Temperature tolerance in the goldfish,Carassius auratus

• 1.Lower and upper temperature tolerances of 240 goldfish, Carassius auratus, were measured at constant acclimation temperatures of 5, 15, 25 and 35 °C via critical thermal methodology.
• 2.Mean critical thermal minima and maxima ranged from 0.3 to12.6 °C and 30.8 to 43.6 ° C, respectively, and were significantly linearly related to acclimation temperature. Acclimation temperature accounted for approximately 90% of the variance in temperature tolerance. Ultimate critical thermal minimum and maximum equaled 0.3 and 43.6 °C, respectively.
• 3.Integrating the temperature tolerance polygon yielded an area of temperature tolerance of 1429 °C², which is approximately 17% larger than the polygon measured via the incipient lethal temperature approach. This difference is explained by methodological differences in these two techniques to quantify temperature tolerance.

     

cDNA cloning and expression of a novel estrogen receptor beta-subtype in goldfish (Carassius auratus)

We have isolated a second goldfish estrogen receptor (ER) beta-subtype (gfER-beta2) cDNA which is distinct from the liver-derived ER-beta (gfER-beta1) cDNA reported previously. The 2650-bp cDNA, isolated from a goldfish pituitary and brain cDNA library, encodes a 610 amino acid (aa) protein which shows only a 53% aa sequence identity with gfER-beta1 in overall structure. RT-PCR analysis showed that mRNA of gfER-beta2, in contrast to that of gfER-beta1, was predominantly expressed in pituitary, telencephalon and hypothalamus as well as in liver of female goldfish. The existence of a second distinct ER-beta subtype opens new dimensions for studying tissue-specific regulation of gene expression by estrogen in the tetraploid goldfish.     

Globins and hypoxia adaptation in the goldfish, Carassius auratus

Goldfish (Carassius auratus) may survive in aquatic environments with low oxygen partial pressures. We investigated the contribution of respiratory proteins to hypoxia tolerance in C. auratus. We determined the complete coding sequence of hemoglobin alpha and beta and myoglobin, as well as partial cDNAs from neuroglobin and cytoglobin. Like the common carp (Cyprinus carpio), C. auratus possesses two paralogous myoglobin genes that duplicated within the cyprinid lineage. Myoglobin is also expressed in nonmuscle tissues. By means of quantitative real-time RT-PCR, we determined the changes in mRNA levels of hemoglobin, myoglobin, neuroglobin and cytoglobin in goldfish exposed to prolonged hypoxia (48 h at Po(2) ~ 6.7 kPa, 8 h at Po(2) ~ 1.7 kPa, 16 h at Po(2) ~ 6.7 kPa) at 20 degrees C. We observed small variations in the mRNA levels of hemoglobin, neuroglobin and cytoglobin, as well as putative hypoxia-responsive genes like lactate dehydrogenase or superoxide dismutase. Hypoxia significantly enhanced only the expression of myoglobin. However, we observed about fivefold higher neuroglobin protein levels in goldfish brain compared with zebrafish, although there was no significant difference in intrinsic myoglobin levels. These observations suggest that both myoglobin and neuroglobin may contribute to the tolerance of goldfish to low oxygen levels, but may reflect divergent adaptive strategies of hypoxia preadaptation (neuroglobin) and hypoxia response (myoglobin).     

Fine-structural study of leucocytes in the goldfish, Carassius auratus

An electron microscopic study was performed on leucocytes from circulating blood of the goldfish. The leucocytes were divided into eight types: neutrophil, eosinophil, large granular leucocyte (LGL), medium-sized granular leucocyte (MGL), small granular leucocyte (SGL), fine granular leucocyte (FGL), lymphocyte, and monocyte. In this report the thrombocyte was excluded from leucocytes, and LGL, MGL, SGL and FGL were tentatively classified based on the size of intracytoplasmic granules possessed by each cell. The existence of goldfish monocytes was electron microscopically demonstrated for the first time in the present report.     

Diurnal variations in the physiology of the goldfish,Carassius auratus

Abstract

Liver glycogen, liver lipid, liver triglycerides, plasma glucose, plasma total lipid, plasma cholesterol, plasma corticoids, hypothalamic serotonin and pituitary pro‐lactin levels were assayed at five times over a 24‐h period in Carassius auratus maintained under a specific photoperiod regime at various times throughout the year. Diurnal variations were observed in all parameters monitored. Daily variations of liver glycogen, plasma glucose, plasma lipid, plasma corticoids and hypothalamic serotonin were affected by time of feeding. Liver glycogen, plasma lipid and plasma corticoid levels were also affected by time of feeding. Diurnal variations of liver glycogen, plasma glucose and plasma lipid were influenced by light‐dark cycles. These data illustrate that feeding time, photoperiod and time of sacrifice are important considerations in the study of metabolic and hormonal parameters in fishes.     

PCR survey of hox genes in the goldfish Carassius auratus auratus

A tetraploidization event took place in the cyprinid lineage leading to goldfishes about 15 million years ago. A PCR survey for Hox genes in the goldfish Carassius auratus auratus (Actinopterygii: Cyprinidae) was performed to assess the consequences of this genome duplication. Not surprisingly, the genomic organization of the Hox gene clusters of goldfish is similar to that of the closely related zebrafish (Danio rerio). However, the goldfish exhibits a much larger number of recent pseudogenes, which are characterized by indels. These findings are consistent with the hypothesis that dosage effects cause selection pressure to rapidly silence crucial developmental regulators after a tetraploidization event.     

Cadmium affects the expression of metallothionein (MT) and glutathione peroxidase (GPX) mRNA in goldfish, Carassius auratus

Cadmium (Cd) is a widespread non-essential heavy metal that enters the aquatic environment as a result of natural and human-caused activities, including industrial effluent, mining, and agricultural runoff. In the present study, we investigated time and dose-related effect of CdCl(2) on metallothionein (MT) and glutathione peroxidase (GPX) mRNA levels in a number of goldfish tissues, in vivo. Basal MT and GPX mRNA levels remained unchanged in the tissues tested throughout the experiment. Injection with CdCl(2) significantly increased MT mRNA levels in the brain, liver, kidney and intestine in a dose-dependant manner at all time tested (6, 12, 24 and 36 h). We isolated the full length GPX cDNA from goldfish kidneys, and found it to contain 785 nucleotides, including an open reading frame, predicted to encode a protein of 142 amino acids. In contrast, injection with CdCl(2) significantly decreased GPX mRNA levels in the liver and kidney in a time-, and dose-, dependant, and became undetectable after 12, 24 and 36 h. The findings provide molecular characterization of MT and GPX in goldfish and suggest that exposure to Cd results in significant physiological changes in goldfish.     

Cd-metallothionein and metal-enzymes interactions in the goldfish Carassius auratus

Goldfish injected with cadmium chloride synthesized metallothionein. Ten days after the first injection, cadmium reached a maximum in the metallothionein peak (2 micrograms/ml) obtained after gel filtration of liver cytosol. Pyruvate kinase activity was inhibited from the beginning of the experiment; after the fourth day, the enzyme activity again started to increase but did not reach the control level. Alkaline phosphatase and fructose biphosphatase did not show any apparent inhibition. From the results here reported, a detoxifying role of metallothionein could be suggested.     

Intestinal peroxisomes of goldfish (Carassius auratus)--examination for hydrolase, dehydrogenase and carnitine acetyltransferase activities.

1. Rate sedimentation and isopycnic centrifugation were used to analyse the subcellular sites of enzymes in homogenates of goldfish intestinal mucosa. 2. The results allowed the following allocations to be made: carnitine acetyl transferase-mitochondrial and peroxisomal, xanthine dehydrogenase and NAD: alpha-glycerophosphate dehydrogenase soluble phase, NADP: isocitrate dehydrogenase soluble phase and mitochondrial, and 2-naphthyl laurate hydrolase microsomal and/or brush border. 3. Histochemistry confirmed the use of alkaline phosphatase and 1-naphthyl acetate esterase as brush border and microsome markers respectively. 4. Urate oxidase, allantoinase, allantoicase, xanthine oxidase and glycollate/lactate oxidase, activities were undetectable, and 1-naphthyl palmitate hydrolase was present only as a contaminant from pancreas.     

Organization of the superficial neuromast system in goldfish, Carassius auratus

Distribution, morphology, and orientation of superficial neuromasts and polarization of the hair cells within superficial neuromasts of the goldfish (Carassius auratus) were examined using fluorescence labeling and scanning electron microscopy. On each body side, goldfish have 1,800-2,000 superficial neuromasts distributed across the head, trunk and tail fin. Each superficial neuromast had about 14-32 hair cells that were arranged in the sensory epithelium with the axis of best sensitivity aligned perpendicular to the long axis of the neuromast. Hair cell polarization was rostro-caudal in most superficial neuromasts on trunk scales (with the exception of those on the lateral line scales), or on the tail fin. On lateral line scales, the most frequent hair cell polarization was dorso-ventral in 45% and rostro-caudal in 20% of the superficial neuromasts. On individual trunk scales, superficial neuromasts were organized in rows which in most scales showed similar orientations with angle deviations smaller than 45 degrees . In about 16% of all trunk scales, groups of superficial neuromasts in the dorsal and ventral half of the scale were oriented orthogonal to each other. On the head, most superficial neuromasts were arranged in rows or groups of similar orientation with angle deviations smaller than 45 degrees . Neighboring groups of superficial neuromasts could differ with respect to their orientation. The most frequent hair cell polarization was dorso-ventral in front of the eyes and on the ventral mandible and rostro-caudal below the eye and on the operculum.     

Cloning and expression analysis of goldfish (Carassius auratus L.) prominin

The integral membrane protein known as prominin was first identified on the apical surface of mouse neural epithelial cells as well as on the surface of human haematopoietic progenitor cells. This report describes a prominin-like sequence and expression analysis of the prominin in the goldfish. The predicted amino acid sequence for goldfish prominin shares all of the hallmark structural characteristics of the prominin family, however the relatedness assessed using the percent amino acid identity indicated that goldfish prominin cannot be placed into the current mammalian dichotomy of type 1 or 2. The real time PCR analyses indicated that prominin was broadly expressed in different tissues with particularly high levels observed in the kidney and gill of the goldfish. Goldfish prominin was also found to be differentially expressed in subpopulations of in vitro-derived goldfish macrophages, with the highest expression observed in progenitor cells.     

Ultrastructure of the swim bladder of the goldfish,Carassius auratus

Summary The swim bladder of the cyprinid Carassius auratus (goldfish) is a two-chambered organ connected to the esophagus by a pneumatic duct. The anterior chamber is lined by a single type of squamous epithelial cell. Two types of epithelial cells are present in the posterior chamber. Flattened cells with differences in the electron density of the cytoplasm line most of the chamber. Darker cells generally contain large amounts of glycogen. Cuboidal epithelial cells also occur in the posterior chamber. A glandular layer external to the muscularis in the posterior chamber is composed of large cells containing little glycogen, an extensive Golgi apparatus, and numerous mitochondria with single large granules. Capillaries and nerves are present in large numbers in this layer. Blood vessels form micro-retia mirabilia in the submuscular layer external to the glandular layer. Vessels are of two distinct types with wide lumina and flattened endothelium characterizing the venous vessels. Arterial vessels have smaller lumina, thick endothelial cells with prominent pinocytotic vesicles, and surrounding pericytes. Collagen is present in three forms in this swim bladder — large tactoids in the tunica externa of the anterior chamber, smaller tactoids in the lamina propria of the posterior chamber, and small fibrils in all other areas.Supported by a Young Investigator Pulmonary Research Grant # 1 R23 HL 19593-01 and by HL 23338-01 from the National Institutes of Health