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Gene regulation and evolution in the chorion locus of Bombyx mori. Structural and developmental characterization of four eggshell genes and their flanking DNA regions.

We report on the detailed structural and developmental characterization of four chorion genes and a truncated pseudogene located within a 9.5 X 10(3) base chromosomal segment. These genes belong to the A and B multigene families and, like previously characterized moth chorion genes, are arranged in tightly linked pairs, which are divergently transcribed (A/B.L11 and A/B.L12). On the basis of their high degree of sequence divergence, the A genes define two distinct subfamilies, while the more homologous B genes represent different copies of the same gene type. The A.L11 and B.L11 introns are much longer, in each case because of a single inserted DNA segment that is missing from A.L12 or B.L12. The 2.1 X 10(3) base insertion in A.L11 is the first retrovirus-like transposable element characterized in Bombyx mori. The very short 5' flanking sequences of A/B.L11 and A/B.L12 (277 and 276 base-pairs) are distinct as shown by hybridization but both recur in additional chorion gene pairs, forming two respective classes that are expressed during distinctly different developmental periods. The divergently transcribed genes of each pair, which border the same 5' flanking sequence, are expressed co-ordinately, during the same developmental period. Detailed comparisons of the 5' flanking regions, and of the corresponding region of the Drosophila s15-1 chorion gene, revealed numerous, very short sequence elements that are shared. One such element, T-C-A-C-G-T, is also associated with all five sequenced Drosophila chorion genes. Some elements are repeated in a dyad symmetrical pattern, i.e. are associated with each of the two genes in a pair, while others, including T-C-A-C-G-T, occur only once per 5' flanking region, and, if functionally important, would presumably act bi-directionally on both genes of the pair.

Animals↗

Organochlorines and eggshell thinning in northern gannets (Sula bassanus) from Eastern Canada, 1968-1984.

The northern gannets (Sula bassanus) nesting on Bonaventure Island, Quebec, were studied from 1968 to 1984 in order to measure environmental contaminant levels and their relationship to productivity. Fresh eggs in 1969 contained a mean DDE level of 18.5 mg kg(-1) and had shells which were 17% thinner than pre-1947 samples. Unhatched eggs in 1969 had no outer calcified layer, a mean thickness 20% less than the pre-1947 mean, and mean DDE concentration of 30.6 mg kg(-1). Significantly higher levels of eight contaminants were measured in unhatched than in fresh eggs collected in 1969. DDE and shell thinning were also intercorrelated with nine other contaminants; these data strongly implicate toxic chemicals, particularly DDE, as the main cause for the low breeding success of Bonaventure gannets from 1966 to 1974. Extensive use of DDT to control forest insects around the Gulf of St Lawrence ceased in 1969. Use of DDT, dieldrin and PCBs was restricted in North America during the early 1970s. Residues of DDT, DDD, DDE, PCBs, dieldrin, HCB and chlordane-related compounds in gannet eggs decreased significantly during this study. Heptachlor epoxide remained constant while alpha-HCH appeared to increase. Estimated half-lives ranged from 3.1 years for DDD to 35.4 years for oxychlordane. The breeding success and population of the colony have coincidentally increased.

Journal Article↗

The effects of DDE, PCB and chlordane on the binding of progesterone to its cytoplasmic receptor in the eggshell gland mucosa of birds and the endometrium of mammalian uterus.

1. The specific binding of progesterone to its cytoplasmic receptor in the mucosa of the shell gland from egg-laying ducks, domestic fowls and from the endometrium of the rabbit uterus was compared. The effect of DDE, PCBs and chlordane added in vitro was also studied. 2. The KD value for progesterone with regard to its receptor was about four times lower in the domestic fowl and the rabbit than in the duck. 3. The chlorinated hydrocarbons decreased the binding of progesterone to its receptor. Their potency was higher in the duck than in the domestic fowl. In rabbit uterine mucosa their effects were similar to those observed in the duck. 4. DDE is an inhibitor of calmodulin: other calmodulin inhibitors as calmidazolium and trifluoperazin also reduced the binding of progesterone in similar concentrations. It is suggested that DDE reduced the progesterone binding by inhibiting calmodulin.

Animals↗

Some ESR observations on bone, tooth enamel and eggshell.

ESR measurements have been made on bone and teeth samples excavated from historical sites on three New Zealand offshore islands and on Taumako, one of the Duff Islands (southeastern Solomons). All of the samples have ages ranging from A.D. 1350 to A.D. 1800. The New Zealand offshore island samples show a very weak natural dating signal, characteristic of recent specimens, virtually obscured by a signal from an organic carbon radical. A detailed analysis of results from these samples suggests that the supralinearity hypothesis (Hennig and Grün, 1983) previously proposed is, in fact, an artifact of the analysis. In addition, all the samples from Taumako show an intense dating signal and evidence of electron trap saturation. The implications of these results are discussed.

Animals↗

Non-invasive measurement of oxygen partial pressure, lateral diffusion and chorioallantoic blood flow under the avian eggshell.

We measured P(O2) under the shell of avian eggs indirectly, by sealing 0.05 mL glass tubes to the shell, sealing them with mercury and using an oxygen microelectrode to measure the contained gas that equilibrates with the gas in the shell membranes. This technique requires a smaller area of contact with the shell and a shorter equilibration period than established techniques, and allows measurements at several locations simultaneously and over a long period of time without endangering the embryo. P(O2) under the shell of chicken eggs decreased to 14.3 kPa on the day before hatching (day 19). P(O2) was unstable during late development and differences up to 3.1 kPa occurred transiently on opposite sides of the equator. By waxing the shell around sampling tubes, we estimated Krogh's coefficient for lateral oxygen diffusion in the shell membranes at 1.1 mmol cm(-1) d(-1) kPa(-1), a value about a third of a previous estimate. Sampling of gas under sufficiently large regions of waxed shell allowed indirect measurements of chorioallantoic venous P(O2), without affecting embryonic respiration. Venous P(O2) was 3.8 kPa on day 19. Assuming 14.3 kPa represents arterialized blood leaving the chorioallantois, it became possible to calculate the effective chorioallantoic blood flow rate, which was 3.5 mL min(-1) on day 19.

Animals↗

Bunched sets a boundary for Notch signaling to pattern anterior eggshell structures during Drosophila oogenesis.

Organized boundaries between different cell fates are critical in patterning and organogenesis. In some tissues, long-range signals position a boundary, and local Notch signaling maintains it. How Notch activity is restricted to boundary regions is not well understood. During Drosophila oogenesis, the long-range signals EGF and Dpp regulate expression of bunched (bun), which encodes a homolog of mammalian transcription factors TSC-22 and GILZ. Here, we show that bun establishes a boundary for Notch signaling in the follicle cell epithelium. Notch signaling is active in anterior follicle cells and is required for concurrent follicle cell reorganizations including centripetal migration and operculum formation. bun is required in posterior columnar follicle cells to repress the centripetal migration fate, including gene expression, cell shape changes and accumulation of cytoskeletal components. bun mutant clones adjacent to the centripetally migrating follicle cells showed ectopic Notch responses. bun is necessary, but not sufficient, to down-regulate Serrate protein levels throughout the follicular epithelium. These data indicate that Notch signaling is necessary, but not sufficient, for centripetal migration and that bun regulates the level of Notch stimulation to position the boundary between centripetally migrating and stationary columnar follicle cells.

Animals↗

Development of preferred orientation in the eggshell of the domestic fowl.

Preferred orientation in the shell of the domestic fowl is shown by x-ray diffractometry to develop gradually throughout the shell, beginning immediately after the start of shell deposition and reaching a maximum at the exterior surface. Only 2 out of 20 shells examined exhibited a single preferred orientation: This was one in which the pole of the (001) plane lies parallel to the shell surface normal. The remaining shells had two preferred orientations present simultaneously, one in which the (001) pole is parallel and the other in which the (104) pole is parallel to the surface normal. Previous work has resulted in conclusions that are in conflict with these; they are discussed in relation to the present work.

Animals↗

The eggshell of Drosophila melanogaster: IX. Synthesis and morphogenesis of the innermost chorionic layer.

Synthesis and morphogenesis of the innermost chorionic layer (ICL) was investigated by conventional EM methods, freeze-fracturing, tissue culture in Robb's medium, and EM autoradiography. Both autoradiography and fine structure results have shown that ICL-components are secreted prior to other chorion proteins. Their secretion starts on stage 12a but the first layer of ICL molecules is visible at stage 12b. Its thickness is gradually increased during the next stages, taking first, a bilaminar form along with the inner endochorion. Later, at the end of choriogenesis, ICL is detached from the endochorion and takes its final thickness and configuration, consisting of a 3-dimensional crystal, about 40 nm thick. The isolated ICL in conditions of air water interface is a monolayer crystal 10 nm thick. Studies on chorion mutants showed that the amount of protein secreted by the follicle cells is independent to the process of crystallisation. These data show how a proteinaceous extracellular substance is gradually assembled to form a 3-D crystal and how it can be organised to perform functions such as the physiological resistance of the insect eggs against water loss or water uptake, whenever they are laid on substrates with extreme environmental conditions. These functions are performed by ICL in conjunction with the underlying wax layer.

Animals↗