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Experiments in vitro have shown that 20-hydroxyecdysone (at a concentration of 0.2 and 2 microM and after 12 and 24 hr of incubation) is able to induce the precocious deposition of chorion materials by the follicular epithelium of young oocytes of the cockroach Blattella germanica. Since previous studies had shown that 20-hydroxyecdysone levels in B. germanica ovaries increase with oocyte maturation to reach a peak just before oviposition, we therefore hypothesize that ovarian ecdysteroids trigger Choriogenesis in this species through an autocrine action.

High-performance liquid chromatography with internal surface reversed-phase packing provides an analytical tool for studying the in vitro and in vivo metabolism of A and E destruxins in the haemolymph and various organs of male adults of Locusta migratoria. A slight amount of injected E destruxin is shown to be hydrated into E-diol destruxin in the haemolymph. The rest of the toxin is recovered unchanged in the fat-body, pericardial tissues and Malpighian tubules, and some further E-diol destruxin formation occurs in these organs. Because E-diol destruxin is only weakly toxic, this appears to be a detoxication process.

In vivo, within the first ovarian cycle of B. germanica, dopamine (1 microgram/cockroach) enhanced oocyte growth when injected on day 1 or 2, just before vitellogenesis, whereas it induced the inverse effect on day 6-7, at the end of vitellogenesis. In vitro, dopamine (10(-4) M) stimulates juvenile hormone production by incubated corpora allata from 2-day-old females, and inhibited it on 6-day-old glands. The physiological significance of these effects is discussed.

Ecdysteroids were studied during the molt cycle of the shrimp Palaemon serratus using radioimmunoassay and HPLC. Continuous presence of ecdysteroids was observed in whole extracts, with two hormonal peaks: a minor peak during stage B and a major one during stage D2. The ecdysteroid titers were generally higher in females than in males. The ecdysteroid peak at stage B was more pronounced in the integument (cuticle and epidermis) than in other tissues and, at this time, 20-hydroxyecdysone was the predominant form. The observed 20-hydroxyecdysone peaks in stages B and D2 are correlatable to the previously reported rise in integumental carbonic anhydrase activity.

Hemolymph ecdysteroid titers in fifth instar larvae of Rhodnius prolixus were determined by radioimmunoassay, and their prothoracic glands were excised and examined by electron microscopy. During the last larval instar, the titer of ecdysteroid increased between the head-critical period until Day 13, at which time the peak titer was 3100 pg 20-hydroxyecdysone equivalents/microliter. The activation of secretory cells at the time of the second period of prothoracicotropic hormone release was correlated with the development of major cellular organelles. The smooth endoplasmic reticulum first appeared at the head-critical period and then proliferated in close relation to the increase in ecdysteroid titer until Day 13, after which time it disappeared. Mitochondria expand and develop tubular cristae. They are closely associated with the smooth endoplasmic reticulum. When insects were decapitated, hemolymph ecdysteroid titer remained below 10 pg/microliter and the prothoracic gland cells failed to develop smooth endoplasmic reticulum. We conclude that in the prothoracic gland cells as well as other steroidogenic tissues the smooth endoplasmic reticulum in association with mitochondria is involved in ecdysone biosynthesis.

The venom of the European Ant Monomorium is composed of five alkaloids: three 2.5 transdialkylpyrrolidines and two 2.5 transdialkylpyrrolines. The venom is synthesized by a glandular complex composed of a biramous external filamentous gland, and a single internal gland invaginated into the reservoir. External glands and the proximal two-thirds of the internal gland are composed of glandular units, with one glandular cell and one associated duct cell. Glandular cells contain numerous giant mitochondria, a well-developed smooth endoplasmic reticulum and enigmatic rod-shaped structures. The secretory product is electron-lucent. The distal third of the internal gland is composed of 38-48 large secretory cells issuing directly into the reservoir. Their secretory product presents the same characteristics as the venom accumulated in the reservoir (opacity, affinity to stains and osmium).

Spontaneous cellular differentiation (glandular units appearance with a well-defined duct) is observed in larval integument of Schistocerca cultured in an hormone free medium. Hormonal stimulations modify the expression of this phenomenon, particularly the frequency of new elaborated duct cells belonging to secretory units. The most effective factor is supplied by the prothoracic glands. Complete secretory units can be observed but the glandular cells are in an undifferentiated and unfunctional state. They appear active if the corpora allata act after the prothoracic glands. Corpora allata alone or synthetic juvenile hormones have an inhibitory effect. The addition of alpha-ecdysone permits only cellular divisions, a preliminary and indispensable condition for ulterior differentiation. Between cuticles deposited with beta-ecdysone, new formed ducts take place in the theorical imaginal exuvia. These observations prove that the elaboration and the activity of imaginal glandular units are conditioned by the larval hormonal context: by contrast, sex dimorphism is only a genetic dependant process.

The cellular localization of the vitamin D-dependent calcium-binding protein (CaBP) in the duodenum of rat was studied using indirect immunofluorescence and immunoperoxidase-staining methods. Specific positive reaction product, indicative of the presence of CaBP, was exclusively located within the villous part of the duodenal mucosa. Moreover, CaBP was detected mainly within the supranuclear region of the cytoplasm of absorptive cells and also at the level of their basal laminae. CaBP was not demonstrable either in the nuclei or associated with the brush border membrane of absorptive cells. Also, CaBP was neither detectable in goblet cells nor in sub-epithelial layers. When the specific anti-CaBP antiserum was replaced by nonimmune rabbit serum or when it was preabsorbed on a CaBP-Sepharose conjugate, no positive immunostaining was seen. Together with recent biochemical data our observations agree well with the view that CaBP may act as an intracellular "buffer" by protecting the cell against too high Ca2+ concentrations.

Corpora allata of Locusta migratoria 5th stage larvae synthesize J.H.1, J.H.2 and J.H.3 in vitro. The C.A. of insects of different ages exbit different rates of J.H. synthesis. J.H.1 and J.H.2 synthesis is less than 1 ng/48 h/gland. During the same time the J.H.3 production may be as much as 25.6 ng/gland. J.H. synthetic activity is the same between right and left C.A. The release of J.H. from the C.A. occurs immediately following synthesis. These results are compared with in vivo haemolymphatic J.H. levels.

The yolk sac from normal pregnant rats contains a vitamin D-dependent calcium-binding protein (CaBP). To localize CaBP within the yolk sac, a rabbit antiserum specifically directed against purified rat CaBP, in indirect immunofluorescence and immunoperoxidase, has been used. Using yolk sac sections, positive reactions indicative of the presence of CaBP were found only in the endodermal cell layer. A method for isolation of pure endodermal cells, using EDTA treatment, was developed. On smears of isolated endodermal cells exposed to anti-CaBP antiserum, CaBP was distributed throughout the endodermal cytoplasm; no CaBP was detected in the nucleus. Our results suggest that the endodermal cells, which are functional absorptive cells of the yolk sac, may play a role in vitamin D-mediated processes.

In the gregarious males of Schistocerca gregaria Forsk., imaginal moulting (mitosis, apolysis, new cuticle synthesis, ecdysis) is associated with the differentiation of numerous glandular units. These units, involved in a sexual, excitatory pheromone secretion are at that time composed only of a basal glandular cell and an apical duct cell. Each glandular unit originates from an isogenic group of cells of which the four elements (tetrade) are disposed on two levels. At each level a principal cell and an accessory one may be recognized. The lower accessory, or ciliary, cell shows, at the time of apolysis, both a strong cytoplasmic protrusion and a typical ciliary formation. This formation associated with a diplosome goes through the duct cell and ends up in the exuvial space. It makes a inner mould; arranged around it are epicuticular materials characteristic of the duct wall; then it disappears. The strong cytoplasmic protrusion also retracts thus allowing a glandular reservoir to form. A glandular cell may be recognized at an early stage owing to its R.E.R. development. The upper accessory cell strengthens the duct cell and secretes junctional cuticle between the duct and general cuticle. Accessory cells, after the imaginal moult do not degenerate but acquire epithelial cell characteristics. The duct has a dual origin : the receptive part is secreted by the ciliary cell and the vector part by the duct cell. The organization and stages of morphogenesis of the glandular unit are discussed and compared to those of other apterygote or pterygote insects.