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Utility of human placental alkaline phosphatase as a genetic marker for cell tracking in bone and cartilage

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Abstract

It was the aim of the current study to evaluate the utility of human placental alkaline phosphatase (hPLAP) as a genetic marker for cell tracking in bone and cartilage, using transgenic Fischer 344 rats expressing hPLAP under the control of the ubiquitous R26 promoter [F344-Tg(R26-hPLAP)]. hPLAP enzyme activity was retained during paraffin and methylmethacrylate (MMA) embedding, and was best preserved using 40% ethanol as fixative. Endogenous alkaline phosphatase activity could be completely blocked by heat inactivation in paraffin and MMA sections, allowing histochemical detection of hPLAP in the complete absence of background staining. In addition, sensitive detection of hPLAP was also possible using immunohistochemistry. F344-Tg(R26-hPLAP) rats demonstrated ubiquitous expression of hPLAP in hematopoietic bone marrow cells and stromal cells such as osteoblasts, osteocytes, and chondrocytes. Osteoclasts only weakly expressed hPLAP. In conclusion, hPLAP provides superb detection quality in paraffin and plastic sections, and constitutes an excellent genetic marker for cell tracking in hard and soft tissues.

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Abbreviations

APES:

3-Aminopropyltriethoxy-silane

BCIP:

5-Bromo-4-chloro-3-indolyl phosphate

EDTA:

Ethylene diamino tetra acetate

F344:

Fischer 344

GFP:

Green fluorescent protein

hPLAP:

Human placental alkaline phosphatase

MMA:

Methylmethacrylate

PFA:

Paraformaldehyde

R26:

0.8 kb piece of the ROSA βgeo 26 promoter

Tg:

Transgenic

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Acknowledgments

The authors thank Claudia Bergow for excellent technical assistance. This work was supported by Deutsche Forschungsgemeinschaft (Er 223/8-1) and by the University of Veterinary Medicine, Vienna.

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Authors and Affiliations

  1. Institute of Physiology, Physiological Chemistry and Animal Nutrition, Ludwig Maximilians University, Munich, 80539, Germany

    Nina J. Unger, Karin Weber & Reinhold G. Erben

  2. Institute of Pathophysiology, Department of Natural Sciences, University of Veterinary Medicine, Veterinaerplatz 1, 1210, Vienna, Austria

    Kathrin I. Odörfer & Reinhold G. Erben

  3. Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Eric P. Sandgren

Authors
  1. Nina J. Unger
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  2. Kathrin I. Odörfer
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  3. Karin Weber
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  4. Eric P. Sandgren
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  5. Reinhold G. Erben
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Correspondence to Reinhold G. Erben.

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Unger, N.J., Odörfer, K.I., Weber, K. et al. Utility of human placental alkaline phosphatase as a genetic marker for cell tracking in bone and cartilage. Histochem Cell Biol 127, 669–674 (2007). https://doi.org/10.1007/s00418-007-0286-6

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  • DOI: https://doi.org/10.1007/s00418-007-0286-6

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