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1 October 2000 Radioimmunoassays for Bull Seminal Plasma Proteins (BSP-A1/-A2, BSP-A3, and BSP-30-Kilodaltons), and Their Quantification in Seminal Plasma and Sperm
Veronica Nauc, Puttaswamy Manjunath
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Three proteins, BSP-A1/-A2, BSP-A3, and BSP-30 kilodaltons (collectively called BSP proteins), represent the major proteins of bovine seminal plasma (BSP). At ejaculation, these proteins bind to the sperm surface and induce molecular changes in the plasma membrane that are deemed to be essential for sperm capacitation. The present study was carried out to develop specific radioimmunoassays (RIAs) for the quantification of each of the BSP proteins in BSP and sperm. RIAs were developed using polyclonal antibodies raised in rabbits against each BSP protein. The purified and iodinated BSP proteins were used as standard and tracer, respectively. The RIAs that were developed were shown to be specific for each protein and the cross-reactivity toward various antigens was negligible (<2%). The average sensitivity limit was 5 ng/ml of sample for BSP-A1/-A2 and BSP-A3, and 40 ng/ml of sample for BSP-30-kDa. The concentration of BSP proteins was determined by analyzing the RIA data with spline function. BSP proteins represented 40% to 57% of seminal plasma total protein (25% to 47% of BSP-A1/-A2, 3% to 5% of BSP-A3, and 3% to 7% of BSP-30 kDa) and 4% to 6% of sperm total protein (2.5% to 4% of BSP-A1/-A2, 0.4% to 0.9% of BSP-A3, and 0.5% to 1% of BSP-30-kDa). We also determined the concentration of BSP proteins that were sperm-bound after semen cryopreservation in Tris-egg yolk-glycerol extender. A significant decrease (70%–80%) in sperm-bound BSP proteins was noted after cryopreservation. The availability of reliable RIA procedures should aid in the further understanding of the role of BSP proteins in sperm function as well as their effect on sperm cryopreservation.

Veronica Nauc and Puttaswamy Manjunath "Radioimmunoassays for Bull Seminal Plasma Proteins (BSP-A1/-A2, BSP-A3, and BSP-30-Kilodaltons), and Their Quantification in Seminal Plasma and Sperm," Biology of Reproduction 63(4), 1058-1066, (1 October 2000).
Received: 26 January 2000; Accepted: 1 May 2000; Published: 1 October 2000
seminal vesicles
sperm capacitation/acrosome reaction
sperm maturation
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