Distribution of Polycomb - GFP fusion proteins in cell nuclei of Drosophila -

Movies and Images


During my time as a Post-Doc in the Lab of Renato Paro at the ZMBH in Heidelberg, Germany, I had the opportunity to study fly lines transgenic for a Polycomb-GFP fusion protein under the microscope. The lines were generated by Hartmut Niemann or Bodo Brückner in the Paro Lab. The results are described in the following article:
 
Dietzel, S., H. Niemann, B. Brückner, C. Maurange and R. Paro (1999).
The nuclear distribution of Polycomb during Drosophila melanogaster development shown with a GFP fusionprotein. Chromosoma , 108(2):83-94. ( Abstract and pdf-Article available at the journal's Web site. (pdf only if your institution subscribes to idealibrary.) Abstract available at " PubMed ")

The material presented here is meant as an addendum to this article. It contains Quicktime movies and images that didn't make it in the paper, mainly due to space restrictions. A very very short introduction is given below. For more detailed information about our findings and our interpretations please read the article.



Polycomb (PC) is a nuclear protein involved in the stable and heritable silencing of a variety of developmentally important genes. For more information about Polycomb see the Web site of the Paro Lab . GFP stands for green fluorescent protein. For more information about GFP see this article . For more information about Drosophila, check the Drosophila virtual library.

The material is grouped in three areas:
 

Visualization of the development of Drosophila embryos with a Polycomb-GFP fusionprotein

In the four movies presented here, PC-GFP is used as a nuclear marker to follow the development of the embryo over time, starting shortly before blastoderm stage. Two movies show the embryo from the blastula stage to the hatching of the larvae while the two others focus on the early stages.

Distribution of Polycomb during blastoderm mitosis

This page contains only one Quicktime movie, but a very nice one. It shows the distribution of the Polycomb-GFP fusion protein during two nuclear cycles in the blastoderm stage in high temporal and spatial resolution. File size of the movie: ~2.4 MBytes.

Distribution of Polycomb during spermatogenesis

Examples for the distribution of Polycomb in larval testis (1 image) and adult testis are given. 
This page is available in two versions: Version 1 contains "only" 8 images in various sizes. Version 2 contains in addition 3 embedded Quicktime movies with a combined (additional) file size of ~2.5 MBytes. Version 1 gives you the possibility to switch to Version two if you decide later that you want the movies. 

Methods

For detailed methods see the paper. The GFP used was EGFP from Clontech , a variety of the S65T mutation (see here for spectrum). Confocal images were recorded with the Leica TCS NT microscope of the ZMBH except otherwise noted. GFP was excited with 488 nm,  a dichroic mirror at 500 nm and a bandpass 500 - 550 nm were used. The images were transferred from the microscope to a Mac, further processed in NIH Image and saved as Quicktime Movies or combined to RGB images in Adobe Photoshop.


Technical note about the drosophila area on this web-site:

(You don't have to read this...)
I first set up this area while I was in Urbana on the local University server. When I moved from Urbana to Munich , I had to reconfigure the whole site. What's much worse: The URLs that some people had bookmarked and links from other Web-sites became suddenly invalid. That was one of the reasons why I got my own domain. However, the available space on this domain is small (1 Mbyte) and I cannot possibly store movies or even images on it. I solve this problem thus that the text pages are stored on dietzellab.de while the actual data files are loaded from the web server of my institution (currently the University of Munich, see homepage). This way a permanent Web-address is available without a big price tag for me.

Last Update: 4. September 2000
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