Lipophorin and its Receptor in Lepidoptera

Lipophorin (Lp) has an approximate native molecular weight of 730 kDa for Bombyx mori and consists of ApoLp‐I and ApoLp‐II with molecular weights of 250 kDa and 90 kDa for B. mori and 230 kDa and 80 kDa for Hyphantria cunea and 230 kDa and 49 kDa for Lymantria dispar, respectively. Lipid in Lp was mostly composed of neutral lipid. Lp of B. mori maintains constant level during larval and pupal stages but greatly increases during adult stage in both male and female. Lp of H. cunea appeared in great amounts in protein yolk bodies of ovary when vitellogenesis is actively taking place and was present in testicular fluid but not in the peritoneal sheath and cysts of testis. ApoLp‐III of B. mori has a molecular weight of 17 kDa and similar amino acid composition as those of other species Lp. H. cunea apoLp‐III has a molecular weight of 18 kDa and was present in all stages and in the protein body of ovary and in the cyst of testis. ApoLp‐III is synthesized in larval and adult fat body. cDNA sequence of Spodoptera litura apoLp‐III encodes a 188 amino acid polypeptide including a 22 amino acid leader peptide. Galleria mellonella Lp receptor has an approximate molecular weight of 97 kDa and 110 kDa under non‐reducing and reducing conditions, respectively and bound HDLp specifically. Lp receptor cDNA of G. mellonella showed th pattern of the VLDL receptor belonging to the LDL receptor family. The variant Lp receptors were expressed in the fat body of G. mellonella; one is a Lp receptor which lacks 84 bp of O linked sugar domain and the other is a full length form of the Lp receptor. The Lp receptor from the fat body of G. mellonella was differently expressed depending on the tissue and the developmental stages with specific abundance in prepupal stage.     

Insect vitellogenin/lipophorin receptors: Molecular structures, role in oogenesis, and regulatory mechanisms

Insect vitellogenin and lipophorin receptors (VgRs/LpRs) belong to the low-density lipoprotein receptor (LDLR) gene superfamily and play a critical role in oocyte development by mediating endocytosis of the major yolk protein precursors Vg and Lp, respectively. Precursor Vg and Lp are synthesized, in the majority of insects, extraovarially in the fat body and are internalized by competent oocytes through membrane-bound receptors (i.e., VgRs and LpRs, respectively). Structural analysis reveals that insect VgRs/LpRs and all other LDLR family receptors share a group of five structural domains: clusters of cysteine-rich repeats constituting the ligand-binding domain (LBD), epidermal growth factor (EGF)-precursor homology domain that mediates the acid-dependent dissociation of ligands, an O-linked sugar domain of unknown function, a transmembrane domain anchoring the receptor in the plasma membrane, and a cytoplasmic domain that mediates the clustering of the receptor into the coated pits. The sequence analysis indicates that insect VgRs harbor two LBDs with five repeats in the first and eight repeats in the second domain as compared to LpRs which have a single 8-repeat LBD. Moreover, the cytoplasmic domain of all insect VgRs contains a LI internalization signal instead of the NPXY motif found in LpRs and in the majority of other LDLR family receptors. The exception is that of Solenopsis invicta VgR, which also contains an NPXY motif in addition to LI signal. Cockroach VgRs still harbor another motif, NPTF, which is also believed to be a functional internalization signal. The expression studies clearly demonstrate that insect VgRs are ovary-bound receptors of the LDLR family as compared to LpRs, which are transcribed in a wide range of tissues including ovary, fat body, midgut, brain, testis, Malpighian tubules, and muscles. VgR/LpR mRNA and the protein were detected in the germarium, suggesting that the genes involved in receptor-endocytotic machinery are specifically expressed long before they are functionally required.     

Studies on the role of protein kinase A in humoral immune response of Galleria mellonella larvae

Protein kinase A (PKA) activity was detected in the fat body of Galleria mellonella larvae by a non-radioactive method using a specific peptide substrate-kemptide. The enzyme activity was stimulated by cAMP and its analogues: BzcMP, 8-Chl-cAMP and 8-Br-cAMP in concentrations of 1-4muM. Cyclic GMP was not effective in PKA activation. A two-fold increase in PKA activity was detected in the fat body of G. mellonella LPS-challenged larvae. Selective, membrane-permeable PKA inhibitors, H89 and Rp-8-Br-cAMPS, inhibited protein kinase A activity in the fat body of G. mellonella larvae in vitro and in vivo. The inhibition of PKA activity in vivo was correlated with a considerable lowering of haemolymph antibacterial activity and a decrease in lysozyme content in the fat body of immune challenged larvae. The use of phospho-motif antibodies recognising PKA phosphorylation consensus site allowed identification of four potential PKA phosphorylation substrates of 79, 45, 40 and 36kDa in G. mellonella fat body.     

Hormonal regulation of Drosophila microRNA let-7 and miR-125 that target innate immunity

The steroid 20-hydroxy-ecdysone (20-HE) and the sesquiterpenoid Juvenile Hormone (JH) coordinate insect life stage transitions. 20-HE exerts these effects by the sequential induction of response genes. In the nematode Caenorhabditis elegans hormones also play a role in such transitions, but notably, microRNA such as let-7 and lin-4 have likewise been found to help order developmental steps. Little is known about the corresponding function of homologous microRNA in Drosophila melanogaster, and the way microRNA might be regulated by 20-HE in the fly is ambiguous. Here we used Drosophila S2 cells to analyze the effects of 20-HE on D. melanogaster microRNA let-7 and miR-125, the homolog of lin-4. The induction by 20-HE of let-7 and miR-125 in S2 cells is inhibited by RNAi knockdown of the ecdysone receptor and, as previously shown, by knockdown of its cofactor broad-complex C. To help resolve the currently ambiguous role of 20-HE in the control of microRNA, we show that nanomolar concentrations of 20-HE primes cells to subsequently express microRNA when exposed to micromolar levels of 20-HE. We then explore the role microRNA plays in the established relationship between 20-HE and the induction of innate immunity. We show that the 3'UTR of the antimicrobial peptide diptericin has a let-7 binding site and that let-7 represses translation from this site. We conclude that 20-HE facilitates the initial expression of innate immunity while it simultaneously induces negative regulation via microRNA control of antimicrobial peptide translation.     

JNK MAP kinase is involved in the humoral immune response of the greater wax moth larvae Galleria mellonella

We investigated the participation of MAP kinases in the response of Galleria mellonella larvae to immune challenge. JNK MAP kinase was activated in fat body 10-15 min after LPS injection in vivo. The level of JNK activation was time- and LPS dosage-dependent. JNK MAP kinase isolated from cell-free extract of fat bodies dissected from immune stimulated larvae phosphorylated c-Jun protein in vitro. The activity of Gm JNK kinase was abolished in the presence of the JNK specific inhibitor SP600125. Our data indicate a correlation between JNK phosphorylation and induction of antimicrobial activity in the insect hemolymph after immune stimulation. Hemolymph from larvae pre-treated with JNK specific inhibitor SP600125 showed a reduced level of antibacterial activity after LPS injection. JNK inhibition by SP600125 abolished antibacterial activity of the in vitro culture of G. mellonella fat body. Finally, we also show a correlation between JNK-dependent immune response of G. mellonella larvae and elevated temperature.     

Venturia canescens parasitizing Galleria mellonella and Anagasta kuehniella: differing suitability of two hosts with highly variable growth potential

Venturia canescens (Grav.) (Hymenoptera: Ichneumonidae) is a solitary larval koinobiont endoparasitoid, ovipositing in several larval instars of different pyralid moth species that are pests of stored food products. After oviposition, the host larva continues to feed and grow for at least several days, the precise time doing so depending on the stage attacked. We examined the relationship between host stage and body mass on parasitoid development in late second to fifth instars of two hosts with highly variable growth potential: the wax moth, Galleria mellonella (L) and the flour moth, Anagasta kuehniella (Zeller)(Lepidoptera: Pyralidae). G. mellonella is the largest known host of V. canescens, with healthy larvae occasionally exceeding 400mg at pupation, whereas those of A. kuehniella rarely exceed 40 mg at the same stage. Parasitoid survival was generally higher in early instars of G. mellonella than in later instars. By contrast, percentage adult emergence in A. kuehniella was highest in late fifth instar and lowest in late second instar. A. kuehniella was the more suitable host species, with over 45% adult emergence in all instars, whereas in G. mellonella we found less than 35% adult emergence in all instars. Adult parasitoid size increased and egg-to-adult development time decreased in a host size- and instar-specific manner from A. kuehniella. The relationship between host size and stage and these fitness correlates was less clear in G. mellonella. Although both host species were parasitized over a similar range of fresh weights, the suitability weight-range of A. kuehniella was considerably wider than G. mellonella for the successful development of V. canescens. However, in hosts of similar weight under 5 mg when parasitized, larger wasps emerged from G. mellonella than from A. kuehniella. Parasitoid growth and development is clearly affected by host species, and we argue that patterns of host utilization and resource acquisition by parasitoids have evolved in accordance with host growth potential and the nutritional requirements of the parasitoid.     

An effect of the microsporidian Vairimorpha ephestiae (Microsporidia: Burenellidae) on activity and spectrum of nonspecific esterases in different tissues of the greater wax moth galleria mellonella (Lepidoptera: Pyralidae) larvae

The effect of the microsporidian Vairimorpha ephestiae Matted (Microsporidia: Burenellidae) on nonspecific esterases was studied in hemolymph, fat body and midgut of the larvae of Galleria mellonella L. (Lepidoptera: Pyralidae). Esterase patterns were analyzed by the polyacrylamide gel electrophoresis, the total esterase activity was detected spectrophotometrically. The increase of total esterase activity was registered in hemolymph of inflected larvae. An overexpression of esterase isozyme in hemolymph was already detected at the 3rd day post infection. No changes in esterases pattern were observed in the fat body's homogenates of the G. mellonella larvae possessing the symptoms of microsporidiosis. The degradation of esterase isozymes and the decrease of total esterase activity in the pattern of the midgut homogenates of infected larvae were registered during parasite sporogony. The greatest esterase activity in hemolymph and midgut tissues was registered during vegetative reproduction of parasite, but the least level of esterase activity was observed during mass sporogony of microsporidia.     

Differential hormone regulation of acid DNase activity in the testes and fat body of Spodoptera litura (Lepidoptera-Insecta).

Acid DNase activity in the testes and fat body is high during the early larval instars which may be correlated with the extensive cell division seen in both the tissues during these stages. The increased enzyme activity, observed in the testes of the pupal stage, might be involved in the in vivo degradation of DNA in a large number of degenerating spermatocysts which occur during this stage. Total activity of acid DNase in the fat body is highest in pupal stage. Like acid phosphatase, this enzyme may also be involved in the process of remodelling of the fat body during metamorphosis. 20-Hydroxyecdysone (20-HE) does not have any effect on acid DNase activity in the testes but it alters the enzyme activity in the fat body. Juvenile hormone-I (JH-I) has no effect on the enzyme activity in the fat body.     

Changes of acid phosphatase content and activity in the fat body and the hemolymph of the flesh fly Neobellieria (sarcophaga) bullata during metamorphosis

Changes in the specific and total activity of the lysosomal marker enzyme acid phosphatase (Acph) and in the amount of enzyme protein were examined in the fat body and the hemolymph from the last larval molt to the larval-pupal apolysis. The specific activity showed minor changes during the last larval period. In contrast, the total activity of the enzyme was low during the feeding period and higher during the wandering stage and strikingly increased at the time of puparium formation. We purified a protein having para-nitrophenyl phosphate phosphatase (Acph) activity and raised antisera against it. The amount of Acph protein in the fat body and hemolymph was examined using an ELISA. The specific Acph content showed little variation, but the total amount of the enzyme protein showed a stepwise increase in both organs during last larval stage and was markedly elevated in the pupal stage in the fat body. In contrast, a considerable decrease in the amount of Acph protein was observed in the hemolymph during this period. These data were in agreement with immunohistochemical observations showing an accumulation of the enzyme protein in fat body cells during the prepupal stage with a concomitant disappearance of the enzyme from the hemolymph. Inhibition of ecdysteroid secretion by water stress prevented the changes both in total enzyme activity and in the amount of Acph protein. However, Acph protein content and enzyme activity could be restored when the water stress was followed by a 20-hydroxyecdysone (20-HE) treatment. Taken together, our data show that Acph is secreted by fat body cells into the hemolymph during the larval stage, where it is stored in an inactive form. Increase in the 20-HE titer at the end of last larval stage reverses this process, and the enzyme is taken up by the fat body cells, where it becomes activated and appears in auto- and heterophagic vacuoles. Arch. Insect Biochem. Physiol. 34:369–390, 1997. © 1997 Wiley-Liss, Inc.     

Ecdysteroid mediated fat body acid phosphatase activity during larval development of rice moth, Corcyra cephalonica (Lepidoptera)

20-hydroxyecdysone (20-HE) stimulates acid phosphatase activity in the fat body of ligated late-last instar larvae. This effect is time dependent and the specific activity of enzyme increases significantly in hormone treated insects. 20-HE also stimulates general protein synthesis. Cycloheximide treatment either in conjunction with 20-HE or after hormone treatment blocks the increase in enzyme activity as well as increase in protein content. However, actinomycin D treatment does not alter the enzyme activity while it blocks the increase in total RNA as well as increase in protein content.     

Modulatory influence of juvenile hormone analogue (JHa) and 20-hydroxyecdysone on lipophorin synthesis in red cotton bug, Dysdercus cingulatus Fabr

Topical supply of methoprene, a juvenile hormone analogue (JHa) caused notable morphological disturbance in insects. Topical supply of methoprene to newly emerged adult female D. cingulatus caused notable disturbance and induced a dramatic reduction in the total haemolymph protein pattern and lipophorin production in tissues like fat body, ovary and haemolymph. Total protein concentration in haemolymph also showed significant reduction in 1 day old insects but increased slightly as age advanced. Application of 20-hydroxyecdysone (20-HE) to 2-day-old adult female stimulated protein synthesis intensively. Lipophorin levels in fat body and ovary also simultaneously increased. Densitometric analysis revealed that methoprene inhibits while 20-HE stimulates lipophorin production in D. cingulatus.     

Receptor-mediated endocytosis of lipid and lipophorin by the larval fat body, adult ovary and testis in the wax moth Galleria mellonella

Lipophorin (Lp) in the hemolymph of insects is known to selectively deliver lipids from sites of absorption or synthesis to sites of storage and utilization, such as the fat body, ovary and testis; however, no study regarding this has been reported in Galleria mellonella. In the present study, we examined the process by which Lp is taken up into the larval fat body, adult ovary and adult testis, and the transfer of lipid by Lp to these tissues in Galleria mellonella. To investigate the involvement of a receptor in Lp endocytosis, the larval fat body, adult ovary and adult testis were incubated for 1 h at room temperature with fluorescein isothiocyanate (FITC)‐Lp, FITC‐Lp plus unlabeled Lp, and FITC‐Lp plus suramin, a receptor endocytic inhibitor. The amounts of FITC‐Lp in the three tissues were significantly decreased in the presence of unlabeled Lp and suramin, indicating that endocytosis of Lp by the tissues is mediated by a receptor. To examine the transfer of lipid by Lp, the tissues were incubated for 1 h at room temperature with 1,1′‐dilinoleyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI)‐Lp, DiI‐Lp plus unlabeled Lp, and DiI‐Lp plus suramin. The transfer of lipid by Lp was inhibited in the presence of unlabeled Lp and suramin, which is consistent with a receptor‐mediated process. Our results show that the transfer process of lipid by Lp and uptake of Lp itself is by receptor‐mediated endocytosis.     

Biological and immune response of Galleria mellonella (Lepidoptera: Pyralidae) to sodium tetraborate

Inorganic insecticides are commonly used in urban pest management because of their low mammalian toxicity. We tested the effects of sodium tetraborate (ST) on life parameters of greater wax moth, Galleria mellonella (L.) (Lepidoptera: Pyralidae), to determine its sublethal toxicity on the insect. Survival, development, adult longevity, and fecundity of the wax moth were investigated by rearing larvae on artificial diets containing ST at concentrations of 0.005, 0.1, 0.2, or 0.3%. Larvae reared on medium at the highest concentration of ST (0.3%) had significantly decreased survival to the seventh instar and prolonged time required to reach the seventh instar. This concentration reduced pupa and adult yields to 12.5%, and it also prolonged development by 5 d. ST did not significantly influence adult longevity. Dietary ST led to significant decreases in fecundity and egg viability. Oviposition of survivors at the highest ST concentration (0.3%) was completely inhibited. Lysozyme content was decreased in larval hemolymph and fat body at high dietary ST concentrations. Fat body lysozyme content was significantly increased two-fold for larvae reared on diet at the lowest concentration of ST (0.005%). However, the highest concentration (0.3%) dramatically decreased fat body lysozyme content from 0.12 +/- 0.013 to 0.006 +/- 0.003 mg/ml in seventh instars. We infer that sublethal levels of dietary ST substantially influence life history parameters and immunocompetence in G. mellonella.     

Induction and inhibition of an apparent neuronal phenotype in Spodoptera frugiperda insect cells (Sf21) by chemical agents

The goal of this research was to induce neuron-like properties in Sf21 cells, an insect ovarian cell line, which could lead to a new high-throughput insecticide screening method and a way to mass produce insect neuronal material for basic research. This study applied differentiation agents to produce viable neuron-like cells. In the presence of the molting hormone 20-hydroxyecdysone (20-HE), or insulin, in the growth medium, a maximum of ca. 30?% of Sf21 cells expressed an apparent neuronal morphology of unipolar, bipolar, or multipolar axon-like processes within 2?C3?days. Maximal differentiation occurred after 2?days in the presence of 50???M 20-HE or 3?days in 10???M insulin. Both 20-HE and insulin displayed time- and concentration-dependent differentiation with biphasic curves, suggesting that two binding sites or processes were contributing to the observed effects. In addition, combinations of 20-HE and insulin produced apparent synergistic effects on differentiation. Caffeine, a central nervous system stimulant, inhibited induction of elongated processes by 20-HE and/or insulin, with an IC₅₀ of 9 nM for 20-HE, and the inhibition was incomplete, resulting in about one-quarter of the differentiated cells remaining, even at high concentrations (up to 1?mM). The ability to induce a neural phenotype simplifies the studies of insect cells, compared to either the use of primary nervous tissue or genetic engineering techniques. The presence of ion channels or receptors in the differentiated cells remains to be determined.     

Lipophorin as a yolk protein precursor in the mosquito, Aedes aegypti

We examined expression of the lipophorin (Lp) gene, lipophorin (Lp) synthesis and secretion in the mosquito fat body, as well as dynamic changes in levels of this lipoprotein in the hemolymph and ovaries, during the first vitellogenic cycle of females of the yellow fever mosquito, Aedes aegypti. Lipophorin was purified by potassium bromide (KBr) density gradient ultracentrifugation and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were produced against individual Lp apoproteins, apolipoprotein-I (apoLp-I) and apolipoprotein-II (apoLp-II), with molecular weights of 240 and 75 kDa, respectively. We report here that in the mosquito A. aegypti, Lp was synthesized by the fat body, with a low level of the Lp gene expression and protein synthesis being maintained in pre- and postvitellogenic females. Following a blood meal, the Lp gene expression and protein synthesis were significantly upregulated. Our findings showed that the fat body levels of Lp mRNA and the rate of Lp secretion by this tissue reached their maximum at 18 h post-blood meal (PMB). 20-Hydroxyecdysone was responsible for an increase in the Lp gene expression and Lp protein synthesis in the mosquito fat body. Finally, the immunocytochemical localization of Lp showed that in vitellogenic female mosquitoes, this protein was accumulated by developing oocytes where it was deposited in yolk granules.     

The cockroach Leucophaea maderae needs more than juvenile hormone, vitellogenin and reserves to make a yolky egg

For the cockroach Leucophaea maderae the developmental profile of lipophorin (Lp) concentrations in the hemolymph was determined through the entire vitellogenic period. At mid-vitellogenesis the concentrations of Lp had risen to 6 times the level at emergence and then declined to 2/3 of such high values at ovulation. The racemic 10R,10S-JH-III bound to Lp with an affinity of K(d) = 5.76 nM and the natural enantiomer 10R-JH-III with a K(d) = 1.60 nM. Injections of anti-Lp into mated females caused a significantly reduced rate of oocyte growth and a substantial degree of oosorption. Injections of gamma-globulin did not significantly reduce oocyte growth and caused only a small number of oocytes to resorb. Starvation after mating had similar effects as treatment with anti-Lp. Because of the high affinity of JH to Lp and since Lp occurs in micromolar concentrations during vitellogenesis one can assume that practically all JH is bound and not available for hydrolysis by the JH esterases. Lp appears to function as an inhibitor of JH metabolism by the JHEs through substrate depletion. One may conclude that a normal rate of egg growth is only achieved when titers of Lp exceed those of JH and remove major portions of this substrate from degradation by the JHEs.     

Hormonal regulation of Drosophila microRNA let-7 and miR-125 that target innate immunity

The steroid 20-hydroxy-ecdysone (20-HE) and the sesquiterpenoid Juvenile Hormone (JH) coordinate insect life stage transitions. 20-HE exerts these effects by the sequential induction of response genes. In the nematode Caenorhabditis elegans hormones also play a role in such transitions, but notably, microRNA such as let-7 and lin-4 have likewise been found to help order developmental steps. Little is known about the corresponding function of homologous microRNA in Drosophila melanogaster, and the way microRNA might be regulated by 20-HE in the fly is ambiguous. Here we used Drosophila S2 cells to analyze the effects of 20-HE on D. melanogaster microRNA let-7 and miR-125, the homolog of lin-4. The induction by 20-HE of let-7 and miR-125 in S2 cells is inhibited by RNai knockdown of the ecdysone receptor and, as previously shown, by knockdown of its cofactor broad-complex C. To help resolve the currently ambiguous role of 20-HE in the control of microRNa, we show that nanomolar concentrations of 20-HE primes cells to subsequently express microRNa when exposed to micromolar levels of 20-HE. We then explore the role microRNa plays in the established relationship between 20-HE and the induction of innate immunity. We show that the 3′UTR of the antimicrobial peptide diptericin has a let-7 binding site and that let-7 represses translation from this site. We conclude that 20-HE facilitates the initial expression of innate immunity while it simultaneously induces negative regulation via microRNa control of antimicrobial peptide translation.Key words: microRNA, ecdysone, innate immunity, let-7, diptericin, inflammation     

猴肝细胞膜脂蛋白(a)受体的研究

猴肝细胞膜蛋白经 SDS- PAGE和蛋白转移电泳后用免疫印迹法测定 ,硝酸纤维素膜分别与抗牛肾上腺皮质 LDL受体的抗体、LDL、Lp( a)和 PLG温育后见有 3条不同的蛋白条带 ,分子量分别为 370 kd、2 90 kd和 80 kd.用酶标法测定猴肝细胞膜蛋白 ,发现经 Lp( a) + PLG温育后用 Lp( a)抗体反应的结果与 Lp( a)温育后用 Lp( a)抗体反应的结果比较无显著差异 ,而经 Lp( a) + PLG温育后用 PLG抗体反应的结果比单独用 PLG温育后用 PLG抗体反应的作用强 ;同样经 Lp( a) +LDL温育后用 Lp( a)抗体反应的结果与经 Lp( a)温育后用 Lp( a)抗体反应的结果比较无显著差异 ,而经 Lp( a) + LDL温育后用 LDL抗体反应比单独经 LDL温育后用 LDL抗体的反应强 ,排除Lp( a)与 PLG和 LDL的交叉反应 ,实验认为猴肝细胞膜上可能同时存在 LDL受体、PLG受体和Lp( a)受体     

Lipophorin as a yolk protein precursor in the mosquito, Aedes aegypti

We examined expression of the lipophorin (Lp) gene, lipophorin (Lp) synthesis and secretion in the mosquito fat body, as well as dynamic changes in levels of this lipoprotein in the hemolymph and ovaries, during the first vitellogenic cycle of females of the yellow fever mosquito, Aedes aegypti. Lipophorin was purified by potassium bromide (KBr) density gradient ultracentrifugation and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were produced against individual Lp apoproteins, apolipoprotein-I (apoLp-I) and apolipoprotein-II (apoLp-II), with molecular weights of 240 and 75 kDa, respectively. We report here that in the mosquito A. aegypti, Lp was synthesized by the fat body, with a low level of the Lp gene expression and protein synthesis being maintained in pre- and postvitellogenic females. Following a blood meal, the Lp gene expression and protein synthesis were significantly upregulated. Our findings showed that the fat body levels of Lp mRNA and the rate of Lp secretion by this tissue reached their maximum at 18 h post-blood meal (PMB). 20-Hydroxyecdysone was responsible for an increase in the Lp gene expression and Lp protein synthesis in the mosquito fat body. Finally, the immunocytochemical localization of Lp showed that in vitellogenic female mosquitoes, this protein was accumulated by developing oocytes where it was deposited in yolk granules.