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Received April 23, 2003
Revised May 27, 2003
Accepted after revision June 12, 2003
1 University of Oslo
* To whom correspondence should be addressed. E-mail: kgunder{at}bio.uio.no.
We here present a new technique with single cell injection of labelled DNA to visualize nuclei in living muscle fibres in the intact animal. This approach allowed us to determine the position of all the nuclei within a sarcolemma without labelling satellite cells. In contrast to what has been found in tissue culture, we found that the nuclei are immobile even when observed over several days. Nucleic density was uniform along the fibre except for higher densities at the endplate, and at some myotendenous junctions. The peri-junctional region had the same number of nuclei as the rest of the fibre. In the EDL muscle the extra-junctional nuclei were elongated and precisely aligned to the length axis of the fibre. In the soleus the nuclei were rounder, and not well aligned. When comparing small and large fibres in the soleus the number of nuclei varied approximately proportionally to cytoplasmic volume, while in the EDL it was proportional to surface area. A statistical analysis revealed that nuclei are not randomly distributed in either EDL or soleus. For each fibre actual distributions were compared with computer simulations where nuclei were assumed to repel each other, which is optimizing the nucleic distribution with respect to minimizing transport distances. The simulated patterns are regular and with clear row like structures when the density of nuclei is low. The non-random and often row like distribution of nuclei observed in muscle fibres may thus reflect regulatory mechanisms where nuclei repel each other aimed at minimizing transport distances.
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