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A Plazaola

Publications and source records attributed to A Plazaola.

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Small ampullate glands of Nephila clavipes.

The small ampullate glands of the orb-web spider, Nephila clavipes, have been studied and compared to other of the silk producing glands from this organism. They exhibit the same gross morphological features of the other glands. Electrophoretic analyses show that the gland's luminal contents migrate as a single band, while the contents of the secretory epithelium reveal a step-ladder array of peptides in addition to the full size product. Previous studies from our laboratory identified these peptides as products generated by translational pauses. This alternate mode of translation is typical of fibroin synthesis in all the spider glands thus far studied as well as in those of the silkworm. The correlation of the peptides to the process of fibroin synthesis is shown through experimental evidence in this paper. The gradual ultrastructural changes in Golgi vesicles elicited by the fibroin synthesis stimulus can be seen in this paper. The response to stimulation is of a higher magnitude in these glands than in any of those previously analyzed. These studies show the small ampullate glands are a promising and certainly exploitable model system for studies on the synthesis of tissue-specific protein product and its control. J. Exp. Zool. 286:114-119, 2000.

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A spider tRNA(Ala) requires a far upstream sequence element for expression.

Within the series of timed differential accumulations of small RNAs we have shown to prelude the synthesis of fibroin in the large ampullate glands of Nephila clavipes (Nc), we are currently directing our attention to the alanine tRNAs. This work reports the subcloning of the members of a tRNAAla gene cluster and the optimization of their transcription in a heterologous cell-free system derived from Bombyx mori (Bm) silkglands. Our data show that the heterologous cell-free system supports the faithful and differential transcription of the individual spider alanine tRNA genes. We are thus making use of the extract to characterize the individual genes with respect to flank-contained regulatory elements through cell-free transcription of gene derivatives. The work has been initiated with pNTA3 because of its high transcriptional activity. Interestingly, the transcription of this gene requires a far upstream sequence, an uncommon modality in tRNA genes.

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Stimulation of fibroin synthesis elicits ultrastructural modifications in spider silk secretory cells.

Electron microscopic studies of unstimulated and stimulated spider fibroin glands show that the fibroin synthesis stimulus evokes visible changes in both the endoplasmic reticulum and Golgi apparatus of their secretory epithelium. Gradual increase in distension of the reticulum accompanies the increase of evoked fibroin synthetic activity. The flattened translucent Golgi vesicles, seen in inactive cells, display a gradual increase in size and number, also with time. The stimulation also elicits a gradual transition in the gland's luminal membrane, during which the microvilli on the lining gradually disappear acquiring an electron dense appearance. Correlations of the observed transitions to the gland's increase in rate of elicited synthetic activity are discussed. The parallelisms between the ultrastructural modifications observed in the spider secretory cells with those described in the silkworm glands during their progression through the fifth instar have been stressed.

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