The influence of dietary protein on ascorbic acid metabolism in rats

1. d-Glucuronolactone reductase, l-gulonolactone oxidase, uronolactonase, dehydroascorbatase, l-gulonate dehydrogenase and l-gulonate decarboxylase have been measured in the tissues of rats fed on diets containing variable amounts of protein. Rats fed on a protein-free or a 2% casein diet for 15 days showed a marked decline in the activities of d-glucuronolactone reductase, l-gulonolactone oxidase, uronolactonase and dehydroascorbatase in the liver, and no change in l-gulonate dehydrogenase and l-gulonate decarboxylase activities in the kidney when compared with rats fed on diets containing 9%, 18% or 25% casein. Giving diets containing 60% or 88% casein to rats did not appreciably alter the activities of uronolactonase, dehydroascorbatase, l-gulonate dehydrogenase and l-gulonate decarboxylase, but inhibited considerably the activities of d-glucuronolactone reductase and l-gulonolactone oxidase in the liver, resulting in decreased synthesis of ascorbic acid. 2. Rats fed on a 25% casein diet showed maximal weight gain, higher tissue reserve of ascorbic acid and higher urinary excretion of both ascorbic acid and glucuronic acid when compared with rats fed on diets containing lower or higher amounts of protein.     

Variation of l-gulonolactone oxidase activity in placental mammals

• 1.1. The activity of l-gulonolactone oxidase (EC 1.1.3.8) in livers of 49 species of eutherian mammals varied intraspecifically among individuals; coefficients of variation were 0.2 to 0.4 in many species.
• 2.2. Differences observed in l-gulonolactone oxidase activity among strains of laboratory rats and domestic rabbits are probably genetically controlled.
• 3.3. Pronounced sex differences in l-gulonolactone oxidase activity were found in some species, particularly in the genera Peromyscus, Reithrodontomys and Onychomys.
• 4.4. Mormota monax exhibited seasonal variation in l-gulonolactone oxidase somewhat like that previously observed in Sylvilagus floridanus; no such seasonal variation was found in Sciurus carolinensis.
• 5.5. Hibernation did not affect l-gulonolactone oxidase activity in Spermophilus tridecemlineatus.
• 6.6. In four species of rodents, Microtus ochrogaster, Tylomys panamensis, Octodon degus and Sigmodon hispidus, l-gulonolactone oxidase activity was not affected by the level of dietary ascorbate.

     

Utilization of U-14C amino acids or U-14C protein by adult Glossina morsitans during in utero development of larva

Administration of U¹⁴C protein hydrolysate in the diet of adult female Glossina morsitans at different times throughout the second reproductive cycle was followed by analysis of the distribution of radioactivity between the adult flies, their excreta, and the fully grown third instar larvae produced by these flies. A constant proportion of the total administered label was recoverable independently of the time lapse between administration and assay. Peak incorporation of labelled material occurred in the larva between the seventh and eighth day of a 9 or 10 day interlarval period, indicating that the larva feeds avidly on recently synthesized maternal uterine gland secretion at this time. Haemocoelic injection of U¹⁴C protein hydrolysate into similar adult females, between feeds, resulted in continued incorporation of labelled material by the larva to within 12 hr of parturition. Results are consistent with the hypothesis that uterine gland secretion and larval feeding continue throughout the intrauterine life of the larva.A constant and low proportion of detectable label remained in the adult fly while increased incorporation by the larva was paralleled by a reduction of detectable label in the adult excreta. This indicates direct competition between the uterine gland cells and those of the Malpighian tubules for free amino acids in the haemolymph.Administration of U¹⁴C protein in the adult diet did not result in incorporation of label by the developing larva, and the bulk was excreted as protein by the adult fly. Apparently the midgut trypsin of G. morsitans is incapable of splitting this labelled protein.Analysis of urine and haemolymph samples from flies in early pregnancy, recently fed on a diet containing U¹⁴C protein hydrolysate or U¹⁴C protein, shows that free labelled amino acids in the diet enter the adult haemolymph almost immediately after feeding, and are excreted along with dietary water during initial diuresis. The labelled protein used in these experiments was not taken up by the haemolymph and consequently did not appear in the urine.Implications are that the adult female G. morsitans possesses little storage capacity for substances in the diet which are destined to provide nutrients for the developing larva. Assuming a 48 hr digestion time, the digestive products of a blood meal ingested on day 5 or 6 of a 9 day interlarval period will provide the bulk of nutrients for larval growth. It is therefore significant that blood meals ingested at this time are larger than those ingested earlier or later in the cycle.     

Influence of thyroid hormones on neuronal death and differentiation in larval Rana pipiens.

The sphingid moth, Manduca sexta, typically passes through five larval instars, a pupal, and an adult stage. The larval labial glands secrete silk in the first instar and a viscous lubricant in the fifth. During metamorphosis the glands develop into salivary organs which produce an invertase-rich secretion. In normal development, the uniform population of cells in the duct of the larval gland transforms into the four sequentially arranged regions of secretory and conductive cells of the adult gland. In order to determine when competence to form the adult gland is established, fragments of labial gland ducts from first through fifth instar larvae were implanted into pupae. These gland fragments underwent metamorphosis with their hosts, passing through the same developmental phases. Glands from as early as the first instar were competent to form histologically and functionally normal adult regions. In later instars, transplants of measured fragments demonstrated that larval cells were programmed in situ to develop into the four adult cell types.     

Effect of dietary cholesterol deficiency upon its distribution in the larvae of the housefly, Musca domestica

The free sterol, total phospholipids and protein content of the various tissues and haemolymph lipoproteins obtained from the larvae of Musca domestica, reared on the diets containing 0.56 μmole cholesterol/g wet weight of diet (normal) and 0.05 μmole cholesterol/g wet weight of diet (deficient) have been determined. The cholesterol in the diet was found to be taken up by the larvae and distributed between all the tissues examined. About 60% of the free sterol in the larvae was recovered from the composite gut fraction and muscle. Cholesterol deficiency reduced both the growth of larvae and the free sterol content of the various tissues and haemolymph when compared to that of normal larvae. Cholesterol deficiency resulted in a slightly higher proportion of sterol and protein of the larval haemolymph being associated with the lipoproteins having slower electrophoretic mobility. Most of the different tissues from the cholesterol deficient larvae contained a much smaller proportion of their normal free sterol content than of their phospholipid or protein; the brain tissue however contained a higher percentage of free sterol and the haemolymph a much lower percentage than would be expected from the lowering of phospholipid and protein content as a result of the deficiency. When the sterol content was expressed relative to the protein, the ratio was higher in the brain tissue of both the normal and deficient larvae than the ratio present in the remaining tissues, apart from the composite gut fraction of the normal larvae. The results suggest that a disproportionate amount of available cholesterol was being concentrated into the nervous system of the cholesterol deficient insect.A rather higher proportion of the total sterol fraction recovered from the various tissues and haemolymph lipoproteins of cholesterol deficient larvae behaved as ‘polar metabolites’ of cholesterol when compared with that of normal larvae.     

Effects of vitamins A and D on the biosynthesis of L-ascorbic acid by rat-liver microsomes.

1. The synthesis of l-ascorbic acid from either d-glucuronolactone or l-gulonolactone by liver microsomes of rats is decreased under conditions of hypervitaminosis A; under hypervitaminosis D the synthesis from d-glucuronolactone is increased and that from l-gulonolactone is not affected. 2. The microsomal conversion of l-gulonolactone into l-ascorbic acid is impaired in liver tissues of rats made deficient with respect to either vitamin A or vitamin D when compared with the controls maintained on stock diet.     

Role of maternal tissue in the synthesis of polyunsaturated fatty acids in response to a lipid-deficient diet during pregnancy and lactation in rats

During pregnancy and lactation, metabolic adaptations involve changes in expression of desaturases and elongases (Elovl2 and Elovl5) in the mammary gland and liver for the synthesis of long-chain polyunsaturated fatty acids (LC-PUFAs) such as arachidonic acid (AA) required for fetal and postnatal growth. Adipose tissue is a pool of LC-PUFAs. The response of adipose tissue for the synthesis of these fatty acids in a lipid-deficient diet of dams is unknown. The aim of this study was to explore the role of maternal tissue in the synthesis of LC-PUFAs in rats fed a low-lipid diet during pregnancy and lactation. Fatty acid composition (indicative of enzymatic activity) and gene expression of encoding enzymes for fatty acid synthesis were measured in liver, mammary gland and adipose tissue in rats fed a low-lipid diet. Gene expression of desaturases, elongases, fatty acid synthase (Fasn) and their regulator Srebf-1c was increased in the mammary gland, liver and adipose tissue of rats fed a low-lipid diet compared with rats from the adequate-lipid diet group throughout pregnancy and lactation. Genes with the highest (P < 0.05) expression in the mammary gland, liver and adipose tissue were Elovl5 (1333%), Fads2 (490%) and Fasn (6608%), respectively, in a low-lipid diet than in adequate-lipid diet. The percentage of AA in the mammary gland was similar between the low-lipid diet and adequate-lipid diet groups during the second stage of pregnancy and during lactation. The percentage of monounsaturated and saturated fatty acids was significantly (P < 0.05) increased throughout pregnancy and lactation in all tissues in rats fed a low-lipid diet than in rats fed an adequate-lipid diet. Results suggest that maternal metabolic adaptations used to compensate for lipid-deficient diet during pregnancy and lactation include increased expression of genes involved in LC-PUFAs synthesis in a stage- and tissue-specific manner and elevated lipogenic activity (saturated and monounsaturated fatty acid synthesis) of maternal tissues including adipose tissue.     

Cellular metamorphosis. II. The larval labial gland duct and its prospective adult fates in the tobacco hornworm.

The larval labial gland of the sphingid moth, Manduca sexta, produces a viscous secretion, presumably a lubricant, facilitating the burrowing which precedes pupation. During metamorphosis, the gland transforms into a salivary organ, producing an invertase-rich digestive secretion. The single-cell type found in the duct of the larval gland transforms into the four structurally and functionally distinct cell types found in the four sequentially arranged secretory and conductive regions of the adult salivary gland. Surgical experiments were performed to study the prospective fates of different parts of the larval gland. The glands were bisected and one or both fragments were left in situ to undergo metamorphosis. In addition, fragments of the larval gland were implanted in pupal hosts and went through metamorphosis free of their prior attachments. The four linearly arrayed adult regions originate from correspondingly positioned areas in the larval duct.     

Regulation of ascorbate oxidase expression in pumpkin by auxin and copper

Ascorbate oxidase expression in pumpkin (Cucurbita spp.) tissues was studied. Specific ascorbate oxidase activities in pumpkin leaf and stem tissues were about 2 and 1.5 times that in the fruit tissues, respectively. In seeds, little ascorbate oxidase activity was detected. Northern blot analyses showed an abundant ascorbate oxidase mRNA in leaf and stem tissues. Fruit tissues had lower levels of ascorbate oxidase mRNA than leaf and stem tissues. Ascorbate oxidase mRNA was not detected in seeds. Specific ascorbate oxidase activity gradually increased during early seedling growth of pumpkin seeds. The increase was accompanied by an increase in ascorbate oxidase mRNA. When ascorbate oxidase activity in developing pumpkin fruits was investigated, the activities in immature fruits that are rapidly growing at 0, 2, 4, and 7 d after anthesis were much higher than those in mature fruits at 14 and 30 d after anthesis. The specific activity and mRNA of ascorbate oxidase markedly increased after inoculation of pumpkin fruit tissues into Murashige and Skoog's culture medium in the presence of an auxin such as 2,4-dichlorophenoxyacetic acid (2,4-D) but not in the absence of 2,4-D. In the presence of 10 mg/L of 2,4-D, ascorbate oxidase mRNA was the most abundant. Thus, ascorbate oxidase is induced by 2,4-D. These results indicate that ascorbate oxidase is involved in cell growth. In pumpkin callus, ascorbate oxidase activity could be markedly increased by adding copper. Furthermore, immunological blotting showed that the amount of ascorbate oxidase protein was also increased by adding copper. However, northern blot analyses showed that ascorbate oxidase mRNA was not increased by adding copper. We suggest that copper may control ascorbate oxidase expression at translation or at a site after translation.     

DOPA and tyrosine decarboxylase activity in tissues of Periplanet a americana in relation to cuticle formation and ecdysis

DOPA decarboxylase activity in haemolymph and integument was low in last instar and early pharate adult Periplaneta americana, but began to increase shortly before ecdysis. Decarboxylation rates of l-DOPA, about 10 times the larval level by the start of ecdysis, reached a peak about 6 hr afterward, coinciding with the main period of cuticular sclerotization. Activity decreased rapidly during the next 18 hr, then decreased gradually for several days. Haemolymph DOPA decarboxylase activity was about four times greater than the integument, based on tissue dry weights. The fat body and gut tissues had low DOPA decarboxylase activity in all ages tested, and this did not increase at ecdysis. Tyrosine decarboxylase activity was significant only in the haemolymph and at consistently low levels.DOPA decarboxylase, therefore, apparently plays a major rôle in production of catecholamine derivatives for cuticular sclerotization in P. americana, while tyrosine decarboxylation is minor. Both haemolymph and integument appear to be important sites of dopamine biosynthesis.     

Comparative analysis of two biliproteins, BP1 and BP2, from haemolymph of cabbage white butterfly, Pieris rapae

Two blue-pigment binding proteins, BP1 and BP2, are present in larval and pupal haemolymph of cabbage white butterfly, Pieris rapae, and fluctuate in expression during development. Both BP1 and BP2 are found in pupal haemolymph in varying proportions as well as in adult haemolymph, while only small amounts of BP2 are found in larval haemolymph. BPs are separated by 75% ammonium sulfate, and then purified effectively by ion exchange column chromatography and preparative gel electrophoresis. It was shown that BP1 and BP2 have molecular masses of 20,244 and 19,878 Da, and isoelectric points of 7.0 and 6.8, respectively. Considering their amino acid compositions and N-terminal amino acid sequences, the two proteins are almost identical except the first N-terminal amino acid. The first amino acid of BP1 is asparagine, whereas the initial residue of BP2 is aspartic acid. Anti-BP1 cross-reacts with BP2, indicating that they have immunological homogeneity. Western blotting analyses revealed that only BP1 was present in the larval tissues such as fat body, integument, muscle, and hindgut. However, BP1 was not found in midgut, Malphigian tubules, and silk gland. BP1 was also present in the protein bodies, and both cuticle and hemocoel sides of larval epidermis cells by the transmission electron microscopic observation. The information in this report will facilitate studies on the molecular biology and biological significance of insect BPs.     

Adipokinetic hormone-induced lipid mobilization and lipophorin interconversions in fifth larval instar locusts

The response of fifth larval instar locusts to injected adipokinetic hormone (AKH) is only poor, as is reflected in both a very moderate elevation of the haemolymph lipid concentration and the slight occurrence of the haemolymph lipophorin interconversions characteristic for adult locusts, resulting in formation of only small quantities of the low density lipophorin (A⁺). However, an additional lipophorin fraction (A′) is induced, which is intermediate in density and size between high and low density lipophorin and which is not identified in adult haemolymph. As in adults, larval A⁺ formation includes association of the resting high density lipophorin with a non-lipid containing protein (C₂), the haemolymph concentration of which is only one-fifth relative to adults. However, the larval haemolymph protein composition is not the primary cause of the incomplete adipokinetic response, as elevation of the concentration of protein C₂ by injection of isolated adult C₂, whether or not in combination with adult high density lipophorin, did not increase lipophorin conversions nor haemolymph lipid elevation.In vitro incubation of larval fat bodies in adult haemolymph showed that competency to both the AKH-induced lipid release and the haemolymph lipophorin conversions of the larval fat body are reduced compared to equal amounts of adult tissue. Reciprocal incubation of adult fat body in larval haemolymph resulted in only a very moderate adipokinetic response, demonstrating that larval haemolymph protein composition is restrictive for full development of hormone action.Both immunoblotting experiments and enzyme-linked immunosorbent assays (ELISA), using monoclonal antibodies specific for the adult lipophorin apoproteins, indicated that the larval lipophorins closely resemble the adult forms. Apparently the structure of locust lipophorins is remarkably constant throughout development despite changes in metabolic functions.     

Chromium and chronic ascorbic acid depletion effects on tissue ascorbate,manganese, and14C retention from14C-ascorbate in guinea pigs

Chromium (Cr) potentiates the effects of insulin and a role for insulin in ascorbic acid transport has been reported. Therefore, the effects of Cr and ascorbate depletion on tissue ascorbic acid and¹⁴C distribution and excretion after a¹⁴C ascorbate dose were investigated in guinea pigs. As utilization of dietary Cr is affected by interaction with other minerals, tissue manganese (Mn), zinc (Zn), copper (Cu), and iron (Fe) were examined. For 20 wk, 40 weanling animals were fed either a Cr-deficient (<0.06 μg Cr/g diet, ?Cr) or a Cr-adequate (2 μg Cr from CrCl₃/g diet, +Cr) casein-based diet and were given 1 mg ascorbate/d (?C) or 10 mg ascorbate/d (+C) for 20 wk. Animals fed the Cr-depleted diet had decreased weight at 20 wk (p<0.01). Six hours before necropsy, animals were dosed by micropipette with 1.8 μCi ofl-[carboxyl-¹⁴C] ascorbic acid and placed in metabolic cages. Ascorbate supplementation increased Fe concentrations in most analyzed tissues, hepatic¹⁴C, tissue ascorbate and Mn concentration in the adrenal and testes, but decreased the concentrations of Cu in the kidney and Mn in the spleen. Liver Mn concentration was higher and kidney Mn concentration was lower in +Cr animals. Interactions between Cr and ascorbic acid affected Mn concentrations in bone and brain. These results indicate that ascorbate and Cr may affect Mn distribution. Chromium supplementation decreased plasma cortisol, brain¹⁴C and the amount of¹⁴C expired as carbon dioxide. These findings suggest that dietary Cr may affect ascorbic acid metabolism and the metabolic response to stress.     

Changes taking place in the electrophoretic haemolymph protein pattern during metamorphosis of Polytela gloriosae (Lepidoptera)

The haemolymph proteins of the larva, pupa and adult of Polytela gloriosae have been fractioned by Polyacrylamide gel disc electrophoresis. In the haemolymph of the fifth instar larval stage a total of ten protein fractions have been detected. The concentration of the protein fractions 2, 3, 4, 9 and 10 shows oscillations in their concentration in the early fifth instar, middle fifth instar and late fifth instar larval stage. In all 11 protein fractionswere detected in the haemolymph of different stages of the pupa. The protein bands 1, 7 and 10 of the pupa appear newly in the haemolymph as these bands were not found in the haemolymph of the larvae. The protein fraction 9 of larva was not found in the pupa. In the haemolymph of adult insect sexual difference was observed in the haemolymph protein pattern. In the haemolymph of adult female a total of 10 protein fractions were detected while from the male haemolymph a total of 8 protein fractions were detected. The pupal band 7 was not found in the adults of both the sexes. In the haemolymph of larva and adult one pigmented protein fraction was observed. No pigmented protein fraction was found in the haemolymph of pupa. Iron - containing protein fraction and the acid mucopolysaccharides were not found in the haemolymph. The protein fractions 3, 4, 5, 6 and 7 of adult haemolymph were darkly stained by the Schiff reagent and, thus, they are the fractions of glycoprotein. One protein fraction of lipoprotein was also found in the haemolymph.     

Characterization of a Monoclonal Antibody Directed against Mytilus spp Larvae Reveals an Antigen Involved in Shell Biomineralization

The M22.8 monoclonal antibody (mAb) developed against an antigen expressed at the mussel larval and postlarval stages of Mytilus galloprovincialis was studied on adult samples. Antigenic characterization by Western blot showed that the antigen MSP22.8 has a restricted distribution that includes mantle edge tissue, extrapallial fluid, extrapallial fluid hemocytes, and the shell organic matrix of adult samples. Other tissues such as central mantle, gonadal tissue, digestive gland, labial palps, foot, and byssal retractor muscle did not express the antigen. Immunohistochemistry assays identified MSP22.8 in cells located in the outer fold epithelium of the mantle edge up to the pallial line. Flow cytometry analysis showed that hemocytes from the extrapallial fluid also contain the antigen intracellularly. Furthermore, hemocytes from hemolymph have the ability to internalize the antigen when exposed to a cell-free extrapallial fluid solution. Our findings indicate that hemocytes could play an important role in the biomineralization process and, as a consequence, they have been included in a model of shell formation. This is the first report concerning a protein secreted by the mantle edge into the extrapallial space and how it becomes part of the shell matrix framework in M. galloprovincialis mussels.     

Regulation of insect prothoracic glands: Existence of a haemolymph stimulatory factor in Manduca sexta

The primary regulator of ecdysone biosynthesis by insect prothoracic glands is the prothoracicotropic hormone. However, it now appears that other factors, secondary regulators, may modulate prothoracic gland activity. One such factor has been isolated from the haemolymph of Manduca larvae. This haemolymph factor stimulates in vitro ecdysone synthesis by larval and pupal prothoracic glands by approx. 5-fold. It has an apparent mol. wt of ～330 kD, is protease-sensitive and is heat labile, the latter clearly distinguishing it from the prothoracicotropic hormone. Further, its steroidogenic effects and those of prothoracicotropic hormone are additive. Treatment of larval or pupal prothoracic glands with both moieties simultaneously effects an approx. 10-fold increase in ecdysone synthesis. The haemolymph titre of the stimulatory factor is low at commitment of the last-larval instar, then increases by approx. 3-fold later in the instar during pharate-pupal development. This increase in the titre is sufficient to effect a significant increase in prothoracic gland activity that could be physiologically important. Thus, it appears that the fluctuating level of this haemolymph stimulatory factor may act in conjunction with prothoracicotropic hormone to regulate the haemolymph ecdysteroid titre by modulating the ecdysone biosynthetic activity of the prothoracic glands.     

Humoral inhibition of the corpora allata in larvae of Diploptera punctata: Role of the brain and ecdysteroids

Juvenile hormone synthesis by adult female corpora allata was inhibited following implantation into final-larval-instar males; inhibition was prevented by decapitation of the larval hosts on day 11 (prior to the head critical period for moulting), but not by decapitation on day 13. Implantation of one larval protocerebrum restored inhibition of implanted corpora allata, demonstrating that the brain releases an inhibitory factor. Corpora allata implanted into larvae decapitated on day 11 were inhibited by injections of 20-hydroxyecdysone. Since treatment of corpora allata with 20-hydroxyecdysone in vitro did not inhibit juvenile hormone synthesis, ecdysteroids probably act indirectly on the corpora allata. Juvenile hormone synthesis and haemolymph ecdysteroid concentration were measured following implantation of corpora allata along with two larval brains into larval hosts. Brain implantation did not affect ecdysteroid concentration, but did inhibit juvenile hormone synthesis, even in animals with low haemolymph ecdysteroid concentration. Incubation with farnesoic acid stimulated juvenile hormone synthesis by corpora allata from males early in the final larval stadium, but not after day 8, showing that one of the final two reactions of juvenile hormone synthesis is rate-limiting in larval corpora allata at this stage. Adult female corpora allata which had been humorally inhibited by implantation into larvae were stimulated by farnesoic acid.     

Structural changes occurring during transformation of the labial gland cells to their adult form,in Manduca sexta

The labial gland of the adult sphingid moth, Manduca sexta, is composed of five distinct regions, each made of a single cellular type. Four of these regions are derivatives of the single specialized cellular population that makes up the caterpillar labial duct. Both the larval labial duct and its derivatives are large, polyploid cells with pleiomorphic nuclei. There is a definite cellular continuity between the larval and adult forms of these cells throughout metamorphosis; no mitoses or cell deaths are seen to occur in the gland during transformation. Cytological studies indicate that in the process of cell transformation the ducts first “dedifferentiate,” elongate, then redifferentiate. Intermediates in this process have well defined structures which should make this system useful in studying covert events in the transformation process.     

A larval haemolymph protein in the eggs of Rhynchosciara americana

In Rhynchosciara americana larvae, a protein referred to as “protein 10” comprises one of the major plasma protein of the last instar. This protein was purified and an antiserum against it shows that an identical protein is present in the eggs from this fly. Protein 10 has an estimated molecular weight of 43,000 and an isoelectric pH of 6.6. Examination of protein 10 from eggs and from haemolymph by limited proteolysis indicates that the two are structurally identical. Protein 10 is synthesized in large amounts by the larval fat bodies up until the end of the feeding stage. Estimation by radial immunodiffusion shows that protein 10 is stored in the larval haemolymph, attaining a maximum level at the end of feeding stage and then decreasing until very little remains in the young adult. By the middle of the pupal stage the ovaries begin to sequester and acummulate the protein 10 which is deposited in the eggs.     

Diet factors responsible for the change of the glucose oxidase activity in labial salivary glands of Helicoverpa armigera

We investigated the change of the glucose oxidase (GOX) activity in labial salivary glands of Helicoverpa armigera larvae fed with the artificial diet or host plant tobacco and the major factors responsible for such a change. Throughout larval development, the labial salivary GOX activities in caterpillars reared on the artificial diet were remarkably higher than those fed with the plant. After fifth-instar plant-fed caterpillars were transferred to the artificial diet, their labial salivary GOX activity increased quickly, which was closely correlated with the time spent feeding on the artificial diet. The total sugar content of the artificial diet was 68 times higher than that of the tobacco leaves. We hypothesized that sugars and secondary metabolites are the possible causes of induction of GOX activity. When fifth-instar caterpillars were fed with tobacco leaves coated with glucose or sucrose, their labial salivary GOX activity was significantly higher than those fed with leaves without sugar coating. Following native PAGE, 1 single band of the labial salivary GOX was observed in all the caterpillars fed with different diets, implying that only the activity of the isoenzyme was changed in response to different diets. Furthermore, the labial salivary GOX activity was determined after caterpillars were fed with artificial diets containing chlorogenic acid, rutin, and quercetin. The results showed that all these phenolic compounds had no effect on the GOX activity. We conclude that sugar in diets was a major factor influencing the labial salivary GOX activity of the larvae. Arch. Insect Biochem. Physiol. 2008.