Identification of heat shock protein 10 within the equine embryo, endometrium, and maternal peripheral blood mononuclear cells.
Abstract: Early pregnancy factor has been identified as a 10-kDa extracellular homolog of heat shock protein 10 (Hsp10). Hsp10 has been detected during early pregnancy in serum of mice, sheep, pigs, horses, cows, and humans by the rosette inhibition test. Hsp10 has also been associated with several neoplastic and autoimmune diseases. The goal of the present study was to determine if Hsp10 could be detected in the early equine embryo through the use of immunohistochemistry and quantitative real-time PCR. Additionally, analysis of systemically harvested peripheral blood mononuclear cells (PBMCs) from both pregnant and nonpregnant mares was evaluated to determine expression levels of HSP10. Embryos were collected from Quarter Horse mares by uterine lavage at either 8 or 25 days after ovulation. Collection and separation of PBMCs occurred on Day 8 for both pregnant and nonpregnant mares. Immunohistochemistry revealed cytoplasmic localization of HSP10 throughout the single layer of ectodermal cells forming the trophoblast in Day-8 embryos. Day-25 embryos demonstrated intense localization focally along the apical border of ectodermal cells forming the trophoblast layer of the developing chorion. There was no nuclear staining in either embryonic population. Quantitative real-time PCR detected the presence of mRNA for HSP10 in both 8- and 25-day equine embryos. Day-25 embryos exhibited an elevated degree of expression (P = 0.006) compared with the 8-day embryos for HSP10. Endometrial samples did not display any significant difference in degree of expression for HSP10 (P = 0.10). Finally, PBMCs from pregnant mares demonstrated elevated (P = 0.03) expression of HSP10 compared to the nonpregnant mares on Day 8 of the estrous cycle. This study confirmed the presence of HSP10 protein and mRNA expression of HSP10 in equine embryos at two maturation stages. Additionally, the presence of increased gene expression within PBMCs of pregnant mares suggests communication, possibly leading to necessary immunomodulatory effects between the embryo and mare.
Published by Elsevier Inc.
Publication Date: 2014-11-26 PubMed ID: 25542459DOI: 10.1016/j.theriogenology.2014.11.020Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article investigates the presence and role of the heat shock protein 10 (Hsp10) in early horse pregnancy. The study finds that Hsp10 can be identified in the early equine embryo and in peripheral blood mononuclear cells (PBMCs) and may play a role in communication between the embryo and mare, perhaps regarding immunomodulatory effects.
Research Focus and Methodology
- The main aim of the study was to identify whether Hsp10 could be detected in the early equine embryo.
- The study used immunohistochemistry and quantitative real-time PCR to detect the presence of Hsp10.
- The embryos were collected from Quarter Horse mares by uterine lavage either on Day 8 or Day 25 after ovulation.
- The collection and separation of peripheral blood mononuclear cells (PBMCs) from both pregnant and nonpregnant mares took place on Day 8.
Results and Findings
- The study found cytoplasmic localization of Hsp10 in the ectodermal cells forming the trophoblast in Day-8 embryos.
- The Day-25 embryos had intense Hsp10 localization along the apical border of ectodermal cells forming the trophoblast layer of the developing chorion, though there was no Hsp10 staining in the nuclei.
- Quantitative real-time PCR showed that Hsp10 gene (HSP10) was present in both 8- and 25-day equine embryos.
- The Day-25 embryos had a higher level of HSP10 gene expression compared to the Day-8 embryos.
- No significant difference in HSP10 gene expression was observed in the endometrial samples.
- PBMCs from pregnant mares had a higher expression of the HSP10 gene compared to nonpregnant mares on Day 8 of the estrous cycle.
Conclusion and Implications
- The research confirmed the presence of Hsp10 protein and HSP10 gene expression in equine embryos at two maturation stages.
- The higher HSP10 gene expression in PBMCs from pregnant mares suggests possible communication between the embryo and mare, which could lead to necessary immunomodulatory effects.
- This finding not only furthers our understanding of early equine pregnancy but also potentially contributes to our understanding and treatment of autoimmune and neoplastic diseases with which Hsp10 has been associated.
Cite This Article
APA
Hatzel JN, Bouma GJ, Cleys ER, Bemis LT, Ehrhart EJ, McCue PM.
(2014).
Identification of heat shock protein 10 within the equine embryo, endometrium, and maternal peripheral blood mononuclear cells.
Theriogenology, 83(5), 832-839.
https://doi.org/10.1016/j.theriogenology.2014.11.020 Publication
Researcher Affiliations
- Equine Reproduction Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA. Electronic address: Patrick.McCue@colostate.edu.
- Animal Reproduction and Biotechnology Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
- Animal Reproduction and Biotechnology Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
- Department of Cell Biology, Stem Cells and Development, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.
- Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA.
- Equine Reproduction Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
MeSH Terms
- Animals
- Chaperonin 10 / genetics
- Chaperonin 10 / metabolism
- Endometrium / metabolism
- Female
- Gene Expression Regulation, Developmental
- Horses / embryology
- Horses / metabolism
- Immunohistochemistry / veterinary
- Leukocytes, Mononuclear / metabolism
- Pregnancy
- Real-Time Polymerase Chain Reaction / methods
- Real-Time Polymerase Chain Reaction / veterinary
- Reverse Transcriptase Polymerase Chain Reaction / methods
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
Citations
This article has been cited 4 times.- Lawson EF, Grupen CG, Baker MA, Aitken RJ, Swegen A, Pollard CL, Gibb Z. Conception and early pregnancy in the mare: lipidomics the unexplored frontier. Reprod Fertil 2022 Jan 1;3(1):R1-R18.
- Mezni A, Khazri A, Khazri O, Limam F, Cosette P, Aouani E. Neuroprotective Activity of Grape Seed and Skin Extract Against Lithium Exposure Using Proteomic Research. Mol Neurobiol 2017 May;54(4):2720-2730.
- Rosato E, Farris M, Bastianelli C. Mechanism of Action of Ulipristal Acetate for Emergency Contraception: A Systematic Review. Front Pharmacol 2015;6:315.
- Lawson EF, Pickford R, Aitken RJ, Gibb Z, Grupen CG, Swegen A. Mapping the lipidomic secretome of the early equine embryo. Front Vet Sci 2024;11:1439550.
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