Collection and proteomic analysis of human tubal fuid

Introduction: The fluid present within the human fallopian tube mediates gamete transport, maturation, fertilization and early embryo development. The collection of tubal fluid is highly problematic, previously being achieved by cannulation for several days, resulting in trauma and inflammation to the tube. The aim of our study was to assess the efficacy of surgical methods for the sterile retrieval of tubal fluid and to perform a proteomic analysis of tubal proteins. Materials and methods: The collection of tubal fluid was performed in 19 infertile women undergoing diagnostic procedures during the preovulatory period. Direct tubal aspiration was performed in seven women undergoing fertiloscopy. Tubal washing with isotonic glycine (1.5ml) was performed in nine women during fertiloscopy and in three women during laparoscopy. The protein concentration of the ̄uid retrieved was measured (Bradford protein assay). Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was performed on suitable samples. Protein spots detected on silver stained gels were characterized in terms of their molecular weight and isoelectric point. Results: Specimens were obtained in five out of nine women who had tubal washing during fertiloscopy; the mean volume was 320 ml. Three specimens out of the five had a mean protein concentration of 31.7 mg/ml and two did not have detectable tubal protein. In the three cases of laparoscopic tubal washing, a mean volume of 633 ml was retrieved; the mean protein concentration was 375 mg/ml. It was not possible to retrieve any fluid by direct tubal aspiration. Proteomic maps were created from the three tubal fluid washings collected during laparoscopy. The proteomic pattern of over 250 individual protein spots was apparent. Conclusions: The findings suggest a low intratubal fluid volume in vivo. Only laparoscopy yielded specimens suitable for 2D-PAGE analysis of tubal protein. We believe that this is the first presentation of the proteomic analysis of tubal proteins by 2D-PAGE and that these findings will provide additional information on the physiology of the tube and the pathogenesis of tubal factor infertility.