Follicular development and ovulatory processes in mammals involve local biochemical changes as a result of substantial modifications in cellular metabolism, the most well known of which is steroid variation. In the present study, the intrafollicular variation of several other components was studied using proton nuclear magnetic resonance ((1)H NMR). This approach made it possible to demonstrate that the intrafollicular biochemical content changes during follicular growth and maturation. Follicular fluid was aspirated by ovarian puncture of the dominant follicle at various physiological stages of its development: early dominant, late dominant and preovulatory. Serum samples were collected during each puncture session. (1)H NMR was used to evaluate intrafollicular and circulating glycoconjugates (sugar chains and N-acetyl groups), lipoproteins (CH(3) and CH(2) groups), glucose metabolites (trimethylamines, acetate and lactate), amino acids (glutamine/glutamate and alanine), creatine/creatinine and polyamines. Follicular fluids were assayed by radioimmunoassay for oestradiol and progesterone contents. The intrafollicular contents of alanine and lipoproteins (CH(3) groups) decreased in the dominant follicle during growth, whereas concentrations of progesterone and oestradiol increased significantly. After injection of gonadotrophin to induce ovulation, follicular maturation was characterized by a decrease in glycoconjugates (sugar chains), trimethylamines and acetate, a decrease in oestradiol concentration, and a further increase in CH(3) groups of lipoproteins and progesterone. The results from the present study showed a clear correlation between the intrafollicular content of alanine and that of oestradiol. A correlation between progesterone and glycoconjugates (sugar chains) was also observed. Therefore, (1)H NMR was shown to be effective for studying specific changes in the biochemical composition of the follicular fluid that occur during follicular development. For the first time, the variation of several compounds (glycoconjugates, lipoproteins, glucose metabolites, amino acids and polyamines) in relation to growth and maturation was demonstrated. Some of these changes could be of crucial importance for follicular maturation and ovulation as well as for oocyte maturation and further fertilization.
N Gerard, S Loiseau, G Duchamp and F Seguin
S. Hamamah, F. Seguin, C. Barthelemy, S. Akoka, A. Le Pape, J. Lansac and D. Royere
Glycerylphosphorylcholine (GPC), glycerylphosphorylethanolamine (GPE), citrate, and lactate content of human seminal plasma was analysed by measuring the peak area of 1H nuclear magnetic resonance (NMR) spectra in samples from four groups of patients: 21 spermatogenic failure subjects; 14 obstructive azoospermic subjects (vasectomized); seven patients presenting very severe oligoasthenozoospermia (OAT) and 18 normozoospermic subjects (control). The peak areas for GPC, citrate and lactate in seminal plasma were significantly smaller for patients with azoospermia than for the control group: 16.79, 8.18 and 2.28 versus 23.38, 10.58 and 4.30, respectively (P < 0.01). The peak area ratios for citrate:lactate and GPC:lactate were significantly different (P < 0.01) between the control group and the spermatogenic failure or obstructive azoospermia groups. A significant difference was also found in GPE:GPC peak intensity ratio between spermatogenic failure and obstructive azoospermia subjects (P < 0.001). These results provide some quantitative markers which may be used for examining infertility by using 1H NMR of seminal plasma samples.