Cellular, ultrastructural and molecular analyses of epidermal cell development in the planarian Schmidtea mediterranea

The epidermis is essential for animal survival, providing both a protective barrier and cellular sensor to external environments. The generally conserved embryonic origin of the epidermis, but the broad morphological and functional diversity of this organ across animals is puzzling. We define the transcriptional regulators underlying epidermal lineage differentiation in the planarian Schmidtea mediterranea, an invertebrate organism that, unlike fruitflies and nematodes, continuously replaces its epidermal cells. We find that Smed-p53, Sox and Pax transcription factors are essential regulators of epidermal homeostasis, and act cooperatively to regulate genes associated with early epidermal precursor cell differentiation, including a tandemly arrayed novel gene family (prog) of secreted proteins. Additionally, we report on the discovery of distinct and previously undescribed secreted organelles whose production is dependent on the transcriptional activity of soxP-3, and which we term Hyman vesicles.

Internalization of Consumed Nanoparticles by a Model Invertebrate Organism

There is littlein vivodata concerning the fate of ingested TiO2nanoparticles (nano-TiO2). We report here experiments aimed at assessing if ingested nano-TiO2accumulates in the digestive gland epithelium or are internalized elsewhere in the body of the terrestrial isopod crustaceans. The animals (Porcellio scaber, Isopoda, Crustacea) fed for 3, 7, or 14 days on food dosed with 100 or 1000 μg nano-TiO2showed no evidence of internalization of Ti measured by microparticle-induced X-ray emission method. The effect of ingested nanoparticles was measured by conventional toxicity measures such as feeding rate, weight change, and mortality and did not indicate any toxicity. However, cell membrane of digestive glands, measured with a modified method for assessing cell membrane stability, was affected already after 3 days of exposure to 1000 μg nano-TiO2per gram dry weight of food indicating cytotoxic potential of ingested nanoparticles. Our results confirmed hypothesis on low toxic potential and no internalization of consumed TiO2nanoparticles by a model invertebrate organism. However, cytological marker unequivocally indicated adverse effect of ingested nano-TiO2. We conclude that the isopod model system could be used for studying the fate and effect of ingested nanoparticles.

Isolation of hnRNP complexes from Drosophila melanogaster.

Nascent RNA polymerase II transcripts, heterogeneous nuclear RNAs (hnRNAs), become associated with nuclear proteins (hnRNP Proteins), and their processing into mRNAs takes place in these hnRNP complexes. hnRNP complexes have previously been purified from vertebrate cells. Here we report the isolation of hnRNP complexes from an invertebrate organism, the fruitfly Drosophila melanogaster. Candidate hnRNP proteins were purified from D. melanogaster embryos by ssDNA affinity chromatography, and mAbs were produced to many of the major proteins. Genuine hnRNP proteins were identified by several criteria, including nucleoplasmic localization, association with nascent transcripts, crosslinking to poly(A)-containing RNA in living cells, and amino acid sequence. In addition, mAbs that cross-react between the fruitfly and human hnRNP proteins were obtained. Most importantly, using hnRNP-specific mAbs we have purified the hnRNP complexes from D. melanogaster cells. These RNAase-sensitive complexes contain at least 10 major proteins designated hrps, the most abundant proteins having apparent molecular masses of 36, 38, 39, 40, 44, 48, 54, 62, 70, and 75 kD. cDNAs and complete sequences for several of these proteins have been obtained and are presented in the accompanying paper (Matunis, E. L., M. J. Matunis, and G. Dreyfuss. 1992. J. Cell Biol. 116:257-269). The purification of D. melanogaster hnRNP complexes will facilitate genetic and cytological studies on the function of hnRNA-binding proteins and on the posttranscriptional regulation of gene expression.

Drosophila nuclei replicate in Xenopus eggs

Nuclei isolated from a permanent cell line derived from Drosophila melanogaster embryos have been injected, along with a radioactive DNA precursor [3H] TTP, into Xenopus laevis eggs. In culture, less than 7 % of the cells were in S phase. After a 90 min incubation, following injection into eggs, 99% of the nuclei were shown by autoradiography to have synthesized DNA. In a similar experiment, a density label BrdUTP was injected into eggs along with the nuclei. Subsequent analysis on caesium chloride gradients showed that this DNA synthesis was semi-conservative replication. Therefore we conclude that signals present in Xenopus egg cytoplasm can initiate and sustain true semi-conservative DNA replication in nuclei from an invertebrate organism.