Brain-corpora Cardiaca Research Articles

Knockdown of a putative insulin-like peptide gene LdILP2 in Leptinotarsa decemlineata by RNA interference impairs pupation and adult emergence

Five insulin-like peptide LdILP genes were identified in Leptinotarsa decemlineata. All of them contained three exons and two introns, with three genes tandemly arrayed and well separated from the other two. Phylogenetic analysis revealed that the three LdILPs from three tandemly-arrayed genes grouped with TcILP1, whereas the other two resembled with TcILP2 and TcILP4 from Tribolium castaneum. Thus, the five LdILP genes were provisionally named LdILP1a, LdILP1b, LdILP1c, LdILP2 and LdILP4. LdILP2 was widely expressed in several tissues such as the brain–corpora cardiaca–corpora allata (BR–CC–CA) complex, gut and fat body. In contrast, LdILP1a and LdILP1b were only transcribed in BR–CC–CA, LdILP4 was in ovaries, and LdILP1c was in both BR–CC–CA and ovaries. Ingestion of double-stranded RNAs (dsRNAs) targeting LdILP2 (dsLdILP2-1 and dsLdILP2-2) specifically knocked down LdILP2 and upregulated the transcription of both LdInR and Ld4EBP, indicating insulin/insulin-like growth factor signaling pathway (IIS) was inhibited. Approximately 50% of the LdILP2 RNAi larvae did not normally pupate and about 50% of the LdILP2 RNAi pupae did not emerge. Moreover, silencing LdILP2 reduced the expression of a juvenile hormone (JH) biosynthesis gene, lowered JH titer and disturbed JH signaling. Finally, knocking down LdILP2 inhibited an ecdysteroidogenesis gene, decreased 20-hydroxyecdysone (20E) titer, and repressed the expression of two 20E-response genes LdHR3 and LdFTZ-F1. Thus, the IIS pathway is involved in larval–pupal metamorphosis by modification of both JH and 20E signaling in L. decemlineata.

Characterization of two juvenile hormone epoxide hydrolases by RNA interference in the Colorado potato beetle

In insect, juvenile hormone (JH) titers are tightly regulated in different development stages through synthesis and degradation pathways. During JH degradation, JH epoxide hydrolase (JHEH) converts JH to JH diol, and hydrolyses JH acid to JH acid diol. In this study, two full length LdJHEH cDNAs were cloned from Leptinotarsa decemlineata, and were provisionally designated LdJHEH1 and LdJHEH2. Both mRNAs were detectable in the thoracic muscles, brain–corpora cardiaca–corpora allata complex, foregut, midgut, hindgut, ventral ganglia, Malpighian tubules, fat bodies, epidermis, and hemocytes of the day 2 fourth-instar larvae, and in female ovaries as well as male reproductive organs of the adults. Moreover, both LdJHEH1 and LdJHEH2 were expressed throughout all larval life, with the highest peaks occurring 32h after ecdysis of the final (fourth) instar larvae. Four double-stranded RNAs (dsRNAs) (dsJHEH1-1, dsJHEH1-2, dsJHEH2-1, dsJHEH2-2) respectively targeting LdJHEH1 and LdJHEH2 were constructed and bacterially expressed. Ingestion of dsJHEH1-1, dsJHEH1-2, dsJHEH2-1, dsJHEH2-2, and a mixture of dsJHEH1-1+dsJHEH2-1 successfully knocked down corresponding target gene function, and significantly increased JH titer and upregulated Krüppel homolog 1 (LdKr-h1) mRNA level. Knockdown of either LdJHEH1 or LdJHEH2, or both genes slightly reduced larval weight and delayed larval development, and significantly impaired adult emergence. Therefore, it is suggested that knockdown LdJHEH1 and LdJHEH2 affected JH degradation in the Colorado potato beetle.

Short neuropeptide F (sNPF) is a stage-specific suppressor for juvenile hormone biosynthesis by corpora allata, and a critical factor for the initiation of insect metamorphosis

Molting and metamorphosis are essential events for arthropod development, and juvenile hormone (JH) and its precursors play critical roles for these events. We examined the regulation of JH biosynthesis by the corpora allata (CA) in Bombyx mori, and found that intact brain-corpora cardiaca (CC)–CA complexes produced a smaller amount of JH than that in CC–CA complexes and CA alone throughout the 4th and 5th (last) instar stadium. The smaller amount of synthesis was due to allatostatin-C (AST-C) produced by the brain. The CC synthesized short neuropeptide F (sNPF) that also suppressed the JH synthesis, but only in day 3 4th stadium and after the last larval ecdysis. For the suppression, both peptides prevented the expression of some of the distinct JH biosynthetic enzymes in the mevalonate pathway. Allatotropin (AT) stimulated sNPF expression in the CC of day 1 5th instar stadium, not of day 3 4th; therefore the stage-specific inhibition of JH synthesis by sNPF was partly due to the stimulative action of AT on the sNPF expression besides the stage-specific expression of the sNPF receptors in the CA, the level of which was high in day 2 4th and day 0 5th instar larvae. The cessation of JH biosynthesis in the last instar larvae is a key event to initiate pupal metamorphosis, and both sNPF and AST-C are key factors in shutting down JH synthesis, along with the decline of ecdysone titer and dopamine.

Development of a real-time PCR assay for measurement of yellow protein mRNA transcription in the desert locust Schistocerca gregaria: A basis for isolation of a peptidergic regulatory factor

A major unresolved issue in insect endocrinology concerns the question of whether or not insects have sex hormones. Conclusive evidence in favor of the presence of such hormones awaits the establishment of appropriate bioassays in males. The cuticle of sexually mature males of the desert locust Schistocerca gregaria turns yellow in gregarious conditions only. Neither females nor isolated males ever turn yellow. The yellowing is due to the deposition in the cuticle of a male-specific Yellow Protein (YP), of which the amino acid sequence is known. In this paper, we describe the partial cloning of the cDNA encoding this Yellow Protein. The tissue distribution and temporal expression of the YP-mRNA is studied in detail using RT-PCR. Furthermore, an RT-PCR based bioassay was developed, which may serve as a reliable tool to help identify the hormones controlling the yellowing process. In addition to juvenile hormone, we have shown that a factor present in the brain–corpora cardiaca is involved in the yellow coloration, as injection of an extract induces the expression of YP-mRNA in isolated gregarious males.

Induction of dauer larvae by application of fenoxycarb early in the 5th instar of the silkworm, Bombyx mori

Fenoxycarb application from 0 h until 60 h of the 5th instar of Bombyx mori induced 100% dauer larvae. Between the 60 and 78 h, the ratio of fenoxycarb-induced dauer larvae decreased, and the ratio of supernumerary instar moulting larvae increased. After application of fenoxycarb at the 48 h of the 5th instar, the haemolymph prothoracicotropic hormone (PTTH) titer was higher in fenoxycarb-treated larvae than in control larvae. Furthermore, brain–corpora cardiaca–corpora allata (Br–CC–CA) complexes from fenoxycarb-treated larvae released higher amounts of PTTH in vitro than the Br–CC–CA complexes of control larvae. Prothoracic glands (PGs) of fenoxycarb-treated larvae at the 48 h of the 5th instar exhibited basal and PTTH-stimulated secretory activities similar to that of control PGs until the 72 h of the 5th instar. After that time point, both basal and PTTH-stimulated secretory activity of PGs from fenoxycarb-treated larvae significantly decreased and remained low for the rest of the investigated period. The combined results suggest that the application of fenoxycarb affects the ability of the PGs to be stimulated by PTTH and the induction of dauer larvae in Bombyx mori is not due to inhibition of PTTH release from Br–CC–CA complexes.

Neuroendocrine control of diapause hormone secretion in the silkworm, Bombyx mori

To clarify the control mechanism of diapause hormone (DH) secretion in the silkworm Bombyx mori a series of anatomical and pharmacological experiments were carried out. The arrangement of `diapause' and `non-diapause' eggs in the ovarioles of the moths was determined by the coloration method to estimate the accumulation of 3-hydroxykynurenine in the eggs. The females destined to lay non-diapause eggs (non-diapause producers) had diapause eggs in their ovaries if their subesophageal ganglions (Sg) had been surgically removed at 2 days after larval–pupal ecdysis or later. In contrast when the surgical extirpation extended to the brain and the corpora cardiaca (CC)–corpora allata (CA) complex in addition to the Sg, the non-diapause producers had no diapause eggs. When the Sg was removed from the females destined to lay diapause eggs (diapause-producers), diapause eggs appeared in response to the treatment at 2 days after larval–pupal ecdysis, but the appearance of diapause eggs was delayed by 2 days when the brain–CC–CA complex was included among the organs removed. These observations suggested that DH is produced in Sg and transferred to the CC–CA complex, and that the secretion of DH from the complex is suppressed in non-diapause producers. The pattern of diapause and non-diapause eggs induced by the transection of the subesophageal connective in diapause and non-diapause producers suggested a regenerative and secretory capacity of the neurosecretory cells after the operation. The appearance of diapause eggs in non-diapause producers with transected protocerebrum of the brain confirmed that there was an inhibitory center in the protocerebrum. Changes in parts of the ovarioles containing diapause and non-diapause eggs with time of injection of γ-aminobutyric acid (GABA) and picrotoxin suggested that a GABAergic inhibitory mechanism in DH secretion may be active in non-diapause producers but inactive in diapause producers throughout the pupal stage.

Development changes in juvenile hormone and juvenile hormone acid titers in the hemolymph and in-vitro juvenile hormone synthesis by corpora allata of the silkworm, Bombyx mori

A simple method was developed to quantify hemolymph juvenile hormone (JH) and JH acid in hemolymph extracts from Bombyx mori with an established radioimmunoassay (RIA) for JH I. When various organic solvent extracts of hemolymph were assayed by RIA, levels of non-specific binding of the labeled ligand in the assay were determined to be greater than 50% of the maximum amount of the label bound by the antiserum. When hemolymph was diluted with methanol:water:8.4 N ammonium hydroxide (10:9:1) and extracted with isooctane, non-specific binding was only 50% higher than control levels obtained with the assay buffer alone. The organic phase contained only JH and aqueous phase, JH acid. Consequently, this extraction method was used to prepare samples for RIA and enabled the separate measurement of JH and JH acid in hemolymph. With this method, changes in the hemolymph titers of JH and JH acid were determined from the third instar through early pupal stage of Bombyx mori. Changes in the in vitro secretory activity of corpora allata and brain–corpora cardiaca–corpora allata complexes from fifth instar larvae were also determined by using JH I RIA of the incubation medium.

An allatostatic factor and juvenile hormone synthesis by corpora allata in Locusta migratoria

The hemolymph juvenile hormone (JH) titer in sexually immature female adults of Locusta migratoria (Ibaraki strain, Japan) was lower than in sexually mature females; nevertheless, JH synthetic activity by the corpora allata (CA) in vitro was considerably higher in immature females than in sexually mature females ( Okuda et al., 1996). We carried out experiments to explain this contradiction. The CA activity of sexually immature female adults was very low when the CA were incubated as a complex together with the corpora cardiaca (CC) and brain. When the same complex was assayed after cutting the nerve cord connecting the CC and CA (NCA1), JH synthesis by the CA was enhanced tenfold. When this pair of CA was incubated in fresh medium without the CC and brain, JH synthesis was further increased. Therefore, the higher in-vitro JH production by CA from immature female adults was the result of isolation of the CA from the brain and CC. A methanolic extract of brain–CC complexes contained a factor that inhibited JH synthetic activity by CA in vitro in both immature and mature insects, and this inhibition was reversible. The factor was heat-resistant but lost allatostatic activity after pronase digestion. These results indicate that the allatostatic factor is probably a heat-stable peptide.

Restricted occurrence of Locusta migratoria ovary maturing parsin in the brain-corpora cardiaca complex of various insect species

Ovary maturing parsin (OMP) is a gonadotrophic molecule previously isolated from the neurosecretory lobes of the corpora cardiaca of Locusta migratoria (acridian Orthoptera). A polyclonal antiserum directed against the two biologically active domains of the L. migratoria (Lom) OMP was used to investigate the occurrence of Lom OMP-like substances in brain-corpora cardiaca complexes of other insect species. Using immunohistochemistry, specimens of 40 different insect species belonging to 13 insect orders were tested. The Lom OMP-like substance was strictly limited to specimens of insect species belonging to the Acridae. It occurred in non-basophilic cells of the pars intercerebralis that project to the corpora cardiaca, as in Locusta. Although the antiserum only detected Lom OMP-like material in the Acridae, it is possible that related molecules exist in other insects. The antiserum may be very specific for domains of the Lom OMP molecule that have not been highly conserved during evolution or possibly these domains are not accessible to the antiserum in other insects.

Presence of Bombyxin-(4K-PTTH)-Like Molecules in Neurosecretory Granules of Brain-Corpora Cardiaca Complexes ofLocusta migratoriaDevelopmental Aspects

Summary Using a monoclonal antibody against a synthetic decapeptide corresponding to the N-terminal of the A-chain of bombyxin (Misoguchi et al., 1987) we show that a significant number of neurosecretory cells of the pars in- tercerebralis of Locusta migratoria contain molecules structurally related to this prothoracicotropic neuropeptide isolated from Bombyx. The immunoreactivity, which is restricted to the neurosecretory granules, can be followed through the axon tracts to the nerve endings in the neurohaemal part of the corpora cardiaca. The strongest reactions are obtained with brain-corpora cardiaca complexes from larvae in the second half of their fourth and fifth instars and from adult females initiating ovarian maturation.

A kinetics analysis of brain-mediated 20-hydroxyecdysone stimulation of the corpora allata of Manduca sexta

An in vitro method for investigating 20-hydroxyecdysone regulation of the corpora allata (CA) has been used to assess the kinetics of stimulation of precommitment day 3 fifth (V) larval instar Manduca sexta CA by 20-hydroxyecdysone. The synthesis of juvenile hormone (JH) I and III acids by 20-hydroxyecdysone-stimulated CA incubated as complexes with the brain-corpora cardiaca (Br-CC) increased similarly over time; the synthesis of JH III acid was greater than that of JH I acid. Maximal stimulation of the CA to synthesize both JH acids occurred when the Br-CC-CA were exposed to 20-hydroxyecdysone for 30-60 min. Following stimulation, the elevated rates of JH I and JH III acid synthesis remained unchanged over an 11 h incubation in the absence of the steroid hormone, suggesting that once stimulated by 20-hydroxyecdysone the CA biosynthetic response is persistent. These kinetics data provide insight into the means by which 20-hydroxyecdysone stimulates the CA via the Br-CC.

Regulation of juvenile hormone biosynthesis by 20-hydroxyecdysone during the fourth larval instar of the tobacco hornworm, Manduca sexta

A previous study of the role of 20-hydroxyecdysone in the regulation of juvenile hormone (JH) biosynthesis during pupal commitment in the last (fifth, V) larval instar of Manduca sexta revealed that the increase in the hemolymph edysteroid titer that elicits commitment also stimulates the corpora allata (CA) to synthesize JHs I and III and/or their acids. This stimulation is exerted indirectly via the brain-corpora cardiaca and results in the postcommitment increase in the JH titer that is important for the molt to a pupa. The possibility that a similar form of interendocrine regulation of JH biosynthesis by 20-hydroxyecdysone is involved in the control of larval molting in Manduca was investigated for the molt from the fourth (IV) to the V larval instar. During the IV instar, the increase in the hemolymph ecdysteroid titer and the capacity of the CA to synthesize JHs I and III in vitro correlated temporally in a manner suggestive of CA regulation by 20-hydroxyecdysone. With an in vitro approach, physiological concentrations of 20-hydroxyecdysone (⩾ 1 μg ml −1 hemolymph) were shown to have a dose-dependent trophic effect on CA activity, specifically stimulating JH I synthesis. Since neither basally active nor stimulated CA contained significant amounts of JH I, it appears that 20-hydroxyecdysone acts at the level of biosynthesis rather than at the level of release. The specificity of 20-hydroxyecdysone stimulation of JH I synthesis was demonstrated by the inability of the biologically inactive ecdysteroids 22-isoecdysone and 5α-ecdysone to activate the glands. 20-Hydroxyecdysone's regulation of IV instar larval CA compared with its regulation of the CA during pupal commitment is discussed.

Interendocrine control by 20-hydroxyecdysone of the corpora allata of Manduca sexta

A positive interendocrine control by 20-hydroxyecdysone regulates the corpora allata (CA) and thus the hemolymph titers of the juvenile hormones (JHs) during the fourth and fifth larval stadia of the tobacco hornworm, Manduca sexta. The effect of 20-hydroxyecdysone is exerted via the brain-corpora cardiaca (Br-CC), and the kinetics of stimulation by the hormone suggest that its control of the CA is indirect, possibly involving neural effectors. Co-incubation of precommitment CA with Br-CC resulted in a stimulation of the synthesis of JH I acid equivalent to that achieved by the incubation of brain-corpora cardiaca-corpora allata (Br-CC-CA) with 20-hydroxyedcysone, suggesting that a diffusible regulatory factor may be involved in the stimulation. During pharate pupal development, a close temporal relationship exists between the drop in the ecdysteroid titer and declines in CA activity and the JH hemolymph titer, suggesting that 20-hydroxyecdysone may now inhbit the CA. Incubations of day 6 Br-CC-CA with physiological concentrations of 20-hydroxyecdysone resulted in an apparent dose-dependent inhibition of CA activity. Thus in Manduca, 20-hydroxyecdysone may exert varying stage-specific interendocrine effects on the CA which direct the postembryonic development of this species.

Ecdysiotropic activity in brains and corpora cardiaca of larvae and adults ofLocusta migratoria:in vitro assays

Summary Under in vitro conditions the excised left and right prothoracic glands of a larva of Locusta migratoria produce similar amounts of ecdysone, as monitored by RIA. When brain-corpora cardiaca extracts from last instar larvae of the same species are added to one of the two glands, its ecdysone production is significantly increased over that of the homologous gland. The stimulation is dose dependent. This in vitro system has been used as an assay to probe the presence of ecdysiotropic factors in brain-corpora cardiaca complexes of larvae and adult females of Locusta migratoria. The results show that the apparent titres of ecdysiotropic factors in these extracts fluctuate during larval development and in adult females. These fluctuations are discussed in relation to our present information on the role of brain and corpora cardiaca in the control of ecdysone synthesis in prothoracic glands and follicle cell epithelia in this species.

Brain-mediated 20-hydroxyecdysone regulation of juvenile hormone synthesis by the corpora allata of the tobacco hornworm, Manduca sexta

In the latter part of the last larval instar (Days 6–7) of Manduca sexta an increase in the hemolymph titer of juvenile hormone (JH) occurs which is involved in directing the pharate pupal development elicited by the large increase in the ecdysteroid titer on Days 7–8 of the instar. A time course analysis on Days 3–4 of the instar of both the first hemolymph ecdysteroid titer peak, which is responsible for wandering behavior and pupal commitment, and of the capacity of the corpora allata (CA) to synthesize JH I and III in vitro revealed a temporal correlation between the increasing ecdysteroid titer and a following increase in CA activity. The temporal arrangement of these increases suggested that ecdysteroids, i.e., 20-hydroxyecdysone, may elicit the increase in CA activity which contributes to the second increase in the JH titer during the latter part of the instar. Regulation of the CA by 20-hydroxyecdysone was investigated in vitro by assessing the effect of this ecdysteroid on JH I and III synthesis by CA incubated either as a complex with the brain-corpora cardiaca or as isolated glands. Dose-response analyses revealed that 20-hydroxyecdysone indeed had a stimulatory effect on JH synthesis by the CA, but only when the glands were complexed with the brain-corpora cardiaca. The effective concentration of 20-hydroxyecdysone was the same as the hemolymph concentration of the hormone at this time. Thus it appears that the increase in CA activity between Days 5 and 7 of the last larval instar is elicited in response to the commitment peak of 20-hydroxyecdysone, acting indirectly via the braincorpora cardiaca. These results constitute the first direct demonstration of interhormonal regulation between the ecdysteroids and juvenile hormones in an insect, and suggest that these hormones act to regulate postembryonic development not only at the level of the target tissue but by regulating the synthesis of the hormones by the endocrine glands themselves.

Interaction Between Ecdysteroid, Eclosion Hormone, and Bursicon Titers inManduca sexta

SYNOPSIS. The end of the molting process in the tobacco hornworm includes the rapid digestion of the old cuticle, molting fluid resorption, ecdysis of the old cuticle, and expansion and hardening of the new cuticle. The coordination of these processes is accomplished by three hormones. Each ecdysis during the life of Manduca appears to be triggered by eclosion hormone. Depending on developmental stage, the hormone comes either from the brain-corpora cardiaca complex or from the chain of ventral ganglia. The neural programs triggered by eclosion hormone include a neuroendocrine event, the release of the tanning hormone, bursicon, thereby ensuring that tanning of the new cuticle must follow ecdysis. Ecdysis, itself, appears to be controlled by the ecdysteroid levels since ecdysteroid injections delay ecdysis at physiological concentrations and in a dose dependent fashion. This delay is due to inhibition of eclosion hormone secretion and to the retardation of the terminal phases of the molt including the digestion of the old cuticle and the onset of sensitivity to eclosion hormone. Thus, eclosion hormone secretion and the ecdysis it triggers are coordinated with the end of development because both are influenced by the same endocrine signal—the decline in the ecdysteroid titer.

Experimental evidence for a neuroendocrine control of ecdysone biosynthesis in adult females of Locusta migratoria

The considerable increase in ecdysteroid concentration which occurs in normal Locusta ovaries at the end of each cycle of oöcyte maturation is prevented if the median neurosecretory cells of the pars intercerebralis are cauterized, or if the corpora cardiaca are excised 24 hr before the onset of ecdysone synthesis in normal females. Implantation of additional brain-corpora cardiaca complexes into young vitellogenic females advances the time of ecdysone synthesis by some 12 hr. Oöcyte growth itself is not affected in these different types of experiments. It is inferred from the data of the present study that ecdysone synthesis in the follicle cell epithelium of maturing Locusta ovaries is stimulated by a neurohormone produced in the median neurosecretory cells of the pars intercerebralis and secreted into the blood via the corpora cardiaca.

The embryonic diapause of Aulocara elliotti (Orthoptera, Acrididiae). Histological and morphometric changes during diapause development and following experimental termination with juvenile hormone analogue.

Progressive hypertrophy of fat body, muscles, gonads and pleuropodial glands was observed with lengthening diapause incubation in embryos of Aulocara elliotti. Embryonic hemolymph stained more darkly and lipoid materials accumulated in the fat body and other tissues. Gradual increase in volumes was observed in the corpora cardiaca, corpora allata and ventral head glands (ecdysial glands). Termination of diapause was achieved in embryos treated with juvenile hormone analogue (JHA) (4 ethyl phenyl 6.7 epoxy geranyl ether--87%, Stauffer Chemical) applied directly to the egg undiluted or when eggs were immersed in a 10% solution in methanol. Treated embryos terminating diapause underwent rapid teratological morphogenesis, achieving pigmentation and sclerotization as in definitive embryos without engulfing their yolk. In treated embryos which remained in diapause, unusual paired masses of pigmented cells, resembling the melanotic pseudotumors of Drosophila, were observed ventral and lateral to the stomodeum and are thought to be subesophageal body cells (pericardial cell homologues). In some embryos these cells were enlarged, lysed or their nuclei rhexic and pycnotic. This response suggests these cells are target organs for JHA, or perhaps for ecdysone or its precursor released by JHA treatment. Endocrine glands in treated embryos had significantly larger volumes (.01) than those of controls or untreated embryos. The ventral head glands of treated embryos appeared to be cytoplasmically depleted. It is suggested that JHA triggered the release of stored ecdysone (or its precursor produced in the ventral head gland) at a rate too rapid for normal morphogenetic processes, such as dorsal closure, to accompany epidermal maturation. The response to JHA cannot be interpreted as juvenilization, but rather must be considered to be precocious maturation. These results indicate that: 1. diapause development involves a transfer of nutrient materials from the yolk into the fat body and other embryonic tissues where they are available to meet the demands of post-diapause morphogenesis; 2. that embryonic diapause in A. elliotti is a consequence of endocrine deficiency which probably results ultimately from the lack of neurosecretory stimulus from the brain-corpora cardiaca complex; and 3. that the subesophageal body cells, and perhaps the pericardial cells, play an essential role in the embryonic physiology, perhaps in the production of carrier protein and/or an essential growth regulator.

Neurosecretory system of the brain-corpus cardiacum complex in diapausing Tyria jacobaeae L. (Lepidoptera : Arctiidae)

The neurosecretory elements of the brain-corpora cardiaca complex were examined in early- (ED) and late-diapausing (LD) pupae of Tyria jacobaeae (Lepidoptera), an insect with obligatory diapause. Neurosecretory granules were present in pars intercerebralis (PI) and lateral (LA) cells of the brain, as well as in the axon component of corpora cardiaca (cc). Variations were observed in the shape, density and distribution of the granules. Differences exist between ED and LD brain cells. Axonal pathways linking PI-LA cells and cc were devoid of granules, but exhibited changes at the microtubular level. This is the first time such observations have been made in an obligatory diapause pupa.

Endocrine effects on the biochemical properties of fat body mitochondria from the cockroach, Blaberus discoidalis

The respiratory rates for fat body mitochondria isolated from adult, male Blaberus discoidalis were determined following long-term cardiacectomy (CC −), allatectomy (CA −), and cardiacectomy-allatectomy (CC − + CA −). All results point to ca. 30 per cent decrease in O 2 uptake following CC − with no effect from CA −. This result was reflected by the activities of the electron transport systems: succinate-cytochrome c reductase and cytochrome c oxidase, which were also reduced 30 per cent following CC − + CA −. Oxidative phosphorylation, respiratory control, and the amount of fat body mitochondrial protein were unaffected by CC − + CA −. Gland extracts had no direct effect on O 2 uptake by fat body mitochondria from CC − + CA − animals. CC − at any time after adult ecdysis resulted in a depressed respiratory rate suggesting the glands exert a continuous effect on the fat body mitochondria. These results indicate the brain-corpora cardiaca neurosecretory complex affects the capacity of fat body mitochondria for respiratory and electron transport activity although respiratory control by ADP and the efficiency of oxidative phosphorylation remain normal. The lack of direct stimulus by corpora cardiaca extracts on O 2 uptake suggests the neurosecretory factors act indirectly on mitochondrial electron transport activities via other biochemical systems and over a relatively long time period. The response of insect fat body mitochondria to long-term CC − is compared to the response of vertebrate mitochondria to thyroidectomy and suggests the neurosecretory factor may have an analogous physiological action.