T J Davies1, R L Gardner. 1. Mammalian Development Laboratory, University of Oxford, Department of Zoology, South Parks Road, Oxford OX1 3PS, UK.
Abstract
BACKGROUND: Marking experiments using phytohaemagglutinin (PHA) and Concanavalin A (ConA) have suggested that the first cleavage plane is related to the point of sperm entry. Because of concerns about the specificity of lectin binding, the distribution of sperm components has been investigated directly. METHODS AND RESULTS: The sperm tail could be identified in cleaving zygotes and early 2-cell stages following their permeabilization and exposure to Oregon Green Paclitaxel. At neither stage did the anterior end of the tail, which lies initially at the site of sperm entry, bear a consistent relationship to the first cleavage plane, even when it had clearly retained its original location. Moreover, using artificial insemination with MitoTracker-labelled sperm, the midpiece was found to remain associated with anterior end of the tail through to the 2-cell stage. Lectins showed no discernible binding to the fertilization cone of mechanically denuded zygotes and very strong binding to the zona pellucida. Moreover, after general labelling of zygotes with either ConA or PHA, persisting surface lectin tended to be concentrated towards the cleavage plane. CONCLUSION: The present findings challenge the claim that the sperm specifies the plane of first cleavage, and also question the methodology on which it was based.
BACKGROUND: Marking experiments using phytohaemagglutinin (PHA) and Concanavalin A (ConA) have suggested that the first cleavage plane is related to the point of sperm entry. Because of concerns about the specificity of lectin binding, the distribution of sperm components has been investigated directly. METHODS AND RESULTS: The sperm tail could be identified in cleaving zygotes and early 2-cell stages following their permeabilization and exposure to Oregon Green Paclitaxel. At neither stage did the anterior end of the tail, which lies initially at the site of sperm entry, bear a consistent relationship to the first cleavage plane, even when it had clearly retained its original location. Moreover, using artificial insemination with MitoTracker-labelled sperm, the midpiece was found to remain associated with anterior end of the tail through to the 2-cell stage. Lectins showed no discernible binding to the fertilization cone of mechanically denuded zygotes and very strong binding to the zona pellucida. Moreover, after general labelling of zygotes with either ConA or PHA, persisting surface lectin tended to be concentrated towards the cleavage plane. CONCLUSION: The present findings challenge the claim that the sperm specifies the plane of first cleavage, and also question the methodology on which it was based.