[Phosphorus absorption by the intestine of the eel (Anguilla anguilla L). 1. Demonstration and characteristics of in vitro phosphate ion transport in the fresh water eel].
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Anguilla↗
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Comparative studies on the chromosomes of the European eel (Anguilla anguilla L.) and the American eel (Anguilla rostrata Le Sueur).
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Anguilla↗
Acetylcholine receptors and myasthenia gravis: the effect of antibodies to eel acetylcholine receptors on eel electric organ cells.
Antisera to acetylcholine receptors purified from Electrophorus electricus were tested for their ability to bind to receptors on electric organ cells, block the depolarizing response of the cells to carbamylcholine, and inhibit binding of 125-I-alpha-bungarotoxin to the cells. It was found that although antibodies could bind to most of the receptors, resulting in substantial inhibition of the depolarizing response, binding of 125-I-alpha-bungarotoxin to the cells was only slightly inhibited. This was consistent with the observation that these antibodies did not compete for the toxin binding site on detergent-solubilized receptor. These results suggest that the antibodies inhibited receptor activity primarily by interfering with the ionophore of the receptor or its regulation by the acetylcholine binding site. However, the possibilities could not be completely eliminated that blockage of a small fraction of the binding sites caused a large inhibition of the depolarizing response or that bound antibody allosterically reduced binding affinity for carbamylcholine without completely inhibiting toxin binding.
Animals↗
Similarities and differences between the massive eel (Anguilla anguilla L.) devastations that occurred in Lake Balaton in 1991 and 1995.
In the past few years, two massive eel (Anguilla anguilla L.) devastations occurred in Lake Balaton, Hungary. In 1991, 300 tons of eel perished in the western basin of the lake, while in the summer of 1995 30 tons of eel died in the eastern part of the lake. Investigations carried out to find the causes of these ecocatastrophes included measurements of certain biochemical parameters: the blood sugar level, and the acetylcholinesterase (AChE, EC 3.1.1.7), lactate dehydrogenase (LDH, EC 1.1.2.3), glutamic-oxaloacetic transaminase (GOT, EC 2.6.1.1) and glutamic-pyruvic transaminase (GPT, EC 2.6.1.2) activities in the blood serum of the collected eels. In both 1991 and 1995, deltamethrin (DM), the active ingredient of the insecticide K-OTHRIN 1 ULV used against mosquitoes, was detected in the eels; in 1995 it was demonstrated in several other animal species, i.e., bream (Abramis brama L.), pike perch (Stizostedion lucioperca L.), and the common gull (Larus canus), and in sediment samples from the lake. Additionally, laboratory experiments were carried out to study the effects of DM on eels. In 1991, eels were collected from the western (the site of the devastation) and eastern basins of the lake. The eels from the eastern basin were used as controls. At that time, the AChE activity in the blood serum of the eels from the western basin was significantly inhibited compared to that in animals from the eastern basin (P < 0.05, Student t test). Eels from the western part of the lake had GOT and GPT levels 20 and 100%, respectively, higher than those of eels from the eastern part of the lake. The blood glucose level was much higher in the eels from the affected area of the lake as compared to those from the eastern part. The brain and liver of the eels contained DM residues at 20 micrograms/kg wet tissue (Gönczy, 1992). Gönczy suspected that one of the causes of the massive eel loss in 1991 was the presence of DM in the fish. In 1995, when the eel devastation occurred in the eastern basin, moribund and surviving eels were collected from this part of the lake. The AChE activity was significantly inhibited in the blood serum of the dying eels as compared to that in surviving animals (P < 0.05, Student t test). The blood glucose content exhibited a difference too: it was 2.5 times higher in the dying eels than in the surviving ones. A huge increase in the LDH level was measured in the dying eels, indicating damage to different muscle tissues to an extent never observed previously. The GOT activities of the serum were twice as high in the dying eels as in the living fish. The GPT was not significantly changed in the serum of dying eels as compared to the surviving animals. DM was detected in different tissue samples of the dying eels: 2.7-18.5 micrograms/kg in the liver, 9.0-31.1 micrograms/kg in the gill, and 3.0 micrograms/kg wet tissue in the muscle. DM residues were found in tissue samples from other animals, in the following concentrations: 0.4 micrograms/kg in bream, 2.1 micrograms/kg in pike perch, 1.1 micrograms/kg wet tissue in dead gulls. The sediment samples collected from different places and at different times contained DM in a concentration of 5.5-30.0 micrograms/kg wet sediment at the time of the eel deaths, while the sediment samples collected from the same places a month later still contained DM at 7.0-8.8 micrograms/kg wet sediment. Laboratory experiments with the insecticide K-OTHRIN 1 ULV revealed that 1.0 microgram/liter of its active ingredient, DM, caused the death of 50% of the eels after an incubation time of 96 hr. In the liver of the dead eels, DM was detected at 2.9-20.0 micrograms/kg wet tissue. All the above-mentioned changes and the DM residue detected in the eels appear to demonstrate the contribution of DM in the severe eel devastation. This finding on the ecological risk of such types of insecticides might be useful in their further application.
Acetylcholinesterase↗
The contribution of a pyrethroid insecticide to the massive eel (Anguilla anguilla) devastation, in Lake Balaton, in 1995.
In the summer of 1995, 30 tonnes of eel (Anguilla anguilla) died in Lake Balaton, Hungary. An investigation was carried out to find the causes of this ecocatastrophe. During this investigation, certain biochemical parameters, i.e. the blood sugar level, the acetylcholinesterase (AChE, EC 3.1.1.7), lactate dehydrogenase (LDH, EC 1.1.2.3), glutamic-oxaloacetic transaminase (GOT, EC 2.6.1.1), and glutamic-pyruvic transaminase (GPT, EC 2.6.1.2) activities in the blood serum of the collected surviving and dying eels were examined. Deltamethrin, the active ingredient of the insecticide K-OTHRIN 1 ULV, used against mosquitoes was detected in different animal species, i.e. eel, bream (Abramis brama), pike perch (Stizostedion lucioperca), and the common gull (Larus canus) and in sediment samples from the lake. Additionally, laboratory experiments were carried out to study the effects of deltamethrin on eels. During the investigation in the field it appeared that the AChE activity was significantly lower in the blood serum of the dying eels as compared to that in living animals (P<0.05, Student's t-test). The blood glucose content exhibited a difference, too: it was 2.5 times higher in the dying eels than in the surviving ones. A huge increase in the LDH level was measured in the dying eels. The GOT activities of the serum were twice as high in the dying eels as in the living fish, while the GPT was not significantly changed. Deltamethrin was detected in different tissue samples of the dying eels: 2.70-18.1 microg/kg in the liver, 9.0-31.1 microg/kg in the gill and 3.0 microg/kg wet tissue in the muscle. Deltamethrin residues were found in tissue samples from other animals, in the following concentrations: 0.44 microg/kg in bream, 2.14 microg/kg in pike perch and 1.06 microg/kg wet tissue in dead gulls. The sediment samples collected from the sites of the devastation contained deltamethrin in a concentration of 5.50-30.00 microg/kg wet sediment at the time of the eel deaths, and in a concentration 7.00-8.75 microg/kg wet sediment a month later. Laboratory experiments with the insecticide K-OTHRIN 1 ULV revealed that 1.00 microg/l of its active ingredient, deltamethrin, caused the death of 50% of the eels after an exposure time of 96 h. During this experiments similar trends could be observed in changes of enzyme activities of the treated eels to those that were detected in filed study during the eel devastation in Lake Balaton. At the end of a one-week treatment with the insecticide at the concentration of 0.5 microg/l of its active ingredient the gills of the treated eels contained deltamethrin at 12.6-44.8 microg/kg wet tissue concentration, while at the 24th hour after the treatment (11.2-42.7 microg/kg wet tissue) deltamethrin concentration in the liver of treated eels could be detected. All the above-mentioned changes and the detected deltamethrin residue in the eels appear to demonstrate the contribution of deltamethrin to the severe eel devastation. This information on the ecological risk of pyrethroid insecticides might be useful in their further application.
Animals↗
Effects of homologous atrial natriuretic peptide on drinking and plasma ANG II level in eels.
The effects of eel atrial natriuretic peptide (ANP) on drinking were investigated in eels adapted to freshwater (FW) or seawater (SW) or in FW eels whose drinking was stimulated by a 2-ml hemorrhage. An intra-arterial infusion of ANP (0.3-3.0 pmol . kg-1 . min-1), which increased plasma ANP level 1.5- to 20-fold, inhibited drinking dose dependently in all groups of eels. The drinking rate recovered to the level before ANP infusion within 2 h after infusate was replaced by saline. The inhibition at 3.0 pmol . kg-1 . min-1 was profound in FW eels and hemorrhaged FW eels, whereas significant drinking still remained after inhibition in SW eels. Plasma ANG II concentration also decreased dose dependently during ANP infusion and recovered to the initial level after saline infusion in all groups of eels. The decrease at 3.0 pmol . kg-1 . min-1 was large in FW eels and hemorrhaged FW eels compared with that of SW eels. Thus the changes in drinking rate and plasma ANG II level were parallel during ANP infusion. Plasma sodium concentration and osmolality decreased during ANP infusion in SW and FW eels, and they were restored after saline infusion. In hemorrhaged FW eels, however, ANP infusion did not alter plasma sodium concentration and osmolality. Hematocrit did not change during ANP infusion in any group of eels. Collectively, ANP infusion at physiological doses decreased drinking rate and plasma ANG II concentration in parallel in both FW and SW eels. It remains undetermined whether the inhibition of drinking is caused by direct action of ANP or through inhibition of ANG II, which is known as a potent dipsogen in all vertebrate species, including eels.
Angiotensin II↗
Endocrine and environmental influences upon plasma cortisol concentrations and plasma renin activity of the eel, Anguilla anguilla L.
The plasma concentrations of cortisol, sodium, potassium and calcium and plasma osmolarity were determined in freshwater silver eels, after intravascular injections of eel renin preparations, mammalian ACTH, mammalian angiotensin II and eel muscle extracts. Arterial blood specimens were taken before and after injection of test substances. Partially purified eel and rat renal renins gave prolonged pressor responses in intact and hypophysectomized eels and in the nephrectomized rat anaesthetized with sodium pentobarbitone. Angiotensin, but not ACTH, produced obvious pressor responses in intact and hypophysectomized eels and in eels without their corpuscles of Stannius. Hypophysectomized eels 4-8 days after operation had reduced plasma cortisol concentrations. No change in cortisol occurred in eels after removal of the corpuscles of Stannius. Eel renin preparations and ACTH gave increased concentrations of plasma cortisol 30 min after injection into hypophysectomized and intact eels. In general, the length of the renin-generated pressor response and the increased cortisol concentration were concomitant occurrences. Angiotensin injected into eels with corpuscles of Stannius removed and into hypophysectomized eels also increased cortisol levels. Control muscle extracts produced no significant changes. There were no acute changes in plasma electrolyte concentrations after the injections. Plasma renin activity measured indirectly by bioassay of angiotensin generated in vitro was more than twice as great in eels adapted to seawater than in eels in fresh water. Plasma renin activity gradually fell when eels were transferred from seawater to fresh water, and increased when the reverse transfer was carried out.
Adaptation, Physiological↗
[Residues of total mercury and methyl mercury in eel products].
Fifty-two samples of broiled eels and broiled eel liver were analyzed for total mercury (total Hg) and methyl mercury. The mean concentrations of total Hg in broiled eels and broiled eel liver were 0.21 ppm and 0.10 ppm, respectively. Meanwhile, the mean concentrations of methyl mercury in broiled eels and broiled eel liver were 0.085 ppm and 0.039 ppm, respectively. The rate of methyl mercury to total Hg mainly ranged from 60 to 80% in broiled eels and from 35 to 65% in broiled eel liver. The total Hg concentrations of 2 samples of broiled eels and one sample of broiled eel liver exceeded the provisional regulation limit (0.4 ppm) of total Hg in fish in Japan. In these samples, the rates of methyl mercury to total Hg were lower than 20%. The muscles and the skin of broiled eels were measured separately. The ratios of skin to muscle concentration of total Hg and methyl mercury were mainly in the range from 1/10 to 1/4. The mean intakes of total Hg from broiled eels and broiled eel liver per individual were 24.6 microg and 3.1 microg, respectively. The mean intakes of methyl mercury from broiled eels and broiled eel liver per individual were 10.4 microg and 1.2 microg, respectively.
Animals↗
Eel ventricular natriuretic peptide: isolation of a low molecular size form and characterization of plasma form by homologous radioimmunoassay.
Ventricular natriuretic peptide (VNP) with 25 amino acid residues was isolated from the low molecular weight fraction of acid extracts of eel cardiac ventricles. No other short forms of VNP were recovered from the fraction. This peptide was named eel VNP(1-25) because it was a C-terminally truncated form of the previously isolated eel VNP(1-36). As observed before with eel VNP(1-36), eel VNP(1-25) had a much higher (146-fold) vasodepressor activity than human atrial natriuretic peptide (ANP) in eels, but was a third to a half as active in rats with respect to vasodepressor and natriuretic activities. Eel VNP(1-25) was generally less potent than eel VNP(1-36) for vasodepressor and natriuretic effects. A specific radioimmunoassay (RIA) has been developed for the measurement of eel VNP. The antiserum, raised against eel VNP(1-36), was highly specific and did not exhibit significant cross-reactivity with eel ANP and C-type natriuretic peptide, even though their amino acid sequences have more than 60% homology with that of eel VNP. The sensitivity of assay was 0.5 fmol/tube for eel VNP(1-36) with more than 99% confidence. Such high sensitivity permitted direct assaying of VNP with only a few microliters of plasma. In fresh water eels, the concentration of VNP in the cardiac ventricle was higher than those in the atrium or brain and that of ANP in the ventricle. Thus, VNP seems to be a ventricular hormone. Although ANP is a major circulating hormone in mammals, the plasma concentration of VNP was threefold higher than that of ANP.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Random amplified polymorphic DNA analysis of eel genome.
Eel family is a huge one, in which many kinds of eels especially some migratory eels, bear strong resemblance to each other, and are therefore difficult to be identified. In this study 29 random primers were used to make RAPD analysis for Japanese eel (Anguilla japonica), European eel (Anguilla anguilla) and Pike eel (Muraenesox cinereus). And totally 299 fragments were counted. Shared or specific fragments were counted and genetic similarity or genetic distance were calculated. The genetic similarity between Japanese eel and Pike eel is 0.68 and the genetic distance between them is 0.32; those between European eel and Pike eel are 0.72 and 0.28 respectively, and between Japanese eel and European eel are 0.74 and 0.25 respectively. The method has been shown to be suitable to molecular identification of eels. It provides an alternative approach to determine the relationship between species.
Animals↗
Development of a heterologous radioimmunoassay for eel (Anguilla anguilla) gonadotropin.
A heterologous radioimmunoassay (RIA) for eel gonadotropin (GTH) was developed, using an antiserum to the beta subunit of carp GTH (cGTHbeta). Partially purified eel GTH preparations or crude eel pituitary extracts produced partial cross-reactions: the inhibition curves reached a plateau before complete inhibition of the binding of the tracer and the slopes were lower than those for cGTH. However, the parallelism between the inhibition curves produced by all the eel samples allowed estimation of their GTH activity relative to an eel GTH standard; under these conditions, the RIA exhibited good reproducibility within and between assays. Three different RIA systems for cGTHbeta using different antisera gave similar estimations of immunoreactive GTH (irGTH) content of eel GTH preparations and pituitary extracts. The specificity of teh heterologous RIA for eel GTH was first assessed by comparing RIA and bioassay data: as with carp pituitary extracts, no significant difference was found between immunological and biological estimations of GTH activity for partially purified or crude pituitary extracts from normal female eel. Further validation of the specificity of the RIA for eel GTH was provided by the variation of the eel pituitary irGTH content under different hormonal treatments: a significant increase in female eel pituitary irGTH content was produced by treatment with estradiol-17 beta or with carp pituitary extract, a result in agreement with previous histological reports of Olivereau and collaborators. The heterologous RIA for eel GTH, in view of its sensitivity, reproducibility, and specificity, is a useful tool for physiological studies in the eel.
Anguilla↗
Effects of 17 beta-estradiol and carp gonadotropin on vitellogenesis in normal and hypophysectomized European silver female eel (Anguilla anguilla L.) employing a homologous radioimmunoassay for vitellogenin.
The European silver eels are at the early stages of vitellogenesis before the marine reproductive migration. Vitellogenesis was induced by 17 beta-estradiol (E2) alone and by a purified carp gonadotropin (cGTH). We studied and compared their effects on plasma vitellogenin (Vg) levels and ovarian yolk contents in female normal (N) and hypophysectomized (H) eels for both treatments. To this purpose an homologous radioimmunoassay (RIA) was established. Eel Vg was purified to homogeneity on 0.1% SDS-Electrophoresis. Native Vg has a molecular weight of 340 +/- 15 kilodalton (kDa) and was partially separated into subunits. The RIA was established with a sensitivity of 1.1 ng and was specific for eel Vg. In control (N and H) silver eels, plasma Vg levels were 0.04 +/- 0.02 microgram/ml and unchanged throughout the experiment. Similarly, yolk was indetectable in control ovarian extracts. E2 treatment increased plasma Vg levels proportionally with time to 783.4 +/- 130.7 micrograms/ml in N eels. The same profile was seen in H eels but terminally the mean value was 36.7 times lower than in N eels (P less than 0.01). Yolk at 0.005 microgram/g in N eels was indetectable in H eels. cGTH treatment gave a biphasic kinetic change: plasma Vg increased within 12 days, peaked at 93.6 +/- 13.0 micrograms/ml at 20 to 24 days, and stabilized to decrease at 40.2 +/- 7.5 micrograms/ml. The gonadosomatic index (GSI) increased alongside the yolk content (980.4 +/- 153.1 micrograms Vg/g). The kinetic profile for H eels was different: a peak was not apparent, rather there was a delayed increase, and at 67 days levels were still 8.23 times lower than in N eels (P less than 0.01). The GSI increased as the yolk content to 202.7 +/- 64.8 micrograms Vg/g ovary showing an ovarian incorporation of Vg in H eels.
Anguilla↗
Purification of follicle-stimulating hormone from immature Japanese eel, Anguilla japonica, and its biochemical properties and steroidogenic activities.
Follicle-stimulating hormone (FSH) was purified, for the first time, from immature Japanese eel, Anguilla japonica, and its biochemical properties were investigated. FSH was extracted from immature eel pituitaries and purified by gel-filtration on Sephadex G-100, and two step anion-exchange chromatography: stepwise elution on DE-52, followed by gradient elution on TSK-gel Super-Q using HPLC. Purification was performed using its molecular mass and the positive reaction with anti-Japanese eel (je) FSHbeta antiserum. Purified eel FSH was detected as a single band after separation by SDS-PAGE under a non-reducing condition, showing positive reaction with both anti-je glycoprotein (GP) alpha and anti-jeFSHbeta antisera. The molecular mass of purified eel FSH was estimated to be approximately 33 kDa. After separation by SDS-PAGE under reducing condition, the intact molecule was detected as distinct proteins, whose N-terminus amino acid sequences coincided with those predicted from cDNA sequences for jeGPalpha and jeFSHbeta mature peptides. Deglycosylation of these subunits led to a decrease in their molecular mass. These results suggest that eel FSH is a heterodimeric molecule which consists of distinct glycoprotein subunits, GPalpha and FSHbeta. Cells reacting with anti-jeFSHbeta antiserum were observed in the proximal pars distalis of an immature eel pituitary, while jeLHbeta-immunoreactive cells were not detected. Gonadotropic activities of eel FSH were demonstrated in vitro by stimulating testosterone and 11-ketotestosterone secretions in immature eel testes. Purified eel FSH stimulated the secretion of both androgens from the immature eel testis in a dose-dependent manner, similar to immature eel pituitary homogenate and recombinant eel FSH produced by yeast. These results show that endogenous and recombinant FSH in this species possess similar activities, presumably stimulating the gametogenesis through the sex steroid secretion during the early stages of gonadal development.
Anguilla↗
Intestinal water absorption through aquaporin 1 expressed in the apical membrane of mucosal epithelial cells in seawater-adapted Japanese eel.
To elucidate the mechanisms associated with water absorption in the intestine, we compared drinking and intestinal water absorption in freshwater- and seawater-adapted Japanese eels, and investigated a possible involvement of aquaporin (AQP) in the absorption of water in the intestine. Seawater eels ingested more water than freshwater eels, the drinking rate being 0.02 ml kg(-1) h(-1) in fresh water and 0.82 ml kg(-1) h(-1) in sea water. In intestinal sacs prepared from freshwater and seawater eels, water absorption increased in time- and hydrostatic pressure-dependent manners. The water absorption rates were greater in seawater sacs than in freshwater sacs, and also greater in the posterior intestine than in the anterior. In view of the enhanced water permeability in the intestine of seawater eel, we cloned two cDNAs encoding AQP from the seawater eel intestine, and identified two eel homologues (S-AQP and L-AQP) of mammalian AQP1. S-AQP and L-AQP possessed the same amino acid sequence, except that one amino acid was lacking in S-AQP and two amino acids were substituted. Eel AQP1 was expressed predominantly in the intestine, and the expression levels were higher in seawater eel than in freshwater eel. Immunocytochemical studies revealed intense AQP1 immunoreaction in the apical surface of columnar epithelial cells in seawater eel, in which the immunoreaction was stronger in the posterior intestine than in the anterior. In contrast, the immunoreaction was faint in the freshwater eel intestine. Preferential localization of AQP1 in the apical membrane of epithelial cells in the posterior intestine of seawater eel indicates that this region of the intestine is responsible for water absorption, and that AQP1 may act as a water entry site in the epithelial cells.
Amino Acid Sequence↗
11-Ketotestosterone induces silvering-related changes in immature female short-finned eels, Anguilla australis.
The developmental transition from a residential, immature 'yellow' eel to a migratory, maturing adult 'silver' eel is accompanied by many morphological changes that appear to be under endocrine control. High circulating levels of the teleost, and usually male-specific, androgen 11-ketotestosterone (11-KT) are found in migrating female short-finned eels, Anguilla australis. We examined the role of this steroid in silvering by implanting immature, female short-finned eels either with blank vehicles or with vehicles containing 11-KT. Six weeks after they had received the implants, eels treated with 11-KT had developed 'chisel-shaped' snouts and black pectoral fins with tapered ends, and the size of their eyes had increased significantly. 11-KT treated eels had a thicker dermis than control eels and an epidermis with fewer or no mucous cells. Ventricular mass at the end of the experiment was two-fold larger than in control eels. 11-KT treated eels also had larger livers and gonads. Ovaries contained predominantly cortical alveolus stage III oocytes, as opposed to the smaller gonads of control eels containing previtellogenic stage II oocytes. All of these changes correspond to changes during the developmental transition from yellow to silver eels in the wild. This demonstrates that silvering in eels is under endocrine control and that the presumed male-specific steroid 11-KT is capable of inducing silvering-related changes in a female teleost. We discuss how species-specific responses to 11-KT may differ depending on tissue-specific androgen receptor abundance and how a dual demand on liver function can explain the apparently positive effects of 11-KT on liver growth.
Animals↗
Atrial natriuretic peptide in eel plasma, heart and brain characterized by homologous radioimmunoassay.
A highly specific and sensitive radioimmunoassay has been developed for the measurement of eel atrial natriuretic peptide (ANP). The antiserum, raised against eel ANP-(1-27) did not cross-react with two other eel natriuretic peptides, i.e. eel ventricular natriuretic peptide and C-type natriuretic peptide (CNP), or with any mammalian ANPs, CNPs or brain natriuretic peptides so far identified. The minimal detectable amount was 0.39 fmol (0.90 pg)/tube with more than 99% confidence. Because of its high sensitivity, the radioimmunoassay makes it possible to measure eel ANP directly with only a few microlitres of plasma without extraction. Using the radioimmunoassay we found high levels of ANP in the atrium (11 +/- 2 pmol/mg wet tissue, n = 8), and much lower levels in the ventricle (56 +/- 8 fmol/mg, n = 8) and the brain (22 +/- 1 fmol/mg, n = 8) of eels. Eel plasma contained a large amount of ANP (247 +/- 66 fmol/ml, n = 8) compared with the levels reported in mammals, although atrial levels are similar between eels and mammals. Gel-permeation chromography revealed that a major form of ANP stored in the eel atrium, ventricle and brain has a molecular mass of approximately 14 kDa but low molecular forms of about 3 kDa are predominant in eel plasma. A detailed analysis with reverse-phase high-performance liquid chromatography showed that a major molecular form circulating in eel plasma is ANP-(1-27). ANP-(1-27) was also detected in small amounts in the eel atrium, ventricle and brain.
Amino Acid Sequence↗
Metamorphosis of the American eel, Anguilla rostrata LeSeur: I. Changes in metabolism of skeletal muscle.
Unequivocal demarcation between immature, nonmigratory yellow eels and migratory silver eels of greater sexual maturity is possible by measuring eye diameter and retinal capillary length, which undergo a 1.5- and 2.3-fold increase during metamorphosis, respectively. Anatomical arrangement of trunk musculature is similar in the two groups except for an increased depth of slow muscle in silver eel. Histochemical analysis reveals a progressive increase in numbers of "displaced" fast fibres within slow muscle of the lateral line triangle in maturing eels, although these are unlikely to affect recruitment pattern of muscle fibre types. Previous studies have suggested greater involvement of fast muscle in locomotion of migratory eels. In contrast, estimates of enzyme activity in fast muscle suggest an inadequate aerobic capacity to fuel sustained activity. Myoglobin content is extremely low, around 0.4 nM g wet wt-1. Prolonged anaerobic metabolism is also discounted as a migratory strategy. Increased energy provision for migration is apparently derived from increased capacity for both aerobic carbohydrate metabolism and mitochondrial fatty acid oxidation within slow muscle of silver eels. Activity of hexokinase (HK) shows a 1.6-fold increase (to 0.51 microM g wet wt-1) and carnitine palmitoyltransferase (CPT) a 3.1-fold increase (to 0.22 microM g wet wt-1 min-1), suggesting a maximal flux through these pathways of 18 and 14 ATP equivalents, respectively. However, the fatty acyl transferase system of skeletal muscle mitochondria displays up to threefold greater activity with palmitoleoyl CoA (C16:1) as substrate than with the usual palmitoyl CoA (C16:0). Slow muscle of silver eel is therefore capable of deriving aerobic energy from free fatty acids and carbohydrate in the ratio 2.3:1. Differences in aerobic enzyme activities are not paralleled by myoglobin content of slow muscle, being 15 and 16 nM g wet wt-1 for yellow and silver eel, respectively. Structural reorganization of muscle fibres during metamorphosis, however, results in a twofold elevation of cytoplasmic myoglobin concentration in silver eel. It would appear that dramatic differences in metabolic capacity between life history stages of eel is required to overcome locomotory inefficiency of yellow eels and to "preadapt" silver eels for migratory activity. This increased locomotory capacity may be amplified by a subsequent training response.
Aging↗
Primary cultures of dispersed pituitary cells from estradiol-pretreated female silver eels (Anguilla anguilla L.): immunocytochemical characterization of gonadotropic cells and stimulation of gonadotropin release.
To analyze the multihormonal control mechanisms of GTH secretion in the eel, primary culture of pituitary cells from control or estradiol-treated female silver eels, a treatment known to stimulate in vivo GTH synthesis, was developed. Dispersed eel pituitary cells obtained by enzymatic (trypsin/DNAse) and mechanical dispersion were cultured in Earles M199, at 18 degrees. Immunoreactive GTH (ir-GTH) cells were characterized by the immunoperoxidase method, using antiserum to carp GTH beta subunit. Ir-GTH cells from control silver eels were small and represented 14% of the dispersed pituitary cells. In contrast, ir-GTH cells from estradiol-treated eels were larger (cell area x 2.5) and represented a higher proportion (21%) of the pituitary cells. Intracellular and medium contents of GTH were measured by radioimmunoassay for the GTH beta subunit. In vivo estradiol-treatment increased more than 100-fold the GTH content of cell cultures. GTH release, studied over 1 to 4 hr, was undetectable in cultures from normal eels. In contrast, GTH release was low (less than 2% of cell content) but measurable in cultures from estradiol-treated eels. Subsequent experiments examined effects of various secretagogues on GTH release from primary cultures of pituitary cells from estradiol-pretreated eels. GTH release was significantly increased (x1.5 to x3 basal release) by 10(-6) M GnRH-A as well as by both native GnRHs in the eel (mammalian GnRH, mGnRH, and chicken GnRH-II, cGnRH-II), at the same concentration. Lower GnRH concentrations had no significant effect, indicating a low sensitivity of gonadotrophs to GnRH, likely to be related to their immature state at the silver stage. The similar efficacy of mGnRH and cGnRH-II suggested that the pituitary GnRH receptor had a low specificity toward various molecular forms, in the eel as in the other nonmammalian species. The protein kinase C (PKC) activator (phorbol ester: PMA) also stimulated GTH secretion, with a maximal effect at 10(-8) M, indicating that the PKC pathway was functional. In contrast, a depolarizing agent (50 mM KCl) had no significant effect on GTH release, suggesting lack of a functional voltagesensitive calcium channel (VSCC) secretory pathway. Perifusion experiments on whole pituitary confirmed the lack of effect of KCl on gonadotrophs from E2-pretreated eels and indicated that an additional in vivo treatment with GnRH agonist and dopamine antagonist could induce the differentiation of a functional VSCC pathway. These characteristics of the transduction mechanisms may be related to the immature state of the eel gonadotrophs at the silver stage.
Anguilla↗