Multiple proteins from ejaculated and epididymal porcine spermatozoa bind glycan motifs found in the oviduct

E. Silva, G. Kadirvel, R. Jiang, N. Bovin, D. Miller

Research output: Contribution to journalArticlepeer-review


Previously, we demonstrated that porcine spermatozoa bind selectively to Lewisx (LeX) and bi-antennary α2-6 sialylated N-acetyllactosamine (bi-SiaLN) glycan motifs found in the oviduct. Recognition of these motifs mediates sperm binding to oviduct epithelial cells to form a reservoir. Here, we characterize proteins that bind to LeX and bi-SiaLN in ejaculated and cauda epididymal spermatozoa. Fluoresceinated 3′-O-sulphated-LeX (suLeX) and bi-SiaLN were used to locate the binding proteins on spermatozoa. Binding to N-acetyllactosamine (LN), a glycan that spermatozoa bind less, was assessed as a control. Over 60% of ejaculated and epididymal spermatozoa bound to suLeX and bi-SiaLN, but only 25% bound to LN. Specific binding to suLeX and bi-SiaLN required divalent cations. Fluoresceinated suLeX bound mostly to the plasma membrane at the apex of the head, whereas bi-SiaLN bound to a broader area of the head. Further characterization of receptors was accomplished by probing, with specific glycans, sperm proteins that had been separated by SDS-PAGE and transferred to blots. Glycan blot assays identified several suLeX-binding bands that migrated as 12-250 kDa. Migration of some of the bands that bound bi-SiaLN was similar to those that bound suLeX. A 12-kDa band protein was detected in seminal fluid and ejaculated spermatozoa, but absent in epididymal spermatozoa. In summary, we determined that: (i) multiple glycan-binding proteins recognize suLeX and bi-SiaLN motifs, (ii) specific binding required divalent cations and (iii) glycan-binding proteins were identified before and after sperm contact with accessory gland fluids. While the identity of these molecules is still unknown, this information predicts characteristics of the authentic oviduct suLeX and bi-SiaLN receptors on spermatozoa.

Original languageEnglish (US)
Pages (from-to)763-771
Number of pages9
Issue number5
StatePublished - Sep 2014


  • Carbohydrates
  • Glycan-binding proteins
  • Glycans
  • Lewis X
  • Oviduct
  • Sialic acid
  • Spermatozoa

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Reproductive Medicine
  • Endocrinology
  • Urology


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