Abstract
Aims/hypothesis
Long-term labelling of mice with halogenated thymidine analogues is an established method for quantifying the contribution of beta cell proliferation to in vivo beta cell mass expansion in (re)generation models. The method is believed to give accurate information on the accrued number of cycling beta cells over a period of time. Multiple thymidine analogue labelling is applied for evaluating the duration of postmitotic quiescence in beta cells. We hypothesise, however, that long-term labelling by thymidine analogues hampers beta cell proliferation.
Methods
Thymidine analogues were administered for 7–14 days via the i.p. route to neonatal mice, or via drinking water to young mice with normal pancreases or adult mice with injured pancreases. The proliferation of insulin-positive cells was assessed by their expression of the proliferation markers Ki67 or phosphorylated histone H3 and by their incorporation of nucleotide analogues.
Results
In the mouse models of beta cell proliferation investigated herein, long-term administration of thymidine analogues decreased the percentage of Ki67+ and phosphorylated histone H3+ beta cells as compared with administration of normal drinking water. Proliferation was restored by washout of the analogue. Labelling with one analogue decreased the subsequent incorporation of another analogue by beta cells.
Conclusions/interpretation
Long-term labelling with halogenated thymidine analogues is a biased method that underestimates the proliferation and re-division potential of mouse beta cells.
Abbreviations
- CldU:
-
5-Chloro-2′-deoxyuridine
- IdU:
-
5-Iodo-2′-deoxyuridine
- PDL:
-
Partial duct ligation
- pHH3:
-
Phosphorylated histone H3
- PPX:
-
Partial pancreatectomy
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Acknowledgements
Special thanks to Ann Demarré, Veerle Laurysens, Vincent Vandenbroucke, Jan de Jonge and Erik Quartier (Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium) for technical assistance.
Funding
Financial support was received from the VUB Research Council (HH, MVDC), Interuniversity Attraction Pole networks (HH), the Fund for Scientific Research Flanders (HH) and Stichting Diabetes Onderzoek Nederland (HH).
Duality of interest
The authors declare that there is no duality of interest associated with this manuscript.
Contribution statement
YC, GL and MVDC provided and analysed data and drafted the manuscript. MVDC and HH designed experiments and critically revised the manuscript. All authors approved the final version of the manuscript.
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Van de Casteele, M., Cai, Y., Leuckx, G. et al. Mouse beta cell proliferation is inhibited by thymidine analogue labelling. Diabetologia 56, 2647–2650 (2013). https://doi.org/10.1007/s00125-013-3049-z
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DOI: https://doi.org/10.1007/s00125-013-3049-z