Adsorption of oviductal fluid proteins by the bovine sperm membrane during in vitro capacitation

Tamara McNutt, Lee Rogowski, Regina Vasilatos‐Younken, Gary Killian

Research output: Contribution to journalArticle

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Abstract

125I‐labeled oviductal fluid (ODF) proteins and antiserum to ODF were used to determine whether ODF proteins associate with the sperm membrane during in vitro capacitation. Luteal and nonluteal pools of ODF were obtained from oviduct catheters during the estrous cycle. Washed sperm (50 × 106 sperm/ml) were incubated up to 4 h in a protein‐free modified Tyrode's medium (MTM), or MTM supplemented with 40% ODF, or 0.5 ng 125I‐labeled ODF proteins. Solubilized sperm membrane proteins and incubatin media containing ODF proteins were separated by gel electrophoresis. Membranes isolated from bovine sperm previously incubated with ODF, absorbed five 125I‐proteins: A doublet at 85–95 kDa, and others at 24, 34, 53, and 66 kDa. The amount of 66 kDA 125I‐protein associated with the sperm decreased during the incubation, whereas the amount of 85 to 95‐kDa protein did not. Western blot analyses also detected the presence of ODF proteins (53, 66, 85–95, and 116 kDa) in solubilized membranes from sperm incubated in ODF. The 85 to 95‐kDa protein in ODF decreased in ODF. The 85 to 95kDa protein in ODF decreased in apparent molecular weight by 5 kDa when associated with the sperm membrane. At 53 kDa ODF proteins which associated with sperm were transformed from two to three separate proteins. These studies indicate that the surface of sperm is modified by adsorption of ODF proteins ot the membrane during in vitro capacitation. (c) 1992 Wiley‐Liss, Inc.

Original languageEnglish (US)
Pages (from-to)313-323
Number of pages11
JournalMolecular reproduction and development
Volume33
Issue number3
DOIs
StatePublished - Nov 1992

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Adsorption
Spermatozoa
Membranes
Proteins
Membrane Proteins
In Vitro Techniques
Estrous Cycle
Oviducts
Corpus Luteum
Electrophoresis
Immune Sera
Catheters
Molecular Weight
Western Blotting
Gels

All Science Journal Classification (ASJC) codes

  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

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title = "Adsorption of oviductal fluid proteins by the bovine sperm membrane during in vitro capacitation",
abstract = "125I‐labeled oviductal fluid (ODF) proteins and antiserum to ODF were used to determine whether ODF proteins associate with the sperm membrane during in vitro capacitation. Luteal and nonluteal pools of ODF were obtained from oviduct catheters during the estrous cycle. Washed sperm (50 × 106 sperm/ml) were incubated up to 4 h in a protein‐free modified Tyrode's medium (MTM), or MTM supplemented with 40{\%} ODF, or 0.5 ng 125I‐labeled ODF proteins. Solubilized sperm membrane proteins and incubatin media containing ODF proteins were separated by gel electrophoresis. Membranes isolated from bovine sperm previously incubated with ODF, absorbed five 125I‐proteins: A doublet at 85–95 kDa, and others at 24, 34, 53, and 66 kDa. The amount of 66 kDA 125I‐protein associated with the sperm decreased during the incubation, whereas the amount of 85 to 95‐kDa protein did not. Western blot analyses also detected the presence of ODF proteins (53, 66, 85–95, and 116 kDa) in solubilized membranes from sperm incubated in ODF. The 85 to 95‐kDa protein in ODF decreased in ODF. The 85 to 95kDa protein in ODF decreased in apparent molecular weight by 5 kDa when associated with the sperm membrane. At 53 kDa ODF proteins which associated with sperm were transformed from two to three separate proteins. These studies indicate that the surface of sperm is modified by adsorption of ODF proteins ot the membrane during in vitro capacitation. (c) 1992 Wiley‐Liss, Inc.",
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Adsorption of oviductal fluid proteins by the bovine sperm membrane during in vitro capacitation. / McNutt, Tamara; Rogowski, Lee; Vasilatos‐Younken, Regina; Killian, Gary.

In: Molecular reproduction and development, Vol. 33, No. 3, 11.1992, p. 313-323.

Research output: Contribution to journalArticle

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T1 - Adsorption of oviductal fluid proteins by the bovine sperm membrane during in vitro capacitation

AU - McNutt, Tamara

AU - Rogowski, Lee

AU - Vasilatos‐Younken, Regina

AU - Killian, Gary

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AB - 125I‐labeled oviductal fluid (ODF) proteins and antiserum to ODF were used to determine whether ODF proteins associate with the sperm membrane during in vitro capacitation. Luteal and nonluteal pools of ODF were obtained from oviduct catheters during the estrous cycle. Washed sperm (50 × 106 sperm/ml) were incubated up to 4 h in a protein‐free modified Tyrode's medium (MTM), or MTM supplemented with 40% ODF, or 0.5 ng 125I‐labeled ODF proteins. Solubilized sperm membrane proteins and incubatin media containing ODF proteins were separated by gel electrophoresis. Membranes isolated from bovine sperm previously incubated with ODF, absorbed five 125I‐proteins: A doublet at 85–95 kDa, and others at 24, 34, 53, and 66 kDa. The amount of 66 kDA 125I‐protein associated with the sperm decreased during the incubation, whereas the amount of 85 to 95‐kDa protein did not. Western blot analyses also detected the presence of ODF proteins (53, 66, 85–95, and 116 kDa) in solubilized membranes from sperm incubated in ODF. The 85 to 95‐kDa protein in ODF decreased in ODF. The 85 to 95kDa protein in ODF decreased in apparent molecular weight by 5 kDa when associated with the sperm membrane. At 53 kDa ODF proteins which associated with sperm were transformed from two to three separate proteins. These studies indicate that the surface of sperm is modified by adsorption of ODF proteins ot the membrane during in vitro capacitation. (c) 1992 Wiley‐Liss, Inc.

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