Myogenesis during holothurian intestinal regeneration

Murray, Gisela; García‐Arrarás, José E.

doi:10.1007/s00441-004-0978-3

Cited by 46 publications

(50 citation statements)

References 29 publications

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“…Western blot techniques were similar to those used previously (Murray and García-Arrará s, 2004). In brief, homogenates were prepared from the intestinal tissue of H. glaberrima specimens using a Polytron (Brinkmann Instruments) and two different extraction buffers: (1) phosphate buffered saline (PBS) with protease inhibitors (Calbiochem) or (2) and a phenol/chloroform method (TriReagent, Molecular Research Center).…”

Section: Western Blotsmentioning

confidence: 99%

Spherulocytes in the echinoderm Holothuria glaberrima and their involvement in intestinal regeneration

García‐Arrarás¹,

Schenk²,

Rodrígues‐Ramírez³

et al. 2006

Developmental Dynamics

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The holothuroid echinoderm Holothuria glaberrima can regenerate its intestine after a process of evisceration. Spherule-containing cells, the spherulocytes, appear to be associated with intestinal regeneration. We have used histochemistry and immunocytochemistry to characterize these cells and their role in the regeneration process. Spherulocytes are 10 -20 m in diameter with an acrocentric nucleus and spherule-like structures within their cytoplasm. They are found in the connective tissue of the intestine and mesentery of noneviscerated and regenerating animals. During the second week of regeneration, the number of spherulocytes in the regenerating intestine increases and a dramatic change in their morphology occurs. Together with the morphological change, the immunohistochemical labeling of the cells also changes; the antibodies not only recognize the spherule structures but also label the cellular cytoplasm in a more homogeneous pattern. Moreover, immunohistochemical labeling also appears to be dispersed within the extracellular matrix, suggesting that the cells are liberating their vesicular contents. Spherulocytes are found in other tissues of H. glaberrima, always associated with the connective tissue component. Our data strongly suggest that spherulocytes are involved in intestinal regeneration but their specific role remains undetermined. In summary, our data expand our knowledge of the cellular events associated with regeneration processes in echinoderms and provide for comparisons with similar processes in vertebrates.

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Section: Western Blotsmentioning

confidence: 99%

Spherulocytes in the echinoderm Holothuria glaberrima and their involvement in intestinal regeneration

García‐Arrarás¹,

Schenk²,

Rodrígues‐Ramírez³

et al. 2006

Developmental Dynamics

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“…This coincides with a disorganization of the muscle layers of the mesentery (Quiñones et al 2002). In addition, selected markers for myocytes, such as a muscle-specific monoclonal antibody and phalloidin labeling (a toxin that labels the actin filaments present within the muscle cell contractile apparatus), are not found within the regenerating structure during the first days of regeneration and eventually appear as the new intestine is formed (Garcia-Arraras and Greenberg 2001; Murray and García-Arrarás 2004). These results suggest that the myocytes close to and within the regenerating structure undergo dramatic changes during the first few weeks of regeneration.…”

Section: Introductionmentioning

confidence: 97%

“…Previous studies have shown that regeneration of the intestinal system begins from a series of thickenings that are formed at the remaining free edge of the lateral and ventral mesenteries after evisceration and proceeds until it is complete in 3-4 weeks (García-Arrarás et al 1998, 1999. The thickenings eventually give rise to a blastema-like structure, and various processes are associated with its formation, including cell division, migration of various cell populations from the esophagus, cloaca, and mesenteries, and changes in the extracellular matrix components of the connective tissue (García-Arrarás et al 1998, 1999García-Arrarás and Greenberg 2001;Quiñones et al 2002;Murray and García-Arrarás 2004).…”

Section: Introductionmentioning

confidence: 98%

Contribution of mesenterial muscle dedifferentiation to intestine regeneration in the sea cucumber Holothuria glaberrima

et al. 2006

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Holothurians (sea cucumbers) have been known from ancient times to have the capacity to regenerate their internal organs. In the species Holothuria glaberrima, intestinal regeneration involves the formation of thickenings along the free mesentery edge; these thickenings will later give rise to the regenerated organ. We have previously documented that a remodeling of the extracellular matrix and changes in the muscle layer occur during the formation of the intestinal primordium. In order to analyze these changes in depth, we have now used immunocytochemical techniques and transmission electron microscopy. Our results show a striking disorganization of the muscle layer together with myocyte dedifferentiation. This dedifferentiation involves nucleic activation, disruptions of intercellular junctions, and the disappearance of cell projections, but more prominently, the loss of the contractile apparatus by the formation and elimination of spindle-like structures. Muscle dedifferentiation can be seen as early as 2 days following evisceration and continues during the next 2 weeks of the regeneration process. Dedifferentiation of myocytes might result in cells that proliferate and give rise to new myocytes. Alternatively, dedifferentiating myocytes could give rise to cells with high nuclear-to-cytoplasmic ratios, with some being eliminated by apoptosis. Our results, together with those in other regeneration models, show that myocyte dedifferentiation is a common event in regeneration processes and that the dedifferentiated cells might play an important role in the formation of the new tissues or organs.

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“…See web site for date of publication (http://physiolgenomics.physiology.org). 11,39), little is known about the genes involved. Previous attempts to identify the genes associated with intestinal regeneration in H. glaberrima have been made in our laboratory using a gene by gene strategy, which is time consuming and provided limited information (37,49,51).…”

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confidence: 99%

Distinct profiles of expressed sequence tags during intestinal regeneration in the sea cucumberHolothuria glaberrima

Rojas‐Cartagena

Ortíz-Pineda

Ramírez-Gómez

et al. 2007

Physiological Genomics

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Repair and regeneration are key processes for tissue maintenance, and their disruption may lead to disease states. Little is known about the molecular mechanisms that underline the repair and regeneration of the digestive tract. The sea cucumber Holothuria glaberrima represents an excellent model to dissect and characterize the molecular events during intestinal regeneration. To study the gene expression profile, cDNA libraries were constructed from normal, 3-day, and 7-day regenerating intestines of H. glaberrima. Clones were randomly sequenced and queried against the nonredundant protein database at the National Center for Biotechnology Information. RT-PCR analyses were made of several genes to determine their expression profile during intestinal regeneration. A total of 5,173 sequences from three cDNA libraries were obtained. About 46.2, 35.6, and 26.2% of the sequences for the normal, 3-days, and 7-days cDNA libraries, respectively, shared significant similarity with known sequences in the protein database of GenBank but only present 10% of similarity among them. Analysis of the libraries in terms of functional processes, protein domains, and most common sequences suggests that a differential expression profile is taking place during the regeneration process. Further examination of the expressed sequence tag dataset revealed that 12 putative genes are differentially expressed at significant level ( R > 6). Experimental validation by RT-PCR analysis reveals that at least three genes (unknown C-4677-1, melanotransferrin, and centaurin) present a differential expression during regeneration. These findings strongly suggest that the gene expression profile varies among regeneration stages and provide evidence for the existence of differential gene expression.

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Myogenesis during holothurian intestinal regeneration

Cited by 46 publications

References 29 publications

Spherulocytes in the echinoderm Holothuria glaberrima and their involvement in intestinal regeneration

Spherulocytes in the echinoderm Holothuria glaberrima and their involvement in intestinal regeneration

Contribution of mesenterial muscle dedifferentiation to intestine regeneration in the sea cucumber Holothuria glaberrima

Distinct profiles of expressed sequence tags during intestinal regeneration in the sea cucumberHolothuria glaberrima

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