Requirements for Glucose Beyond Sperm Capacitation During In Vitro Fertilization in the Mouse1
Alexander J. Travis, Levent Tütüncü, Carolina J. Jorgez, Teri S. Ord, Brian H. Jones, Gregory S. Kopf, Carmen J. Williams
Abstract
Alexander J. Travis, Levent Tütüncü, Carolina J. Jorgez, Teri S. Ord, Brian H. Jones, Gregory S. Kopf, Carmen J. Williams
Abstract
In both the mouse and the human, it is a point of controversy whether glucose is necessary for in vitro fertilization. Some of this controversy has resulted from a failure to distinguish between requirements for glucose during sperm capacitation versus requirements during the multistage process of fertilization. Using the mouse as a model, we performed a series of experiments designed to identify specific processes that might require glucose. We observed a positive correlation between increasing glucose concentrations during capacitation and fertilization, and increasing fertilization of zona pellucida (ZP)-intact eggs. These data supported a requirement for glucose in the fertilization medium even when sperm were first capacitated in the presence of 5.5 mM glucose. This glucose requirement was observed for both ZP-intact and ZP-free eggs. During ZP-free in vitro fertilization, some binding and fusion between the plasma membrane of the sperm and egg occurred in the absence of glucose and at concentrations less than 1 mM, suggesting that this substrate is not absolutely required. However, glucose concentrations of 1 mM or higher greatly facilitated both binding and fusion under these conditions. These subtle distinctions suggest that during ZP-free in vitro fertilization, 1 mM glucose represents a threshold level that facilitates binding and fusion. Taken as a whole, the data suggest requirements for glucose during both capacitation and fertilization under normal physiologic conditions.
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In both the mouse and the human, it is a point of controversy whether glucose is necessary for in vitro fertilization. Some of this controversy has resulted from a failure to distinguish between requirements for glucose during sperm capacitation versus requirements during the multistage process of fertilization. Using the mouse as a model, we performed a series of experiments designed to identify specific processes that might require glucose. We observed a positive correlation between increasing glucose concentrations during capacitation and fertilization, and increasing fertilization of zona pellucida (ZP)-intact eggs. These data supported a requirement for glucose in the fertilization medium even when sperm were first capacitated in the presence of 5.5 mM glucose. This glucose requirement was observed for both ZP-intact and ZP-free eggs. During ZP-free in vitro fertilization, some binding and fusion between the plasma membrane of the sperm and egg occurred in the absence of glucose and at concentrations less than 1 mM, suggesting that this substrate is not absolutely required. However, glucose concentrations of 1 mM or higher greatly facilitated both binding and fusion under these conditions. These subtle distinctions suggest that during ZP-free in vitro fertilization, 1 mM glucose represents a threshold level that facilitates binding and fusion. Taken as a whole, the data suggest requirements for glucose during both capacitation and fertilization under normal physiologic conditions.
Key concepts: Capacitation, Human fertilization, Biology, Sperm, Zona pellucida, In vitro fertilisation, In vitro, Andrology