Molecular cloning and characterisation of heparanase mRNA in the porcine placenta throughout gestation
Jeremy R. Miles A B , Jeffrey L. Vallet A , Brad A. Freking A and Dan J. Nonneman AA US Department of Agriculture – Agricultural Research Service, Roman L. Hruska US Meat Animal Research Center, Clay Center, NE 68933, USA.
B Corresponding author. Email: jeremy.miles@ars.usda.gov
Reproduction, Fertility and Development 21(6) 757-772 https://doi.org/10.1071/RD09041
Submitted: 25 February 2009 Accepted: 10 May 2009 Published: 1 July 2009
Abstract
Heparanase (HPSE) is an endoglycosidase that specifically degrades heparan sulfate, which is an abundant glycosaminoglycan of the pig placenta. The aim of the present study was to clone cDNA encoding porcine HPSE and characterise the expression level and localisation of HPSE mRNA in porcine placentas throughout gestation. Placental tissues were collected from litters on Days 25, 45, 65, 85 and 105 of gestation. Three transcript variants similar to HPSE were identified in the pig placenta. In addition, the HPSE gene was mapped to pig chromosome 8 in close proximity to quantitative trait loci for litter size and prenatal survival. Real-time polymerase chain reaction and in situ hybridisation were used to characterise the expression of two HPSE variants, namely HPSE v1 and v2, in the pig placenta throughout gestation. The expression of HPSE v1 and v2 was elevated (P < 0.01) in placentas during very early gestation (Day 25) as well as during late gestation (Days 85 and 105). Finally, HPSE v1 and v2 mRNA were localised to the cuboidal trophoblast cells of the folded bilayer located nearest to the maternal endometrium. These findings illustrate that HPSE likely plays a role in the development and modification of the pig placenta, which has implications for litter size and prenatal survival.
Additional keywords: extracellular matrix, gene expression, glycosaminoglycans.
Acknowledgements
The authors thank Susan Hassler and Troy Gramke for technical assistance in the collection and processing of samples, David Sypherd for assistance with in situ hybridisation, Alan Kruger for assistance with microscopy, Susan Hauver for assistance with radiation hybrid mapping, the USMARC swine crew for animal husbandry, the USMARC abattoir crew for assistance with killing of the pigs and Drs David Guthrie (US Department of Agriculture – Agricultural Research Service, Beltsville Agricultural Research Center) and Leah Zorrilla (US Environmental Protection Agency, National Health and Environmental Effects Research Laboratory) for critical review of this manuscript. This research was supported by USDA-ARS, CRIS Project No. 5438–31000–084–00D.
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