Vitellogenin levels in hemolymph, ovary and hepatopancreas of the freshwater crayfish Cherax quadricarinatus (Decapoda: Parastacidae) during the reproductive cycle

The freshwater crayfish Cherax quadricarinatus is a tropical species of great interest for aquaculture. Vitellogenin (Vg), a lipoprotein precursor of the vitellum accumulated in spawned eggs, can be synthesized in the ovary and/or hepatopancreas of most crustaceans, being the hemolymph the way for transporting Vg throughout the reproductive cycle. Concentration of Vg in hemolymph, ovary and hepatopancreas of Cherax quadricarinatus adult females was measured by means of ELISA, specifically developed after purifying the native Vg. Measurements were made at four periods of the reproductive cycle: pre-reproductive, mid-reproductive, late reproductive and post-reproductive. Besides, both hepatosomatic (HSI) and gonadosomatic (GSI) indexes were determined in each period. Significant variations in Vg levels were detected in both hemolymph and hepatopancreas, being the highest values observed during the mid-reproductive period. Besides, such variations were positively correlated to the HSI. A positive correlation between Vg levels in hepatopancreas and ovary was also seen. These results support previous evidences about the central role of the hepatopancreas as a site of Vg synthesis in the studied species, together with the relevancy of hemolymph for transporting Vg from the hepatopancreas to the ovary. For aquaculture purposes, Vg monitoring in hemolymph could be used as a non-injurious method, to check the reproductive activity of C. quadricarinatus females.

La langosta de agua dulce Cherax quadricarinatus es una especie tropical de gran interés para la acuicultura. Se midió la concentración de vitelogenina (Vg) en hemolinfa, ovario y hepatopáncreas de hembras adultas de esta especie, por medio de ELISA. Las mediciones fueron hechas en los cuatro períodos del ciclo reproductivo: pre-reproductivo, reproductivo medio, reproductivo tardío y post-reproductivo. Se detectaron variaciones significativas en los niveles de Vg tanto en hemolinfa como en hepatopáncreas, se observó el mayor valor durante el período reproductivo medio. Además, tales variaciones se correlacionaron positivamente con el índice hepatosomático. Se observó además una correlación positiva de los niveles de Vg entre hepatopáncreas y ovario. Estos resultados apoyan evidencias previas sobre el papel central del hepatopáncreas como sitio de síntesis de Vg, en esta especie, y también enfatizan la importancia de la hemolinfa para el transporte de la Vg del hepatopáncreas al ovario. Para propósitos de acuicultura, la medición de Vg en hemolinfa podría ser utilizada como un método no lesivo, con el fin de constatar la actividad reproductiva de hembras de C. quadricarinatus.

Histological ultrastructural aspects fat body virgin physogastric queens Melipona quadrifasciata anthidioides lepeletier, 1836 (HYMENOPTERA, APIDAE, MELIPONINI)

Variations in the morphology and biochemical content of insect fat body have been associated withmetabolic activity and the reproductive cycle (synthesis of vitellogenin). In social insects such as bees,the functional traits of fat body also differ between workers and queens. In this work, we used light andtransmission electron microscopy to examine the morphological features of fat body trophocytes of virginand physogastric mated queens of the stingless bee Melipona quadriafasciata anthidioides before and duringvitellogenesis. Virgin queens had few, small fat body cells in which lipid deposits predominate, and showedno evidence of biosynthetic activity or the uptake of exogenous substances. In contrast, the fat body cells ofphysogastric queens were almost completely devoid of lipids, exhibit a well-developed rough endoplasmicreticulum with an obvious intraluminal product, and contained Golgi stacks that release numerous vesicles.These ultrastructural findings were suggestive of proteosynthesis. However, there was no evidence for theaccumulation of synthesized material in the form of secretory granules. We conclude that the trophocytes ofvirgin and physogastric queens differ basically in their switch from a storage role in the former to a syntheticrole in the latter. In addition, the high level of vitellogenesis seen in egg-laying queens suggests that themain material synthesized is vitellogenin.

Oogenesis and egg development in triatomines: a biochemical approach

Em triatomíneos, assim como em outros insetos, o acúmulo de vitelo é um processo no qual um tecido extraovariano, o corpo gorduroso, produz proteínas que são empacotadas no interior de um ovo. A principal proteína, sintetizada pelo corpo gorduroso, que é acumulada no interior de um ovócito, é a vitelogenina. Este processo é também conhecido por vitelogênese. Existem crescentes evidências em triatomíneos, que além do corpo gorduroso, o ovário também produz proteínas de vitelo. A forma como estas proteínas de vitelo entram nos ovócitos será aqui comentada. O vitelo é um material complexo composto por proteínas, lipídeos, carboidratos e outros compostos minoritários que são empacotados de uma maneira organizada no interior dos ovócitos. A fertilização dispara a embriogênese, um processo que culmina com o desenvolvimento do embrião. Durante a embriogênese o vitelo será utilizado para a construção de um novo indivíduo, a ninfa de primeiro estádio. O desafio para a próxima década é entender onde e como estas proteínas de vitelo são utilizadas junto com os seus componentes não protéicos, em compasso com o programa genético do embrião, que comanda a diferenciação celular (fase inicial da embriogênese) e diferenciação do embrião (fase final da embriogênese) no interior do ovo.

Characters that differ between diploid and haploid honey bee (Apis mellifera) drones

Diploid males have long been considered a curiosity contradictory to the haplo-diploid mode of sex determination in the Hymenoptera. In Apis mellifera, 'false' diploid male larvae are eliminated by worker cannibalism immediately after hatching. A 'cannibalism substance' produced by diploid drone larvae to induce worker-assisted suicide has been hypothesized, but it has never been detected. Diploid drones are only removed some hours after hatching. Older larvae are evidently not regarded as 'false males' and instead are regularly nursed by the brood-attending worker bees. As the pheromonal cues presumably are located on the surface of newly hatched bee larvae, we extracted the cuticular secretions and analyzed their chemical composition by gas chromatograph-mass spectrometry (GC-MS) analyses. Larvae were sexed and then reared in vitro for up to three days. The GC-MS pattern that was obtained, with alkanes as the major compounds, was compared between diploid and haploid drone larvae. We also examined some physical parameters of adult drones. There was no difference between diploid and haploid males in their weight at the day of emergence. The diploid adult drones had fewer wing hooks and smaller testes. The sperm DNA content was 0.30 and 0.15 pg per nucleus, giving an exact 2:1 ratio for the gametocytes of diploid and haploid drones, respectively. Vitellogenin was found in the hemolymph of both types of imaginal drones at 5 to 6 days, with a significantly lower titer in the diploids.

Structures and properties of vitellin isolated from Triatominae species

Se realizaron estudios comparativos de los pesos moleculares composicion quimica e identidad inmunológica de la vitelina (VN) de distintas especies de triatominos. Se determinó que la VN es una lipoglicofosfoproteina con 12.76% de hidratos de carbono 12.53% de lipidos y 0.6% de fósforo unido a mas aproteica. La VN nativa muestra por filtración en gel una única banda de 310 KDa. La VN delipidizada muestra por PAGE SDS una banda principal de 175 KDa coincidente con la banda hallada en la vitellogenina (VG). La composición de aminoácidos fue similar en las cuatro especies ensayadas

The effect of pH and metal chelators upon the sedimentation velocity of Odontophrynus americanus vitellogenin

In the tetraploid amphibian Odontophrynus americans the selective precipitation of vitellogenin by Mg2+ from plasma treated with ethylene diaminetetraacetic acid (EDTA) or ethylene bis (oxyethylenenitrilo)-tetraacetic acid (EGTA) is a pH-dependent phenomenon. Utilizing sucrose gradient centrifugation of whole plasma we have shown that under standardconditions (pH 7.0) the estimated apparent sedimentation coefficient of vitellogenin is 17s.At pH 8.0 and in the presence of EDTA or EGTA there is a decrease of the vitellogenin sedimentation coefficient.This behavior is restricted to vitellogenin as othewr plasma proteins show no alteration in their sedimentation coefficient after similar treatment. The treatment with EDTA at pH 8.0 also induces changes in the vitellogenin molecule which can be detected by partial proteolysis with chymotripsin A

The vitellogenin gene family of Aedes aegypti

We have been interested in identifying genes that play a role in reproduction of the mosquito Aedes aegypti. Our interests are currently focused on the vitellogenin genes which in the mosquito are expressed only in the fat body in response to the insect steroid hormone, 20-hydroxyecdysone. Four of the five vitellogenin genes in the genome have been cloned. We have examined the relationships between these genes and find that they form a small gene family exhibiting different levels of relationship.

Vitellin and vitellogenin characterization of Triatoma infestans and related species

La vitelogenina-vitelina (VG-VN) de Triatoma infestans es una glicolipoproteína con un peso molecular de 220 000, es una lipoproteína de alta densidad (1.18-1.2 gm/ml) con poca movilidad electroforética a pH 8.2. Hembras, machos, ninfas y huevos comparten numerosas proteínas pero ninguna de ellas es glicolipoproteica. La proteína común más importante posee un peso molecular de 43 000, es anódica y se encuentra a lo largo de todo el gradiente salino. Machos y hembras poseen una glicolipoproteína con una movilidad electroforética similar a l VG-VN, pero es una proteína de baja densidad. Otra VG-VN, catódica, no glicolipoproteica, se encuentra en huevos y hemolinfa de hembra. La VG-VN principal puede ser aislada por ultracentrifugación en gradiente de BrNa o por cromatografía en DEAE-celulosa. Varias especies dentro del género Triatoma demostraron identidad inmunológica entre sus VG-VN. Las hembras y los huevos de Panstrogylus megistus y las hembras, los huevos y los machos de Rhodnius prolixus poseen identidad inmunológica parcial con las VG-VN de T. infestans

Desarrollo y maduración de los ovocitos en triatominos / Development and maturation of ovocytes in triatominae

Se estudia al desarrollo y maduración de los ovocitos de triatominos, describiéndose la anatomía y fisiología del aparato reproductor femenino de estos insectos, y analizándose resultados y principales investigaciones realizadas sobre la caracterización, identificación y control hormonal de la síntese de vitelogeninas y vitelinas

Avances en la línea entomológica y control integrado / Advances in the line entomologycal and integrated control

Discute los resultados obtenidos en estudios sobre triatominos, realizados en la línea entomológica y de control integrado, del Progrma Nacional de Investigación en Enfermedades Endémicas (Argentina), dentro de las áreas de: bioquímica, toxicología y control de triatominos (metabolismo de lípidos y de hormonas, caracterización de vitelogeninas, toxicodinámica de insecticidas, y, metodologías de control químico); morfología, sistemática, biología y ecología de triatominos y sus enemigos naturales); morfología de diversas estructuras cuticulares vinculadas con órganos sensoriales y estridulatorios, composición de substancias volátiles liberadas por glándulas epidérmicas; y, aspectos del comportamiento de insectos