Étude préliminaire des variations circadiennes des protéines de l'hémolymphe de Penaeus japonicus Bate

The protein contents of the haemolymph in the decapod crustacean Penaeus japonicus Bate exhibit a tricircadian rhythm characterized by two maxima during the night and another one during the day. During the night the total protein concentration in haemolymph is higher in males. Maximal values in males are approximately four times greater (100 mg/ml) than minimal ones (25 mg/ml). Variations are less in females: maximal values are approximately two and a half times higher (75 mg/ml) than minimal ones (30 mg/ml).Electrophoresis in polyacrylamide gradient showed that the different protein constituents in general exhibit similar circadian variations. Time of acrophases depends on the sex and the stage in the intermoult cycle. On the other hand, the mean concentration of each protein constituent calculated from values measured at different times of the day are similar whatever the sex and moulting stage. Between 0 and 6 h certain protein fractions - two in females and one in males - are no longer detectable by electrophoresis. At 12 h, concentration of haemocyanin and its subunits increase simultaneously in both sexes.     

Variation in the free amino acid content of ovary, hepatopancreas and hemolymph of Penaeus schmitti during ovarian maturation (Crustacea, Decapoda, Penaeidae)

Quantitative and qualitative variations of free-amino-acids contents (FAA) of ovaries, hepatopancreas and haemolymph have been studied in the course of ovarian maturation in Penaeus schmitti. In the ovary, FAA content, expressed as microM/g ovary fresh weight, decreases from 56.2 to 23.6 microM between gonadosomatic index (GSI) 0.8 and GSI 7.1. There is an increase of hepatopancreas FAA content of 33.4% till GSI 4. Haemolymph FAA content increases from 0.32 to 1.76 microM/ml, with maximal values of 3.74 microM/ml when GSI are between 3 and 4. FAA content of ovaries, expressed as microM/g of animal, is 3.2 fold higher at the end of maturation, with a main increase in the first stages of ovogenesis. Maturation process has a quantitative effect on all FAA, but, on a qualitative point of view, it concerns mainly lysine, arginine and glycine. The main variations of AA composition (expressed as percentage of total FAA) are observed in haemolymph.     

Variations de la composition lipidique tissulaire au cours de la vitellogenese chez la crevette Penaeus indicus Milne Edwards

Changes in lipid composition are observed in the ovary, the hepatopancreas, the haemolymph and the muscle of the female Penaeus indicus Milne Edwards, during the increase of the gonad-somatic index (GSI). Vitellogenesis is characterized by a large accumulation of lipids in the ovary, which seems to be practically linear when expressed as a function of GSI, if there is no interference from moulting. Phospholipids and triacylglycerols are the major lipid fractions in the ovary, but some classes of minor importance, like wax esters, hydrocarbons, and glyceryl ethers, are also incorporated. These last components may play a rôle in the maturation process.The lipid reserves of the hepatopancreas contribute partially to the formation of vitellin, but direct input of lipids from feeding seems to be preponderant. An intraovarian synthesis of lipids may be possible, but probably restricted, considering the low concentrations of free fatty acids and partial acylglycerols found in the ovary. Phospholipids are the main components involved in the increase in lipid concentration of the haemolymph during vitellogenesis. This increase, and the potential rise in the turnover of circulating lipids, suggest a deep alteration in the lipid metabolism of the females during the breeding period.     

Neuroendocrine control of blood lipids in the locust, Locusta migratoria

In order to determine the best conditions, the influence of various parameters on the haemolymph lipid concentration were studied. These parameters are the age, the sex and the feeding of the animals, the time and the number of the haemolymph sample-taking and the temperature of the locust culture. A large in vivo increase in haemolymph lipid concentration was obtained in locusts which received extracts of the whole CC and of their glandular or neurohemal lobes. Reversely, a decrease in this concentration was obtained in locusts operated 7 days before (cardiacectomy or glandular lobe removal). Moreover the pars intercerebralis extracts increased the level of haemolymph lipids. We conclude that adipokinetic factors are present, both in the glandular lobes of the CC and in their neurohemal lobes. It is likely that the latter partly originate from the pars intercerebralis. Results of allatectomy and injections of corpora allata extracts led to the conclusion that corpora allata contain an adipokinetic factor, the juvenile hormone. and factors that inhibit the haemolymph lipid concentration. Finally, from different injections of neurotransmitters and drugs it is argued that it is mainly octopamine which is involved in the mechanism governing the increase of the level of haemolymph lipids.     

Total lipids and phospholipids in buffalo semen (Bubalus bubalis)

Spermatozoal and seminal plasma concentrations of total lipids from 50 ejaculates and phospholipids and their fractions from 30 ejaculates were quantified in the semen of five Murrah buffalo bulls. Sperm lipid content ranged from 0.93 to 1.72 mg/10(9) cells with an overall average 1.32 +/- 0.03 mg/10(9) cells. Its concentration in seminal plasma varied from 1.39 to 2.22 mg/ml with overall average of 1.75 +/- 0.03 mg/ml. Spermatozoal total phospholipid content ranged from 0.44 to 0.94 mg/10(9) cells with overall mean being 0.64 +/- 0.02 mg/10(9) cells. The corresponding values for seminal plasma were 0.53 and 0.88 mg/ml with an overall mean of 0.69 +/- 0.02 mg/ml. Phosphatidyl choline constituted the major fraction both in the spermatozoa and and seminal plasma.     

Partial characterization and amino acid composition of a high molecular weight peptide with salmon calcitonin immunoreactivity in a crustacean: Nephrops norvegicus

Anti-salmon calcitonin antibodies were used to follow the purification of a high molecular weight peptide present both in the haemolymph and in the hepatopancreas of the Norway lobster Nephrops norvegicus. An apparent molecular weight of 22 kDa has been measured in electrophoresis on SDS gels and amino acid composition compared to salmon calcitonin. The amount determined by the immunoreactivity assay corresponds to about 1/40 and 1/140 of that which is based on direct protein measurement for the hepatopancreas and the haemolymph respectively. The total amount of this peptide could be estimated as 3.5 mg/g fresh weight for the hepatopancreas and 140 ug/ml for the haemolymph. The function of this peptide is still unknown.     

Variations in protein levels in the hemolymph, hepatopancreas and ovary of Penaeus schmitti during ovarian maturation (Crustacea, Decapoda, Peneidae)

Variations of protein concentrations in the haemolymph, hepatopancreas and ovaries were studied during the ovarian maturation of the shrimp Penaeus schmitti. The main variations observed during this physiological process are the increase of protein and the decrease of water contents in the ovary, more pronounced at the first stages of gonadosomatic index (GSI:0-4). During ovarian maturation, water content of the ovaries and hepatopancreas decrease from 10.7 and 3.9% respectively. In relative values, protein content increases from about 40.0% in the ovary, 47.5% in the haemolymph and 15.4% in the hepatopancreas. In absolute values, the protein concentrations of the hepatopancreas showed no significant variation. In the opposite, ovary protein content is 14.7 fold higher at the end than the early stages of vitellogenesis. Relations between haemolymph, hepatopancreas and ovary with eventual transfers of protein material are discussed.     

The human red cell glucose transporter in octyl glucoside. High specific activity of monomers in the presence of membrane lipids

Human red cell membranes were stripped of peripheral proteins and partially solubilized with 50-260 mM octyl glucoside at 2-14 mg protein/ml, to find conditions that afford a high concentration of active glucose transporter after purification on DEAE-cellulose. Transporter-egg yolk phospholipid vesicles were prepared by gel filtration. The specific D-glucose equilibrium exchange activities increased with increasing dilution of the glucose transporter. At 260 mM octyl glucoside the glucose transporter became partially denaturated. At 225 mM detergent the DEAE-cellulose chromatography showed one main and one minor fraction of active glucose transporter. Nucleoside transport activity was enriched in the minor fraction. Solubilization with 75 mM octyl glucoside at 8 mg protein/ml gave a maximal concentration of purified transporter, 0.8 mg/ml, probably corresponding to complete solubilization. The phospholipids were partially retarded on the DEAE-cellulose. The specific D-glucose equilibrium exchange was high, up to 200 nmol glucose/micrograms transporter in two min at 50 mM glucose. High performance gel filtration in octyl glucoside indicated that the transporter formed dimers during the fractionation. These eluted at Mr 125,000, partially separated from the phospholipids, which appeared at Mr 55,000 (cf. Mascher, E. and Lundahl, P. (1987) J. Chromatogr. 397, 175-186). The D-glucose transport activity was low in the main fraction and high in the transporter-phospholipid fraction. Mixing of these fractions did not increase the activity. The glucose transporter is probably dependent on one or more specific membrane lipid(s). Presumably the transporter dimerizes and loses activity upon removal of these lipids.     

Biochemical aspects of Balanus hameri haemolymph,together with some comparative haemolymph data for Balanus balanus and Lepas anatifera

• 1.1. The concentration of protein in the haemolymph of Balanus hameri ranged from 2.0 to 17.3 mg/ml, and the lipid from 1.4 to 7.7 mg/ml; the haemolymph protein and lipid levels increased significantly prior to cross-fertilization.
• 2.2. The protein and lipid concentrations in Balanus balanus haemolymph were 8.1 and 1.7 mg/ml respectively.
• 3.3. The lipid concentration of Lepas anatifera haemolymph was 1.2 mg/ml.
• 4.4. The neutral lipid and phospholipid components of B. hameri and L. anatifera haemolymph were the same, with the major components of the phospholipid fraction being phosphatidyl ethanolamine and phosphatidyl choline.
• 5.5. The osmolarity (970.4 mOsm), chloride ion concentration (501.3 m-eq/l) and pH (7.29) of B. hameri haemolymph were also determined.

     

Characteristics of changes in lipids of granulation fibrous tissue of rats subjected to different doses and modes of melatonin administration

Melatonin influences the time-course of changes of lipids in granulation fibrous tissue in rats. Its effect depends on a dose, modes of administration (intraperitonial, subcutaneous or local) and duration of treatment. Intraperitonial administration of a single dose of melatonin (4 mg/kg) did not influence lipid content in the granulation fibrous tissue, while repeated injections of this hormone limited the increase in contents of lipids and phospholipids on the 5th and 8th days of regeneration. Long-term subcutaneous injections of melatonin caused distinct changes of lipids: at the dose of 0.3 mg/kg it prevented, and at the dose of 4 mg/kg it promoted the increase of lipid content in the granulation fibrous tissue on the fifth day of this study. Local application of melatonin solution (1.5 mg/ml) in early periods of regeneration caused insignificant changes of total lipids and total phospholipids in the granulation fibrous tissue. However, the higher concentration (15 mg/ml) of melatonin caused the decrease of total lipids due to reduced content of cholesterol and triglycerides and the increase of total phospholipids and some of their fractions.     

Seasonal variations of plasma lipids and lipoproteins in the hedgehog, an animal model for lipoprotein (a) metabolism: relation to plasma thyroxine and testosterone levels

We describe a study of the seasonal variations of hedgehog plasma lipids and lipoproteins and their correlation with changes in the activities of the thyroid and testis. In ten male hedgehogs, plasma concentrations of lipids, thyroxine and testosterone were assayed each month for 1 year beginning in September, while plasma lipoproteins from five of these animals were analyzed at the same dates using density gradient ultracentrifugation. All classes of plasma lipids (cholesterol, total glycerol and phospholipids) exhibited statistically significant seasonal variations in their respective concentrations, with simultaneous maxima (cholesterol: 207 +/- 39 mg/100 ml; total glycerol: 50 +/- 9 mg/100 ml; phospholipids: 266 +/- 25 mg/100 ml) during late fall-early winter, i.e., during the period of the year when plasma levels of both thyroxine and testosterone were minimal. Plasma lipids subsequently decreased to minimal levels either in early summer (cholesterol: 129 +/- 18 mg/100 ml; phospholipids: 178 +/- 20 mg/100 ml) or in late winter (total glycerol: 22 +/- 9 mg/100 ml). Very low density lipoproteins (d less than 1.015 g/ml) were found at low levels (less than 15 mg/100 ml) during the cold months, and then became detectable as trace components only. The total concentration of the mixed lipoprotein population (i.e., low density lipoproteins, Lp(a), and high density lipoprotein (HDL)-like particles) in the d 1.015-1.065 g/ml interval decreased by almost 50% from January to February (from 164.3 to 89.2 mg/100 ml), i.e., following a 10-fold increase in the level of plasma testosterone, and immediately before the rapid doubling in plasma thyroxine concentration. The staining intensity of the electrophoretic band with migration characteristics corresponding to those of Lp(a) decreased considerably during winter. At the same period of the year, lower density (1.032-1.055 g/ml) HDL-like particles disappeared. The concentration of lipoproteins with d 1.065-1.162 g/ml, which included Lp(a) particles in addition to typical HDL, equally underwent seasonal variations. These variations consisted of two successive maxima in late fall (426.4 mg/100 ml) and late winter (458.3 mg/100 ml) with two subsequent decreases leading to minima in February (327.8 mg/100 ml) and August (257.1 mg/100 ml). Finally, very high density lipoproteins (d 1.162-1.259 g/ml) were heterogeneous, containing both cholesterol-rich (d 1.162-1.227 g/ml) and phospholipid-rich (d 1.194-1.259 g/ml) subpopulations.(ABSTRACT TRUNCATED AT 400 WORDS)     

Lipid mobilization and locomotor stimulation in Gryllus bimaculatus by topically applied adipokinetic hormone

Abstract.  Walking activity of 3-day-old adult female Gryllus bimaculatus (de Geer) (Ensifera, Gryllidae) was measured over 24 h. A high level of locomotor activity during the scotophase was found, which was two- to three-fold higher than that during the photophase. The titre of lipid in the haemolymph was relatively low 2 h after lights on, increased significantly 2 h after lights off, although, 2 h after lights on in the next photophase, the lipid titre had decreased to the basal level. Topical application of homologous Grybi-adipokinetic hormone (AKH) (100 pmol in 20% 2-propanol) led to a significant increase in haemolymph lipids, comparable with the maximal increase caused by injection of AKH (3 pmol in water). Topical application of AKH also stimulated locomotor activity in crickets (maximal stimulation 1.8-fold with 100 pmol Grybi-AKH). The results suggest that AKH penetrates the cuticle quickly. It is assumed that AKH stimulates locomotory activity at least in part via the increase of haemolymph lipid titres; however, the stimulation of locomotor activity via a direct neuromodulatory effect of AKH cannot be excluded.     

Assessment of sheep blastocyst effects on neutral lipids in the uterine caruncular epithelium

Neutral lipids in the maternal uterine caruncular epithelium were studied by histochemical localization with Oil Red O. Results using a scoring system of 1 (negligible lipid) to 5 (maximal lipid) showed that intraepithelial lipid stores were minimal until Days 7-8 of the oestrous cycle and then increased to have a mean score of 4.4 on Day 14-15. In early pregnancy, although relatively high with a mean score of 3.2 at Day 15-16, such neutral lipids were significantly lower than those present at a comparable stage in the oestrous cycle. Thereafter, levels declined to a mean score of 1 on Days 21-23 of pregnancy. Such neutral lipid loss appears to be one of the first signs of the maternal response to the implanting embryo and precedes morphological evidence of transformation of either maternal or fetal tissues.     

Composition and synthesis of cellular lipids in Neurospora crassa during cellular differentiation.

The synthesis of cellular lipids of Neurospora crassa was measured during growth on low (2% sucrose)- and high (15% glucose)-carbohydrate supplementation. The amount of lipid per dry weight of cells does not change during the germination and early logarithmic growth periods, but the percentage of phospholipid in the lipid does increase, reaching a maximal value of 90% at 4 to 5 h after inoculation, at which time the phospholipid content of the cells is approximately 60 mumol/g (dry weight). The content of the anionic phospholipids, as a percentage of the lipid fraction, is relatively constant during the growth period, but the contents of the zwitterionic phospholipids phosphatidylcholine and phosphatidylethanolamine change in a reciprocal fashion. During the first 8 h of growth, phosphatidylcholine falls from 53% of the phospholipid to 43%, whereas phosphatidylethanolamine rises from 29 to 38%. The total of these two phospholipids is approximately 83% during the growth period studied. The synthesis of cellular phospholipids, measured either by [32P]H3PO4 or [14C]glucose incorporation, reached maximal levels between 3 and 5 h of growth. The effect of the high-carbohydrate supplement on cellular lipids was minimal. Inclusion of 15% glucose decreased the labeling of phospholipid by [32P]H3PO4, but did not affect lipid composition. This observation is in contrast to the effects of high glucose on mitochondrial phospholipid synthesis.     

Metal binding compounds in hepatopancreas and haemolymph of porcellio scaber (isopoda) from contaminated and reference areas

1. In order to evaluate the role of metal-binding proteins in the tolerance mechanism of Porcellio scaber to heavy metals, a comparative study was made using isopods from three locations: a zinc-lead mine (Plombiéres), a zinc smelter (Budel) and a reference wood (Spanderswoud). The Cu, Zn and Cd concentrations and the protein composition were determined in the haemolymph and hepatopancreas from the isopods.2. A constant Cu/Zn molar ratio of about 5 was found in the haemolymph of all populations and no correlation was found between hepatopancreas and haemolymph Cu and Zn content.3. Using fast-performance liquid chromatography (FPLC), most of the haemolymph Cu and Zn appeared to be associated with a single UV absorbing peak corresponding with an apparent molecular weight of ± 70 kD; this peak is probably the monomer of hemocyanin.4. The hepatopancreas Zn and Cd concentration were elevated compared to the hepatopancreas of the smelter and mine isopods; after homogenization and centrifugation 70–80% of the metals were found in the supernatant.5. In all populations the hepatopancreas Cu-, Zn- and Cd-binding compounds eluted in separate peaks of low molecular weight, suggesting the absence of an MT-like compound in Porcellio scaber.6. The similarity of the protein profiles in haemolymph, and the similar distribution of the metals over the fractions in haemolymph and hepatopancreas suggests that inducible metal binding compounds are not involved in metal tolerance differences between populations.     

Change in the composition of phospholipids and antioxidative activity of lipids of mouse organs after administration of adriamycin

The phospholipid composition and the antioxidative activity (AOA) of lipids from liver, heart and brain of F1(CBA X C57Bl) mice change significantly after intraperitoneal injection of adriamycin (7 mg/kg). All tissues studied are characterized by a drastic fall in the phosphatidylethanolamine (PEA) content and the AOA of lipids with a minimum on the 2nd-4th days and a subsequent return to normal values on the 10th-14th days. These results are consistent with the hypothesis on the intensification of adriamycin-induced lipid peroxidation and the predominant expenditure of readily oxidized phospholipids, including PEA.     

Incorporation of palmitic acid-1-14C into lipids of pharate adult tissues of Bombyx mori

Incorporation of palmitic acid-1-¹⁴C into pharate adult tissues and their lipid components of Bombyx mori was investigated. Rapid incorporation of radioactivity took place predominantly in fat body and haemolymph lipids, and partially in ovarian lipids immediately after the injection at the middle stage of pharate adult development. The major parts of the radioactivities in fat body, haemolymph and ovary were distributed in triglycerides and phospholipids, diglycerides, and triglycerides, respectively. The patterns of time course of incorporation of radioactivity into lipid components of pharate adult tissues suggest that the major form of lipid released from fat body may be diglycerides and the diglycerides in haemolymph are probably the main source of ovarian triglycerides.     

Trehalose turnover and the coexistence of trehalose and active trehalase in cockroach haemolymph

In the cockroaches Periplaneta americana, Periplaneta australasiae, Leucophaea maderae, and Nauphoeta cinerea, undiluted haemolymph, undiluted haemolymph to which 10% solid trehalose was added, and haemolymph diluted 100 or more times with 1% trehalose solution showed approximately equal trehalase activities (3 to 8 mg/ml per hr). No evidence for the presence of a trehalase inhibitor was found.Freshly drawn haemolymph of Periplaneta americana contained 14 to 16 mg trehalose/ml, which on standing was hydrolyzed to glucose at a rate of 4 to 8 mg/ml per hr. In this cockroach, the rate of haemolymph trehalose turnover was only 1.3 mg/ml per hr. This means that in vitro trehalose is hydrolyzed by undiluted haemolymph at several times the rate at which it is replaced in the haemolymph of the intact insect. The mechanism through which trehalose and trehalase can coexist in the haemolymph of the intact cockroach remains therefore unexplained.     

Seasonal changes in leaf lipids of Diapensia lapponica, with special reference to storage lipid bodies

Seasonal fluctuations in storage lipids in the cushion plant Diapensia lapponica , growing in Northern Finland (70°N 27°E), were studied by microscopy and chemical analysis. Lipid bodies in the mesophyll cells were stained with Sudan Black for quantitative observation by light microscope. Electron microscope observations were made using aldehyde prefixed and osmium tetroxide postfixed sections of leaf blades. Thin layer and gas capillary chromatographic techniques were used to analyse total lipids and total fatty acids in green shoots of Diapensia . Free sugars and starch were extracted separately and determined by the anthrone method.
A mesophyll cell was characterized by a large lipid body (storage lipid) in summer but by several small spherules in winter. Total surface area of the cross-sectioned lipid globules was at its lowest from April to September; the maximal value was in March. The amount of total lipids in the leafy tops of D. lapponica was 91–200 mg g^-1 dry weight. Values were lowest at the end of June, when the total carbohydrate level was at its highest. Accordingly, the decrease in the total lipid level in the early growing season, when new leaves were developing, can be attributed primarily to the increase in the level of carbohydrates, particularly starch. The amount of total fatty acids varied from 21 to 30 mg g^-1 dry weight. The level increased in the early growing season and remained elevated throughout the summer. Like the total lipids, the total fatty acids are derived from structurally different parts of the sclerophyllous leaves, including the well-developed cuticle and epicuticular wax layer. The discrepancies in the results obtained from microscopic and chemical analyses are discussed.     

中华绒螯蟹对Pb和Cd的富集与释放特性

应用生物富集双箱动力学模型模拟了中华绒螯蟹(Eriocheir sinensis)分别在Pb浓度为0.25、0.50、0.75mg/L,Cd浓度为0.025、0.050、0.075mg/L的单一水环境中暴露时,蟹鳃、肝胰腺、肌肉和血淋巴对Pb和Cd的生物富集与释放实验,并通过非线性拟合得到中华绒螯蟹对Pb和Cd的富集速率常数k1、排出速率常数k2、生物富集系数BCF、生物半衰期B1/2、富集平衡时生物体内Pb和Cd含量CAmax等动力学参数。结果表明:(1)中华绒螯蟹对Pb和Cd具有明显的富集,蟹鳃、肝胰腺和肌肉中Pb和Cd的含量与富集时间和水环境中Pb和Cd暴露浓度表现出了很好的正相关,血淋巴在富集阶段没有明显的规律。理论平衡状态下鳃、肝胰腺和肌肉中Pb和Cd含量CAmax随着暴露浓度的增大而增大,且成正相关。(2)Pb和Cd在中华绒螯蟹组织器官中的富集具有选择性,开始实验前,Pb在中华绒螯蟹体内的的分布规律为:肝胰腺>鳃>肌肉>血淋巴;Cd的分布规律为:鳃>肝胰腺>血淋巴>肌肉。在实验浓度的Pb和Cd水环境中暴露16d后,Pb的分布规律为:鳃>肝胰腺>肌肉>血淋巴;Cd的分布规律为:肝胰腺>鳃>肌肉>血淋巴。(3)中华绒螯蟹对Pb和Cd的生物富集和释放都较缓慢。经过16d的生物富集,各组织器官中Pb和Cd的含量均未达到稳态平衡。Pb和Cd在组织器官中的生物富集系数(BCF)范围分别为5-51和6-3148,中华绒螯蟹对Cd的富集能力明显高于Pb(P1/2)范围分别为4-9d和8-57d,中华绒螯蟹对Cd的排出能力明显低于Pb。