Juvenile hormone titre and regulation in the cockroachDiploptera punctata

Summary Titres of juvenile hormone (JH) have been determined in both hemolymph and whole body extracts of femaleDiploptera punctata during the first gonotrophic cycle using a method employing gas chromatography/mass spectrometry for qualitative and quantitative analysis. JH III is the sole JH found in both adult and last instarD. punctata. Maximum values of 1500 ng/ml (6M) were observed at the middle of the gonotrophic cycle, when basal oocyte growth rate was greatest. Changes in rates of JH release in vitro by corpora allata paralleled closely the changes in JH titre, suggesting that biosynthesis is a major regulator of titre. JH levels per animal were calculated from observed JH titres, and at certain time points in the gonotrophic cycle JH levels obtained from analysis of whole bodies were significantly greater than those predicted from hemolymph titres. These results suggest the existence of a nonhemolymph JH pool inD. punctata. Decay in JH titre after allatectomy of 5 day females has also been studied. Following a rapid initial decline, the rate of decay slowed appreciably 4 h post-operation. Thus, use of a first-order rate constant to estimate half-life of JH significantly underestimated the longevity of the hormone. The apparent persistence of JH following allatectomy may be due to the existence of a nonhemolymph JH pool.     

Forced synthesis of trace amounts of juvenile hormone II from propionate by corpora allata of a juvenile hormone III-producing insect

Corpora allata of the cockroach Diploptera punctata normally synthesize only the isoprenoid juvenile hormone III (JH III). Only under extreme in vitro conditions (absence of carbon sources other than propionate) do they produce trace amounts of the homoisoprenoid JH II in addition to JH III. The specificity of the in vitro synthesis of JH III by D. punctata is thus consistent with the observed lack of homoisoprenoid JHs in this insect.     

Changes in binding of JH-III in hemolymph of adult femaleLocusta migratoria

Summary Hemolymph from adult femaleLocusta migratoria migratorioides was analyzed for binding of juvenile hormone III (JH-III) after allatectomy and transection of thenervus corporis allati 1 (NCA-I). These operations did not affect the apparent dissociation constant of the binding (Kd=3.3 10^–8 M). The concentration of binding sites exhibited fluctuations in relation to age and type of operation: an increased concentration of binding sites in females with disconnected corpora allata and a decreased concentration in allatectomized females. The changes in concentration of binding sites was not due to differences in water content or hemolymph volume in operated animals. The hemolymph protein concentration was reduced after NCA-I transection and even more after allatectomy. However, variations in protein concentration did not correlate with changes in concentration of JH-III binding sites. The changes in binding site concentration were related to changes in JH-titer.     

Tissue distribution of juvenile hormone hydrolytic activity inGalleria mellonella larvae

Summary Juvenile hormone (JH) hydrolytic activity was determined in different tissues of day-4 last instar larva ofGalleria mellonella. Midgut, gonad, imaginal wing discs and fat body contain higher JH hydrolytic activity than hemolymph, while silk gland and body wall have lower activity. JH esterase activity in imaginal wing discs exhibits a pattern of age-related changes different from that of the hemolymph.We acknowledge the support of this research by the National Institutes of Health (GM 22429) and the Johnson Wax Foundation. Address to which reprint requests may be sent.     

Juvenile hormone I is the principal juvenile hormone in a hemipteran insect,Riptortus clavatus

Juvenile hormone I (JH I) was identified by combined gas chromatography/mass spectrometry as the predominant JH in the hemolymph of female adults of the bean bug,Riptortus clavatus (Thunberg) (Hemiptera: Alydidae). Among JH I, II, and III, JH I was the most effective hormone for inducing the synthesis of yolk proteins in diapause adults.     

The transfer of juvenile hormone from male to female during mating in the Cecropia silkmoth

Summary The juvenile hormone (JH) stored in the accessory sex glands (ASG) of adult maleHyalophora cecropia (L.) originates both from sequestration of circulating hormone and from JH synthesized de novo in the ASG from JH acid taken up from the hemolymph. The secretions present in the lumina of the ASG contain most of the accumulated JH. During mating, endogenous JH, labeled biosynthetically via injected [³H-methyl]-methionine, is transferred along with the other seminal material to the bursa copulatrix of the female. The physiological significance of the JH transfer remains unknown.Research was supported by a grant from the National Science Foundation (PCM72-01892) and Organized Research Funds from Texas A&M University.This work was done as partial fulfillment for the degree of Doctor of Philosophy.     

The alteration of juvenile hormone titre inSpodoptera litura (F.) due to a baculovirus infection

Summary The haemolymph juvenile hormone levels ofSpodoptera litura were remarkably low (540 Galleria units (GU)/ml) at the last larval moult as well as prior to pupation (194 GU/ml). During the last intermoult period this was 2600 GU/ml for a 24 h-period. On the other hand, the JH level in the haemolymph of NPV-infected last instar larvae was initially 1740 GU/ml but was maintained at 2400–2600 GU/ml during the next 48 h. Finally, the JH titre fell to 1393 GU/ml, but only prior to death. The failure of the diseased larvae to undergo the larval pupal moult is ascribed to the alteration of the JH titre in the haemolymph.     

Identification of JH III as the princpal juvenile hormone inLocusta migratoria

Summary JH titers in the hemolymph of nymphal and adult femaleLocusta migratoria migratorioides (R. and F.) were determined using a selective mass spectrosc opic detection technique. Only JH III could be found in either stage, with no detectable JH I (or II). Titers observed were 10–1000-fold lower than those found via a recently reported radioimmunoassay procedure.     

Phosphoramidon enhances allatostatin-mediated inhibition of juvenile hormone biosynthesis in the corpora allata of the cockroach, Diploptera punctata.

Use of enkephalinase inhibitor phosphoramidon in the in vitro radiochemical assay for juvenile hormone biosynthesis enhanced allatostatin-mediated inhibition of hormone production by corpora allata of the cockroach, Diploptera punctata. Significant increases in inhibition in day 2 virgin female CA by AST 1 (at 10(-7) M) and AST 4 (10(-8)-10(-7) M) were observed in the presence of phosphoramidon (10(-5) M or greater). No significant increase in inhibition were seen in CA from day 6 mated females with AST 4 (10(-9)-10(-7) M) and phosphoramidon combined. Phosphoramidon alone had no effect on JH biosynthesis. Analysis of allatostatin content of the CA, as determined by ELISA, revealed that addition of phosphoramidon to the medium increased the endogenous allatostatin content in CA of virgin and mated females. The similarity in primary structure between allatostatins and enkephalin-like peptides and their similar distribution makes it probable that phosphoramidon acts by preventing breakdown of allatostatins within the CA.     

Forced synthesis of trace amounts of juvenile hormone II from propionate by corpora allata of a juvenile hormone III-producing insect

Summary Corpora allata of the cockroachDiploptera punctata normally synthesize only the isoprenoid juvenile hormone III (JH III). Only under extreme in vitro conditions (absence of carbon sources other than propionate) do they produce trace amounts of the homoisoprenoid JH II in addition to JH III. The specificity of the in vitro synthesis of JH III byD. punctata is thus consistent with the observed lack of homoisoprenoid JHs in this insect.     

Hormonal levels and protein variations during sexual maturation ofSchistocerca gregaria; effect of rearing temperature

Summary As a result of the decrease of diurnal rearing temperature from 33 to 28°C the following phenomena were induced in adults ofSchistocerca gregaria: a) In females, a delay in the appearance of maximal levels of JH III and ecdysteroids in the hemolymph, a slowing down of oocyte growth, and an accumulation of hemolymph proteins; b) in males, a decrease of hemolymphatic JH III levels without changes in protein levels.     

Head critical period and juvenile hormone titre during the last larval instar of the cockroach,Periplaneta americana

From neck ligation experiments with last instar larvae of the cockroachPeriplaneta americana it was concluded that the head critical period is reached around day 17, which corresponds to 59% of the last larval stage. At the same stage the juvenile hormone III titre in the hemolymph dropped to undectable levels.     

Juvenile hormones inThermobia domestica females: Identification and quantification during biological cycles and after precocene application

Summary JH I and JH III immunoreactive substances were detected in the hemolymph of imaginal females of the primitive insectThermobia domestica. Periodic changes in the levels of these hormones were investigated in correlation with molting and reproductive cycles in inseminated, virgin and precocene-treated females. The presumed influence of JH III on the various phases of vitellogenesis is discussed, also taking into account the periodic changes of the hemolymphatic ecdysteroid levels.     

Autoinhibition of juvenile hormone production. The case of the cockroachBlattella germanica (L.)

In 6-day-old females ofBlattella germanica, the activity of corpora allata (CA) was inhibited in vitro by juvenile hormone III (JH III). Effective doses (281.5 and 375.4 M in the medium) were somewhat higher than (although of the same order of magnitude as) the estimated intraglandular concentration of JH III at this age, and they induced about 45% inhibition of hormonal release and a significant intraglandular accumulation of JH III and methyl farnesoate. The results suggest that autoinhibitory mechanisms operate in the CA to constrain the upper limit of JH III production at the end of the gonadotrophic cycle.     

In vitro biosynthesis of juvenile hormone III by the corpora allata ofCalliphora vomitoria and its role in ovarian maturation and sexual receptivity

Summary JH III is the only JH detected by GLC-MS in medium from in vitro incubations of corpora allata of adult females ofCalliphora vomitoria. When corpora allata were removed from females at various times during the reproductive cycle and the JH III produced by the glands in vitro measured by a JH III radioimmunoassay, an increase in the level of synthesis was found to occur before previtellogenesis (0–24 h). A second increase appeared at the onset of vitellogenesis (72–83 h) and continued until the end of vitellogenesis (96 h) and the occurrence of chorionation (120 h). Since sexual receptivity develops with vitellogenesis, the significantly higher levels of JH III biosynthesis in vitro at this time supports a possible role for JH in the acquisitive of receptivity.     

Specificity of binding of juvenile hormone III to hemolymph proteins ofLeptinotarsa decemlineata andLocusta migratoria

Summary Binding specificity of juvenile hormone (JH) III enantiomers and analogs to hemolymph proteins ofLeptinotarsa decemlineata andLocusta migratoria was investigated by competitive displacement tests. The order of binding affinity was 10R-JH-III>10R, 10S-JH-III10S JH-III> methylfarnesoate for analogs of the epoxide group and diazo-JHA-IV>EFDA for analogs of the methylester. Both the epoxide and ester groups are important for the interaction of JH-III with its binding protein.18 November 1986     

Reversible juvenile hormone inhibition of ecdysteroid and juvenile hormone synthesis by the ring gland of Drosophila melanogaster

Juvenile hormone bisepoxide (JHB3) and juvenile hormone III (JH III) both inhibited the in vitro production of ecdysteroids by ring glands and brain-ring gland complexes from third instar post-feeding larvae of Drosophila melanogaster in a reversible manner, although JHB3 had greater efficacy. The JH III and JHB3 precursor, methyl farnesoate, did not affect ecdysteroid production. The in vitro synthesis of total detectable JH (JHB3 + JH III + methyl farnesoate) by the corpus allatum portion of the isolated ring gland was also inhibited reversibly in the presence of exogenous JHB3 and JH III, but not by methyl farnesoate. These data indicating negative feedback are in agreement with the accepted dogma of endocrine gland regulation.     

Titers of ecdysone, 20-hydroxyecdysone and juvenile hormone III throughout the life cycle of a hemimetabolous insect,the ovoviviparous cockroachNauphoeta cinerea

Summary Titers of ecdysone, 20-hydroxyecdysone and juvenile hormone III were measured in whole body extracts or hemolymph of embryos, first, penultimate and last stadium nymphs, and adult females ofNaupoheta cinerea. We used a gas-chromatography/mass spectrometry method for quantifying juvenile hormone and a radio-immunoassay for ecdysteroid determination. Juvenile hormone III is particularly abundant in the embryonic stage (up to 960 ng/g), at a low level in first and penultimate stadium nymphs (2–10 ng/ml) and almost absent in the last nymphal stadium; in the adult female the juvenile hormone titer rises to 180 ng/ml in hemolymph during rapid oocyte growth. The titers of ecdysone and 20-hydroxyecdysone undergo similar fluctuations in the embryonic and nymphal stages, being highest at the time of cuticle formation in the embryo and a few days before the nymphal and adult molts (around 100–200 ng/ml for exdysone and 2–4 g/ml for 20-hydroxyecdysone).Acknowledgments. We thank Mrs A. Tschan for rearing the cockroaches, Mr M. Kaltenrieder for drawing the graphs, Mr G.C. Jamieson and Mrs C. Reuter for GC/MS analyses. We are also grateful to the Swiss National Science Foundation (grant no. 3.291-0.82 to B. Lanzrein) and the United States National Science Foundation (grant no. PCM 82-08665 to D.A. Schooley) for their financial support.     

Reversible juvenile hormone inhibition of ecdysteroid and juvenile hormone synthesis by the ring gland ofDrosophila melanogaster

Juvenile hormone bisepoxide (JHB₃) and juvenile hormone III (JH III) both inhibited the in vitro production of ecdysteroids by ring glands and brain-ring gland complexes from third instar post-feeding larvae ofDrosophila melanogaster in a reversible manner, although JHB₃ had greater efficacy. The JH III and JHB₃ precursor, methyl farnesoate, did not affect ecdysteroid production. The in vitro synthesis of total detectable JH (JHB₃+JH III+methyl farnesoate) by the corpus allatum portion of the isolated ring gland was also inhibited reversibly in the presence of exogenous JHB₃ and JH III, but not by methyl farnesoate. These data indicating negative feedback are in agreement with the accepted dogma of endocrine gland regulation.