Hormonal control of diapause in the tick Argas arboreus

During the mid-diapause period of the female tick Argas arboreus, nerve ganglion (G) extract (GE) from nondiapause (ND) females injected into diapause (D) female haemocoel after feeding, but not before feeding, terminated diapause in 50% of the ticks. ND haemolymph (H) failed to terminate diapause. Reciprocal injections of D GE, and H induced diapause in most ND females. During the late diapause season, ND GE, and H terminated diapause in D females and reciprocal injections of D GE, and H failed to induce diapause in ND females. The results suggest that in A. arboreus the facultative diapause results from gonadotropic hormone (GTH) deficiency in the presence of a diapause inducing factor (DIF). We suggest that the DIF is formed under the influence of short photoperiod in unfed females. The DIF interferes with normal release of the GTH formed in ganglion after feeding and also with the effect of GTH in the H by direct inhibition and/or by preventing the target organs (ovary and/or gut cells) from responding to GTH. The DIF is degraded slowly during short photoperiod and rapidly during long photoperiod.     

Argas (Persicargas) arboreus: juvenile hormone analog termination of diapause and oviposition control

The juvenile hormone analog acetaldehyde, 2-(2-ethoxyethoxy) ethyl p-(methylthio) phenyl acetal, topically applied in a concentration of 10 μg/5 μl acetone/female, terminated diapause and induced vitellogenesis and oviposition in fed female Argas (Persicargas) arboreus Kaiser, Hoogstraal, and Kohls (Argasidae). This is the first reported hormonal termination of diapause in argasid ticks. A full bloodmeal and mating were critical factors controlling oviposition in nondiapausing females, and may enhance juvenile hormone activity. Experiments suggest that tick bodies possess sites of hormonal activities similar to those in other arthropods.     

Juvenile hormone III produced in male accessory glands of the longhorned beetle,Apriona germari,is transferred to female ovaries during copulation

We report on juvenile hormone (JH) biosynthesis in vitro by male accessory glands (MAGs) in the longhorned beetle, Aprionona germari, accompanied by the transfer of JH from males to females during copulation. JH was extracted from the MAGs and separated by reversed‐phase high‐performance liquid chromatography. JH III was identified as the major JH by gas chromatography–mass spectrometry. A radiochemical assay and a non‐radioactive method were used to measure the in vitro rate of JH biosynthesis by the MAGs. After 4 h of incubation with ³H‐methionine in the medium, the radioactivity in the MAGs substantially increased. In a separate assay, incubation of the MAGs with non‐radioactive methionine for 4 h resulted in a 39% increase in JH III. Seven‐day‐old males were injected with medium 199 containing ³H–methionine and 24 h later they were mated with virgin females. Hemolymph and the MAGs were collected from the mated males and hemolymph, ovaries and eggs were collected from the mated females for assaying radioactive JH. The radioactivity incorporated into JH in the MAGs was transferred to the females during copulation and later transferred into their eggs. Assayed 1 h after copulation, JH III level in the MAGs decreased 42% and the content of JH III in the male hemolymph did not change, whereas the content of JH III in the female hemolymph and ovaries both increased. © 2010 Wiley Periodicals, Inc.     

Biochemical and physiological studies of certain ticks (Ixodoidea): specific gravity of hemolymph and coxal and gut fluids of Argas (Persicargas) persicus and Argas (Persicargas) arboreus (Argasidae)

Specific gravity (sp. gr.) of cell-free hemolymph and gut and coxal fluids was determined at different states of the gonotrophic cycle (unfed + 15 days, engorgement day before and after coxal fluid emission, engorgement + 1 day, oviposition day, and oviposition completion + 1 day) of female Argas (Persicargas) persicus and A. (P.) aboreus (Argasidae). The patterns of hemolymph and gut fluid sp. gr. change differed from each other during the gonotrophic cycle, but both patterns were similar in the 2 Argas species. Hemolymph sp. gr. decreased to a minimum one day after feeding (1.0085 and 1.0081 for persicus and arboreus, respectively), and increased through oviposition to a maximum on oviposition + 1 day (1.0187 and 1.0221). Minimum gut fluid sp. gr. occurred on engorgement day before coxal fluid emission (1.0565 and 1.0697). Afterward, gut fluid sp. gr. increased to a maximum on engorgement day + 1 for persicus (1.1089) and on oviposition day for arboreus (1.0973), and then decreased during oviposition in both species. In each tested state of each species, the sp. gr. was consistently higher in gut fluid than in hemolymph. In each species, coxal fluid and hemolymph sp. gr. were the same on engorgement day.     

Release of egg development neurosecretory hormone in Aedes aegypti and Aedes taeniorhynchus induced by an ovarian factor

Ovariectomized Aedes aegypti do not synthesize vitellogenin after a blood meal, unless an ovary from a blood-fed donor is implanted. Decapitation, however, prior to implantation inhibits vitellogenin synthesis. A female ovariectomized and decapitated 6 hr after a blood meal, synthesizes vitellogenin if an ovary from a blood-fed donor is implanted. On the other hand, females that are fed on blood and immediately decapitated can not be stimulated to synthesize vitellogenin with implanted ovaries removed from blood-fed donors. These experiments led to the hypothesis that the blood meal stimulates the ovary to secrete a corpus cardiacum stimulating factor, that in turn promotes release of egg development neurosecretory hormone stored in the corpus cardiacum.Injection of 20-hydroxy-ecdysone or ovarian extract prepared from ovaries removed from unfed females does not release egg development neurosecretory hormone. Thus corpus cardiacum stimulating factor is not 20-hydroxy-ecdysone, and ovaries removed from unfed females do not store it.The rate of inactivation of egg development neurosecretory hormone released from the corpus cardiacum after a blood meal was investigated by implanting an ovary into females that were blood fed for various intervals than decapitated and ovariectomized. Seventy per cent of implants grow when the operation is done 18 hr after feeding, and 30% when the operation is done between 18 and 24 hr after feeding, indicating that egg development neurosecretory hormone is stable for the first 18 hr after a blood meal.Aedes taeniorhynchus females ovariectomized 24 hr after adult emergence do not synthesize vitellogenin. When such a female is implanted with an ovary removed from a sugar-fed or blood-fed Aedes aegypti donor vitellogenin synthesis is initiated, and the implant grows. Decapitation prior to implantation inhibit vitellogenin synthesis and implants do not grow. These results indicate that corpus cardiacum stimulating factor is not species specific.     

Regulation of pheromone inhibition in mated females of Choristoneura fumiferana and C. rosaceana

In the spruce budworm, Choristoneura fumiferana, and the obliquebanded leafroller, C. rosaceana, mating significantly depressed pheromone production after 24 h. On subsequent days, the pheromone titre increased slightly in C. fumiferana, but not in C. rosaceana. No pheromonostatic activity was associated with male accessory sex gland (ASG) extracts, 20-hydroxy-ecdysone or hemolymph taken from mated females. However, pheromone production in mated females was not suppressed when the ventral nerve cord (VNC) was transected prior to mating, indicating that an intact VNC is required to permanently switch off pheromone production after mating. As suggested for other moth species, the presence of sperm in the spermatheca probably triggers the release of a signal, via the VNC, to inhibit pheromone production. The fact that in both species the brain-suboesophageal ganglion (Br-SEG) of mated females contains pheromonotropic activity and that their pheromone glands may be stimulated by the synthetic pheromone-biosynthesis-activating-neuropeptide (PBAN) or a brain extract supports the hypothesis that the neural signal prevents the release of PBAN into the hemolymph rather than inhibiting its biosynthesis. Therefore, we speculate that following the depletion of sperm in the spermatheca, the neural signal declines and is less effective in preventing the release of PBAN, thereby stimulating the resumption of pheromone production, as seen in mated C. fumiferana females. In a previous study, mating was shown to induce a significant rise in the juvenile hormone (JH) titre of both Choristoneura female moths, suggesting that post-mating pheromone inhibition may be under hormonal regulation. However, following topical applications or injections of the juvenile hormone analogue (JHA) and JH II into virgins, the pheromone only declined significantly 48 h after treatment in C. rosaceana. This suggests that the significant rise in the hemolymph JH titre after mating in C. rosaceana females plays a role in keeping the pheromone titre consistently low throughout their reproductive life. These findings will be discussed in relation to the different life histories of the two Choristoneura species.     

Hepatopancreas but not ovary is the site of vitellogenin synthesis in female fresh water crab, Oziothelphusa senex senex

The objective of the present study was to explore the site of synthesis of vitellogenin (Vtg) in fresh water edible crab, Oziothelphusa senex senex. Vtg cDNA fragments were isolated from the hepatopancreas of female crabs using RT-PCR method, and the deduced amino acid sequence of O. senex senex showed more than 60% identity with other brachyuran Vtg sequences. RT-PCR analysis showed that Vtg mRNA can be detected only in hepatopancreas of female Oziothelphusa but not in other tissues including eyestalks, Y-organs, mandibular organs, thoracic ganglion, hypodermis and ovary. Antibodies were raised against vitellin purified from the ovary of O. senex senex. Immunoprecipitation analysis revealed the presence of Vtg in the hepatopancreas of vitellogenic stage I females and in the hemolymph, hepatopancreas and ovary extracts from vitellogenic stage II females but absent in hemolymph and hepatopancreas extract of males. These results suggest that Vtg is synthesized only in hepatopancreas but not in the ovaries of O. senex senex. In addition, Vtg synthesized in hepatopancreas is transported to ovary through hemolymph.     

In vivo role of 20-hydroxyecdysone in the regulation of the vitellogenin mRNA and egg development in the American dog tick, Dermacentor variabilis (Say)

Injection of the hormone 20-hydroxyecdysone (20-E) into partially fed (virgin) female adults of the American dog tick, Dermacentor variabilis, while they are attached and feeding on the rabbit host, initiated the expression of the vitellogenin (Vg) gene, and Vg protein secretion and uptake by the ovary. The induction of egg production by 20-E in this bioassay was dose dependent in the range of 1-50 times the concentration normally found in a replete, vitellogenic female. Ticks examined 4 d after the 50 x treatment were still attached to the host, had numerous enlarged vitellin-filled (brown) oocytes in their ovaries, but had not engorged to repletion. The ovaries reached weights similar to those found in untreated, replete (mated) females (pre-oviposition) while solvent-injected controls demonstrated no increase in oocyte size or increase in ovary weight. An increase in the levels of a putative Vg protein was observed in hemolymph samples collected 1, 2 and 3d post-20-E injection but was not observed in the corresponding solvent controls as determined by native PAGE. Analysis of the ecdysteroid-induced protein by tryptic digestion-mass fingerprinting and BLASTP found that the putative Vg had the strongest match to GP80 (U49934), the partial sequence for the vitellogenin protein from Boophilus microplus. A partial Vg cDNA was cloned and sequenced from replete females of D. variabilis with a high similarity to GP80. Using this message as a probe, Northern blots conducted with RNA collected from partially fed, virgin females 1, 2 and 3d post-20-E injection showed upregulation of the Vg mRNA on all 3 days. Controls injected with solvent only showed no Vg mRNA. Injections with juvenile hormone III did not stimulate Vg expression, oocyte growth or full engorgement. These studies indicate that ecdysteroids and not JH can initiate expression of the Vg gene, Vg protein synthesis and release into hemolymph, and Vg uptake into developing oocytes under bioassay conditions mimicking normal feeding on the host.     

Biological activities of juvenile hormone III skipped bisepoxide in last instar nymphs and adults of a stink bug, Plautia stali

Juvenile hormone III skipped bisepoxide (JHSB₃), methyl (2R,3S,10R)-2,3;10,11-bisepoxyfarnesoate was recently determined as a novel juvenile hormone (JH) in a stink bug, Plautia stali. To further confirm the biological function of JHSB₃ in this insect, its juvenilizing, reproduction-stimulating and diapause-terminating activities and the presence in the hemolymph were examined. Topical application of JHSB₃ to last instar nymphs inhibited their metamorphosis in a dose-dependent fashion. In allatectomized and diapausing adults, JHSB₃ application exerted stimulatory effects on the development of ovaries and ectadenia in females and males, respectively. JHSB₃ was detected from the hemolymph of reproductively active females by gas chromatography-mass spectrometry analysis while its titer in the hemolymph collected from diapausing adults was too low to be detected. These results demonstrated that JHSB₃ has biological function as a JH in P. stali. Topical application of JHSB₃, its stereoisomers and 10R-JH III also indicated that compounds with the 2R,3S-configuration were more potent than those with the 2S,3R-configuration and 2,3-double bond.     

OVARIAN ECDYSONE ELICITS RELEASE OF A MYOTROPIC OVULATION HORMONE IN RHODNIUS (INSECTA: HEMIPTERA)

The hemolymph titres of ecdysteroids in both mated and virgin females peak 5 days after a blood meal. Ovariectomy prevents both the peak in ecdysteroids and the release of a myotropic ovulation hormone from the neurosecretory cells of the brain. Injection of ecdysterone into ovariectomized females results in the release of ovulation hormone in the mated, but not the virgin, female.     

Expression of allatostatins in the Mediterranean field cricket, Gryllus bimaculatus de Geer (Ensifera, Gryllidae)

The allatostatin (AST) type A gene of the cricket Gryllus bimaculatus encodes a hormone precursor including at least 14 putative peptides with a common C-terminus Y/FXFGL/Iamide. By RT-PCR we have analyzed the expression of the allatostatin precursor in various tissues of 0-21 days old adult virgin and mated females. In 3-day-old virgin females, the gene is strongly expressed in the brain (oesophageal ganglion), the suboesophageal ganglion and the caecum, but to a lower extent in other parts of the digestive tract (ileum, midgut, colon), and in various other tissues such as the fat body, ovaries and female accessory reproductive glands. In the brain and ovaries of virgin females, the AST expression is rather constant throughout adult life, whereas in brains of mated animals, expression is low until day 7, but increases sharply from day 8 onwards to reach values triple those before day 7. In ovaries of mated animals AST gene expression is also age-dependent, with high expression rates during the first 4 days after imaginable moult, a second but smaller peak from day 15 to 21, and very low values in between. In the fat body of virgin crickets allatostatin expression is high during the first 9 days after ecdysis and declines thereafter, whereas in mated animals two peak values, day 1 and day 6, are observed, and a third peak in older animals.     

Vitellogenin and egg production in the moth,Heliothis virescens

The characteristics of vitellogenin (Vg) and the relationship between Vg production and egg production in the tobacco budworm, Heliothis virescens, were studied. The relationship between Vg production and juvenile hormone (JH) and the impact of mating on Vg and egg production were also investigated. Vg appears in the hemolymph of H. virescens about 6 h after moth eclosion. Vg may be separated into two apoproteins (ApoVg-I and ApoVg-II) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights were calculated to be 156,065 ± 800 for ApoVg-I and 39,887 ± 323 for ApoVg-II. SDS-PAGE analysis revealed that the female hemolymph Vg polypeptides appear to be identical to those from eggs but are absent in male hemolymph. Vg concentration was significantly higher in mated females than in virgin females of the same age at 48 h after emergence. Rates of egg production increased as Vg production increased; rates of egg production in mated females were significantly higher than those of virgin females at 48, 72, 96, and 120 h postemergence. Vg production is dependent on JH, because hemolymph from decapitated females lacked Vg while that of decapitated females treated with synthetic JH had Vg at levels comparable to similarly aged, normal H. virescens females. Hemolymph JH titers in mated females were significantly higher compared with those in virgin females at all sampling periods. The high JH level in mated females may explain the high Vg and egg production in mated H. virescens. Arch. Insect Biochem. Physiol. 34:287–300, 1997. © 1997 Wiley-Liss, Inc.     

The molting hormone activity in Sarcophaga buttata in relation to metamorphosis and reproduction

Ecdysteroid levels were determined by radioimmunoassay, using an antiserum with higher affinity for ecdysterone than for ecdysone. In both sexes, the ecdysteroid level remains low until day 3 after eclosion. When fed liver on day 3, the ecdysteroid titre declines within 4 h in both sexes, stays low in males, but increases again after 12 h in females. The content in non-liver-fed animals, specially females, was significantly higher than if given liver. The level in liver-fed whole animals increased during oogenesis from the onset of follicular growth until the 4B—4C stage, and decreased thereafter.

During embryogenesis, a new high level was found in females, due to the developing embryos which contained a high titre. In isolated ovaries, the ecdysteroid content increased during follicular growth, with a steep increase during the last phase (4C—M and from M to uterine eggs). The ecdysteroid content in hemolymph of 7-day-old females, fed liver or not, was around 10 ng/ml, but in hemolymph of males of the same age no activity was found.

Ovariectomized females contained 12.4 ng/g ecdysteroid. The level during metamorphosis was also determined, mainly to be able to compare our radioimmunoassay results with known bioassay data. Our results support the statement that the ovary cannot be the only site of synthesis of molting hormone in adult S. buttata.     

Fine structure of the hemocytes and nephrocytes of Argas (Persicargas) arboreus (Ixodoidea: Argasidae)

The fine structure of the hemocytes and nephrocytes in Argas (Persicargas) arboreus is described and compared with that of similar cells in other tick species and insects. The hemocytes are of three types: prohemocytes, with a relatively undifferentiated cytoplasm lacking granular inclusions and probably serving as progenitors of the other hemolymph cell types; plasmatocytes, containing abundant mitochondria, cisternae of rough endoplasmic reticulum (RER), and free ribosomes, as well as some small granular inclusions; granulocytes, the predominant cell type in the hemolymph, containing numerous granules of variable electron density and maturity, and pseudopodia-like processes on the cell surface. Plasmatocytes and granulocytes are phagocytic and possibly also have other functions in the tick body. Cells with intermediate features appear to be in a stage of transition from plasmatocyte to granulocyte. Nephrocytes contain vacuoles enclosing fibrillar material, some electrondense granules, and moderate amounts of the active organelles—mitochondria, RER, and ribosomes. The nephrocyte is surrounded by a basal lamina and its plasma membrane infolds to form many deep invaginations coated by a fine fibrillar material. Openings to these invaginations are closed by membranous diaphragms. Coated tubular elements connect the surface invaginations with large coated vesicles, which appear to be specialized for internalization of proteins from the hemolymph. The dense granules may represent an advanced stage of condensation of ingested protein and thus may be lysosomal residual bodies, or they may develop by accumulation of secretory products.     

Ultrastructure of the midgut and blood meal digestion in the adult tickDermacentor variabilis

Digestive cells in the midgut of male and femaleDermacentor variabilis (Say) took up the blood meal in coated vesicles and smooth flask-shaped vesicles, and deposited it in endosomes which were digested via heterophagy. Iron was concentrated in residual bodies.Digestion occurred in three distinct phases in mated females: (1) continuous digestion (initiated by feeding) occurred during slow engorgement; (2) reduced digestion (initiated by mating) occurred in mated females during the period of rapid engorgement; (3) a second continuous digestion phase (initiated by detachment from the host) occurred throughout the post-feeding periods of preoviposition and oviposition.It is proposed that the stem cells in the midguts of unfed females were progenitors of digestive, replacement, and presumed vitellogenic cells in midguts of mated feeding females. Digestive cells were present in all three digestion phases. Only during the first continuous digestion phase did digestive cells fill up with residual bodies, rupture and slough into the lumen, or did whole cells slough into the lumen. During the other two digestion phases no sloughing of digestive cells was observed. At the end of oviposition the digestive cells were filled with residual bodies. Replacement cells were present only during the first continuous-digestion phase. Presumed vitellogenic cells were present only during the reduced-digestion phase and during the second continuous-digestion phase. Stem cells in unfed males developed only into digestive cells in feeding males. Fed males and fed unmated females had only the first continuous-digestion phase. After being hand-detached from the host, unmated 13-day-fed females went through cellular changes associated with the reduced-digestion phase and second continuous-digestion phase of fed mated females, then began ovipositing. Maximum development of the basal labyrinth system and lateral spaces matched the known time of maximum water and ion movement across the midgut epithelia.Spectrophotometric analyses of lumen contents and midgut cells, sampled after detachment from the host, showed that concentrations of protein and hemoglobin at day 1 post-detachment decreased by one-half at the beginning of oviposition, while hematin increased about twofold by the end of oviposition. This supported the idea of the presence of a second continuous-digestion phase.     

Juvenile hormone-like substances can induce vitellogenesis in the tick Ornithodoros moubata (Acarina: Argasidae)

Topical application of different juvenile hormone analogs (JHA) or of a mixture of stereoisomers of insect juvenile hormone (JH) 1 and 3 to fed virgin female Ornithodoros moubata immediately after feeding induced vitellogenesis and egg-laying in up to 70% of treated females. In controls only 13.7% oviposited. The eggs were sterile, with abnormal shape, but their number versus the weight of engorged females was normal or sometimes greater than in mated females. However, preoviposition period was longer than in mated females.

It was more difficult to induce egg-laying by similar topical applications 100 days after feeding of virgin females. A maximum of 58% of ovipositing females was obtained with a very high dosage of JH mixture (500 fig). Injection of this mixture into the females was more potent; 15 to 50 fig induced oviposition in about 60% of the females. The preoviposition period was also longer than in control females.

Our results suggest the presence of a JH-like substance which is involved in the hormonal control of vitellogenesis. However, since natural isomers of JH were much less efficient than isomeric mixtures or JHA, we suppose that the natural tick hormone does not correspond to JH, but rather to a JH-like substance.     

Mating Effect on Sex Pheromone Production of the Oriental tobacco budworm,Helicoverpa assulta

This study was undertaken to clarify the suppression phenomenon of sex pheromone production after mating and its relationship to the physiological mechanism in adult females of Helicoverpa assulta, and determine the mating factor from males causing depletion of sex pheromonc production. Sex pheromone production of H. assulta females was mostly terminated in 3 hours after mating. Mated females maintained with a low titer of sex pheromone until 3 days when it started to increase again, which showed a characteristic of species mating more than once. The mated female again produced pheromone upon injection of pheromone biosynthesis activating neuropeptide (PBAN) or extracts of brain-suboesophageal ganglion complexes (Br-Sg) of mated female, which were shown similar pheromonotropic activities as compared with virgin females. These results indicated that the mating did not inhibit the receptivity of pheromone gland itself and PBAN biosynthesis in suboesophageal ganglion of the mated females. And it seems to support that the depletion of sex pheromone production is responsible for blocking of PBAN release from head. To investigate the mating factor from adult males, when extracts of reproductive organs of male were injected into hemocoel of virgin females evoking depletion of sex pheromone production as shown in mated female. The results suggest that a chemical substance(s) from the male reproductive organs could be responsible for the loss of sex pheromone biosynthesis in H. assulta.     

柑桔爆皮虫卵巢发育动态

本研究利用室内饲养成虫,对柑桔爆皮虫的蛹及正常取食和交配、正常取食但未交配以及正常交配但未取食三种处理的雌成虫的卵巢发育进行了系统观察。结果显示：该虫具有1对卵巢,每侧有5根卵巢管。前两种处理的雌虫卵巢管均能正常发育,成虫寿命在30天左右; 没有取食的雌虫寿命只有7天左右,在其卵巢管的生长区和成熟区均无卵形成。根据卵巢的形状、卵的产生过程、卵巢萼内有无卵粒以及卵黄沉积情况等将卵巢发育程度分为6个级别,即发育初期（0级）、卵黄沉积前期(Ⅰ级)、卵黄沉积期(Ⅱ级)、成熟待产期(Ⅲ级)、产卵盛期(Ⅳ级)和产卵末期(Ⅴ级)。每头雌虫最高怀卵量在140粒左右,根据雌虫怀卵量变化趋势,推测正常取食和交配的柑桔爆皮虫雌虫在出孔后10天左右开始产卵,产卵历期可达22天左右。据此提出该虫卵巢管发育到成熟待产期(Ⅲ级)之前（即羽化出孔后10天之内）为出孔成虫的防治适期。     

The Regulation of yolk polypeptide synthesis in Drosophila ovaries and fat body by 20-hydroxyecdysone and a juvenile hormone analog

In adult female Drosophila melanogaster an increase in the synthesis and secretion of three yolk polypeptides (YPs) occurs during the first 24 hr after eclosion. During organ culture, these same polypeptides are synthesized and secreted into the medium by both fat body and ovaries. Two hormones, 20-hydroxyecdysone (20-HE) and a juvenile hormone analog (ZR-515) stimulate synthesis and secretion of YPs into the hemolymph of isolated female abdomens. The present experiments were undertaken to compare synthesis of YPs in normal females with YP synthesis in preparations deprived of anterior endocrine glands, and to find which hormone stimulates synthesis in the different organs. Separation of hemolymph proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that at eclosion incorporation of [³⁵S]methionine into YP1 and YP2 was low and was barely detectable in YP3. Over the next 24 hr the rate of label incorporation increased for all the YPs. Isolation of female abdomens at eclosion prevented this increase in label incorporation but did not entirely abolish YP synthesis. Application of either ZR-515 or 20-HE to isolated abdomens stimulated up to ninefold label incorporation into three polypeptides which comigrated with YPs from normal vitellogenic females. The response of isolated abdomens to ZR-515 or 20-HE was first detectable between 90 and 135 min after hormone application. The stimulated bands were confirmed to be YPs by a comparison of peptide digests of each of the three labeled polypeptides with those of the yolk polypeptides from intact vitellogenic females. The hypothesis that the two hormones might act on different organs was tested by treating isolated female abdomens with various concentrations of either ZR-515 or 20-HE and then culturing the stimulated organ in vitro with [³⁵S]methionine. The fat body responded to both hormones by synthesizing and secreting into the culture medium polypeptides which comigrated with the YPs found in hemolymph, whereas the ovary produced similar polypeptides only after ZR-515. These secreted polypeptides were confirmed to be YPs by repeating the experiment using organs from heterozygotes for both YP2 and YP3 electrophoretic variants. Such organs synthesized five polypeptides which comigrated with the corresponding yolk polypeptides. These findings are discussed in relation to a hypothesis for the action of the two hormones.     

Effect of mating on the maturation of ovarian follicles in Anopheles atroparvus Van Thiel

Blood fed virgin females of Anopheles atroparvus did not develop ovarian follicles to maturation. In these females, the ovaries were characterized by small follicular size and little yolk deposition. Only the ovaries of blood fed mated females completed development. Thus, the mating permits the complete maturation of the eggs in these mosquitoes.