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1 Anatomy, University of Zuerich, Zuerich, Zuerich, Switzerland
2 Anatomy, University Zuerich, Zuerich, Zuerich, Switzerland
3 Institute of Anatomy, University of Zurich, Zurich, Switzerland
* To whom correspondence should be addressed. E-mail: lehir{at}anatom.unizh.ch.
We searched for evidence for a contribution of stem cells in growth of the proximal S3 segments of healthy rats. According to the stem cell model stem cells are undifferentiated and slow-cycling; the bulk of cycling cells are transit amplifying rapidly cycling cells. We show (1) by continuous application of a thymidine analogue (ThA) for 7 days that S3 proximal epithelial cells in healthy kidneys display a high cycling rate; (2) that slow-cycling cells, identified by lack of ThA uptake during 14 days of continuous ThA application up to sacrifice and by expression of the cell cycle protein Ki67 at sacrifice, have the same degree of differentiation as quiescent cells. (3) To detect rapidly cycling cells rats were sacrificed at various time points after injection of a ThA. Double immunofluorescence for ThA and for a cell cycle marker was performed, co-localization indicating successive divisions. During one week after division daughter cells display a very low proliferation rate, indicating the absence of rapidly cycling cells. (4) Labelling with cyclin D1 showed that this low proliferation rate is due to cycle arrest. (5) More than 50% of the S3 cells entered the cell cycle 36 hours after a potent proliferative stimulus (lead acetate injection). We conclude that generation of new cells in the proximal tubule relies on division of differentiated, normally slow cycling cells. These may rapidly enter the cycle under an adequate stimulus.
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