dorsal, anal and caudal fins were cut off and dipped in 50 ppm iodine solution (Clem et al, 1961;Law et al, 1978); however, it was described by Nicholson et al (1987) that additional NaCl was not required for many marine fish cell cultures. In our study, FF-11 was successfully cultured without adding NaCl.…”

Section: Primary Culture and Passagementioning

confidence: 99%