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doi: 10.1242/10.1242/dev.00167


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The Schmidtea mediterranea database as a molecular resource for studying platyhelminthes, stem cells and regeneration

Alejandro Sánchez Alvarado*,{dagger},§, Phillip A. Newmark*,{ddagger}, Sofia M. C. Robb{dagger} and Réjeanne Juste

Carnegie Institution of Washington, Department of Embryology, Baltimore, MD 21210, USA
{dagger} Present address: University of Utah School of Medicine, Department of Neurobiology and Anatomy, Salt Lake City, UT 84132-3401, USA
{ddagger} Present address: University of Illinois at Urbana-Champaign, Department of Cell and Structural Biology, Urbana-Champaign, IL 61801, USA



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Fig. 1. Bioinformatics and categorization of S. mediterranea sequences. (A) Organigram depicting the steps performed to produce a non-redundant collection of S. mediterranea cDNA sequences (SmedDb). Bioinformatic analyses compared the database against itself to identify and remove redundant clones before sending sequences to the public databases. The GenBank protein and nucleotide collections as well as the EST database (dbEST) were queried using the BLAST algorithm. Sequences returning significant matches (E<=10-4) were subjected to annotation into functional categories based on the identity/function of the GenBank match. (B) Distribution of informative sequences by functional categories. For simplicity the cell signaling category includes the cell/cell communication and internal signaling categories. Metabolism is an amalgamation of the general metabolism, mitochondria and protein metabolism categories. Visit http://planaria.neuro.utah.edu for a detailed categorization profile and access to sequences. (C) Distribution by percentage of planarian genes displaying highest similarities with members of either the vertebrates or invertebrates.

 


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Fig. 2. Representative results of high-throughput, whole-mount in situ hybridization. (A-C) Probe identity corresponding to the images is given from top to bottom with each clone ID in parentheses as follows. (A) Gene expression patterns within the nervous system of S. mediterranea: synaptotagmin (H.6.7h); quinoid dihydropteridine reductase (H.9.5b); pax6 (H.109.7h) and degenerin (H.112.3c). (B) Gene expression patterns in organ systems: gastrovascular system (D.14; unknown function), dorsal epithelium (H.7.1e; gp25L/p24 family), excretory system (H.14.9d; carbonic anhydrase); and pharynx (H.14.11f; unknown function). (C) Gene expression in discrete cell types: matrix metalloproteinase (A115) in central secretory cells; epithelial cells (H.12.11a; intermediate filament); subepidermal marginal adhesive gland cells (H.1.3b; zonadhesin); and free-mesenchymal cells (neoblasts) expressing piwi (H.2.12c). (D) Ventral view (left) of clone H.8.1f (unknown function), and lateral view of the same specimen (right) demonstrate a dorsoventral segregation of differentiation. The red asterisk indicates the position of the pigmented photoreceptor. Anterior is to the left in all panels.

 


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Fig. 3. Regulation of cell number in planarians of different lengths. (A) In situ hybridization of clone H112.3c, showing distribution of putative chemoreceptive neurons underlying anterior margin. Nomarski DIC view. Scale bar: 100 µm. (B) Number of H112.3c-positive cells/side in organisms of different lengths. Mean number of H112.3c-positive cells (±s.d.) is indicated (for 1 mm, n=14; 2 mm, n=9; 6 mm, n=10; 8 mm, n=7).

 

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