Smed-SmB, a member of the LSm protein superfamily, is essential for chromatoid body organization and planarian stem cell proliferation

Fernandéz-Taboada, Enrique; Moritz, Sören; Zeuschner, Dagmar; Stehling, Martin; Schöler, Hans R.; Saló, Emili; Gentile, Luca

doi:10.1242/dev.051847

Cited by 17 publications

(26 citation statements)

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“…Reactions were aliquoted using a Corbett Robot (Corbett Robotics) and analysed with a Rotor Gene 6000 (Corbett, Qiagen). qPCR oligos were used as already described (Eisenhoffer et al, 2008;Fernandez-Taboada et al, 2010), except for: ODC-1F: 5'-AGTGAAGTGCAATCAGTTGG-3' ODC-1R:…”

Section: Real Time Pcrmentioning

confidence: 99%

Decreased neoblast progeny and increased cell death during starvation-induced planarian degrowth

González‐Estévez

Felix²,

Rodríguez-Esteban³

et al. 2012

Int. J. Dev. Biol.

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The development of a complex multicellular organism requires a careful coordination of growth, cell division, cell differentiation and cell death. All these processes must be under intricate and coordinated control, as they have to be integrated across all tissues. Freshwater planarians are especially plastic, in that they constantly replace somatic tissues from a pool of adult somatic stem cells and continuously undergo growth and degrowth as adult animals in response to nutrient availability. During these processes they appear to maintain perfect scale of tissues and organs. These life history traits make them an ideal model system to study growth and degrowth. We have studied the unique planarian process of degrowth. When food is not available, planarians are able to degrow to a minimum size, without any signs of adverse physiological outcomes. For example they maintain full regenerative capacity. Our current knowledge of how this is regulated at the molecular and cellular level is very limited. Planarian degrowth has been reported to result from a decrease in cell number rather than a decrease in cell size. Thus one obvious explanation for degrowth would be a decrease in stem cell proliferation. However evidence in the literature suggests this is not the case. We show that planarians maintain normal basal mitotic rates during degrowth but that the number of stem cell progeny decreases during starvation and degrowth. These observations are reversed upon feeding, indicating that they are dependent on nutritional status. An increase in cell death is also observed during degrowth, which is not rapidly reversed upon feeding. We conclude that degrowth is a result of cell death decreasing cell numbers and that the dynamics of neoblast self-renewal and differentiation adapt to nutrient conditions to allow maintenance of the neoblast population during the period of starvation.

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Section: Real Time Pcrmentioning

confidence: 99%

Decreased neoblast progeny and increased cell death during starvation-induced planarian degrowth

González‐Estévez

Felix²,

Rodríguez-Esteban³

et al. 2012

Int. J. Dev. Biol.

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“…In recent years, various methods have been used to dissociate planarian tissues -mechanical (Fernandez-Taboada et al, 2010), trypsin-based (Reddien et al, 2005;Hayashi et al, 2006) and collagenase-based dissociation (Wagner et al, 2011). Papain is a gentle protease that has been successfully used to isolate cells with rather complex morphologies, such as visual neurons from adult rats (Huettner and Baughman, 1986).…”

Section: Planarian Cell Dissociation With Papain Improves Both Yield mentioning

confidence: 99%

“…Previously, we utilized mechanical dissociation (Fernandez-Taboada et al, 2010), which produces a good yield of viable cells, but is time consuming and laborious. We therefore tested whether papain could be a suitable protease for the dissociation of planarian cells.…”

Section: Planarian Cell Dissociation With Papain Improves Both Yield mentioning

confidence: 99%

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Heterogeneity of planarian stem cells in the S/G2/M phase

Moritz¹,

Stöckle²,

Ortmeier³

et al. 2012

Int. J. Dev. Biol.

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The planarian adult stem cell (pASC) population has a specific molecular signature and can be easily visualized and isolated by flow cytometry. However, the lack of antibodies against specific surface markers for planarian cells prevents a deeper analysis of specific cell populations. Here, if we describe the results of the immunoscreening of pASC plasma membrane proteins (PMPs). A novel papain-based method for planarian cell dissociation enabling both high yield and improved cell viability was used to generate single cell preparations for PMP purification. PMPs were used for intraperitoneal immunization of mice and thus about 1000 hybridoma clones were generated and screened. Supernatants collected from the hybridoma clones were first screened by ELISA and then by live immuno-staining. About half of these supernatants stained all the planarian cells, whereas the other half specifically labeled a subfraction thereof. A detailed analysis of two hybridoma supernatants revealed that large subfractions of the X1, X2 and Xin populations differentially express specific membrane markers. Quantitative PCR data disclosed a correlation between the immunostaining results and the expression of markers of the early and late progeny, also for those pASCs in the S/G2/M phase of the cell cycle (X1 population). Thus, about two thirds of the cycling pASCs showed a specific membrane signature coupled with the expression of markers hitherto considered to be restricted to differentiating, post-mitotic progeny. In summary, a library of 66 monoclonal antibodies against planarian PMPs was generated. The analysis of two of the clones generated revealed that a subset of cells of the X1 population expresses early and late progeny markers, which might indicate that these cells are committed while still proliferating. The findings demonstrate the usefulness of our PMP antibody library for planarian research.

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“…Dugesia japonica, a freshwater planarian, has been traditionally used in regenerative [20][21][22], developmental biology [22][23][24], toxicological [25][26][27] and neuroscience research [28,29]. In contrast, only a few papers have reported immunological research with this evolutionarily important animal [30,31].…”

Section: Introductionmentioning

confidence: 99%

Purification, characterization and induction of a C-type lectin in the freshwater planarian Dugesia japonica

et al. 2012

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Lectins are important components of the immune defense system of invertebrates. Given their important functions, numerous investigations have been carried out on the characterization and function of lectins in invertebrates. However, lectin studies with the freshwater planarian, an evolutionarily important animal, are rare. In this paper, we demonstrate agglutination of glutaraldehyde treated erythrocytes by a lectin with preference for rabbit erythrocytes. The result of hemagglutinating activity inhibition assays with several carbohydrates showed the most potent inhibitor was maltose. A natural lectin from the crude homogenates of freshwater planarian Dugesia japonica was purified by single step affinity chromatography using amylose-coupled agarose. The purified protein appeared as one band with a molecular mass of 350 kDa in PAGE, and as one band, approximately 56 kDa, in SDS-PAGE. The purified lectin showed dependence on calcium. The activity of the purified lectin was inhibited at temperatures greater than 50°C and showed a pH optimum between 5–8. The purified lectin also has binding activity to the Gram-negative bacteria E. coli, and the Gram-positive bacteria B. subtilis. Furthermore, the purified lectin obtained from injured and bacteria-induced planarians showed increased agglutinating activity against rabbit erythrocytes. These results suggest that the purified lectin may play an important role in the innate immunity of the freshwater planarian.

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Smed-SmB, a member of the LSm protein superfamily, is essential for chromatoid body organization and planarian stem cell proliferation

Cited by 17 publications

References 0 publications

Decreased neoblast progeny and increased cell death during starvation-induced planarian degrowth

Decreased neoblast progeny and increased cell death during starvation-induced planarian degrowth

Heterogeneity of planarian stem cells in the S/G2/M phase

Purification, characterization and induction of a C-type lectin in the freshwater planarian Dugesia japonica

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