Abstract

Recent evidence supports involvement of the acute phase protein haptoglobin in numerous events during mammalian reproduction. The present study represents an in-depth investigation of haptoglobin expression and secretion in the porcine oviduct and uterus, and assesses its effect on porcine in vitro embryo production. A systematic study was made of sows in different oestrous stages: late follicular, early luteal and late luteal stages. Relative haptoglobin mRNA abundance was quantified by RT-qPCR. In addition, expression of the protein was analysed by immunohistochemistry and the results were complemented by Western-blot and proteomic analyses of the oviductal and uterine fluids. In vitro porcine fertilization and embryo culture were carried out in the presence of haptoglobin. The results indicate that haptoglobin mRNA expression in the porcine oviduct and uterus is most abundant during the late luteal stage of the oestrous cycle. By means of Western blot and proteomic analyses haptoglobin presence was demonstrated in the oviduct epithelium and in the oviductal and uterine fluids in different stages of the oestrous cycle. The addition of haptoglobin during gamete co-incubation had no effect on sperm penetration, monospermy or efficiency rates; however, compared with the control group, blastocyst development was significantly improved when haptoglobin was present (haptoglobin: 64.50% vs. control: 37.83%; p < 0.05). In conclusion, the presence of haptoglobin in the oviduct and uterus of sows at different stages of the oestrous cycle suggests that it plays an important role in the reproduction process. The addition of haptoglobin during in vitro embryo production improved the blastocyst rates.

Highlights

  • Recent evidence supports involvement of the acute phase protein haptoglobin in numerous events during mammalian reproduction

  • Haptoglobin has traditionally been considered as an acute phase p­ rotein[32,33,34,35] that acts within the physiological context of an inflammatory response, since it is present during infection, inflammation, traumatic damage and malignant proliferation

  • In light of the above, the aim of this study was to analyze the following in porcine: (1) the relative abundance of mRNA codifying for haptoglobin protein in the oviduct and uterus, (2) the presence of the protein in oviductal fluid and uterine fluid in different phases of the oestrous cycle, (3) the localization of haptoglobin protein in the oviduct during the oestrous cycle, (4) the haptoglobin function by means of functional analyses during in vitro embryo production

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Summary

Introduction

Recent evidence supports involvement of the acute phase protein haptoglobin in numerous events during mammalian reproduction. The presence of haptoglobin in the oviduct and uterus of sows at different stages of the oestrous cycle suggests that it plays an important role in the reproduction process. Additional information is provided by knock-out mice for this gene These animals are viable and fertile, there is a small but significant adverse effect on the postnatal viability of mice, suggesting that the hemoglobin-haptoglobin complex plays an important role in the amelioration of tissue damage caused by lipid p­ eroxidation[37]. The physiological function of haptoglobin within the reproductive tissues remains u­ nclear[41]

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