Reversible metabolic depression in lamprey hepatocytes during prespawning migration: dynamics of mitochondrial membrane potential

The lamprey (Lampetra fluviatilis L.) is an extant representative of the ancient vertebrate group of Agnathans. During the prespawning migration (the river period of life from autumn until spring) lamprey hepatocytes exhibit widely different energy states: a high-energy state in autumn and spring, corresponding to a normal physiological standard, and a low-energy state in winter, which is provoked by prolonged starvation and profound metabolic arrest. In spring the restoration of energy status (return to an active state) is associated with hormonally induced lipolysis of the lipid droplets stored in the cells. Lamprey hepatocytes demonstrate an aerobic metabolism based on oxidation of free fatty acids. The dynamics of mitochondrial membrane potential (MMP) were measured throughout the prespawning migration. Pharmacological inhibition of the electron transport chain decreased the MMP and caused extensive depletion of cellular ATP without loss of cell viability. The potential molecular mechanisms responsible for winter metabolic depression in lamprey hepatocytes are discussed.     

饥饿和再投喂对虎斑乌贼幼体存活、生长和消化酶活力的影响

本文探究了饥饿胁迫与饥饿后再投喂对虎斑乌贼幼体存活率、生长、行为、肝体比、摄食率以及消化酶活力的影响.在室内控制条件下开展了幼体(初始体质量为4.95±0.48 g)的饥饿(0、1、2、3、4、5、6 d)和再投喂(15 d)试验.结果表明: 不同饥饿时间对虎斑乌贼的幼体存活率、体质量降低率、肝体比和消化酶活力影响显著.随着饥饿胁迫时间的增加,其存活率、肝体比呈下降趋势,其中饥饿3 d后,存活率开始明显下降,体质量降低率明显增大,幼体出现喷墨、互相残杀等异常行为;4种消化酶活力呈先下降后上升的趋势,淀粉酶活力以饥饿4 d组最低 (0.07±0.02 U·mg^-1·prot^-1);脂肪酶活力以饥饿2 d组最低(18.47±2.07 U·g^-1·prot^-1),饥饿6 d组最高(57.60±3.98 U·g^-1·prot^-1),胃蛋白酶活力和胰蛋白酶活力以饥饿5 d组(1.98±0.59 U·mg^-1·prot^-1)和饥饿4 d(186.68±20.72 U·mg^-1·prot^-1) 最低.饥饿处理结束后,经15 d再投喂,各试验组存活率、特定生长率、肝体比和摄食率差异显著,幼体的存活率、特定生长率、肝体比和摄食率均与饥饿处理时间呈负相关;饥饿1和2 d组与对照组的存活率、特定生长率和肝肝体比无显著差异,而饥饿3～6 d组显著低于对照组;饥饿1～2 d组的摄食率明显高于对照组,而饥饿6 d组的摄食率明显小于对照组;各组淀粉酶与脂肪酶活力无显著差异,胃蛋白酶与胰蛋白酶活力差异显著,均以对照组最高(胃蛋白酶活力7.06±0.64 U·mg^-1·prot^-1,胰蛋白酶活力914.67±26.54 U·mg^-1·prot^-1),饥饿6 d组最低(胃蛋白酶活力3.21±0.57 U·mg^-1·prot^-1,胰蛋白酶活力660.04±37.92 U·mg^-1·prot^-1).说明虎斑乌贼的幼体饥饿不可逆点(PNR)为第6天,且不能补偿生长.     

Exceptional long-term starvation ability and sites of lipid storage of the Arctic pteropod <Emphasis Type="Italic">Clione limacina</Emphasis>

The Arctic pteropod Clione limacina was collected in Kongsfjorden, Svalbard, in mid June 2004, to study the lipid metabolism within the sites of lipid storage structures during long-term starvation. Animals survived in an aquarium without any food for nearly 1 year (356 days). Size, number of lipid droplets, dry and lipid mass, lipid class and fatty acid compositions of C. limacina were determined and separately analysed for the digestive gland and the remaining integument. During the starvation period, animals shrunk from 22.4 to 12 mm in length on average, and the number of lipid droplets decreased from 1,600 to 1,000 per animal. Dry mass (DM) and total lipid mass both dropped by about 80% from day 200 to the end. The lipid content as percentage DM of the total organism did not decrease significantly ranging from 43.8 to 32.3%DM. The lipid content of the trunk was moderate with about 20%DM. The digestive gland was very rich in lipids with more than 70%DM throughout the experiment and is the major site of lipid metabolism and storage. Triacylglycerols (TAG) decreased, in the total organism, from high initial levels of 62.6 to 43% of total lipid at the end. In contrast, the proportions of 1-O-alkyldiacylglycerols [diacylglycerol ethers (DAGE)] remained almost constant, varying between 20.4 and 28.4%. In the digestive gland, TAG ranged from 60.3 to 64.8% and DAGE from 23.6 to 32.2% from day 200 to the end of the experiment. TAG and DAGE of the trunk were most likely located in the lipid droplets and were almost depleted at the end of starvation. Besides their function as lipid deposit DAGE may also act as protecting substance against bacterial and fungal infections. During the first 200 days of starvation, the fatty acid compositions showed only small variations. Thereafter, fatty acids typical for storage lipids decreased in all body compartments. In adaptation to long periods of food scarcity, C. limacina has evolved various strategies as body shrinkage, utilisation of body constituents not essential for survival, a very low metabolism and slow lipid consumption.     

pH值对中国龙虾消化酶活力的影响

采用酶学分析方法研究了pH对中国龙虾胃蛋白酶、类胰蛋白酶、淀粉酶、纤维素酶和脂肪酶活力的影响。结果表明,在设定的pH范围内,中国龙虾各消化酶的活力均随着pH的升高呈现先升后降的变化趋势。其中,胃、肠、肝胰腺内胃蛋白酶最适pH均为2.2,类胰蛋白酶最适pH分别为8.8-9.2、8.4、8.8,淀粉酶最适pH分别为7.0、7.0、7.4,纤维素酶最适pH分别为4.2、4.2-4.6、5.4,脂肪酶最适pH分别为7.2-7.6、7.2、6.8-7.2。同时测得中国龙虾胃、肠、肝胰腺内的生理pH分别为5.33、6.93、6.60。中国龙虾的消化酶活力存在器官特异性。在最适pH下,胃蛋白酶活力顺序为胃>肠>肝胰腺,类胰蛋白酶、纤维素酶、脂肪酶的活力顺序均为肝胰腺>肠>胃,淀粉酶的活力顺序为肠>肝胰腺>胃。     

Freshwater and coastal migration patterns in the silver‐stage eel Anguilla anguilla

The unimpeded downstream movement patterns and migration success of small female and male Anguilla anguilla through a catchment in north‐west Europe were studied using an acoustic hydrophone array along the River Finn and into the Foyle Estuary in Ireland. Twenty silver‐stage A. anguilla (total length, L_T, range: 332–520 mm) were trapped 152 km upstream from a coastal marine sea‐lough outlet and internally tagged with acoustic transmitters of which 19 initiated downstream migration. Migration speed was highly influenced by river flow within the freshwater (FW) compartment. Anguilla anguilla activity patterns were correlated with environmental influences; light, tidal direction and lunar phase all influenced the initiation of migration of tagged individuals. Migration speed varied significantly between upstream and lower river compartments. Individuals migrated at a slower speed in transitional water and sea‐lough compartments compared with the FW compartment. While 88·5% survival was recorded during migration through the upper 121 km of the river and estuary, only 26% of A. anguilla which initiated downstream migration were detected at the outermost end of the acoustic array. Telemetry equipment functioned efficiently, including in the sea‐lough, so this suggests high levels of mortality during sea‐lough migration, or less likely, long‐term sea‐lough residence by silver A. anguilla emigrants. This has important implications for eel management plans.     

The impact of fasting on adipose tissue metabolism

Fasting and starvation were common occurrences during human evolution and accordingly have been an important environmental factor shaping human energy metabolism. Humans can tolerate fasting reasonably well through adaptative and well-orchestrated time-dependent changes in energy metabolism. Key features of the adaptive response to fasting are the breakdown of liver glycogen and muscle protein to produce glucose for the brain, as well as the gradual depletion of the fat stores, resulting in the release of glycerol and fatty acids into the bloodstream and the production of ketone bodies in the liver. In this paper, an overview is presented of our current understanding of the effects of fasting on adipose tissue metabolism. Fasting leads to reduced uptake of circulating triacylglycerols by adipocytes through inhibition of the activity of the rate-limiting enzyme lipoprotein lipase. In addition, fasting stimulates the degradation of stored triacylglycerols by activating the key enzyme adipose triglyceride lipase. The mechanisms underlying these events are discussed, with a special interest in insights gained from studies on humans. Furthermore, an overview is presented of the effects of fasting on other metabolic pathways in the adipose tissue, including fatty acid synthesis, glucose uptake, glyceroneogenesis, autophagy, and the endocrine function of adipose tissue.     

Digestive enzyme profiles reveal digestive capacity and potential energy sources in fed and starved spiny lobster (Jasus edwardsii) phyllosoma larvae

The impact of starvation on the digestive enzyme (protease, trypsin, lipase and amylase) activities of Stage I and IV Jasus edwardsii phyllosoma larvae was used to identify the nutrients metabolised or conserved during food deprivation, highlighting the most critical energy reserves. Protease activities increased significantly in both Stages I and IV phyllosoma, suggesting that protein catabolism provided energy during food deprivation. Lipase activity decreased significantly in starved Stages I and IV larvae indicating that lipid may be spared for fuelling later developmental moults. The use of protein, while sparing lipid, may provide immediate energy but not at the expense of long-term lipid energy stores which are known to be important during their lengthy larval phase. The preferential use of protein during short-term periods of starvation suggests that particular attention must be given to providing sufficient protein in artificial diets at all times. Amylase activity in starved Stage I larvae was lower than in fed animals, suggesting that the starved animals are not gaining sufficient carbohydrate. However, amylase activity was similar in fed and starved Stage IV larvae, possibly due to the catabolism of accumulated glycogen stores that were not sufficiently developed in Stage I animals.     

Life history of Coilia nasus from the Yellow Sea inferred from otolith Sr:Ca ratios

The habitat use and migratory patterns of the estuarine tapertail anchovy, Coilia nasus, from three sites near- or off-shore of the Yellow Sea, were studied by examining the environmental signatures of Sr and Ca in otoliths using electron probe microanalysis (EPMA). All the samples analyzed were offsprings of typical anadromous C. nasus. The fish grew in freshwater habitat nearly for 1?year, and then entered the brackish and sea water which could be as far as ca. 300?km from the Chinese coast line. The Sr:Ca ratios at the edge of otoliths appeared to correlated with water salinities in sampling sites.     

红螯螯虾胚胎发育期主要消化酶和同工酶的活性变化

采用生物化学方法测定了红螯螯虾(Cherax quadricarinatus)胚胎发育各期主要消化酶(胃蛋白酶、胰蛋白酶、淀粉酶、纤维素酶和脂肪酶)的比活力及主要同工酶(乳酸脱氢酶、苹果酸脱氢酶、葡萄糖-6-磷酸脱氢酶和酯酶)的活力。结果显示,5种消化酶各自表现出不同的变化模式,胃蛋白酶和胰蛋白酶的比活力早期均逐渐上升,到发育后期胃蛋白酶出现快速下降,而胰蛋白酶却仍保持较高水平;淀粉酶比活力呈“V”字型变化趋势,晚期活性较高;纤维素酶和脂肪酶的比活力则均较低。4种同工酶酶谱随胚胎的发育渐趋复杂,酶活性也随之增强。结果表明,消化酶和同工酶活力的高低均受其基因的调控,并随胚胎发育适时表达,为胚胎组织、器官和系统的形成以及未来仔虾的开口摄食提供物质保证。     

Nutritional condition of Anguilla anguilla starved at various salinities during the elver phase

The effects of food deprivation and environmental salinity (<1, 10 and 20) on survival, fish morphology, organization of the digestive system and body lipid reserves in European eel Anguilla anguilla during the transition from glass eel to elver, were evaluated. Fasted elvers kept in fresh water were able to withstand starvation for >60 days, while those in brackish environments (salinity 10 and 20) reached the level of irreversible starvation at 37 and 35 days, respectively. The high level of lipid reserves contained in liver inclusions and the abdominal cavity (perivisceral deposits) in elvers might explain their long resistance to starvation and differences in fasting tolerance under different salinities. Fasting resulted in a significant reduction of the elvers' condition factor and body depth. There were severe histopathological changes in the digestive system and musculature, such as the alteration of the liver organization, and hepatic glycogen and lipid content, shrinkage of enterocytes and reduction of their height, pancreas degeneration, autolysis of the oesophageal and intestinal mucosa and disarrangement of myofibrils and degeneration of trunk musculature. Degeneration of the oesophageal and intestinal mucosa as a consequence of fasting might have impaired digestive and osmoregulatory functions in feed‐deprived fish, directly affecting the tolerance to starvation and survival. Length of food deprivation was associated with a significant increase in mortality, coefficient of variation, cannibalism and point of no return at high salinities. Mortality was dependent on food deprivation and salinity concentrations. Environmental salinity directly influenced the ability of elvers to withstand starvation; once glass eels metamorphosed into elvers, they tolerated starvation better in fresh water than in brackish environments.     

Molecular cloning and characterization of a lipase from the honeybee Apis mellifera

Lipids perform essential and important functions, such as serving as an energy source for growth, development, reproduction, flight, starvation, and diapause in insects. Lipases are key enzymes involving in lipid metabolism and have been reported in several insect species. However, the molecular characterization of the pancreatic triacylglycerol lipase-like genes in honeybees has not been elucidated. Here, we describe the cDNA cloning and characterization of lipase from the honeybee Apis mellifera (Am-Lipase). Am-Lipase consists of 321 amino acids, including a Ser-Glu-His catalytic triad and a consensus active site motif GXSXG. Recombinant Am-Lipase protein degrades triglycerides, preferentially catalyzing the hydrolysis of long-chain fatty acids. Furthermore, the expression pattern of Am-Lipase seems to be a fat body-specific lipase, shows higher expression in forager bees, and is decreased under starvation conditions. Thus, our results suggest that Am-Lipase plays a role in the utilization of lipids stored in the fat body for lipid metabolism.     

Long-Term Starvation and Posterior Feeding Effects on Biochemical and Physiological Responses of Midgut Gland of Cherax quadricarinatus Juveniles (Parastacidae)

We investigated the effect of long-term starvation and posterior feeding on energetic reserves, oxidative stress, digestive enzymes, and histology of C. quadricarinatus midgut gland. The crayfish (6.27 g) were randomly assigned to one of three feeding protocols: continuous feeding throughout 80 day, continuous starvation until 80 day, and continuous starvation throughout 50 day and then feeding for the following 30 days. Juveniles from each protocol were weighed, and sacrificed at day 15, 30, 50 or 80. The lipids, glycogen, reduced glutathione (GSH), soluble protein, lipid peroxidation (TBARS), protein oxidation (PO), catalase (CAT), lipase and proteinase activities, and histology were measured on midgut gland. Starved crayfish had a lower hepatosomatic index, number of molts, specific growth rate, lipids, glycogen, and GSH levels than fed animals at all assay times. The starvation did not affect the soluble protein, TBARS, PO levels and CAT. In starved juveniles the lipase activity decreased as starvation time increased, whereas proteinase activity decreased only at day 80. The histological analysis of the starved animals showed several signs of structural alterations. After 30 days of feeding, the starved-feeding animals exhibited a striking recovery of hepatosomatic index, number of molts, lipids and glycogen, GSH, lipase activity and midgut gland structure.     

Energetics of fattening and starvation in the long-distance migratory garden warbler,Sylvia borin,during the migratory phase

Garden warblers (Sylvia borin) were subjected to starvation trials during their autumnal migratory phase in order to simulate a period of non-stop migration. Before, during and after this treatment the energy expenditure, activity, food intake and body mass of the subjects were monitored. Assimilation efficiency was constant throughout the experiments. The catabolized (during starvation) and deposited body tissue (during recovery) consisted of 73% fat. Basal metabolic rate was decreased during the starvation period and tended to a gradual increase during the recovery period. The reduced basal metabolic rate can possibly be attributed to a reduced size/function of the digestive system, which is consistent with the sub-maximal food intake immediately after resuming the supply of food to the experimental birds. The observed reductions in basal metabolic rate during starvation and activity during recovery can be viewed as adaptations contributing to a higher economization of energy supplies. The experimental birds were unable to eat large quantities of food directly after a period of starvation leading to a comparatively low, or no increase in body mass. Such a slow mass increase is in agreement with observations of migratory birds on arrival at stop-over sites.Abbreviations BM body mass - BMR basal metabolic rate - LBM lean body mass - RQ respiratory quotient     

Silver eel,Anguilla anguilla (Linnaeus, 1758), migration patterns in lowland rivers and lagoons in the North‐Eastern region of their distribution range

Escapement success and migration patterns of silver eels Anguilla anguilla (L.) was studied by acoustic telemetry in three natural free‐flowing and one dammed river and in Curonian Lagoon in Lithuania. Mean downstream migration speed and escapement success were almost the same in the shorter 210 km dammed river (52%, 13.6 km/day) and the considerably longer 300–480 km free‐flowing rivers (53%, 10.7 km/day). Despite the similarity between migration speed in the Curonian Lagoon (14.6 km/day) to that in rivers, migration success was significantly higher (71%) in the Lagoon. Although a majority of silver eels in Lithuania start migrating downstream in spring, the peak of eel migration into the Baltic Sea was observed during late fall. Overall migration success in the rivers and the Lagoon was 35%. Relatively low escapement may have negative consequences on the success on eel stock restoration and must be addressed when strategically planning for the production of spawners.     

Lipophagy maintains energy homeostasis in the kidney proximal tubule during prolonged starvation

Macroautophagy/autophagy is a self-degradation process that combats starvation. Lipids are the main energy source in kidney proximal tubular cells (PTCs). During starvation, PTCs increase fatty acid (FA) uptake, form intracellular lipid droplets (LDs), and hydrolyze them for use. The involvement of autophagy in lipid metabolism in the kidney remains largely unknown. Here, we investigated the autophagy-mediated regulation of renal lipid metabolism during prolonged starvation using PTC-specific Atg5-deficient (atg5-TSKO) mice and an in vitro serum starvation model. Twenty-four h of starvation comparably induced LD formation in the PTCs of control and atg5-TSKO mice; however, additional 24 h of starvation reduced the number of LDs in control mice, whereas increases were observed in atg5-TSKO mice. Autophagic degradation of LDs (lipophagy) in PTCs was demonstrated by electron microscopic observation and biochemical analysis. In vitro pulse-chase assays demonstrated that lipophagy mobilizes FAs from LDs to mitochondria during starvation, whereas impaired LD degradation in autophagy-deficient PTCs led to decreased ATP production and subsequent cell death. In contrast to the in vitro assay, despite impaired LD degradation, kidney ATP content was preserved in 48-h starved atg5-TSKO mice, probably due to increased utilization of ketone bodies. This compensatory mechanism was accompanied by a higher plasma FGF21 (fibroblast growth factor 21) level and its expression in the PTCs; however, this was not essential for the production of ketone bodies in the liver during prolonged starvation. In conclusion, lipophagy combats prolonged starvation in PTCs to avoid cellular energy depletion.     

高盐胁迫对凡纳滨对虾消化及免疫相关酶活力的影响

为探讨高盐对凡纳滨对虾(Litopenaeus vannamei)消化及免疫相关酶活力的影响,实验设置了30、40、50、60共4个盐度梯度。对虾体长(7.84±0.68)cm,养殖密度333尾/m~3,每个梯度设3个平行,实验周期30d。取血淋巴、肌肉、肝胰腺等组织,检测其超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、碱性磷酸酶(AKP)和酸性磷酸酶(ACP)及蛋白酶、脂肪酶、淀粉酶活力。结果表明,盐度显著影响凡纳滨对虾肝胰脏中胃蛋白酶、脂肪酶、淀粉酶的活力(P0.05);随着盐度增加,消化相关酶活力均不断下降,处理间差异显著(P0.05);盐度对凡纳滨对虾不同组织的免疫指标产生影响,表现为随着盐度升高,血淋巴中,AKP活力逐渐升高,ACP、CAT和SOD活力均表现为先升高后降低;肌肉中,AKP、ACP和SOD活力呈现先升高后降低的变化趋势;肝胰脏中,AKP活力呈现先降低后升高再降低的变化趋势,ACP活力高盐处理间差异不显著(P0.05),CAT活力先降低后升高,SOD活力盐度40后逐渐降低。实验结果初步说明,高盐显著影响凡纳滨对虾的消化及免疫相关酶活力,且盐度对不同组织中免疫酶活力影响存在一定的组织特异性,50以上的高盐胁迫对对虾消化和免疫相关酶活力的影响尤为显著。