Forskolin (FORSK), octopamine (OA) and adipokinetic hormone (AKH) stimulate the production of diacylglycerol (DG) in fat body of the locust, Locusta migratoria and the release of DG from fat body into hemolmph. The three effectors also increase the level of cAMP in fat body, but the cAMP content is not proportional to DG production. AKH stimulates the uptake of Ca2+ by fat body cells and requires the presence of extracellular Ca2+ to increase cAMP and DG levels in fat body. The production of DG seems to be an energy-dependent process. The uptake of DG by lipophorin (LP) from fat body is also energy-dependent but does not require extracellular Ca2+. 相似文献
To understand the hormonal control of embryonic diapause, juvenile hormone analogs (JHAs), methoprene and hydroprene, and 20-hydroxyecdysone (20E) were applied onto diapause eggs of Locusta migratoria and Oxya yezoensis. These insects enter diapause at the mid-stage of embryogenesis prior to blastokinesis. Topical application of JHAs significantly facilitated diapause termination in both species but JHA-treated embryos underwent abnormal morphogenesis, pigmentation and sclerotization without dorsal closure. The Locusta eggs immersed in the 20E solution for 24h terminated diapause in a dose-dependent manner. We also investigated phosphorylation of extracellular signal-regulated kinase (ERK), a member of mitogen-activated protein kinase (MAPK), during diapause-terminating process of Locusta migratoria and found that ERK was activated either by cold exposure or JHA treatment. The possible involvement of the hormones and ERK in embryonic diapause and the possibility of ecdysteroids synthesis by prothoracic glands of diapause embryo were proposed. 相似文献
A high-molecular-weight protein, Mr 500,000, has been isolated and characterized from the hemolymph of the migratory locust, Locusta migratoria. It is composed of six seemingly identical subunits of apparent Mr 78,000. It contains low concentrations of carbohydrate and lipid, but high percentages of aspartate and glutamate as well as high proportions of hydrophobic amino acid residues. An antiserum, developed against this purified hemolymph protein, does not react in the double-diffusion test or after immunoblotting with purified lipophorin or cyanoprotein, two other major proteins in locust hemolymph. The concentration of this larval specific protein in the hemolymph of Locusta was examined during the last larval instar and in adult males by quantitative rocket immunoelectrophoresis. Its concentration increases in the second half of the fifth instar, concommitant with an increase in total protein. The protein is detectable by immunological techniques in adults, although its concentration is very low at this stage. 相似文献
Heat shock induced by an increase in temperature from 30°C to 47°C led to changes in protein synthesis in wing pads of the fifth larval instar of Locusta migratoria. Synthesis of heat shock proteins in the molecular weight range of 85,000, 70,000 and 18,000–22,000 was first detected at a threshold temperature of 45°C and was found to be highest at 47°C. A marked decline in the synthesis of many other proteins was also evident at 47°C. Recovery of general protein synthesis was observed when wing pads were shifted back to 30°C after a 2-h heat shock at 47°C. Heat shock protein patterns in Locusta and Drosophila were compared. 相似文献
Dopa decarboxylase (DDC) which converts dopa to dopamine is important for cuticular melanization and sclerotization in insects. An antibody to Drosophila DDC was found to precipitate both DDC activity and a 49-kDa polypeptide synthesized by the epidermis of molting Manduca larvae. Using the Drosophila DDC gene, we isolated the Manduca DDC gene which on hybrid selection produced a 49-kDa translation product precipitable by the Drosophila DDC antibody. The 3.1-kb DDC mRNA appeared 12 hr after head capsule slippage (HCS) and reached maximal levels 7 hr later. Peak expression was twofold higher in melanizing allatectomized larvae and could be depressed to normal levels by application of 0.1 micrograms juvenile hormone I at HCS. Infusion of 1 microgram/hr 20-hydroxyecdysone (20-HE) for 18 hr beginning 2 hr after HCS or addition of 1 microgram/ml 20-HE to the culture medium for 24 hr prevented the normal increase in DDC mRNA. When Day 2 fourth instar epidermis was explanted before the molting ecdysteroid rise and cultured with 1-3 micrograms/ml 20-HE for 17 hr and then for 24 hr in hormone-free medium, DDC expression was three- to fourfold higher than that in epidermis cultured in the absence of hormone. Twelve or more hours of incubation with 20-HE was required for an increase in DDC mRNA, but continuous exposure to 20-HE prevented the increase. In all cultures an initial rapid increase in DDC mRNA was observed which decayed with time in vitro and apparently was associated with the wound response. Thus, ecdysteroid during a larval molt is necessary to program the later expression of DDC, but the subsequent decline of the ecdysteroid is required for this expression to occur. 相似文献
The roles of cyclic AMP, cyclic AMP-dependent protein kinase and the phosphorylation of specific proteins in the regulation of cardiac contractility are briefly reviewed. Criteria for determining whether changes in cyclic AMP and protein phosphorylation are involved in a physiological response are discussed. Although cyclic AMP-dependent phosphorylation of the voltage-operated Ca2– channel, phospholamban, troponin-I and C-protein have all been implicated in the response of the heart to inotropic agents which elevate cyclic AMP, none of these phosphorylations satisfy all of the criteria completely. Evidence is presented that there are compartments of cyclic AMP in heart which are coupled to different functional responses.Abbreviations cAMP
3,5 cyclic adenosine monophosphate
- PDE
cyclic nucleotide phosphodiesterase
- cA-PrK
cAMP-dependent protein kinase
- SR
sarcoplasmic reticulum
- PGE1
prostaglandin E1
- Tn-I
troponin I 相似文献
ABSTRACT. . Final instar nymphs of the oligophagous acridid Locusta migratoria (L.) and larvae of the polyphagous noctuid Spodoptera littoralis (Boisduval) were fed for 4, 8 or 12 h, the conditioning period, on one of four artificial diets. Of these, diet PC contained 20% protein and 10% digestible carbohydrate; another, P, contained 20% protein but with the digestible carbohydrate component replaced by cellulose; a third, C, had the protein component substituted by cellulose, and the fourth, O, had both protein and digestible carbohydrate replaced. After this conditioning period, insects were given a choice of two diets, P and C, and hence an opportunity to select for the nutrients, if any, which were lacking in their previous food. Amounts eaten and selection behaviour were then recorded in detail for a total of 9 h. This paper deals with total amounts of diet eaten during the conditioning and choice periods. Spodoptera larvae were more sensitive than the locusts to being fed a nutritionally inadequate conditioning diet, and ate only small quantities of the P, C and O diets as compared with the PC diet, irrespective of the duration of conditioning. Locusts, on the other hand, when restricted to the P diet continued to eat relatively large amounts of it throughout a 12 h conditioning period. Those nymphs fed the C diet ingested large quantities (more than of the PC diet) up until 8 h, after which intake fell. When offered a choice, both species selected for the nutrients missing from the conditioning diet, even if the conditioning period had been as short as 4 h. During the first hour of choice locusts selected the P diet if they had been previously fed C and the C diet if previously fed P. Those deprived of both nutrients increased consumption of both P and C diets. Spodoptera larvae were more sensitive to prior deprivation of digestible carbohydrate than of protein. During the first hour of choice they selected the C diet if previously fed P or O but did not choose the P diet if previously fed C. In the subsequent 8 h of choice, however, a strong selection for the P diet after previous deprivation became apparent. In the locust, the selection for nutrients missing from the conditioning diet continued for the following 8 h of choice but became masked by a tendency, shown by all nymphs, to select C over P. The functional significance and possible physiological basis of all these responses is discussed. 相似文献
In fifth instar larvae of Locusta migratoria the haemolymph lipid concentration is elevated after injection of adipokinetic hormone (AKH). This hyperlipaemic response in larvae remains substantially lower than in adults; over 75% of the mobilized lipid consists of diacylglycerol. In addition, unlike adult locusts, fifth instar larvae also exhibit a consistent, though moderate, hypertrehalosaemic response to AKH. The increases of both lipid and carbohydrate concentrations in larvae are dose-dependent, showing a significant linear regression on log dose in the range 0.2–20 pmol AKH.Glycogen phosphorylase in the fat body of fifth instar larvae as well as young adults is activated on injection of AKH, the percentage active phosphorylase increasing linearly with log dose in the range 0.04–20 pmol AKH. For a given response, a somewhat higher dose of AKH is needed in larvae than in young adults.Fat body glycogen phosphorylase is strongly activated during the period of the larval-adult ecdysis, when active phosphorylase accounts for almost half of the total enzyme, which is approximately ten times more than it is two days before, and two days after the ecdysis.The corpora cardiaca of fifth larval instar locusts already possess the potencies to elevate carbohydrate and lipid concentrations in larval haemolymph, and to activate fat body glycogen phosphorylase. 相似文献
Cellular (non-cuticular) polypeptides were isolated from mid-instar larval epidermis incubated in the presence or absence of 20-hydroxyecdysone (20HE) for 14–16 h in vitro. Polypeptides synthesized during the last few hours of incubation were labelled with [35S]methionine, separated in cell lysates by two-dimensional polyacrylamide gel electrophoresis and detected fluorographically. Cells incubated with hormone incorporated 28% more label into polypeptide. The synthesis of over 250 polypeptides was detected in total cell lysates. Of these, 54 showed an altered level of synthesis in response to 20HE treatment. The synthesis of most (33) was depressed. The relative synthetic rate of most “down-regulated” 20HE-sensitive polypeptides began to drop at 10−3 μg/ml whereas that of most “up-regulated” polypeptides increased only at concentrations above 10−1 μg/ml. An early response to 20HE, involving the de novo synthesis of a 43 kDa polypeptide, was first detectable after 2 h of exposure to hormone, and peaked after 4 h. The synthesis of this 43 kDa polypeptide was selectively enhanced by the addition of 10−2 μg/ml cycloheximide to medium containing 20HE. The long-term effect (12–16 h) of 20HE on polypeptide synthesis in subcellular fractions of the epidermis was also examined. Polypeptide synthesis found in the nuclear, mitochondrial-lysosomal, microsomal and soluble fractions changed in response to 20HE. It appears that 20HE influences epidermal behaviour predominantly through its ability to modulate the synthetic rate of many cellular polypeptides, rather than by turning off the synthesis of a few pre-existing ones and switching on that of a few new ones. 相似文献
The ultrastructure of the colon of Locusta migratoria is described. The colon is lined by a thick cuticle that, for the most part, adheres to the underlying epithelium. The cuboid epithelial cells are characterized by moderate invaginations of the apical and, to a lesser extent, basal plasma membranes; the lateral plasma membranes are relatively flat. The bulk of the mitochondria are located in the apical region of the cell and are not particularly associated with any of the plasma membranes. The basal region of the cells contains much rough endoplasmic reticulum, glycogenlike granules, and a predominance of spherical, electron-dense bodies of various sizes. Where muscle fibers make contact with the epithelium, the cells are much reduced; the cytoplasm is usually less electron-dense, and, typically, the nucleus has a thick layer of granular material associated with the inner nuclear membrane. The apical and basal plasma membranes of the reduced epithelial cells contain numerous hemidesmosomes. The apical hemidesmosomes occur in pairs around an extracellular space that contains electron-opaque material. The latter forms tonofibrillae that extend into the endocuticle. Bundles of microtubules are associated with the hemidesmosomes. The tubules traverse the cell from the apical to the basal region. The possible significance of these findings is discussed. 相似文献
The genetic skin disease psoriasis has been examined as a model system that may provide an understanding of the control of normal epidermal specialization (differentiation) and the perturbed regulatory processes in proliferative diseases. The excessive glycogen accumulation, increased proliferation and decreased tissue specialization characteristic of psoriasis involve cellular processes that have been shown to be regulated by cyclic AMP in other cells and tissues. It has also been suggested that cyclic GMP is a cellular effector that may be involved in promoting cell proliferation and other events that oppose those believed to be mediated by cyclic AMP. It was postulated, therefore, that the epidermis of the psoriasis lesion might exhibit an imbalance in the cellular concentrations of these two cyclic nucleotides. In this study the levels of cyclic AMP were measured in the involved epidermis (IE) and uninvolved epidermis (UE) from 25 psoriasis patients. The concentrations of cyclic AMP were found as reported previously using a different analytical procedure, to be significantly lower in IE based on protein and DNA. A comparison of the levels of cyclic GMP in IE versus UE of 12 other psoriasis patients showed the levels of this cyclic nucleotide to be significantly increased in IE based on protein, DNA and wet weight. We suggest that this imbalance in the ratio of these two cyclic nucleotides may have pathophysiological relevance to the initiation and/or the maintenance of the psoriasis lesion. 相似文献
We have observed that stimulation of human natural killer cells with dibutyryl cAMP (Bt2cAMP) reproduced the effects of ADP ribosylation of the GTP binding protein RhoA by Clostridium botulinum C3 transferase: both agents induced similar morphological changes, inhibited cell motility and blocked the cytolytic function. We demonstrate here that cAMP-dependent protein kinase A (PKA) phosphorylates RhoA in its C-terminal region, on serine residue 188. This phosphorylation does not affect the ability of recombinant RhoA to bind guanine nucleotides, nor does it modify its intrinsic GTPase activity. However, treatment of cells with Bt2cAMP results in the translocation of membrane-associated RhoA towards the cytosol. Experiments using purified membrane preparations indicated that Rho-GDP dissociation inhibitor, which can complex phosphorylated RhoA in its GTP-bound state, was the effector of this translocation. Taken together, these data suggest that PKA phosphorylation of RhoA is a central event in mediating the cellular effects of cAMP, and support the existence of an alternative pathway for terminating RhoA signalling whereby GTP-bound RhoA, when phosphorylated, could be separated from its putative effector(s) independently of its GTP/GDP cycling. 相似文献
The digestion of various carbohydrates and synthetic substrates by the gut of Locusta migratoria was analysed quantitatively. Maltose, starch, and sucrose were found to be hydrolysed most rapidly, whereas the splitting of cellobiose, trehalose, lactose, and melecitose took place at much slower rates.The absolute carbohydrase activities in foregut and midgut are nearly equal. However, specific activities are much higher in the foregut. Only low activities were found in extracts from the hindgut and salivary glands. The latter show a pattern of sugar splitting which is different from that found in gut preparations.The distribution of carbohydrase activities between the epithelia and lumina of the foregut, midgut, and hindgut and between soluble and particulate fractions were studied. The midgut epithelium is shown to have a particularly high content of enzymes, although some carbohydrases are rather active also in the epithelium of the hindgut. During hunger periods the relative enzymatic activities of the epithelium are distinctly increased.The isolation and purification of the carbohydrases were attempted and a partial separation of individual enzymes was obtained by gel-filtration. These results indicate the presence of at least seven distinct carbohydrases in the locust gut. The molecular weights of the enzymes were estimated by gel-filtration, and KM values and pH-optima are reported. 相似文献
Protein phosphorylation was studied in human peripheral lymphocytes. The cells were preincubated with , exposed to phytohemagglutinin (PHA), N6 monobutyryl cAMP (MBcAMP) or 8 bromo cGMP (BcGMP), homogenized and analyzed by SDS-polyacrylamide gel electrophoresis. Both PHA and MBcAMP produced early increases in the [32P]content of multiple proteins in the 30,000–100,000 molecular weight range. After further incubation with PHA there was a shift in the phosphorylation response to smaller molecular weight proteins. BcGMP (1 mM-10 pM) had no effect on protein phosphorylation. These results suggest a role for cAMP in the early action of PHA on human peripheral lymphocytes. 相似文献
1. The injection of azadirachtin into eight-day-old 5th instar nymphs of Locusta migratoria elicits different morphological effects dependent upon the timing of the injection relative to the major haemolymph ecdysteroid peak.2. Ecdysteroid levels can be drastically reduced, delayed and extended or not significantly affected by azadirachtin treatment dependent upon the timing of injection.3. Air swallowing to inflate the gut during the moult is significantly impaired in azadirachtin-treated insects resulting in both a reduced rate of inflation and a reduction in total air volume.4. The significance of azadirachtin treatment to the attainment of a “critical body mass”, the release of eclosion hormone and successful ecdysis is discussed. 相似文献