Biomedical subjects

Reinhard W Köster

Publications and source records attributed to Reinhard W Köster.

5 recordsLinked to original sources

PubMed2006

FGF signaling mediates regeneration of the differentiating cerebellum through repatterning of the anterior hindbrain and reinitiation of neuronal migration.

To address the regenerative capability of the differentiating hindbrain, we ablated the cerebellum in wild-type and transgenic zebrafish embryos. These larvae showed no obvious locomotive malfunction several days after the ablation. Expression analysis and in vivo time-lapse recording in GFP (green fluorescent protein)-transgenic embryos indicate that cerebellar neuronal cells can regenerate from the remaining anterior hindbrain. The onset of regeneration is accompanied by repatterning within the anterior hindbrain. Inhibition of FGF signaling immediately after cerebellar ablation results in the lack of regenerating cerebellar neuronal cells and the absence of cerebellar structures several days later. Moreover, impaired FGF signaling inhibits the repatterning of the anterior hindbrain and the reexpression of rhombic lip marker genes soon after cerebellar ablation. This demonstrates that the hindbrain is highly plastic in recapitulating early embryonic differentiation mechanisms during regeneration. Moreover, the regenerating system offers a means to uncouple cerebellar differentiation from complex morphogenetic tissue rearrangements.

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PubMed2006

Multicolor in vivo time-lapse imaging at cellular resolution by stereomicroscopy.

Intravital time-lapse imaging has altered significantly many long-standing rules of biological mechanisms, but being apparatus-intense and laborious, time-lapse imaging remained mostly restricted to specialized labs. We show that recently introduced, fully automated fluorescence stereomicroscopes represent cost-effective but powerful means of imaging dynamic events ranging from observing embryogenesis over several days to detailed tissue rearrangements and fast blood cell rolling in vivo. When combined with deconvolution approaches, even subcellular resolution in several colors can be achieved. Using three-dimensional image recording, we show the spatial reconstruction of expression patterns. Furthermore, by combining three-dimensional image recording over time with subsequent deconvolution analysis, we demonstrate that subcellular dynamics such as axonal pathfinding can be resolved. These findings promise that time-lapse imaging using a stereomicroscope will become a hands-on standard method for phenotype analysis in many fields of biology.

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PubMed2005

Quantum dots are powerful multipurpose vital labeling agents in zebrafish embryos.

Recently, inorganic fluorescent contrast agents composed of semiconductor materials have been introduced to biological imaging approaches. These so-called quantum dots provide unique and promising properties unreached by organic fluorophores, but their use as contrast agents within live organisms has been limited, probably due in part to concerns about their in vivo tolerance. Using transparent zebrafish embryos, we challenged quantum dots with a series of intravital imaging problems. We show that quantum dots provide a high fluorescent yield within targeted tissues, possess immense photostability, can be targeted to specific subcellular compartments, remain within targeted cells as lineage tracers, are easily separable from conventional organic fluorescent dyes, and are fixable, allowing them to be used in combination with immunohistochemistry after live recordings. Thus, quantum dots combine the specific advantages of different organic fluorescent contrast agents and promise to become the first fluorophore feasible for long-lasting intravital time-lapse studies. Finally, we show by co-labeling blood vessels of the vasculature and major axon tracts of the nervous system that, for establishing these networks, the same guidance cues might be used in some body parts, whereas in others, both networks appear to develop independently from one another. Thus, the bright fluorescence of quantum dots will help to unravel many open questions in the fields of embryology, cell biology, as well as phenotyping and disease diagnosis.

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PubMed2004

Time-lapse microscopy of brain development.

Zebrafish embryos represent an ideal vertebrate model organism for noninvasive intravital imaging because of their optical clarity, external embryogenesis, and fast development. Many different labeling techniques have been adopted from other model organisms or newly developed to address a wealth of different developmental questions directly inside the living organism. The parallel advancements in the field of optical imaging let us now observe dynamic processes at the cellular and subcellular resolution. Combined with the repertoire of available surgical and genetic manipulations, zebrafish embryos provide the powerful and almost unique possibility to observe the interplay of molecular signals with cellular, morphological, and behavioral changes directly within a living and developing vertebrate organism. A bright future for zebrafish is yet to come, let there be light.

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PubMed2003

Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis.

The pattern of blood flow in the developing heart has long been proposed to play a significant role in cardiac morphogenesis. In response to flow-induced forces, cultured cardiac endothelial cells rearrange their cytoskeletal structure and change their gene expression profiles. To link such in vitro data to the intact heart, we performed quantitative in vivo analyses of intracardiac flow forces in zebrafish embryos. Using in vivo imaging, here we show the presence of high-shear, vortical flow at two key stages in the developing heart, and predict flow-induced forces much greater than might have been expected for micro-scale structures at low Reynolds numbers. To test the relevance of these shear forces in vivo, flow was occluded at either the cardiac inflow or outflow tracts, resulting in hearts with an abnormal third chamber, diminished looping and impaired valve formation. The similarity of these defects to those observed in some congenital heart diseases argues for the importance of intracardiac haemodynamics as a key epigenetic factor in embryonic cardiogenesis.

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