Biomedical subjects

S Reber-Müller

Publications and source records attributed to S Reber-Müller.

6 recordsLinked to original sources

PubMed2001

Integrin and talin in the jellyfish Podocoryne carnea.

We have isolated an integrin-beta and -alpha subunit from Podocoryne carnea (Cnidaria, Hydrozoa) and studied their expression in the life-cycle and during cell migration, in vitro transdifferentiation and regeneration. Comparison of the integrin expression pattern with a Podocoryne talin homologue by RT-PCR demonstrates that all three genes are maternal messages and continuously expressed in the life-cycle, in medusa development and in all medusae tissues. In situ hybridisation experiments confirm co-expression of both integrin subunits in the different life-stages. Integrin expression was furthermore studied in isolated striated muscle induced to transdifferentiate to new cell types, or grafted on ECM where the muscle adheres and migrates. Integrin expression was maintained continuously throughout both processes. These results suggest that in Podocoryne carnea processes such as cell migration and differentiation are not controlled by up- or downregulation of alternative integrin subunits, but by a single integrin heterodimer which activates different downstream signalling cascades.

Amino Acid Sequence↗

PubMed1999

Cell-substrate interactions in cnidaria.

Studies on morphogenesis and regeneration in cnidarians have a long history, and the importance of cell-ECM (extracellular matrix) interactions for these processes has been well recognized and studied since the middle of the 20th century. Cnidarians have a life cycle with a larva, a polyp, and often a medusa generation. In the medusa, the ECM (mesoglea) is very prominent and essentially shapes the animal. In the larva and the polyp, the ECM is a thin layer. Some of the ECM components known from vertebrates have been identified in cnidarians by immunohistochemistry, electron microscopy, rotary shadowing, biochemistry, and molecular cloning. In vivo and in vitro experiments suggest that the cnidarian ECM plays a role in cell migration and morphogenesis comparable to that known from other developmental systems. In the fresh water polyp Hydra, regeneration of body patterns and migration of nematocytes seems to require the presence of ECM ligands and the corresponding cell receptors. In hydrozoan medusae, DNA replication and the stability of the differentiated state of isolated tissue can be influenced by altering the properties of the ECM substrate. When cultured, most cnidarian cells survive only when attached to ECM substrates, they rarely divide and die within short times.

Animals↗

PubMed1995

An extracellular matrix protein of jellyfish homologous to mammalian fibrillins forms different fibrils depending on the life stage of the animal.

A monoclonal antibody generated against the isolated extracellular matrix (ECM) of the medusa Podocoryne carnea M. Sars (Coelenterata, Cnidaria, Hydrozoa) stains a fibrillar component of the Podocoryne ECMs in immunohistochemical preparations. The antigen shows a different staining pattern according to the type of ECMs from the animals life cycle. In ontogeny the epitope first appears after gastrulation in the planula larva as single widely dispersed small fibrils, which later accumulate to form a dense meshwork in the larval ECM. The distribution of the antigen strongly suggests an important role of the molecule to cover the biomechanical needs of the animal. In immunoblots one band with a size of 330 kDa is detectable in the polyp ECM, whereas in the outer ECM of the medusa a 340-kDa band is observed. Both the 330- and the 340-kDa bands appear when probed on the inner ECM of the medusa or on ECMs of the larva. The antibody was used to isolate a cDNA clone from an expression library. The deduced amino acid sequence of this cDNA fragment reveals a molecular structure composed of tandemly repeated epidermal growth factor-like repeats interrupted by a second cystein-rich motif first found in the latent transforming growth factor beta binding protein. Comparison of the sequence to the data bases indicates < 40% identity to human fibrillins. The presence of fibrillin-like beaded microfibrils in the ECM of P. carnea is furthermore demonstrated by electron microscopy after rotary shadowing. Our results demonstrate for the first time the existence of this noncollagenous interstitial ECM protein in invertebrates and suggest that the structure and the function of fibrillin have been conserved during evolution.

Amino Acid Sequence↗

PubMed1995

Transdifferentiation of isolated striated muscle of jellyfish in vitro: the initiation process.

Fragments of striated muscle tissue of Anthomedusae can be isolated and cultured. Without further treatment the isolated muscle fragments maintain the differentiated state. When treated with enzymes degrading the adhering extracellular matrix, drugs activating protein kinase C or substances destroying the actin cytoskeleton, dedifferentiation and DNA replication are initiated and transdifferentiation to several new cell types occurs. Initiation of DNA replication seems to be correlated with a disturbance of cell-ECM interactions. If muscle fragments are combined with isolated ECMs, cell migration onto the grafted ECMs occurs and DNA-replication and transdifferentiation are initiated in those cells which adhere to both, the native and the grafted ECM. If, however, the cells can stretch into a monolayer and adhere entirely to either the native or the grafted ECM, DNA-replication is inhibited. Carbohydrate moieties seem to be involved in mediating these cell-substrate interactions.

Animals↗

PubMed1993

Mechanochemical interactions between striated muscle cells of jellyfish and grafted extracellular matrix can induce and inhibit DNA replication and transdifferentiation in vitro.

Striated muscle tissue of jellyfish was isolated with its adhering extracellular matrix (ECM) and cultured. Without further treatment the cultured muscle cells maintain their differentiated state. If, however, the isolated tissues are combined with cell-free ECM from the jellyfish or its polyp, DNA replication and proteolytic activity are induced followed by transdifferentiation into RF-amide-positive nerve cells. Changes in the mechanochemical interactions between the cells and the grafted ECM seem to induce the signals which lead to transdifferentiation. If the isolates are combined with small floating pieces of ECM most cells will leave their own ECM and overgrow the ECM graft. All cells in the combinations will then transdifferentiate. If the isolates are grafted onto large pieces of ECM kept permanently stretched on glass, a majority of cells will migrate onto the grafted ECM where they form a flat monolayer. In this case, however, DNA replication and transdifferentiation occurs mainly in those cells which have remained on or near their own ECM. Labeling experiments with [3H]-thymidine demonstrate that initiation of DNA replication occurs first in those cells which bridge from the native ECM to the grafted ECM. On the other hand inhibition of DNA replication and transdifferentiation is generally suppressed whenever tissues are allowed to form a monolayer of well-stretched cells. From these observations we conclude that mechanochemical interactions between the muscle cells and their substrate are responsible for both activation and inhibition of DNA replication and transdifferentiation.

Animals↗

PubMed1993

Life stage specific expression of a myosin heavy chain in the hydrozoan Podocoryne carnea.

In order to study life stage specific gene-expression and also to investigate the molecular components of jellyfish muscle we cloned a tissue specifically expressed cDNA of the metagenetic hydrozoan Podocoryne carnea. A monoclonal antibody was made that reacts with the filaments of striated muscle of the medusa stage. The mAb detects a protein of 220 kDa in Western blots. The antibody was used to clone a cDNA from an expression library. Sequence analysis showed that the clone codes for the rod part of a myosing heavy chain (MHC). Surprisingly, the sequence is more similar to MHCs from striated muscle than to smooth muscle or non-muscle MHCs from either invertebrates or vertebrates. A particularly well conserved region, which may be correlated to a function in striated muscle, was found. Expression studies showed that the corresponding RNA is only present in the striated muscle cells of the medusa. During medusa development, the transcript became detectable at the same time as muscle differentiation becomes visible in the electron microscops.

Amino Acid Sequence↗