Localization of parathyroid hormone-related protein in the preimplantation mouse embryo is associated with events of blastocyst hatching.
OBJECTIVE
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.
Department of Animal Sciences, University of Illinois, 310 Animal Science Laboratory, 1207 W. Gregory Drive, Urbana, IL, 61801, USA.
Gregory T
GT
Erbach
John D
JD
Biggers
Peter C
PC
Manning
Romana A
RA
Nowak
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D047108:Embryonic Development; D051379:Mice; D044162:Parathyroid Hormone-Related Protein / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D015044:Zona Pellucida / Q000378:metabolism / Q000648:ultrastructure
D044162:Parathyroid Hormone-Related Protein