FISH in whole-mount <i>Drosophila</i> embryos. RNA: activation of a transcriptional locus, DNA: gene architecture and expression

Gemkow, Mark J; Buchenau, Peter; Arndt‐Jovin, Donna J.

doi:10.1002/1361-6374(199606)4:2<107::aid-bio8>3.0.co;2-q

Cited by 5 publications

(1 citation statement)

References 65 publications

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“…In early developing Drosophila embryos the chromosomes are positioned inside the nucleus with a defined centromere-telomere polarity following a rule first described by Rabl (1885;Swedlow et al, 1993). However, during gastrulation this orientation largely disappears, and homologous associations are formed (Foe and Alberts, 1983;Campos-Ortega and Hartenstein, 1985;Hiraoka et al, 1993;Dernburg et al, 1996;Gemkow et al, 1996). In many other species or cell types one can observe only a territorial delineation with no defined polarity or homologous pairing of the chromosomes (Cremer et al, 1994).…”

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The Dynamic Nuclear Redistribution of an hnRNP K-homologous Protein during Drosophila Embryo Development and Heat Shock. Flexibility of Transcription Sites In Vivo

Buchenau

Saumweber

Arndt‐Jovin

1997

The Journal of Cell Biology

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The Drosophila protein Hrb57A has sequence homology to mammalian heterogenous nuclear ribonucleoprotein (hnRNP) K proteins. Its in vivo distribution has been studied at high resolution by confocal laser scanning microscopy (CLSM) in embryos injected with fluorescently labeled monoclonal antibody. Injection of antibody into living embryos had no apparent deleterious effects on further development. Furthermore, the antibody-protein complex could be observed for more than 7 cell cycles in vivo, revealing a dynamic redistribution from the nucleus to cytoplasm at each mitosis from blastoderm until hatching. The evaluation of two- and three-dimensional CLSM data sets demonstrated important differences in the localization of the protein in the nuclei of living compared to fixed embryos. The Hrb57A protein was recruited to the 93D locus upon heat shock and thus serves as an in vivo probe for the activity of the gene in diploid cells of the embryo. Observations during heat shock revealed considerable mobility within interphase nuclei of this transcription site. Furthermore, the reinitiation as well as the down regulation of transcriptional loci in vivo during the recovery from heat shock could be followed by the rapid redistribution of the hnRNP K during stress recovery. These data are incompatible with a model of the interphase nucleus in which transcription complexes are associated with a rigid nuclear matrix.

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confidence: 96%

The Dynamic Nuclear Redistribution of an hnRNP K-homologous Protein during Drosophila Embryo Development and Heat Shock. Flexibility of Transcription Sites In Vivo

Buchenau

Saumweber

Arndt‐Jovin

1997

The Journal of Cell Biology

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A strand-specific switch in noncoding transcription switches the function of a Polycomb/Trithorax response element

et al. 2014

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Polycomb/Trithorax response elements (PRE/TREs) can switch their function reversibly between silencing and activation, by mechanisms that are poorly understood. Here we show that a switch in forward and reverse noncoding transcription from the Drosophila vestigial (vg) PRE/TRE switches the status of the element between silencing (induced by the forward strand) and activation (induced by the reverse strand). In vitro, both ncRNAs inhibit PRC2 histone methyltransferase activity, but in vivo only the reverse strand binds PRC2. Over-expression of the reverse strand evicts PRC2 from chromatin and inhibits its enzymatic activity. We propose that interactions of RNAs with PRC2 are differentially regulated in vivo, allowing regulated inhibition of local PRC2 activity. Genome-wide analysis shows that strand switching of ncRNAs occurs at several hundred PcG binding sites in fly and vertebrate genomes. This work identifies a novel and potentially widespread class of PRE/TREs that switch function by switching the direction of ncRNA transcription.

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A comparison of image restoration approaches applied to three‐dimensional confocal and wide‐field fluorescence microscopy

Verveer

Gemkow

Jovin

1999

Journal of Microscopy

125

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SummaryWe have compared different image restoration approaches for fluorescence microscopy. The most widely used algorithms were classified with a Bayesian theory according to the assumed noise model and the type of regularization imposed. We considered both Gaussian and Poisson models for the noise in combination with Tikhonov regularization, entropy regularization, Good's roughness and without regularization (maximum likelihood estimation). Simulations of fluorescence confocal imaging were used to examine the different noise models and regularization approaches using the mean squared error criterion. The assumption of a Gaussian noise model yielded only slightly higher errors than the Poisson model. Good's roughness was the best choice for the regularization. Furthermore, we compared simulated confocal and wide-field data. In general, restored confocal data are superior to restored wide-field data, but given sufficient higher signal level for the wide-field data the restoration result may rival confocal data in quality. Finally, a visual comparison of experimental confocal and wide-field data is presented.

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FISH in whole-mount Drosophila embryos. RNA: activation of a transcriptional locus, DNA: gene architecture and expression

Cited by 5 publications

References 65 publications

The Dynamic Nuclear Redistribution of an hnRNP K-homologous Protein during Drosophila Embryo Development and Heat Shock. Flexibility of Transcription Sites In Vivo

The Dynamic Nuclear Redistribution of an hnRNP K-homologous Protein during Drosophila Embryo Development and Heat Shock. Flexibility of Transcription Sites In Vivo

A strand-specific switch in noncoding transcription switches the function of a Polycomb/Trithorax response element

A comparison of image restoration approaches applied to three‐dimensional confocal and wide‐field fluorescence microscopy

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