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.     

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.     

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.     

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.     

Juvenile hormone titre and regulation in the cockroach Diploptera punctata

Titres of juvenile hormone (JH) have been determined in both hemolymph and whole body extracts of female Diploptera 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 instar D. punctata. Maximum values of approximately 1500 ng/ml (approximately 6 microM) 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 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 in D. 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.     

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.     

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.     

Juvenile hormone catabolism inManduca sexta: Homologue selectivity of catabolism and identification of a diol-phosphate conjugate as a major end product

We studied time-dependent metabolism of (10R)-[³H] juvenile hormone (JH) III and (10R, 11S)-[³H]JH I injected intoManduca sexta larvae; the hormones are metabolized to polar metabolites, expecially the JH acid-diol, and an unknown. Products were analyzed using a reversed-phase liquid chromatography assay. (10R)-JH III is metabolized much more rapidly than (10R, 11S)-[³H]JH I, whether injected seperately or as a mixture of hormones. The unknown metabolites of JH I and JH III were identified as phosphate conjugates of JH I and JH III diol by tandem mass spectral analysis of isolated samples. The phosphate conjugate of JH I diol is the principle end product of JH I metabolism.     

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.     

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     

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.     

Enhancement of boll weevilAnthonomus grandis Boh. (Coleoptera: Curculionidae) pheromone biosynthesis with JH III

Summary The juvenile hormone JH III, when incorporated at 1.0 ppm in the diet of adult male boll weevils (Anthonomus grandis Boh.), increased the biosynthesis of its 4 pheromone compounds by 3 times. The biosynthesis at lower and higher levels of JH III was less. JH I was not active at any of the concentrations tested.In cooperation with the Miss. Agr. and For. Exp. Sta., Miss. State, MS 39762. Mention of a proprietary product does not necessarily imply endorsement of this product by the USDA.     

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.     

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.     

Identification of in vitro release products of corpora allata in female and male loreyi leafworms,Leucania loreyi

The in vitro release of juvenile hormones (JH) by female, and of JH acids (JHA) by male corpora allata (CA) ofLeucania loreyi was identified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Separation and quantification were accomplished by HPLC and GC, respectively. JH II and JH III were the major components released by CA of females. Four JHA analogues were identified as the release products of male CA, i.e. JHA III, Iso-JHA II, JHA II and JHA I. JHA III and Iso-JHA II were reported for the first time as the major release products of CA of adult male Lepidoptera. Iso-JHA II is a new member of the insect juvenile hormone analogue family.     

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.     

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.     

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.     

Action of juvenile hormone on the follicle cells ofRhodnius prolixus: Evidence for a novel regulatory mechanism involving protein kinase C

Summary Juvenile hormone (JH) is known to act on the membranes of the follicle cells ofRhodnius, activating a specific Na⁺, K⁺-ATPase. This leads to a decrease in volume of the cells and the appearance of spaces between them (patency). The addition of an inhibitor of protein kinase C, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), to the medium in vitro inhibits the action of JH on the follicle cells. PDBU (phorbol-12,13-dibutyrate) mimics the action of JH in vitro and the response of the follicle cells to, PDBU is blocked by ouabain. It is concluded that the activation of protein kinase C is a required step in the chain of events leading to activation of the JH-dependent ATPase and set in train by the binding of JH to the membrane.