Biomedical subjects

Felicitas Pröls

Publications and source records attributed to Felicitas Pröls.

11 recordsLinked to original sources

PubMed2006

Myogenin (Myf4) upregulation in trans-differentiating fibroblasts from a congenital myopathy with arrest of myogenesis and defects of myotube formation.

Congenital myopathies often have an unclear aetiology. Here, we studied a novel case of a severe congenital myopathy with a failure of myotube formation. Polymerase chain reaction-based analysis was performed to characterize the expression patterns of the Desmin, p21, p57, and muscle regulatory factors (MRFs) MyoD, Myf4, Myf5 and Myf6 in differentiating skeletal muscle cells (SkMCs), normal human fibroblasts and patient-derived fibroblasts during trans-differentiation. The temporal and spatial pattern of MRFs was further characterized by immunocyto- and immunohistochemical stainings. In differentiating SkMCs, each MRF showed a characteristic expression pattern. Normal trans-differentiating fibroblasts formed myotubes and expressed all of the MRFs, which were detected. Interestingly, the patient's fibroblasts also showed some fusion events during trans-differentiation with a comparable expression profile for the MRFs, particularly, with increased expression of Myf4 and p21. Immunohistochemical analysis of normal and patient-derived skeletal musculature revealed that Myf4, which is downregulated during normal fetal development, was still present in patient-derived skeletal head muscle, which was also positive for Desmin and sarcomeric actin. The abnormal upregulation of Myf4 and p21 in the patient who suffered from a severe congenital myopathy suggests that the regulation of Myf4 and p21 gene expression during myogenesis might be of interest for further studies.

Cell Differentiation↗

PubMed2006

Avian stanniocalcin-2 is expressed in developing striated muscle and joints.

The glycoprotein hormone stanniocalcin (STC) has originally been described in the teleost kidney. Since then, STC homologs have been identified in various genomes including human, mouse, rat, Xenopus and zebrafish. In mammals, two STC genes, STC1 and STC2, are known. We cloned a chicken STC homolog to analyze its expression pattern during chick development. Sequence analyses revealed a high sequence similarity of the chicken STC (cSTC) clone to mammalian STC2. Interestingly the expression pattern of cSTC2 largely resembles those of murine STC1: we found expression of cSTC2 in the nephric tubules, in the myocardium, in skeletal muscle cells from the onset of differentiation, and in synovial joint anlagen of the limbs.

Amino Acid Sequence↗

PubMed2005

Ectodermal Wnt-6 promotes Myf5-dependent avian limb myogenesis.

Limb muscles of vertebrates are derived from precursor cells that migrate from the lateral edge of the dermomyotome into the limb bud. Although several signaling molecules have been reported to be involved in the process of limb myogenesis, none of their activities has led to a consolidate idea about the limb myogenic pathway. Particularly, the role of ectodermal signals in limb myogenesis is still obscure. Here, we investigated the role of the ectoderm and ectodermal Wnt-6 during limb muscle development. We found that ectopic expression of Wnt-6 in the limb bud specifically extends the expression domains of Pax3, Paraxis, Myf5, Myogenin, Desmin and Myosin heavy chain (MyHC) but inhibits MyoD expression. Ectoderm removal results in a loss of expression of all of these myogenic markers. We show that Wnt-6 can compensate the absence of the ectoderm by rescuing the expression of Pax3, Paraxis, Myf5, Myogenin, Desmin and MyHC but not MyoD. These results show that, in chick, at least two signals from the limb ectoderm are necessary for muscle development. One of the signals is Wnt-6, which plays a unique role in promoting limb myogenesis via Pax3/Paraxis-Myf5, whereas the other putative signaling pathway involving MyoD expression is negatively regulated by Wnt-6 signaling.

Animals↗

PubMed2005

Paradoxical effects of hypoxia-mimicking divalent cobalt ions in human endothelial cells in vitro.

Divalent cobalt ions (Co2+) induce the expression of hypoxia responsive genes and are often used in cell biology to mimic hypoxia. In this in vitro study we compared the effects of hypoxia and Co2+ on human endothelial cells and examined processes that are stimulated in hypoxia in vivo (proliferation and angiogenesis). We analyzed the expression of the hypoxia-inducible factor-1alpha (HIF-1alpha) under different hypoxic conditions (3% and nearly 0% O2) and Co2+ -concentrations (0.01-0.7 mM). As in hypoxia, the amount of HIF-1alpha protein was enhanced by exposure to Co2+ (did not correlate with mRNA amount). however, contrary to the results of hypoxia, in vitro-angiogenesis was inhibited after exposure to even low Co2+-concentrations (> or =0.01 mM). This led to the conclusion that although hypoxia signaling after Co2+ -exposure took place, further yet unknown Co2+ -induced event(s) must have occurred.

Cell Line↗

PubMed2004

PCNA in situ hybridization: a novel and reliable tool for detection of dynamic changes in proliferative activity.

In order to investigate developmental processes, several methods have been established that allow the visualization of local proliferation zones and to follow their dynamics during morphogenesis. In this study we present a detailed description of transitory and continuous proliferation zones in the developing chick embryo. By tracing the S-phase marker proliferating cell nuclear antigen (PCNA) at the mRNA level we were able to identify the initiation and termination of proliferation programs. This approach provides additional information in comparison to the well-known BrdU incorporation or the PCNA immunostaining, which exclusively labels cells that contain PCNA protein. By means of PCNA in situ hybridization we analyzed the normal expression pattern in the 2- to 5-day-old chick embryo. We furthermore monitored the effects on PCNA expression after various manipulations such as removal of the apical ectodermal ridge (AER), the zone of polarizing activity (ZPA), and the surface ectoderm. In addition, we applied morphogens, such as fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), and retinoic acid (RA), and subsequently analyzed changes in the pattern of PCNA expression. While ablation of ZPA, AER, or ectoderm are known to reduce cell proliferation and were paralleled by loss of PCNA expression, neither BMP-2 nor BMP-4 affected PCNA expression. Upregulation of PCNA expression could be achieved by application of RA or FGFs, factors known to induce cell proliferation during limb bud outgrowth. The PCNA in situ hybridization data presented here clearly show that this method offers a novel, very sensitive tool for tracing cell proliferation and for visualizing the dynamic patterns arising due to the initiation and termination of the proliferation program.

Amino Acid Sequence↗

PubMed2004

Liquid 2-P-HEMA for endovascular tumor therapy: in vivo feasibility study.

BACKGROUND: To evaluate a new liquid polymer developed for selective arterial tumor embolization. MATERIAL/METHODS: In six rabbits we embolized the renal artery with pure 2-P-HEMA. Three rabbits were killed immediately after embolization, one after 12 h, and two animals underwent follow-up angiography after two weeks. In a second part of the study we embolized the left renal artery in three mini-swine with pure 2-P-HEMA. The kidneys were examined post mortem by soft-tissue radiography and kidney-tissue section microscopy. RESULTS: Complete occlusion of small distal arteries, most of the arterioles together with the precapillary vascular bed, was achieved in all animals. The speed of injection influenced the grade of distal occlusion; repeated injections through the same microcatheter were possible. The relatively fast injection speed of 0.3 ml/min led to an optimal distribution of the liquid embolic. There was no reperfusion in the two follow-up angiographies two weeks after embolization. Histopathologically, there was no angionecrosis and subintimal bleeding. No marked inflammatory reaction in the vessel wall or perivascular tissue was observed in the embolized arteries. CONCLUSIONS: This study suggests that liquid 2-P-HEMA may be feasible for the treatment of tumors.

Animals↗

PubMed2004

Expression and regulation of ROR-1 during early avian limb development.

ROR-1 is a member of the ROR family of tyrosine kinase like orphan receptors and is highly conserved among various species. We have isolated the chick ROR-1 ( cROR-1) and show that cROR-1 expression is high and restricted to the proximal limb region until HH-stage 25. At later stages, expression spreads towards the distal limb region. In order to determine the signals that control cROR-1 expression, factors known to be involved in limb patterning (FGFs, BMPs, SHH, retinoic acid) were applied to the developing limb. Whereas neither FGFs, BMPs, nor SHH affected cROR-1 expression, upregulation could be achieved by ectopic application of retinoic acid to the distal limb region. As retinoic acid also upregulated retinoic acid receptor beta ( Rar-beta), we assume that cROR-1 upregulation is mediated by Rar-beta. We conclude that ROR-1 signaling is an independently regulated pathway, which is involved in late rather than early limb development.

Amino Acid Sequence↗

PubMed2003

High levels of the molecular chaperone Mdg1/ERdj4 reflect the activation state of endothelial cells.

Mdg1/ERdj4, a mammalian chaperone that belongs to the HSP40 protein family, has been reported to be located in the endoplasmic reticulum (ER), is induced by ER stress, and protects ER stressed cells from apoptosis. Here we show that under normal physiological conditions, Mdg1/ERdj4 is expressed at various levels in the vasculature due to different activation states of the endothelium. To elucidate the stimuli that induce ER stress and thus upregulate Mdg1/ERdj4, we investigated the effect of several endothelium specific stressors on its expression. Mdg1/ERdj4 mRNA is induced by activated macrophages, by nitric oxide (NO) and heat shock, and during terminal cell differentiation, whereas shear stress does not affect Mdg1/ERdj4 expression levels. While the mRNA stability of BiP/GRP78 is unaffected in ER stressed cells, the stability of Mdg1/ERdj4 mRNA is prolonged during ER stress resulting in rapid increases and high levels of Mdg1/ERdj4 mRNA. Mdg1/ERdj4 protein is localized in the ER under control conditions. While heat shock induces a rapid translocation of Mdg1/ERdj4 to the nucleoli, no translocation could be observed during ER stress. This indicates that Mdg1/ERdj4 protein has diverse mechanisms to protect stressed cells from apoptosis.

Animals↗

PubMed2003

Spatial and temporal pattern of Wnt-6 expression during chick development.

The WNT family of proteins is composed of several members. In the present study we isolated the full length chick Wnt-6 cDNA and analyzed its expression pattern by in situ hybridization during chick development. Wnt-6 expression is observed in the ectoderm from HH-stage 4 onwards. At HH-stages, 7-16 expression can be seen in the ectoderm overlying the segmental plate and the epithelial somite, while the ectoderm overlying the compartmentalized somite is Wnt-6 negative. Expression is also observed at the heart outflow tract and in the ectoderm overlying the pharyngeal arches. From HH-stages 17 to 27, expression is also observed at limb level, both in the dorsal and ventral ectoderm and a stronger expression in the dorsoventral boundary. Furthermore, expression in the ectoderm delimiting the somitic boundaries in the anteroposterior and mediolateral axis at limb level was observed, as well as in the ventral body wall. Expression becomes evident in the inner ear. From HH-stage 30 onwards, expression is restricted to the feather buds and to the gastrointestinal tract.

Amino Acid Sequence↗

PubMed2003

Revisiting vimentin expression in early chick development.

To identify somite-specific antigens exhibiting novel expression patterns in the developing chick embryo, epithelial somites were isolated and used for intrasplenic immunization. Immunohistochemical screening of chick embryos of various stages focussed our attention on a monoclonal antibody, Som5H5, which stained somitic derivatives, spinal nerves, and neural tubes as well as the costameres of adult skeletal muscle. Western blot and mass spectrometric analysis revealed the Som5H5 antigen to be vimentin, the distribution of which has been well characterized before. In addition to the described vimentin pattern, Som5H5 stained a region in the proximal portion of the developing limb bud. This novel expression domain was confirmed by in situ hybridization using vimentin riboprobes. Signaling molecules (Shh and BMP-2), known to play a role in limb development, did not influence vimentin expression. Thus, no functional or cellular correlate to this vimentin-positive region could be determined.

Amino Acid Sequence↗

PubMed2002

BMPs induce dermal markers and ectopic feather tracts.

Bone morphogenetic protein (BMP) signaling is known to be involved in multiple inductive events during embryogenesis including the development of amniote skin. Here, we demonstrate that early application of BMP-2 to the lateral trunk of chick embryos induces the formation of dense dermis, which is competent to participate in feather development. We show that BMPs induce the dermis markers Msx-1 and cDermo-1 and lead to dermal proliferation, to expression of beta-catenin, and eventually to the formation of ectopic feather tracts in originally featherless regions of chick skin. Moreover, we present a detailed analysis of cDermo-1 expression during early feather development. The data implicate that cDermo-1 is located downstream of BMP in a signaling pathway that leads to condensation of dermal cells. The roles of BMP and cDermo-1 during development of dermis and feather primordia are discussed.

Animals↗