Protein metabolism by the salivary glands and other organs of the larva of the blowfly, Calliphora erythrocephala

Protein metabolism in salivary glands, gut, haemolymph, and fat body during the last larval instar of the blowfly, Calliphora erythrocephala, has been investigated. In salivary glands, protein release, protein synthesis, amylase, and pepsin-like protease activity were maximal in 6 day larvae, this being at a time when the larvae had finished feeding. All these functions declined in glands from the rounded-off white puparial stage (R.O.) while acid phosphatase activity rose throughout the third instar to a maximum at the R.O. stage, Glands from 6 and 7 day larvae released protein which on disk gel electrophoresis separated into four minor bands and two major bands one of the latter possessing protease activity.In the gut, pepsin-like protease activity was maximal in 4 day larvae after which it fell rapidly thus following the feeding pattern of the larva in contrast to that in the salivary glands which did not.In vitro experiments showed that protease was released from 6 day glands through the basal membrane of the cells and not via the duct. A pepsin-like protease was also found in the haemolymph and fat body, the activity in the fat body rising rapidly during the latter part of the third instar, a rise which is attributed to the fat body sequestering protease from the haemolymph. Acid phosphatase activity in the fat body was maximal in 5 day larvae indicating that this enzyme was synthesized early in the third instar. It was shown that fat body sequestered ¹⁴C-labelled protein synthesized by and released from the salivary glands, most of the ¹⁴C activity being associated with a 600 g precipitable, acid-phosphatase rich fraction.It is proposed that in late third instar larvae the salivary glands function as glands of internal secretion, releasing protease into the haemolymph, which is then sequestered by the fat body (and perhaps other tissues) and is subsequently used in the lysis of the tissues at the time of metamorphosis.     

Plant growth-promoting factor in the salivary gland of the bug, Lygus disponsi

In the salivary gland of Lygus disponsi, the factor promoting the activity of 3-indoleacetic acid (IAA), or the factor inhibiting the activity of IAA-oxidase, was present. It showed a maximum activity in fraction III of the salivary gland solution, 1·6 times as high as that of the IAA control. The significance of the factor in the salivary gland is discussed with respect to injury (malformation). Auxin was not found in the salivary gland.     

The chemical nature of the defensive larval secretion of the citrus swallowtail, Papilio demodocus

The eversible cervical gland or osmeterium of the larva of the African citrus swallowtail, Papilio demodocus Esper, produces a secretion containing isobutyric acid and 2-methyl-butyric acid as well as small quantities of the methyl and ethyl esters of these acids. The secretion of final instar larvae was found to differ from that of younger larvae.     

Cell differentiation during the ontogeny of larval salivary glands of the fly, Telmatoscopus albipunctatus

Using the larvae, pharate pupa, and pharate adults of the moth fly, Telmatoscopus albipunctatus, histological and ultrastructural features of the salivary glands were investigated. The gland lumen contains a milky secretion from the first instar. This secretion continues to ccur at all subsequent developmental stages; with the onset of the pharate pupal stage, however, the secretion becomes transparent and rather viscous. Histochemical tests revealed that it is mainly proteinaceous. Glands from the same developmental stage may respond differently to PAS-reaction.Various cell organelles were compared at consecutive stages of larval development and of secretory activity of the salivary glands. In first and second instar larvae autophagic vacuoles are virtually absent in the salivary gland cells. They were occasionally found in the third instar, when they appear to be engaged in the process of organelle turnover. Histolysis of the larval glands is initiated towards the close of the fourth instar when the number of autophagic vacuoles starts to increase. Simultaneously, the cytoplasm, previously full of ribosomes and endoplasmic reticulum, starts losing these structures. At the beginning of the pharate adult stage, the cytoplasm becomes practically devoid of all structures other than those engaged in autophagy.Polyteny of the chromosomes during ontogeny of the larval salivary glands is also discussed.     

Factors influencing the developmental capacity of Galleria larval epidermis

The nature of the cuticle secreted by integument from a day-1 penultimate instar larval Galleria when cultured in vivo in the abdomen of a last instar larva varied with the age of the host. When placed in a day-5 last instar larva, the implanted integument secreted a pupal cuticle at the time the host metamorphosed and became a pupa. However, when placed in a day-7 last instar larva the implant, from the same stage donor, secreted a larval cuticle at the time the host pupated. Experimental studies involving implantation of the integument for a 24 hr period, into various developmental stages of normal and ligated last instar larvae, pupae, and pharate adults, prior to placing it in a day-7 last instar larva suggest that a non-hormonal factor present in day-4 and -5 last instar larvae is important to initiate pupal syntheses.     

Hormonal effects on carotenoid uptake by the silk gland in the silkworm, Bombyx mori

The carotenoid uptake by the silk gland of the silkworm (Bombyx mori), which occurs only during the middle to late period of the last (fifth) instar in the natural condition, was studied in relation to the hormonal controls. During certain stages of the fourth and last instars, the corpus allatum hormone (JH) was found to inhibit the activation of the absorbing function of the silk gland. The absorbing activity was inactivated, if the activated silk gland was implanted into larva at the late stage of the fourth instar in the presence of the moulting hormone (MH). As more ponasterone-A (ecdysone-analogue) was injected into decapitated larvae, the pigmentation of the silk gland was increased; but injection of a high titre inhibited its activity. It seems that, through serial transplantations, the silk gland inactivated experimentally at the late stage of the fourth instar is reactivated in the presence of MH during the middle to late period of the last instar. The results indicate that MH and JH at each stage control the activity of the carotenoid uptake.     

Puffing patterns in the salivary gland chromosomes of the melonfly,Dacus cucurbitae

The puffing pattern changes in the salivary gland chromosomes of the third instar larvae of the melonfly, Dacus cucurbitae are described. Three classes of puffs were noticed over a period of development of 120 hrs. Class (1) are those which are more or less constantly found; class (2) are those which oscillate, i.e. appear and disappear at irregular time intervals; and class (3) are those that are linked to a specific developmental event. Also, 3 peaks of puffing activity have been noticed during the present study; one in the 120 hr old larva, the second in the 168 hr old larva and the third in the 240 hr old larvae. The significance of these 3 classes of puffs and the 3 peaks in puffing activity has been discussed. The puff RNA has a high rate of synthesis and incorporates ³H-cytidine within 30 secs after being offered. There is a high degree of variation in the incorporation of labelled precursors into the different nuclei of the same gland, such a variation is not noticed in the diploid and embryonic cells.     

Brain neurosecretory cell and ecdysiotropin activity of the non-diapausing,pre-diapausing and diapausing southwestern corn borer, Diatraea grandiosella Dyar

The location and number of brain neurosecretory cells were studied in the larval southwestern corn borer. One posterior, two median and two lateral groups of paraldehyde-fuchsin positive cells were found in each cerebral hemisphere.Implantation of brain parts containing different groups of neurosecretory cells revealed that the median neurosecretory cells contained higher ecdysiotropic activity than the other cell groups. In vitro culture of ecdysial gland with brain or brain-parts extract showed also that the median neurosecretory cells contained much higher ecdysiotropic activity than other neurosecretory cells. To estimate the ecdysiotropic activity of pre-diapausing 6th instar larvae, their brain or brain extract was incubated in culture medium containing an ecdysial gland from a day-4 last-instar non-diapausing larva. Data showed that the ecdysiotropic activity in the pre-diapausing larvae was far lower than in non-diapausing and diapausing larvae.     

The activity of Balbiani rings 1 and 2 in salivary glands of Chironomus tentans larvae under different modes of development and after pilocarpine treatment

The activity of the Balbiani rings 1 and 2 (BR1 and BR2) in the salivary gland was followed during development of fourth instar larvae of Chironomus tentans under different modes of development, with or without a previous pilocarpine treatment. The activity was determined in parallel by two different methods, by incorporation of [3H]uridine into BR-RNA (75 S) and by morphometry of BR1 and of BR2. In glands of untreated larvae BR2 does not change dramatically except for a depression of activity during oligopause (resting phase). BR1 is completely inactive during this phase but exhibits a pronounced activity maximum in the middle of the prepupal period, in subitaneously developing (i.e., uninterrupted) as well as in postoligopause cultures. After pilocarpine treatment the activity of BR2 (rather than of BR1) is generally increased. The extent of this stimulation, however, is strongly development dependent. A most striking activity difference is observed in postoligopause between animals of stage 5 and of stage 6. The relationship between BR2 activity and degree of emptying of the salivary gland lumen was investigated. A model is proposed in which BR2 activity is conceived as being regulated by two parameters: by the degree of filling of the gland lumen and by the stage and mode of development of the larva.     

琥珀蚕丝腺转录因子基因AaSGF-1的克隆、原核表达及组织表达分析

[目的]本研究旨在克隆琥珀蚕Antheraea assama丝腺转录因子基因AaSGF-1,分析其序列特征及表达模式并制备多克隆抗体,为探讨该基因的生理功能奠定基础.[方法]采用RT-PCR和RACE技术从琥珀蚕丝腺中克隆AaSGF-1的cDNA序列,并进行生物信息学分析;利用qPCR检测AaSGF-1在琥珀蚕5龄第4...     

Mandibular glands in the endoparasitic larva of Uropsylla tasmanica Rothschild (Siphonaptera : Pygiopsyllidae)

An histological study of flea larvae was carried out in order to compare free-living, larvae with the unique endoparasitic larva of Uropsylla tasmanica Rothschild (Siphonaptera : Pygiopsyllidae), a species confined to dasyurid hosts in Tasmania and Victoria, Australia. The free-living species examined were Ctenocephalides felis Bouché (Siphonaptera : Pulicidae) and Odontopsyllus quirosi Gil Collado (Siphonaptera : Leptopsyllidae). Mandibular glands are present in 1st and 3rd instar U. tasmanica, but are absent from 2nd and 3rd instar O. quirosi and all larval instars of C. felis. Such glands in flea larvae have not beeb described previously and they appear to be unique to the endoparasitic U. tasmanica larva. Their presence during the 3rd instar as well as in the 1st instar suggests that although it is possible they play a role in the initial penetration of host skin by the newly hatched larva, they are active in secretion also during the time when the larvae are feeding on host tissue within the dermis.     

The activity of platelet activating factor-acetyl hydrolase (PAF-AH) in the salivary glands of Rhodnius prolixus

In this work, we investigated the activity of the platelet activating factor acetyl hydrolase (PAF-AH) in the salivary gland homogenates and saliva of Rhodnius prolixus. PAF-AH activity in the salivary gland homogenates was lower than in the saliva. Preliminary characterization of the enzyme demonstrated that it hydrolyzed the substrate 2-thio-PAF, was detectable just in 1 pair of salivary gland homogenates in 0.5 ml buffer, and was stable under different conditions. PMSF, TPCK, TLCK, pepstatin A and p-BPB all inhibited the PAF-AH activity. Enzyme specific activity in salivary gland homogenates diminished immediately after feeding of 5th-instar larvae, and increased before feeding by adult insects. 2-Thio-PAF induced platelet-aggregation that was inhibited by previous incubation of the substrate with salivary gland homogenates or saliva. The relevance of PAF-AH for providing Rhodnius with a feeding mechanism for facilitating the sucking of a high volume of blood meal in a short period is discussed.     

Inhibitory effects of starvation on prothoracic gland cell DNA synthesis during the last larval instar of the silkworm, Bombyx mori

Activation of DNA synthesis in prothoracic gland cells of the silkworm, Bombyx mori, during the middle stages of the last larval instar appears to be nutrition dependent, with starvation on day 3 of the last larval instar inhibiting its dramatic increase. The possible cellular mechanism causing the inhibition of DNA synthesis owing to starvation was further examined by determining changes in the growth-promoting activity of the hemolymph and the responsiveness of gland cells to starvation. The results showed that on starvation, the activity of the growth-promoting factor in the hemolymph did not greatly decrease until 2 days after starvation had begun. However, the dramatic increase in the responsiveness of gland cells (the ability to synthesize DNA when exposed to the hemolymph growth factor), which normally occurs on day 4 of the last instar, was not observed when starvation was begun on day 3. A dramatic increase in gland cell size was observed in control larvae during later stages of the last larval instar. However, with starvation beginning on day 3, gland cell size was maintained at lower levels compared with those of control larvae, indicating that the inhibition of DNA synthesis is indeed related to the inhibition of cell size. From these results, it was assumed that alterations in growth factor receptors and downstream signaling may be related to the inhibition of DNA synthesis by starvation and that the deficiency in growth-promoting factor signaling may guarantee that the growth of endoreplicative tissues in these larvae ceases.     

Specific repression of a puff in the salivary gland chromosomes of Drosophila virilis after injection of glucosamine

Five hours after the injection of glucosamine into the hemolymph of Drosophila virilis larvae, puff 55E in the salivary gland chromosomes is significantly reduced in its activity. This observation in connection with the circumstances under which the activity of puff 55E decreases during normal development led to the proposition that its activity is involved in mucoprotein synthesis in the salivary glands during the third larval instar. Factors that may regulate the activity of puff 55E are discussed.     

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.     

TnaA,an SP-RING Protein,Interacts with Osa,a Subunit of the Chromatin Remodeling Complex BRAHMA and with the SUMOylation Pathway in Drosophila melanogaster

Tonalli A (TnaA) is a Drosophila melanogaster protein with an XSPRING domain. The XSPRING domain harbors an SP-RING zinc-finger, which is characteristic of proteins with SUMO E3 ligase activity. TnaA is required for homeotic gene expression and is presumably involved in the SUMOylation pathway. Here we analyzed some aspects of the TnaA location in embryo and larval stages and its genetic and biochemical interaction with SUMOylation pathway proteins. We describe that there are at least two TnaA proteins (TnaA₁₃₀ and TnaA₁₂₃) differentially expressed throughout development. We show that TnaA is chromatin-associated at discrete sites on polytene salivary gland chromosomes of third instar larvae and that tna mutant individuals do not survive to adulthood, with most dying as third instar larvae or pupae. The tna mutants that ultimately die as third instar larvae have an extended life span of at least 4 to 15 days as other SUMOylation pathway mutants. We show that TnaA physically interacts with the SUMO E2 conjugating enzyme Ubc9, and with the BRM complex subunit Osa. Furthermore, we show that tna and osa interact genetically with SUMOylation pathway components and individuals carrying mutations for these genes show a phenotype that can be the consequence of misexpression of developmental-related genes.     

Physiological and Biochemical Analysis of Factors in the Female Venom Gland and Larval Salivary Secretions of the Ectoparasitoid Wasp Eupelmus orientalis

The stinging adult female and the biting newly-hatched larva of the solitary ectoparasitoid wasp Eupelmus orientalis can both cause permanent paralysis and stop the development of Callosobruchus maculatus host larvae. These two processes of host envenomation appeared to be independent and complementary in primary parasitism or in hyperparasitism of a distantly related hymenopteran host species. In contrast, the development of larvae as hyperparasites on members of their own species or genus depended completely on the prior injection of female venom. The venoms of the female and the first instar larva had similar effects on the cellular metabolism of the primary hosts. Protein synthesis was blocked in C. maculatus hosts envenomated by a female or a first instar larva of E. orientalis, but the absence of DNA breakdown indicated that these paralysed hosts were alive and quiescent. The venomous secretions injected by adult females and first instar larvae of E. orientalis had distinct electrophoretic profiles. The immunoreactive features of proteins from female venom and larval secretions were also examined. There is evidence for antigenic conservation between some venom proteins of E. orientalis and Apis mellifera. Lastly, the hyaluronidase, phospholipase and lipase activities in the female venom gland and in larval-derived secretions of E. orientalis were assayed. No lipase activity was detected. Phospholipase activity was found in both the female venom and the larval secretions of E. orientalis, whereas hyaluronidase was specific to the female venom.     

The acceleration of apolysis by the brain hormone in the silkworm, Bombyx mori

The rôle of the brain hormone as an activation hormone of the prothoracic glands in silkworm larvae was examined using decapitated larva of the fourth instar. The gland was activated by the brain hormone injected and induced early larval or larval-pupal apolysis.     

THE ENDOPARASITOID Campoletis chlorideae INDUCES A HEMOLYTIC FACTOR IN THE HERBIVOROUS INSECT Helicoverpa armigera

Although lysis of invading organisms is a major innate form of immunity used by invertebrates, it remains unclear whether herbivorous insects have hemolysin or not. To address this general question, we tested the hemolytic (HL) activity of the hemolymph and tissue extracts from various stages of the polyphagous insect Helicoverpa armigera (Hübner) against the erythrocytes from chicken, duck, and rabbit. An HL activity was identified in the hemolymph of H. armigera larvae. Further studies demonstrated that the HL activity is proteinaceous as it was precipitable by deproteinizing agents. Hemolysins were found in Helicoverpa egg, larva, pupa, and adult, but the activity was higher in feeding larvae than in molting or newly molted larvae. Hemolysins were distributed among a variety of larval tissues including salivary gland, fat body, epidermis, midgut, or testes, but the highest activity was found in salivary gland and fat body. Relative to nonparasitized larvae, parasitization of H. armigera larvae by the endoparasitoid Campoletis chlorideae Uchida induced a 3.4‐fold increase in the HL activity in the plasma of parasitized host at day two postparasitization. The present study shows the presence of a parasitoid inducible HL factor in the parasitized insect. The HL activity increased significantly in H. armigera larvae at 12 and 24 h postinjection with Escherichia coli. We infer the HL factor(s) is inducible or due to de novo synthesis, which means that the HL factor(s) is associated with insect immune response by inhibiting or clearance of invading organisms.     

Effects of juvenile hormone on some phase characteristics in the common cutworm, Spodoptera litura

Phase characters of the common cutworm, Spodoptera litura, were influenced by different rearing densities from the 4th-larval instar. Primarily the final feeding period of isolated larvae was 1 day longer than that of crowded larvae causing an increase in pupal weight. Applications of juvenile hormone I, II, or methoprene to crowded larvae caused an increased feeding period similar to that of isolated larvae when the juvenile hormones were applied within 1 day after the last-larval ecdysis. Allatectomy of isolated Spodoptera during the moult to the final-larval instar decreased the duration of the final feeding period to that of intact crowded larvae. These results suggested that one of the characters of phase variation, pupal weight, is influenced by the differences in the regulation and activity of the corpora allata during the last-larval instar. Other characteristics of phase variation such as behaviour (feigned death) and colour were not affected by alteration in juvenile hormone levels after the last larva ecdysis.