A Possible Role for the Pentose Phosphate Pathway: MATERIALS AND METHODS(1)

MATERIALS AND METHODS(1)

Chemicals and Media

The basic culture medium used in all experiments was M16 containing 23.3 mM lactate, 0.33 mM pyruvate, and 5.56 mM glucose, and supplemented with 15 mg/ml type V BSA. Cytochalasin B, phloretin, and iodoacetate were dissolved as stock solutions (1000 X concentrated in dimethyl sulfoxide) and stored at -20°C. Dilutions in culture medium were made before use. All chemicals were obtained from Sigma Pharmaceuticals (Buchs, Switzerland). The antibodies anti-GLUT3 and anti-GLUT1 were kindly provided by Dr. G.W. Gould (University of Glasgow, Scotland) and Dr. B. Thorens (University of Lausanne, Switzerland), respectively. buy cipro

Oocyte Preparation

Oocytes were obtained from 3- to 4-wk-old female B6D2F1 mice (IFFA-CREDO, L’Arbresle, France), which received injections of 5 IU eCG (Folligon; Veterinaria, Zurich, Switzerland) followed 48 h later by 5 IU hCG (Chol-uron; Veterinaria). Fourteen to 16 h after administration of hCG, oocytes surrounded by their cumulus cells were collected. After cumulus digestion by 0.2 mg/ml hyaluronidase (Sigma), zonae pellucidae were mechanically removed as we have previously described.

This entry was posted in Spermatozoa and tagged Gamete Fusion, Pentose Phosphate Pathway, Spermatozoa.