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Localization of parathyroid hormone-related protein in the preimplantation mouse embryo is associated with events of blastocyst hatching

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To determine the pattern of expression of parathyroid hormone-related protein (PTHrP) and its receptor, parathyroid hormone receptor 1 (PTHR1), in mouse embryos in different stages of preimplantation development.

Methods

Embryos were cultured from the pronuclear zygote stage and harvested as 2-cell, 4-cell and 8-cell embryos, morulae and blastocysts. RT-PCR was carried out on mRNAs of these and of trophoblast outgrowths for detection of PTHrP and PTHR1. Whole mounted embryos intact or stripped of zonae pellucidae were immunofluorescently stained for PTHrP and PTH receptor and observed with confocal microscopy.

Results

PTHrP mRNA was present in the pronuclear zygote, not present in 2-cell, 4-cell and uncompacted 8-cell embryos, present in the 8-cell compacting embryo, and not detected in 16-cell morulae or blastocysts. The mRNA was present in trophoblasts growing on fibronectin beds. mRNA for PTHR1 was detected in the pronuclear zygote, then undetected until the compacted 8-cell stage and thereafter. PTH receptor protein was observed in 2-cell embryos, morulae and in the inner cell mass and trophectoderm of blastocysts. PTHrP was observed dispersed in the cytoplasm of 2-cell, 4-cell and uncompacted 8-cell embryos, and in distinct foci near the nuclei of morulae. In blastocysts, PTHrP appeared on the apical surface of only trophoblast cells which had extruded from the zona pellucida. Fully hatched blastocysts expressed the protein on the apical side of all trophoblasts. When morulae were prematurely stripped of their zonae, PTHrP was observed on the embryos’ outer surface.

Conclusions

PTHrP protein is expressed throughout early embryo development, and its receptor PTHR1 is expressed from the morula stage. Embryo hatching is associated with translocation of PTHrP to the apical plasma membrane of trophoblasts. PTHrP may thus have autocrine effects on the developing blastocyst.

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Acknowledgements

The authors thank Dr. Lynda McGinnis for advice and technical assistance, and Mr. Michael Raffin for help with confocal microscopy.

This work was supported by a March of Dimes Grant (RAN), NIH HD30496 (RAN) and NIH UO1-HD21988 (JDB).

GTE and RAN recognize with special gratitude and admiration the guidance and support of John D. Biggers, Ph.D., D.Sc. He has throughout his career exemplified rigorous adherence to the scientific method, the degrees of creativity and imagination with which it can be applied, and its value in informing all other scholarship. We are honored to be associated with him, and hope to continue to prove his influence in our endeavors.

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Authors and Affiliations

  1. Department of Animal Sciences, University of Illinois, 310 Animal Science Laboratory, 1207 W. Gregory Drive, Urbana, IL, 61801, USA

    Gregory T. Erbach & Romana A. Nowak

  2. Department of Cell Biology, Harvard Medical School, Boston, MA, USA

    John D. Biggers & Peter C. Manning

Authors
  1. Gregory T. Erbach
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  2. John D. Biggers
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  3. Peter C. Manning
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  4. Romana A. Nowak
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Corresponding author

Correspondence to Romana A. Nowak.

Additional information

Capsule Parathyroid hormone-related protein (PTHrP) is expressed by preimplantation embryos and its localization is markedly altered in trophectoderm cells during blastocyst hatching.

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Erbach, G.T., Biggers, J.D., Manning, P.C. et al. Localization of parathyroid hormone-related protein in the preimplantation mouse embryo is associated with events of blastocyst hatching. J Assist Reprod Genet 30, 1009–1015 (2013). https://doi.org/10.1007/s10815-013-0094-y

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