Qualitative and quantitative strategies of thermal adaptation of grass carp (Ctenopharyngodon idella) cytoplasmic malate dehydrogenases

Malate dehydrogenase isozymes of grass carp,Ctenopharyngodon idella, were identified by mitochondrial preparation and thermal denaturation. The structural and kinetic characteristics of chromatographically separated thermostable and thermolabile cMDHs were different in (1) half-life at 42°C, 10 min and 24 h, respectively, (2) optimal substrate, oxaloacetate and malate, concentrations, and (3) the apparent Michaelis-Menten constants of NADH and oxaloacetate.Total MDH activity in white muscle of 11°C-acclimated fish was about twice that of the 30°C-acclimated group. In addition, the ratio of the thermostable to thermolabile cMDH activity in white muscle of 30°C-acclimated fish was significantly higher than that of 11°C-acclimated fish. These results suggest that temperature acclimation can induce temperature compensation in MDH activity and differential expression of thermostable and thermolabile cMDH isozymes in freshwater fish.     

Effects of metomidate anaesthesia or transfer to pur sea water on plasma parameters in ammonia-exposed Atlantic salmon (Salmo salar L) in sea water

Atlantic salmon (Salmo salar L) postsmolts weighing 150 ± 53 g were exposed to 14–15 mg l^–1 TA-N (total ammonia-N) in sea water in 1 m³ tanks for 24h. Blood samples were then taken A) immediately after the fish were netted from the exposure tanks and stunned by a blow to the head; B) 2–20 min after the fish were transferred to 15 l of an anaesthetic solution of metomidate in ammonia-free sea water; or C) 2–20 min after the fish were transferred to 15 l of ammonia-free sea water. Plasma TA-N level was 18% lower in the anaesthetised fish compared to in the fish sampled directly from the exposure tanks (p 0.05), and accordingly 16% lower in the fish transferred to pure sea water although this difference was not significant (p = 0.07). Plasma glucose level was higher in the fish transferred to pure sea water than in the fish receiving the two other treatments (p 0.05), but plasma urea, osmolality, Na^{+, Cl–, Ca2+ or Mg2+} levels did not vary significantly between the different treatments. Plasma TA-N level increased with time in the fish in the metomidate solution (p 0.02).     

Photoperiodic manipulations of the reproductive cycle of sea bass (Dicentrarchus labrax) and their effects on gonadal development, and plasma 17ß-estradiol and vitellogenin levels

The annual profile of plasma vitellogenin (VTG) and 17ß-estradiol (E₂) levels, as well as gonadal development and spawning characteristics were investigated in captive female sea bass (Dicentrarchus labrax). Endocrine and gonadal changes were studied in fish reared under natural conditions or exposed to manipulated photothermal cycles. In natural conditions of photoperiod and temperature, sea bass spawned from February through March (East coast of Spain, 40°N 0°E). One or two months of constant long-days (15L/9D) in a constant short-day photoperiod regime (9L/15D) all-year-round, given early in the year (March and March–April), advanced spawning by 3 months. The same treatment applied later in the year (September–October) delayed spawning by 1 month, compared to controls.In all groups, changes in plasma VTG levels were correlated with E₂ levels, oocyte growth and spawning time. In control females, VTG was low (<100 ng ml^-1) during the summer, until its first surge in plasma 4 months before the beginning of spawning. The VTG (3.1 ± 0.3 mg ml^-1) and E₂ (4.1 ± 0.5 ng ml^-1) levels showed a single annual peak during late vitellogenesis, the time of the highest proportion of vitellogenic oocytes in the ovary. Constant high levels of VTG (1–1.4 mg ml^-1) and E₂ (1.6–1.9ng ml^-1) were maintained during the entire spawning time, together with the presence of vitellogenic oocytes, suggesting the existence of several waves of oocyte growth in the ovary and thus, several spawns per female. Endocrine profiles and oocyte development in fish exposed to constant photoperiods were similar to controls, but were shifted in time in relation to the displacement of the spawning time. In the fish showing advanced spawns, the duration of the gametogenic proces was compressed when compared to controls. The differences observed in the evolution of the reproductive-related factors in the advanced groups, which were exposed to a reduction in temperature to 15°C, suggest an influence of the temperature in the early stages of the reproductive cycle in sea bass.     

Whole animal transepithelial potential (TEP) of coho salmon during the parr-smolt transformation and effects of thyroxine,prolactin and hypophysectomy

Whole animal transepithelial potentials (TEP) of yearling coho salmon (Oncorhynchus kisutch) in fresh water and after transfer to seawater were recorded throughout parr-smolt transformation (smoltification) from February to August 1984, along with plasma Na⁺ and Cl^– concentrations and osmolality. Based on plasma ion regulation in seawater, the yearling coho in this study completed smoltification and attained sea-water adaptability in April. TEP in freshwater fish decreased (became inside-negative) after smoltification, and the TEP increased significantly (P < 0.01) after seawater transfer. When fish were transferred into seawater, thyroxine increased TEP of the transferred smolts by approximately 30% over the control level (P < 0.01) in April, but this did not occur when freshwater postsmolts were transferred in July and August. Hypophysectomy increased TEP (P < 0.01) in fresh water; it did not affect the TEP of the fish after seawater transfer. Ovine prolactin (3 g/g body weight) implanted into seawater-adapted fish caused a reduction in TEP (P < 0.01) when fish were exposed to fresh water. Whole-animal TEP appears to provide a valuable index of the completion of smoltification (April–May) and a useful tool for investigating the endocrine control of salmonid osmoregulation.     

Thyroid hormones down-regulate thyrotropin β mRNA level in vivo in the turbot (Psetta maxima)

Thyroid stimulating hormone (TSH) is a vertebrate pituitary heterodimeric hormone that stimulates the thyroid gland to produce the thyroid hormones, T3 and T4. We report here the cloning, by PCR on reverse-transcribed pituitary RNAs, of a 180 bp fragment of the cDNA encoding TSH subunit in the turbot (Psetta maxima). The deduced amino acid sequence displayed 66 and 75% identity with the corresponding sequence from the European eel (Anguilla anguilla) and the rainbow trout (Oncorchyncus mykiss), respectively. This cDNA was then used as a probe for densitometric analysis of individual pituitary Northern blots. TSH mRNA levels were quantified in turbot where circulating thyroid hormones were modified by dietary treatments or hormone supplementation. Recombinant rainbow trout growth hormone had no effect on circulating thyroid hormone levels or on pituitary TSH mRNA level. In turbot fed heat-treated rapeseed meal, plasma T4 levels were lowered and TSH mRNA increased more than two fold. In contrast, when turbot were fed a standard fish-meal supplemented with T3, circulating T3 levels were elevated and there was a dramatic decrease in TSH mRNA level. It is concluded that both thyroid hormones are able to down-regulate TSH mRNA level in vivo in the turbot. These results are discussed in the context of the evolution of the TH feed-back on TSH production.     

The tilapia prolactin cell: A model for stimulus-secretion coupling

The tilapia prolactin (PRL) cell responds rapidly (10–20 min) to small physiological changes in medium osmotic pressure (OP), releasing increasing quantities of hormone as medium OP is reduced. This release is rapidly (≤ 10 min) inhibited by somatostatin (SRIF). There is now extensive evidence that tilapia PRL cell function is regulated through the second messengers Ca⁺⁺ and cAMP. Our studies have shown that PRL release is augmented by treatments that lead to increased levels of intracellular Ca⁺⁺ or cAMP. On the other hand, PRL release is blocked when tissues are incubated in Ca⁺⁺-depleted medium or upon the addition of Co⁺⁺, an inhibitor of Ca⁺⁺-mediated processes. The use of⁴⁵Ca⁺⁺ to characterize the movement of Ca⁺⁺ into PRL cells has provided evidence that an increase in the influx of extracellular Ca⁺⁺ may participate in PRL release upon exposure to hyposmotic medium. Our studies have also shown that SRIF suppresses the increase in⁴⁵Ca⁺⁺ accumulation that is brought about when OP is reduced. We have also examined the effects of OP and SRIF on cAMP levels. The reduction of medium OP did not alter cAMP metabolism during 20 min of incubation. By contrast, cAMP accumulation in the presence of IBMX was enhanced at 1 hr of incubation in reduced OP. Thus, an increase in cAMP turnover may play a role in maintaining PRL release under sustained stimulation. SRIF reduced the accumulation of cAMP during 10 min of incubation with IBMX and also reduced the forskolin-stimulated increase in cAMP. Thus, SRIF may suppress adenylate cyclase activity. Finally, our studies have revealed that the forskolin-stimulated increase in cAMP levels is not dependent upon medium Ca⁺⁺. The presence of Ca⁺⁺ in the medium is required, however, for PRL release even when the cAMP messenger system has been activated. Moreover, cAMP accumulation was augmented when intracellular Ca⁺⁺ was increased. This raises the possibility that reduced OP may stimulate an increase in cAMP turnover indirectly through its action(s) on cytosolic Ca⁺⁺.     

Dopamine inhibits gonadotropin secretion in the Chinese loach (Paramisgurnus dabryanus)

The effects of dopamine on gonadotropin (GtH) secretion in sexually mature Chinese loach were investigated. Spontaneous secretion of GtH was inhibited within 1 h following an intramuscular injection of dopamine (100 g/g body wt). Similarly, dopamine (50 and 100 g/g body wt) caused a significant reduction in serum GtH in fish with elevated GtH levels as a result of pretreatment with gonadotropin-releasing hormone (GnRH) analogs either alone or in combination with the dopamine receptor antagonist domperidone. In summary, the present study provides direct evidence that dopamine functions as a gonadotropin-release inhibitory factor in the Chinese loach by blocking spontaneous and GnRH-stimulated GtH release.     

怒江裂腹鱼鳃的显微和超微结构

鳃为鱼类重要的呼吸器官,是鱼类进行离子交换、酸碱调节和含氮废物排泄的重要结构基础,也是鱼类重要的外周黏膜免疫器官之一,在抵御病原微生物侵染过程中发挥重要的免疫屏障作用。当前,硬骨鱼类鳃黏膜免疫反应是研究热点之一。本文首先对硬骨鱼类鳃的结构和特点进行分析,之后综述了抗菌肽、干扰素、白细胞介素、Toll样受体、补体等先天性免疫相关分子以及T细胞受体和免疫球蛋白等适应性免疫相关分子在硬骨鱼类鳃黏膜中的表达规律、分子功能,最后探讨了化学因素 (重金属、杀虫剂等)、生物因素 (细菌、病毒、真菌、和寄生虫等)以及营养物质和疫苗等对硬骨鱼类鳃黏膜结构的影响,以期为深入研究鳃在鱼类黏膜免疫反应中的角色和应答机制提供指导,为硬骨鱼类病原性疾病的免疫防控策略的制定提供理论基础。     

DIAGNOSTIC ULTRASOUND EXAMINATION AND ECHOCARDIOGRAPHY IN ATLANTIC SALMON (Salmo salar)

Diagnostic ultrasound examination was performed on 53 mature, tranquillized, Atlantic salmon. Wild fish and farmed fish were equally represented. The purpose was to find a rapid method of examination that could be used to identify and separate fish with cardiomyopathy syndrome while fish are tranquillized for other procedures. Three methods of examination were studied. The non-elastic pericardium and absence of lungs results in cardiac function which is clearly different from that in mammals. M-mode echocardiography was complex, technical and time consuming and calculated fractional shortening was not reliable. Subjective evaluation of the heart and liver was valuable and easy to perform. Measurement of the transverse dimension of the atrium and ventricle on a ventro-dorsal axis through the cranial limit of the A-V valve gave acceptable objective data for comparison of the ventricular and atrial size. Diagnostic ultrasound is both practical and valuable for the clinical study of disease in fish.