Caste-specific modulation of juvenile hormone III content and ecdysteroid titer in postembryonic development of the stingless bee,Scaptotrigona postica depilis

Summary Juvenile hormone III content and ecdysteroid titer were analyzed for larval and pupal development of the stingless bee,Scaptotrigona postica depilis. Castespecific differences in juvenile hormone III content were detected at three developmental phases: at the transition from the fourth to the fifth larval stadium, in the spinning phase of the fifth larval stadium, and shortly after the imaginal moult. During the fifth larval stadium, juvenile hormone content closely reflects corpora allata activity. Juvenile hormone synthesis may thus be responsible for the elevated hormone titer in spinning-phase queen larvae, a phase of known sensitivity for induction of queen characters by exogenous juvenile hormone. For ecdysteroids, two phases of caste-specific differences were found: in the pre-pupal phase, and shortly after the imaginal moult. In both periods the titer in queens is distinctly higher compared to workers.Abbreviations Im imago 1 day after eclosion - L3, L4, L5 larval instars 3, 4, and 5 - L5F1, L5F2 substages of feeding phase in fifth larval instar - L5S1, L5S2, L5S3 substages of spinning phase in fifth larval instar - PP1, PP2 substages of prepupal phase - Pw white eyed pupa - Pp pink eyed pupa - Pr red eyed pupa - Pd dark eyed pupa - Pdl, Pdm, Pdd dark eyed pupa with progressive tanning of cuticle - RIA radioimmunoassay     

The juvenile hormone titer of Trichoplusia ni and its potential role in embryogenesis of the polyembryonic wasp CopidosomaFloridanum

The hemolymph juvenile hormone (JH) titer of third through fifth stadia Trichoplusia ni parasitized by the polyembryonic parasitoid, Copidosoma floridanum, was measured by radioimmunoassay and compared to the titers of unparasitized larvae. The JH titer of parasitized larvae fluctuated from 28 pg/μl to undetectable levels. Maximum levels of hormone were present at ecdysis to the fourth and fifth stadium, and at the prepupal stage. Qualitatively, similar fluctuations were observed in unparasitized larvae. However, the titers in unparasitized larvae were much lower than those of parasitized larvae in the third and early fourth stadia, and the titer fell to undetectable levels in the fifth stadium 24 h earlier (48 h) than in parasitized larvae (72 h). Preventing the JH titer from falling during the fourth and fifth stadia by topical application of (RS)-methoprene or JH II had a juvenilizing effect on parasitized T. ni, and inhibited C. floridanum embryo morphogenesis. The effect of exogenous methoprene and JH on C. floridanum development depended on timing of application and dosage. Application of 100 pmol per day of methoprene beginning at 2 h of the host fourth stadium, prior to the large drop in the endogenous JH titer, inhibited morphogenesis in the majority of C. floridanum embryos. Application of methoprene at later times of host development did not inhibit morphogenesis although other developmental alterations were observed. The potential significance of host JH and ecdysteroid titers on polyembryonic development are discussed.     

A role of unliganded thyroid hormone receptor in postembryonic development in Xenopus laevis

A fascinating feature of thyroid hormone (T3) receptors (TR) is that they constitutively bind to promoter regions of T3-response genes, providing dual functions. In the presence of T3, TR activates T3-inducible genes, while unliganded TR represses these same genes. Although this dual function model is well demonstrated at the molecular level, few studies have addressed the presence or the role of unliganded TR-induced repression in physiological settings. Here, we analyze the role of unliganded TR in Xenopus laevis development. The total dependence of amphibian metamorphosis upon T3 provides us a valuable opportunity for studying TR function in vivo. First, we designed a dominant negative form of TR-binding corepressor N-CoR (dnN-CoR) consisting of its receptor interacting domain. We confirmed its dominant negative activity by showing that dnN-CoR competes away the binding of endogenous N-CoR to unliganded TR and relieves unliganded TR-induced gene repression in frog oocytes. Next, we overexpressed dnN-CoR in tadpoles through transgenesis and analyzed its effect on gene expression and development. Quantitative RT-PCR revealed significant derepression of T3-response genes in transgenic animals. In addition, transgenic tadpoles developed faster than wild type siblings, with an acceleration of as much as 7 days out of the 30-day experiment. These data thus provide in vivo evidence for the presence and a role of unliganded TR-induced gene repression in physiological settings and strongly support our earlier model that unliganded TR represses T3-response genes in premetamorphic tadpoles to regulate the progress of development.     

Juvenile hormone and regulation of dichotomous spermatogenesis during the larval diapause of the codling moth

Spermatogenesis ceases during diapause and resumes after diapause. A possible causal relationship between the regulation of these phenomena and the fluctuations of the juvenile hormone titre connected with the diapause was examined in the codling moth. Spermatocyte lysis and spermatogenesis arrest are directly and reversibly related to these fluctuations. They are induced in the last-larval instar by sole application of the juvenoid Altosid to the penultimate-instar larvae predetermined not to enter diapause. Conversely, allatectomy of early last-instar larvae predetermined to diapause induces renewal of spermatogenesis and disruption of diapause. However, the continuation of the arrest of the spermatogenesis during diapause is not directly related to the presence of a high titre of juvenile hormone. The arrest extends throughout diapause which may continue for months, although no juvenile hormone could be detected in the species after 35 days of larval diapause.     

The role of cell-cell interactions in postembryonic development of the Caenorhabditis elegans germ line.

This review addresses the role of cell-cell interactions in the development of the Caenorhabditis elegans germ line: specifically, the relative contributions of germ-line-soma interactions versus autonomous processes are considered. Current knowledge of the interacting cell types and the genes essential for various aspects of germ-line development is discussed.     

The role of juvenile hormone and juvenile hormone esterase in wing morph determination in Modicogryllus confirmatus

The role of juvenile hormone (JH) and juvenile hormone esterase (JHE) in regulating wing morph determination was studied in the cricket Modicogryllus confirmatus. JHE activities were significantly higher in nascent long-winged (LW) vs short-winged (SW) crickets during the latter half but not during the first half of the last stadium. The magnitude and direction of the activity differences were similar to those previously documented between wing morphs of the cricket, Gryllus rubens. In contrast, activities of general esterase, an enzyme or group of enzymes with no demonstrated role in regulating the JH titer in insects, showed no or only minor differences between morphs. The magnitude and direction of the JHE activity variation is consistent with a regulatory role for this enzyme in some aspect of wing dimorphism. However, the timing of the differences (exclusively during the last half of the last stadium) argue against a role in regulating wing length development per se. Single or multiple applications of juvenile hormone-III to nascent LW individuals during the first few days of the last stadium significantly redirected development from long to short wings. Multiple applications of acetone, by itself, also increased the production of short-winged adults. For most treatments, all individuals with shortened wings also had undeveloped flight muscles. These data suggest that JH may play a role in wing morph determination in M. confirmatus but that it affects a different aspect of the polymorphism from JHE.     

Modulation of the ecdysteroid-induced cell death by juvenile hormone during pupal wing development of Lepidoptera

Females of the tussock moth Orgyia recens have only vestigial wings, whereas the males have normal wings. We previously found that ecdysteroid induces both apoptotic events and phagocytotic activation in sex-specific and region-specific manners. To investigate whether different responses to ecdysteroid are controlled at the receptor level, we cloned ecdysteroid receptor isoforms, EcR-A and EcR-B1, in O. recens. In both male and female wings, EcR-A signal was detected in the distal region of the bordering lacuna (BL), whereas EcR-B1 signal was detected in the proximal region of the BL. The similar expression patterns of both EcR isoforms suggested that molecules other than EcR should be involved in different ecdysteroid responses between male and female of O. recens. We next tested juvenile hormone (JH) effects on pupal wing morphogenesis in O. recens. Interestingly, both JH and 20E addition induced wing degeneration not only in females but also in males. In addition, higher concentration of JH pre-treatment of the pupal wings of the silkworm, Bombyx mori, also caused wing degeneration under ecdysteroid treatment. These results indicate that JH modulates the ecdysteroid action to induce the cell death on pupal wings, generally in Lepidoptera.     

The role of allatostatic and allatotropic neuropeptides in the regulation of juvenile hormone biosynthesis in Lacanobia oleracea (Lepidoptera: Noctuidae)

In the sphinghid moth Manduca sexta, two allatoactive neuropeptides appear to be responsible for regulating juvenile hormone (JH) production by the corpora allata (CA). These peptides (M. sexta allatostatin, Mas-AS, and M. sexta allatotropin, Mas-AT) respectively inhibit and stimulate in vitro JH biosynthesis by CA in this insect. However, although Mas-AS inhibits CA in both larval and adult insects, Mas-AT is active only in adult M. sexta. The situation in other lepidopteran species is less clear-cut and, although both peptides have been detected (usually by immunologic and/or molecular techniques) in several other moths (including noctuids), their function as regulators of JH production remains uncertain. In the tomato moth Lacanobia oleracea (Lepidoptera: Noctuidae), we have previously demonstrated the occurrence of Mas-AS and/or Mas-AT in extracts of CA, brain and other organs, and have shown that both peptides are present in larval and adult forms. However, in L. oleracea, although Mas-AS inhibits larval and adult CA in vitro, it does so only at relatively high concentrations, and to a maximum of only approximately 70%. By contrast, Mas-AT (which is also present in larval and adult L. oleracea) stimulates larval and adult CA, but is substantially more potent ( approximately 100 fold) than the allatostatin. In this paper we present the results of paired, concurrent measurements (using ELISA) of levels of Mas-AS and Mas-AT in brains, CA and hemolymph (plasma and hemocytes) of L. oleracea at times when there are marked changes in JH titers. We also present data on the in vitro rates of JH biosynthesis by isolated CA, and on hemolymph JH esterase activity measured at the same critical developmental times, and discuss all of these data in relation to the putative allatoregulatory roles of the M. sexta allatotropic and allatostatic neuropeptides in L. oleracea.     

The role of abiotic factors in the pupation of Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) in the soil

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褐飞虱体内保幼激素滴度变化及其与翅型分化的关系

用高压液相色谱及放射化学法,分别测定了褐飞虱Nilaparvata lugens 1～5龄若虫体内的保幼激素滴度与保幼激素酯酶活性变化,并用保幼激素类似物(ZR-515)进行体表点滴加以验证。结果表明,褐飞虱雌、雄4龄若虫期及雄虫的5龄若虫初期,短、长翅型间体内保幼激素滴度差异明显,可以认为该阶段是其翅型分化的关键时期。     

Ultrastructural study of the postembryonic development of the neural lamella of Galleria mellonella L. (Lepidoptera)

Summary The neural lamella encapsulating the brain of the wax moth Galleria mellonella develops from a very thin (80–120 nm) layer in the first larval instar, resembling the basal lamina, to a thick (1–4 m) sheath composed of two zones in the seventh (last) instar. After its breakdown at the time of larval-pupal ecdysis the neural lamella is reconstructed in the pupa, 2–3 days before pupaladult ecdysis.The cells of the perineurium seem to be responsible for the formation of the neural lamella, both in the larva and pupa, even though its ultrastructure differs at these stages.This paper is dedicated to the memory of doc. dr hab. Andrzej B. Dutkowski who proposed this studyPreliminary results of these studies were presented at the XII-th Congress of the Polish Zoological Society, Pozna (Osiska, 1979)The authors previous surname was Dybowska. She wishes to express her gratitude to Prof. Aleksandra Przeecka for valuable assistance in the preparation of this paper. She is also greatly indebted to Joanna Koodziejczyk, M.Sc. for technical aid     

Qualitative and quantitative decline in spermatogenesis of the follicle-stimulating hormone receptor knockout (FORKO) mouse

Sertoli cells express functional receptors for FSH, one of the two pituitary hormones that regulate spermatogenesis in mammals. We recently produced genetic mutant (FORKO) mice that lack FSH receptor, in order to examine the effects on testicular function and fertility. Mutant males exhibited weight loss of testis, epididymis, and seminal vesicle as well as low levels of testosterone. Except for reduced seminiferous tubular diameter, no gross changes were apparent upon histological examination. Analysis of testicular germ cells by flow cytometry revealed a significant increase in the percentage of 2C cells (spermatogonia and non-germ cells) and a significant decrease in the percentage of HC cells (elongated spermatids) of FORKO males. The absolute number of homogenization-resistant elongated spermatids was also significantly reduced in the mutant males. A 2-fold increase in c-kit-positive 2C cells was recorded in the mutant males. Elongated spermatids of FORKO males showed a dramatic increase in propidium iodide binding suggesting reduced nuclear compaction. The increase in size of the sperm head in mutants, as well as susceptibility to dithiothreitol-induced decondensation, suggests the inadequate condensation of sperm chromatin. Sperm chromatin structure assay, a technique that reflects DNA stability, revealed that sperm from FORKO males are susceptible to acid denaturation, indicating the poor quality of sperm. These data allow us to conclude that genetic disruption of FSH receptor signaling in the rodent induces major changes that might contribute to reduced fertility.     

Involvement of a putative allatostatin in regulation of juvenile hormone titer and the larval development in Leptinotarsa decemlineata (Say)

Juvenile hormone III (JH III) plays primary roles in regulation of metamorphosis, reproduction and diapause in Leptinotarsa decemlineata, a notorious defoliator of potato. The neurosecretory cell-borne substance(s) negatively affects the final two steps in JH biosynthesis, catalyzed respectively by an epoxidase CYP15A1 and a juvenile hormone acid methyltransferase (JHAMT). In a few insect species other than L. decemlineata, the inhibitory substance is allatostatin (AS) neuropeptide. In this study, two putative AS genes encoding LdAS-C and LdAS-B precursors were cloned. Both LdAS-C and LdAS-B were expressed in the egg, larvae, pupae and adults, and highly expressed in the brain and the gut. Dietary introduction of double-stranded RNAs (dsRNAs) targeting LdAS-C and LdAS-B successfully knocked down respective target genes. Ingestion during 3 and 6 consecutive days of dsLdAS-C significantly increased the LdJHAMT mRNA levels by 3.8 and 9.9 fold respectively. In contrast, ingestion of dsLdAS-B only slightly increased the LdJHAMT expression level by 1.1 and 1.7 fold. Moreover, after one, two and three days' ingestion of dsLdAS-C, the relative JH levels in the hemolymph of treated larvae were 2.5, 4.2 and 1.9 fold higher than those in control beetles. Furthermore, ingestion of dsLdAS-C and dsLdAS-B significantly affected larval growth and delayed larval development. Thus, we provide a line of experimental evidence in L. decemlineata to support the concept that AS-C acts as an allatostatin and inhibit JH biosynthesis.     

The role of piRNAs in mouse spermatogenesis.

The Argonaute proteins, which are the direct partners of the small RNAs involved in RNA interference mechanisms, can be divided into two subfamilies, the Argonautes and the Piwis. In animals, the Argonaute subfamily binds 21-22 nucleotide small interfering RNAs (siRNAs) and microRNAs (miRNAs), which direct cleavage and translational inhibition of their target RNAs respectively. The partners of the Piwi proteins are 24-30-nucleotide small RNAs called Piwi-interacting RNAs or piRNAs. In Drosophila, Piwi proteins and piRNAs protect the genome of the germline against selfish elements. Recent studies suggest that this function is conserved in mammals.La famille des Argonautes, les partenaires directs des petits ARNs dans les mécanismes d'interférence par l'ARN, se divise en deux sous-groupes : les Argonautes et les Piwis. Chez les animaux, le sous-groupe des Argonautes se lie aux petits ARNs interférents (siARNs) et aux microARNs (miARNs) qui mesurent 21-22 nucléotides et sont responsables du clivage et de l'inhibition traductionnelle des ARNs cibles respectivement. Les protéines Piwis ont pour partenaires de petits ARNs de 24-30 nucléotides appelés Piwi-interacting RNAs ou piARNs. Chez la drosophile, les protéines Piwi et les piARNs protègent le génome de la lignée germinale contre les éléments mobiles. Des analyses récentes suggèrent que cette fonction est conservée chez les mammifères.     

The role of juvenile hormone in competition and cooperation by burying beetles

Few studies have addressed the physiological mechanisms that modulate aggression in insects. In some social insects, there is a correlation of JH and aggression in colony defense and in the establishment of dominance, but only a few studies demonstrate a causal relationship. Burying beetles aggressively defend a breeding resource, a carcass, and juvenile hormone (JH) hemolymph titers increase rapidly upon the discovery of a carcass. In this study, I show that treatment with the JH analog, methoprene, in the absence of a carcass increases the probability of injuries from aggressive interactions, but treatment to one member of a pair of competing Nicrophorus orbicollis females does not increase the probability that she will win control of the resource. In addition, higher JH levels are not associated with greater competitive ability in communally breeding Nicrophorus tomentosus females. Treatment of one female N. tomentosus does not increase her share of the communal brood. Methoprene seems to make a less competitive female more persistent and less willing to concede, which, although maintaining her share of reproduction, results in her exclusion from the brood chamber.