Cloning and molecular characterization of an ovarian-derived (brain-like) P450 aromatase cDNA and development of a competitive RT-PCR assay to quantify its expression in the fathead minnow (Pimephales promelas)     

S. Halm  M. Rand-Weaver  J.P. Sumpter  C.R. Tyler 《Fish physiology and biochemistry》2001,24(1):49-62

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Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)     

Dra. Eva Danulat  Stephan Kempe 《Fish physiology and biochemistry》1992,9(5-6):377-386

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.  相似文献   

Pituitary and plasma AVT content in the flounder (Platichthys flesus)     

K.E. Harding  J.M. Warne  S. Hyodo  R.J. Balment 《Fish physiology and biochemistry》1997,17(1-6):357-362

Radioimmunoassay measurement of pituitary AVT content and plasma AVT concentration indicated comparable levels in fully adapted sea water (SW) and fresh water (FW) flounders. Circulating AVT represented less than 0.1% of the pituitary AVT reserve. The urophysis contained AVT but the total content was only 2 or 3 fold that of circulating AVT. In fish adapted to hypertonic media, there was a close correlation between plasma AVT concentration and plasma Na⁺ concentration or osmolality. The present study examined the effects of acute osmotic challenge, associated with FW to SW transfer, and the influence of extracellular fluid volume status on AVT secretion. Short-term transfer of fish from FW to SW (up to 3 days) did not evoke a clear change in plasma AVT levels, though pituitary content was reduced at 24 h. During the first 3 days after transfer to SW, only small increments in plasma tonicity were apparent. The sensitivity of AVT secretion to osmotic stimuli may only be expressed when plasma osmolality has exceeded a specific threshold, which was probably not reached in these transfer studies. Fish in hypotonic media showed no relationship between plasma osmolality/tonicity and plasma AVT concentration. Acute extracellular fluid volume expansion of SW adapted fish also abolished the normally observed relationship between plasma osmolality and AVT concentration in these hypertonic media fish. This trend indicates that volume status may modulate the sensitivity of AVT secretion to osmotic stimuli as occurs in tetrapods.  相似文献   

Control of ion transport by the sperm duct epithelium of brook trout (Salvelinus fontinalis)     

William S. Marshall  Sharon E. Bryson  David R. Idler 《Fish physiology and biochemistry》1989,7(1-6):331-336

The sperm duct epithelium of brook trout (Salvelinus fontinalis), mountedin vitro in Ussing-style epithelial chambers actively absorbs Na⁺ (measured as the short-circuit current, I_sc) and secretes K⁺ (measured using⁸⁶Rb⁺ as tracer). Dibutyryl-cyclic-adenosine monophosphate (db-cAMP) and 3-isobutyl-1-methylxanthine (IMX) produce a rapid, sustained stimulation of both ion transport processes, but the hormone connected to the response is unknown. Purified sockeye salmon CON A2 gonadotropin (GtH) produces a dose-dependent, rapid and sustained rise in Na⁺ uptake and K⁺ secretion. The time course, electrophysiological and transport characteristics are similar to those evoked by IMX. Carbohydrate-poor (chum salmon CON A1) GtH is ineffective. Pretreatment of fish with 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-P) significantly increases milt volume but is without effect on resting or stimulated (IMX + db-cAMP) levels of sperm duct ion transport. This is the first indication of a direct, rapid action of GtH on ion transport by the vertebrate blood-testis barrier. The results suggest direct involvement of GtH in control of later stages of sperm maturation.  相似文献   

Oxygen delivery to the fish eye: blood flow in the pseudobranchial artery of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)     

W.P.?WaserEmail author  N.?Heisler 《Fish physiology and biochemistry》2004,30(1):77-85

Similar to gas secretion into the swimbladder, some teleosts also exhibit ocular gas secretion to supply their thick and avascular retina with oxygen. The process in the eye, however, is complicated by the fact that the retina is supplied with arterial blood only indirectly through the pseudobranch, a reduced gill arch of still unknown function. Approaches for closer analysis of the physiological implications of this serial blood supply arrangement and the associated process of O₂ release include complete unilateral vascular isolation of eye and/or pseudobranch while maintaining normal blood flow characteristics for undisturbed operation of the counter-current rete mirabile.Blood flow velocity in the afferent pseudobranchial artery (aPA; essentially equal to ophthalmic artery flow) of rainbow trout was measured by application of ultrasonic Doppler techniques. Accurate determination of flow rate in the untouched vessel was facilitated by mounting the Doppler crystal with a custom-made holder on the bone of the jaw and raw signal analysis by Fast Fourier Transformation for improved signal/noise ratio. Normal unilateral aPA blood flow was 746 l min^–1 kg^–1 body weight, (1.6% of cardiac output), the tissue-weight-specific blood flow was 3.6 ml min^–1 g^–1 for pseudobranch and 2.3 ml min^–1 g^–1 for the retina. Only about 28% of the oxygen potentially available with maximal activation of the Root effect is actually consumed in pseudobranch and retina. This generous supply of blood and the implicit safety margin may serve minimization of pH disturbances of retinal tissues, but may also come suitable during unilateral occlusion of supplying arteries for continued oxygen supply to the retinae. During contralateral occlusion, flow through the ipsilateral aPA increased to 136%. According to the normally low O₂ extraction, a minimal flow of 36% through bilateral anastomosis and contralateral ophthalmic artery may maintain complete contralateral oxygen supply.  相似文献   

Development of an enzyme-linked immunosorbent assay (ELISA) for vitellogenin measurement in greenback flounder Rhombosolea tapirina     

B. Sun  N.W. Pankhurst  M. Watts 《Fish physiology and biochemistry》2003,29(1):13-21

Vitellogenin (Vtg) was purified from the plasma of 17-estradiol (E₂)-injected male greenback flounder,Rhombosolea tapirina. The molecular weight of the native Vtg was estimated by gel filtration as 540 kD. SDS-PAGE and Western blotting analyses indicated that this protein consisted of three bands with molecular weights of 155, 104, 79 kD, respectively. A polyclonal antibody against the highest molecular weight band of putative Vtg was generated in sheep and an indirect antibody-capture competitive enzyme-linked immunosorbent assay (ELISA) was developed. The assay was validatedfor plasma Vtg measurement in greenback flounder. Serial dilutions of plasma from vitellogenic females parallelled the standard Vtg curve, whereas no cross-reaction was observed with the plasma of males in the ELISA. The Vtg ELISA was used to assess the induction of Vtg by E₂ in vivo in males. The induction of Vtg in greenback flounder showed a time- and dose-dependent response as in other species. In E₂-treated fish, detectable levels of Vtg were first found at 48 h, and reached a peak at 96 h post-injection. Plasma levels of Vtg increased as the E2 dose increased with a threshold of 0.1 mg kg^–1.  相似文献   

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)     

J.M. Puente-Novoa  P. Barja 《Fish physiology and biochemistry》1999,20(2):101-114

PKC activity was detected in spleen extracts from the turbot, Scophthalmus maximus, a teleost flatfish that is farmed commercially in several countries, in assays with the substrate EGF- R651–658 as phosphate acceptor. The activity was purified about 700-fold by a three-step chromatographic procedure (DEAE-cellulose, phenyl-Sepharose and threonine-Sepharose). Maximal activity was obtained in the presence of the typical PKC cofactors Ca2+ (0.1 mM) PtdS (20 g ml–1) and either DAG (2 g ml–1) or PMA (2 g ml–1). Activity was dose-dependently inhibited by H7 and by the PKC-specific inhibitors PKC19–36 and N-myristoylated PKC19–31. The rate of phosphorylation was highest with the PKC-specific substrate MARCKS161–175. In immunoblotting, MC5 (a mouse monoclonal antibody raised against bovine PKC) recognized bands of 80 and 100 kDa. Immunoblotting with antibodies raised against mouse PKC isozymes (, , , , , , and ) indicated the presence of all these isozymes in turbot spleen.  相似文献   

Circulating levels and in vitro production of two maturation-inducing hormones in teleost: 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20β,21-trihydroxy-4-pregnen-3-one, in a daily spawning wrasse, Pseudolabrus japonicus     

M. Matsuyama  K. Ohta  S. Morita  M.M. Hoque  H. Kagawa  A. Kambegawa 《Fish physiology and biochemistry》1998,19(1):1-11

The female bambooleaf wrasse, Pseudolabrus japonicus, spawns daily during the spawning season, and exhibits a diurnal rhythm of ovarian development. In the present study, we have investigated: (1) circulating levels of 17a, 20-dihydroxy-4-pregnen- 17,20-P) and 17,20,21-trihydroxy-4-pregnen-3-one (20-S) in females sampled at different times of the day during spawning season in captivity, and (2) in vitro production of 17,20-P and 20-S by follicle-enclosed oocytes at seven different develo tal stages. In addition, we developed a microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17,20-P. Serum levels of 17,20-P and 20-S showed similar diurnal changes; substantial increases in these levels occurred around the time of germinal vesicle breakdown (GVBD). In vitro experiments showed that massive production of 17,20-P and 20-S occurred in follicles collected just before or during GVBD. Further, acute decreases in 17,20-P and 20-S production were found in the ovarian follicles just prior to ovulation, suggesting inactivation of the maturation-inducing hormone (MIH). These results, taken together with our previous data on the occurrence of GVBD in vitro, suggest a role for both 17,20-P and 20b-S as MIHs in the bambooleaf wrasse.  相似文献   

Effects of (D-Ala6, Pro9N ethylamide) — LHRH on plasma levels of gonadotropin, 17α,20β-dihydroxy-4-pregnen-3-one and testosterone in male goldeye (hiodon alosoides Rafineque)     

N. W. Pankhurst  G. van der Kraak  R. E. Peter  B. Breton 《Fish physiology and biochemistry》1986,1(3):163-170

Male goldeye were treated with (D-Ala⁶, Pro⁹-N ethylamide) — LHRH (LHRH-A) in saline or a silastic pellet (100 µg.kg^–1 body weight) and changes in plasma gonadotropin (GtH), 17,20-dihydroxy-4-pregnen-3-one (17,20P) and testosterone (T) levels were determined by radioimmunoassay. LHRH-A increased plasma GtH levels, with the response to LHRH-A in saline being of much greater magnitude and duration than the response to silastic pellet implants. Seasonal differences were found in the response to LHRH-A. Spermiated fish were the most responsive, recrudescing fish the least, and regressed fish displayed an intermediate response. Plasma 17,20P levels were elevated in response to LHRHA in fish of all sexual stages although the magnitude of the increase was not related to the magnitude of the increase in GtH levels. Treatment with LHRH-A also resulted in a transitory increase in plasma T levels. The endocrine control of GtH and steroid secretion in goldeye is discussed in relation to studies in cyprinids and salmonids. Address for reprint requests: Department of Zoology, University of Alberta, Edmonton, Alberta T6G 2E9 Canada.  相似文献   

Endocrine and gonadal changes during the annual reproductive cycle of the freshwater teleost,Stizostedion vitreum     

Jeffrey A. Malison  Lynne S. Procarione  Terence P. Barry  Anne R. Kapuscinski  Terrence B. Kayes 《Fish physiology and biochemistry》1994,13(6):473-484

The annual reproductive cycle of walleye (Stizostedion vitreum) was characterized by documenting changes in gonadal development and serum levels of estradiol-17β (E₂), testosterone (T), 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P), and 11-ketotestosterone (11-KT) in wild fish captured from upper midwestern lakes and rivers throughout the year. Fish from the populations used in this study spawn annually in early- to mid-April. Walleye showed group synchronous ovarian development with exogenous vitellogenesis beginning in autumn. Oocyte diameters increased rapidly from ∼ 200 μm in October to ∼ 1,000 μm in November, and reached a maximum of 1,500 μm just prior to spawning. Changes in gonadosomatic indices (GSIs) paralleled changes in oocyte diameters. Serum E₂ levels in females increased rapidly from low values in October (< 0.1 ng ml⁻¹) to peak levels of 3.7 ng ml⁻¹ in November, coinciding with the period of the most rapid ovarian growth. Subsequently, E₂ levels decreased from December through spawning. Serum T levels exhibited a bimodal pattern, increasing to 1.6 ng ml⁻¹ in November, and peaking again at 3.3 ng ml⁻¹ just prior to spawning. We detected 11-KT in the serum of some females at concentrations up to 5.6 ng ml⁻¹, but no seasonal pattern was apparent. In this study (unlike our results in a related study) 17,20-P was not detected. In males, differentiation of spermatogonia began in late August, and by January the testes were filled (> 95% of germ cells) with spermatozoa. Mature spermatozoa could be expressed from males from January through April. GSIs ranged from 0.2% (post-spawn) to 3.2% (pre-spawn). Serum T levels rose from undetectable levels in post-spawn males to 1.6 ng ml⁻¹ by November, remained elevated throughout the winter, and peaked at 2.8 ng ml⁻¹ I prior to spawning. Levels of 11-KT in males remained low (< 10 ng ml⁻¹, from post-spawning through January, then increased significantly by March and peaked just prior to spawning at 39.7 ng ml⁻¹. Our results indicate that vitellogenesis and spermatogenesis are complete or nearly so, in walleye by early winter, and suggest that it may be possible to induce spawning in this species several months prior to the normal spawning season by subjecting fish to relatively simple environmental and hormonal treatments.  相似文献