JH Synthesis Research Articles

Insulin Receptor Substrate-1 (IRS1) Regulates Oogenesis and Vitellogenesis in Propylea japonica by Mediating the FOXO Transcription Factor Expression, Independent of JH and 20E Signaling Pathways.

The insulin receptor substrate (IRS), as the core cytoplasmic adapter protein in the insulin/insulin-like signaling (IIS) pathway, is an important mediator of cellular signaling. However, it is still unknown how IRS crosstalk with hormone signaling regulates insect growth, development, and reproduction. In this study, we demonstrated that knockdown of IRS1 significantly inhibited oogenesis, vitellogenesis, and the development of nurse cells and follicular epithelial cells. In addition, qRT-PCR results showed that FOXO transcription factors significantly responded to silencing of the IRS1 gene. However, IRS1 silencing had no significant effect on the expression of juvenile hormone/20-hydroxyecdysone (JH/20E)-signaling genes, JH synthesis, and degradation enzyme-related genes and the JH/20E titers. Our results suggested that the IIS pathway regulated ovarian development and Vg production through FOXO, independent of JH and 20E signaling pathways. This study revealed the reproductive regulation mechanism in Propylea japonica, which provides a theoretical basis for large-scale expansion of P. japonica as an environment-friendly biological control strategy.

Identification and function of ETH receptor networks in the silkworm Bombyx mori

Insect ecdysis triggering hormones (ETHs) released from endocrine Inka cells act on specific neurons in the central nervous system (CNS) to activate the ecdysis sequence. These primary target neurons express distinct splicing variants of ETH receptor (ETHR-A or ETHR-B). Here, we characterized both ETHR subtypes in the moth Bombyx mori in vitro and mapped spatial and temporal distribution of their expression within the CNS and peripheral organs. In the CNS, we detected non-overlapping expression patterns of each receptor isoform which showed dramatic changes during metamorphosis. Most ETHR-A and a few ETHR-B neurons produce multiple neuropeptides which are downstream signals for the initiation or termination of various phases during the ecdysis sequence. We also described novel roles of different neuropeptides during these processes. Careful examination of peripheral organs revealed ETHRs expression in specific cells of the frontal ganglion (FG), corpora allata (CA), H-organ and Malpighian tubules prior to each ecdysis. These data indicate that PETH and ETH are multifunctional hormones that act via ETHR-A and ETHR-B to control various functions during the entire development—the ecdysis sequence and associated behaviors by the CNS and FG, JH synthesis by the CA, and possible activity of the H-organ and Malpighian tubules.

Hormonal Regulation of Reproductive Diapause That Occurs in the Year-Round Mass Rearing of Bombus terrestris Queens.

Adult reproductive diapause is an adaptive strategy under adverse environments for insects and other arthropod species, including bumblebees, which enables queens to survive through a harsh winter and then build new colonies in the following spring. Little research has been done on the molecular regulatory mechanism of reproductive diapause in Bombus terrestris, which is an important pollinator of wild plants and crops. Our previous research identified the conditions that induced reproductive diapause during the year-round mass rearing of B.terrestris. Here, we performed combined transcriptomics and proteomics analyses of reproductive diapause in B.terrestris during and after diapause at three different ecophysiological phases, diapause, postdiapause, and founder postdiapause. The analyses showed that differentially expressed proteins/genes acted in the citrate cycle, insect hormone biosynthesis, insulin and mTOR signaling pathway. To further understand the mechanisms that regulated the reproductive diapause, genes involved in the regulation of JH synthesis, insulin/TOR signal pathway were determined. The BtRheb, BtTOR, BtVg, and BtJHAMT had lower expression levels in diapause queens. The JH III titer levels and the activities of the metabolic enzymes were significantly up-regulated in postdiapause queens. Also, after the microinjection of insulin-like peptides (ILPs) and JH analogue (JHA), hormones, cold-tolerance metabolites, metabolic enzymes, and reproduction showed significant changes. Together with results from other related research, a model of the regulation of reproductive diapause during the year-round mass rearing of B.terrestris was proposed. This study contributes to a comprehensive insight into the molecular regulatory mechanism of reproductive diapause in eusocial insects.

Methoprene-Induced Genes in Workers of Formosan Subterranean Termites (Coptotermes formosanus Shiraki)

Termites have a distinct polyphenism controlled by concise hormonal and molecular mechanisms. Workers undergo double molts to transform into soldiers (worker–presoldier–soldier). Juvenile hormone analogs, such as methoprene, can induce workers to transform into presoldiers. However, the molecular mechanism underlying the worker-to-presoldier transformation in Coptotermes formosanus Shiraki is still not clear. We sequenced the transcriptome of workers four days after they had fed on methoprene-treated filter paper and control group workers, which fed on acetone-treated filter paper. The transcriptome of C. formosanus was assembled using the de novo assembly method. Expression levels of unigenes in the methoprene-treated group and the control group were compared. The differentially expressed genes were further analyzed by Gene Ontology (GO) term enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Tetrapyrrole binding, oxidoreductase activity, and metal ion binding were the only three enriched GO terms. Juvenile hormone synthesis was the first ranked enriched pathway. Carbohydrate, amino acid, and lipid metabolism pathways were also enriched. These three pathways may be related to fat body development, which is critical for presoldier formation. Our results have demonstrated the significance of JH synthesis pathways, and pathways related to fat body development in the artificial induction of presoldiers.

20-hydroxyecdysone stimulation of juvenile hormone biosynthesis by the mosquito corpora allata

Juvenile hormone III (JH) is synthesized by the corpora allata (CA) and plays a key role in mosquito development and reproduction. JH titer decreases in the last instar larvae allowing pupation and metamorphosis to progress. As the anti-metamorphic role of JH comes to an end, the CA of the late pupa (or pharate adult) becomes again “competent” to synthesize JH, which plays an essential role orchestrating reproductive maturation. 20-hydroxyecdysone (20E) prepares the pupae for ecdysis, and would be an ideal candidate to direct a developmental program in the CA of the pharate adult mosquito. In this study, we provide evidence that 20E acts as an age-linked hormonal signal, directing CA activation in the mosquito pupae. Stimulation of the inactive brain-corpora allata-corpora cardiaca complex (Br-CA-CC) of the early pupa (24 h before adult eclosion or −24 h) in vitro with 20E resulted in a remarkable increase in JH biosynthesis, as well as increase in the activity of juvenile hormone acid methyltransferase (JHAMT). Addition of methyl farnesoate but not farnesoic acid also stimulated JH synthesis by the Br-CA-CC of the −24 h pupae, proving that epoxidase activity is present, but not JHAMT activity. Separation of the CA-CC complex from the brain (denervation) in the −24 h pupae also activated JH synthesis. Our results suggest that an increase in 20E titer might override an inhibitory effect of the brain on JH synthesis, phenocopying denervation. All together these findings provide compelling evidence that 20E acts as a developmental signal that ensures proper reactivation of JH synthesis in the mosquito pupae.

Juvenile Hormone Biosynthesis in Insects: What Is New, What Do We Know, and What Questions Remain?

Our understanding of JH biosynthesis has significantly changed in the last years. In this review I would like to discuss the following topics: (1) the progresses in understanding the JH biosynthesis pathway. Access to genome sequences has facilitated the identification of all the genes encoding biosynthetic enzymes and the completion of comprehensive transcriptional studies, as well as the expression and characterization of recombinant enzymes. Now the existence of different flux directionalites, feed-back loops and pathway branching points in the JH biosynthesis pathways can be explored; (2) the new concepts in the modulation of JH synthesis by allatoregulators. The list of putative JH modulators is increasing. I will discuss their possible role during the different physiological states of the CA; (3) the new theoretical and physiological frameworks for JH synthesis analysis. I will discuss the bases of the flux model for JH biosynthesis. JH plays multiple roles in the control of ovary development in female mosquitoes; therefore, the CA presents different physiological states, where JH synthesis is altered by gating the flux at distinctive points in the pathway; (4) in the final section I will identify new challenges and future directions on JH synthesis research.

Functional characterization of an allatotropin receptor expressed in the corpora allata of mosquitoes

Allatotropin is an insect neuropeptide with pleiotropic actions on a variety of different tissues. In the present work we describe the identification, cloning and functional and molecular characterization of an Aedes aegypti allatotropin receptor (AeATr) and provide a detailed quantitative study of the expression of the AeATr gene in the adult mosquito. Analysis of the tissue distribution of AeATr mRNA in adult female revealed high transcript levels in the nervous system (brain, abdominal, thoracic and ventral ganglia), corpora allata–corpora cardiaca complex and ovary. The receptor is also expressed in heart, hindgut and male testis and accessory glands. Separation of the corpora allata (CA) and corpora cardiaca followed by analysis of gene expression in the isolated glands revealed expression of the AeATr primarily in the CA. In the female CA, the AeATr mRNA levels were low in the early pupae, started increasing 6h before adult eclosion and reached a maximum 24h after female emergence. Blood feeding resulted in a decrease in transcript levels. The pattern of changes of AeATr mRNA resembles the changes in JH biosynthesis. Fluorometric Imaging Plate Reader recordings of calcium transients in HEK293 cells expressing the AeATr showed a selective response to A. aegypti allatotropin stimulation in the low nanomolar concentration range. Our studies suggest that the AeATr play a role in the regulation of JH synthesis in mosquitoes.

An anti-juvenile hormone agent, ethyl 4-(2-benzylhexyloxy)benzoate, inhibits juvenile hormone synthesis through the suppression of the transcription of juvenile hormone biosynthetic enzymes in the corpora allata in Bombyx mori

Ethyl 4-[(S)-2-benzylhexyloxy)]benzoate (KF-13S), derived from ethyl 4-[2-( tert-butylcarbonyloxy)butoxy]benzoate (ETB), has strong anti-juvenile hormone (JH) activity which causes precocious metamorphosis in Bombyx mori, and the mode of action of this compound was studied. Application of KF-13S inhibited JH biosynthesis by the corpora allata (CA) in a reversible manner, and in vitro culture experiments showed that this inhibition was due to the direct action of this compound on the CA. When mRNA expression of the JH biosynthetic enzymes were studied, KF-13S strongly suppressed those of HMG Co-A synthase and HMG Co-A reductase. mRNA levels of other mevalonate enzymes and JH acid O-methyltransferase were also suppressed but were less sensitive to the compound. These studies showed that KF-13S prevents the transcription of many of the JH biosynthetic enzymes so that JH synthesis is suppressed.

Remodeling of the corpora cardiaca and the corpora allata during adult metamorphosis in Bombyx mori: identification of invisible corpora cardiaca by the expression of adipokinetic hormone

It is problematic to identify the corpora cardiaca (CC) of pupal and adult stages of Bombyx mori because of the presence of tissues similar in appearance near the associated corpora allata (CA). To find the CC, we performed in situ hybridization experiments using the adipokinetic hormone II (AKH-II) gene as a probe, as AKH-II expression occurs only in the CC. Shortly after pupation, the CC and CA began to move toward the esophagus and had attached there tightly by 4 days; during this period, the distance between the CC and CA increased from 65 to 507 μm. The CC became flattened during the remodeling period, although the CA stayed globular until adult ecdysis. In situ hybridization analyses, after the CC had been correctly identified during adult development, showed that the mRNA expression of all the mevalonate enzymes and juvenile hormone acid O-methyltransferase involved in JH synthesis occurred only in CA cells, indicating that the fluctuations of enzyme mRNA amounts in CC-CA complexes (Kinjoh et al. in Insect Biochem Mol Biol 37:808–818, 2007) were derived solely from those in the CA.

Molecular and functional characterization of a juvenile hormone acid methyltransferase expressed in the corpora allata of mosquitoes

A juvenile hormone acid methyltransferase (JHAMT) was isolated as an abundant EST in a library of the corpora allata of the adult female mosquito Aedes aegypti. Its full length cDNA encodes a 278-aa protein that has 43% amino acid identity with BmJHAMT, a juvenile hormone acid methyltransferase previously cloned from Bombyx mori. Heterologous expression produced a recombinant protein that metabolizes farnesoic acid (FA) into methyl farnesoate, as well as juvenile hormone acid into juvenile hormone III (JH III) with exquisite stereo specificity. Real time PCR experiments showed that JHAMT mRNA levels are not an unequivocal indicator of JH III synthesis rates; the A. aegypti JHAMT gene, silent in female pupae, was transcriptionally activated just 4–6 h before adult eclosion. Radiochemical methyltransferase assays using active and inactive corpora allata glands (CA) dissected from sugar and blood-fed females respectively, clearly indicated that significant levels of JHAMT enzymatic activity are present when the CA shows very low spontaneous rates of JH III synthesis. Having the last enzymes of the JH synthetic pathway readily available all the time might be critical for the adult female mosquito to sustain rapid dynamic changes in JH III synthesis in response to nutritional changes or peripheral influences, such as mating or feeding. These results suggest that this gene has different roles in the regulation of JH synthesis in pupal and adult female mosquitoes, and support the hypothesis that the rate-limiting steps in JH III synthesis in adult female mosquitoes are located before entrance of FA into the synthetic pathway.

Biochemical and molecular characterization of allatotropin and allatostatin from the Eri silkworm, Samia cynthia ricini

In a previous study, allatotropic and allatostatic activities were observed in brain extract from the Eri silkworm, Samia cynthia ricini ( Samcri) [Li, S., Jiang, R.-J., Cao, M.-X., 2002b. Allatotropic and allatostatic activities in brain extracts of the Eri silkworm, S. cynthia ricini, and the effects of Manduca sexta allatotropin and M. sexta allatostatin on juvenile hormone in vitro. Physiol. Entomol. 27, 322–329]. In the present study, the HPLC purified Samcri-allatotropin (AT) and -allatostatin (AST) factors were shown to have the same retention time as those of M. sexta ( Manse)-AT and -AST, respectively. Moreover, the amino acid sequences of mature Samcri-AT and -AST deduced from their encoding cDNAs are identical to the Manse-AT and -AST amino acid sequences. Both Samcri-AT and -AST genes were expressed in brain, nerve cord, and midgut, with Samcri-AT also detected in gonads and epidermis, suggesting their pleiotropic physiological functions. The expression levels of Samcri-AT and -AST genes correlated well with the allatoregulatory activities during the period of adult emergence indicating the two peptides tightly control JH synthesis, in a contradictive and cooperative manner. Our biochemical and molecular data of Samcri-AT and -AST and other studies demonstrate that these two peptides regulate JH synthesis by corpora allata in Lepidoptera and have pleiotropic physiological effects.

Allatostatin in ovaries, oviducts, and young embryos in the cockroach Diploptera punctata

The quantity and localization of -Phe-Gly-Leu-amide allatostatins (-F-G-L-amide AST) was determined by ELISA and immunohistochemistry in ovaries and oviducts and in pre-dorsal closure embryos. AST in the cytoplasm of basal oocytes gradually increased from 4 to 35 fmol/ovary pair from the start (day 2) to the completion of vitellogenesis (day 6), then rapidly increased to 121 fmol/ovary pair during choriogenesis. In oviducts, AST-immunoreactivity was found in nerves to the muscle layer and in epithelial cells. AST-immunoreactivity in oviduct epithelial cells increased during vitellogenesis. A marked increase in quantity of AST in oviduct tissue between completion of chorion formation and immediately after ovulation appears to result from AST released from oocytes as they travel down the oviducts because AST content of newly ovulated eggs was 40% lower than late stage chorionated oocytes, and these oocytes released AST when incubated in saline. AST in embryos, localized in yolk cells, decreased as embryos approached dorsal closure. That this material in ovaries and embryos is AST was confirmed by its ability to inhibit JH synthesis in vitro and identification by MALDI–TOF mass spectrometry of a peptide with a mass corresponding to that of a Diploptera punctata AST. These findings indicate likely novel functions for ASTs: facilitation of ovulation and utilization of yolk.

1,5-Disubstituted imidazoles inhibit juvenile hormone biosynthesis by the corpora allata of the mosquito Aedesaegypti

We investigated the effect of fifteen 1,5-disubstituted imidazoles (1,5-dis) on juvenile hormone III (JH III) and methyl farnesoate (MF) biosynthesis by the corpora allata (CA) of the mosquito Aedes aegypti in vitro. Four compounds (TH-35, TH-83, TH-62 and TH-28) significantly decreased JH biosynthesis in the CA dissected from 3-day old sugar-fed females. The decrease of JH synthesis was not always associated with increased MF. TH-30 and TH-83 increased MF levels, while TH-85 and TH-61 significantly decreased MF levels. Five compounds (TH-26, TH-60, TH-83, TH-35 and TH-30) significantly inhibited JH biosynthesis in the CA dissected from females 15 h after a blood meal. Four 1,5-dis (TH-30, TH-26, TH-28 and TH-66) caused MF increases in CA from blood-fed females. 1,5-Disubstituted imidazoles had higher inhibitory activity on JH synthesis when substituted at position 5 by a 3-benzyloxyphenyl group and at position 1 by a benzyl group (such as TH-35). Inhibition of JH and MF biosynthesis by TH-35 was age-dependent and influenced by nutritional status; inhibition differed when evaluated in the CA dissected from sugar-fed females at different days after emergence and in the CA dissected from females at different hours after a blood meal. Inhibition was always higher when the CA was more active. The addition of TH-35 significantly reduced the stimulatory effect of Aedes-allatotropin and farnesoic acid on JH synthesis. This is the first report of an inhibitory effect of 1,5-disubstituted imidazoles on JH synthesis in Diptera.

Variation in JH synthesis rate in mature honeybees and its possible role in reprogramming of hypopharyngeal gland function

Variation in JH synthesis rate in mature honeybees and its possible role in reprogramming of hypopharyngeal gland function

A factor causing stable stimulation of juvenile hormone synthesis by Diploptera punctata corpora allata in vitro

Co-incubation of corpora allata (CA) from the cockroach, Diploptera punctata, with ovaries, fat body or muscle but not brain or testis, leads to a substantial increase in juvenile hormone synthesis. Incubation of the glands in medium pre-conditioned with ovaries also stimulates JH synthesis. The ovary was used as a convenient source of stimulatory factor for a detailed analysis of its physiological effects on the CA. The increase in JH synthesis is stable, maintained over 24 h after exposure to the stimulatory factor. Stimulation is dose-dependent, and the corpora allata show an exquisite relationship between sensitivity to this factor and developmental stage. Day 0 and day 1 glands, as well as glands from post-vitellogenic females, are sensitive to stimulation, whereas glands from vitellogenic females are not sensitive. Corpora allata attached to the brain do not respond to the stimulatory factor, and denervation in vivo leads to an increase in JH synthesis by the glands and a loss in sensitivity to the factor. These data suggest that glands from pre- and post-vitellogenic females are inhibited by their nervous connection to the brain. In contrast, glands from vitellogenic females are normally responding to the endogenous stimulatory factor and are thus no longer stimulated in vitro. Co-incubation of CA with allatostatin and conditioned medium still leads to a stimulation of JH synthesis, suggesting that the restraining effect of the nervous connections to the brain is not caused by allatostatin. The CA cell number increases between emergence and day 2, then remains stable until after oviposition. The stimulatory factor accelerates the increase in cell number in young adult females. The results are interpreted as providing evidence for a constitutive change in CA activity caused by a humoral factor produced by various tissues including the ovary, and modulated by nervous connections to the brain.

Juvenile hormone synthesis in relation to corpus allatum development in embryos of the viviparous cockroach Diploptera punctata

Summary Few studies have addressed endocrinology of the corpora allata (CA) in insect embryos. We now report on development and biosynthetic activity of CA in embryos of a viviparous cockroach, Diploptera punctata. When newly-eclosed adult females of D. punctata were mated, they oviposited and gave birth, respectively, about 8 and 73 days later; thus, gestation and corresponding embryogenesis lasted approximately 65 days. Dorsal closure, which coincides with differentiation of the CA, was concluded when embryos were about 13 days old and had completed 20% of embryogenesis. Reverse phase-high performance liquid chromatography revealed that embryonic CA released predominantly juvenile hormone III (JH) in vitro. Furthermore, an in vitro radiochemical assay showed that between day 28 of embryogenesis (43% of embryonic development completed) and hatch rates of JH synthesis rose, plateaued and then fell. When CA activity was increasing or was high, from day 28 to 54 (83% development), mitosis occurred at low and constant rates within embryonic CA, and corpus allatum cell number increased gradually. Between days 56 (86% development) and 60 (92% development), CA activity fell to a low level, rates of mitosis peaked, and corpus allatum cell number rose rapidly. Throughout embryogenesis, CA volume increased in parallel with CA cell number, suggesting that glandular growth was due largely to cell proliferation. Although CA activity and volume changed considerably in embryos, the diameter of corpus allatum cells, as measured from enzymatically dissociated CA, remained surprisingly constant at 11–12 μm on days 32, 46 and 60 (49, 71 and 92% development). Ultrastructural observations confirmed the large size of cells in low-activity CA of 60-day-old embryos and also showed that these cells, like those in highly active CA of 46-day-old embryos, contained abundant cytoplasm, ribosomes, microtubules and mitochondria.

Apparent inhibition of in vitro juvenile hormone biosynthesis by the corpus cardiacum of adult crickets (Gryllus bimaculatus andAcheta domesticus) is due to juvenile hormone esterase

When measuring the in vitro JH III-biosynthesis by corpora allata (CA) from adult female crickets in the presence of corpora cardiaca (CC), the amount of JH III in the medium decreased in a dose dependent manner. The CC of a 4-day-old female Gryllus bimaculatus contain 42 pmol.pair CC-1 Grb-AKH, 0.62 pmol.pair CC-1 octopamine, and a JH-esterase activity of 9.8 pmol JH.h-1.pair CC-1. Comparable values for Acheta domesticus are 21 pmol.pair CC-1 Grb-AKH, 0.53 pmol.pair CC-1 octopamine, and 6.5 pmolJH.h-1.pair CC-1 of JH-esterase activity. Even if the entire octopamine content of the CC were released into the medium, the concentration would be below the 10(-5) M threshold for octopamine inhibition of JH synthesis. An in vitro AKH inhibition of JH III synthesis was observed, but only at a relatively high concentration (10(-5) M). If the entire AKH content (10(-6) M) of the CC were released into the medium, the AKH concentration would approach JH synthesis inhibiting levels. However, the rate of release of AKH in vitro was very low, and, therefore, AKH from the CC could not affect JH synthesis. In contrast, a specific JH-esterase, released by isolated CC into the medium, was sufficiently high in both cricket species to account for the observed decrease in JH III present. OTFP-sulfone (10(-5) M) restored apparent JH synthesis of the CA to the control level. There was no reduction in the amount of JH released when CA were incubated with heat treated CC. The CA themselves contained almost no JH-esterase activity.

Allatotropin released by the brain controls larval molting in Galleria mellonella by affecting juvenile hormone synthesis.

Monoclonal antibodies were raised against a protein fraction which was purified from larval brains of Galleria mellonella and which was shown to stimulate the corpora allata to synthesize juvenile hormone in vivo as well as in vitro. Immunoblot analysis revealed that one polypeptide band of 20 kDa was specifically recognized. Two pairs of median neurosecretory cells of the brain and the cells of the corpus cardiacum were demonstrated to be immunoreactive to the antibodies. Our results sustain the hypothesis that the larval brain directly governs insect larval molting by controlling JH synthesis via an allatrophic factor.

Control of cell proliferation in the corpora allata during the reproductive cycle of the cockroachDiploptera punctata

Unlike in Blettelta gcrmanica and Supella longipalpa, the corpora allata (CAI of Diploptera punctafa exhibited cyclic changes in cell number during the reproductive cycle. In mated females, a wave of DNA synthesis followed by mitosis resulted in a significant increase in CA cell number from about 9,000 cells on day 0 to 12,000 cells at ovulation on day 8. Subsequently, the number of cells per CA underwent a gradual decline to about 10,000 cells by day 64. During this long period of gestation, mitotic activity was undetectable (by colchicine arrest) and pycnotic nuclei were frequently observed by transmission electron microscopy. Just before parturition on day 72 another mitotic wave was detected and CA cell number increased again. The early wave of CA cell proliferation could be postponed by delaying mating or abolished by maintaining females as virgins. Neural disconnection of the CA from the brain mimicked the effect of mating, suggesting that enhanced cell proliferation is permitted by the removal of inhibitory signals from cerebral neurosecretory cells. The proliferative activities after mating were neither abolished by ovariectomy, which suppressed the normal increase in JH synthesis, nor elevated by unilateral allatectomy, which doubled the rates of JH synthesis in the remaining CA. These data corroborate previous results (Szibbo and Tobe, 1981 a; Tobe et al., 1984; Johnson et at., 1993) and suggest that waves of cell proliferation and JH synthesis, though simultaneous, are regulated independently by inhibitory signals from cerebral neurosecretory cells. D 1996 Witey-Liss, Inc.

Control of cell proliferation in the corpora allata during the reproductive cycle of the cockroach Diploptera punctata

Unlike in Blettella germanica and Supella longipalpa, the corpora allata (CA) of Diploptera punctata exhibited cyclic changes in cell number during the reproductive cycle. In mated females, a wave of DNA synthesis followed by mitosis resulted in a significant increase in CA cell number from about 9,000 cells on day 0 to 12,000 cells at ovulation on day 8. Subsequently, the number of cells per CA underwent a gradual decline to about 10,000 cells by day 64. During this long period of gestation, mitotic activity was undetectable (by colchicine arrest) and pycnotic nuclei were frequently observed by transmission electron microscopy. Just before parturition on day 72 another mitotic wave was detected and CA cell number increased again. The early wave of CA cell proliferation could be postponed by delaying mating or abolished by maintaining females as virgins. Neural disconnection of the CA from the brain mimicked the effect of mating, suggesting that enhanced cell proliferation is permitted by the removal of inhibitory signals from cerebral neurosecretory cells. The proliferative activities after mating were neither abolished by ovariectomy, which suppressed the normal increase in JH synthesis, nor elevated by unilateral allalectomy, which doubled the rates of JH synthesis in the remaining CA. These data corroborate previous results (Szibbo and Tobe, 1981a; Tobe et al., 1984; Johnson et al., 1993) and suggest that waves of cell proliferation and JH synthesis, though simultaneous, are regulated independently by inhibitory signals from cerebral neurosecretory cells. © 1996 Wiley-Liss, Inc.