Investigation of spermatozoa and seminal plasma by fourier transform infrared spectroscopy.

Fourier transform infrared (FT-IR) spectra of human spermatozoa and seminal plasma were recorded and analyzed. The procedure that was established for sample preparation enabled acquisition of reproducible spectra. The parameter I(1087)/I(966) for controlling spectra reproducibility was defined. The assignment of bands was carried out using an empirical approach and the origin of the "sperm specific doublet", the bands at 968 cm(-1) and 981 cm(-1), was determined. The principal component regression (PCR) algorithm was used to define the specific spectral regions correlating to characteristics of spermatozoa, such as concentration, straight-line velocity (VSL), and beat cross frequency (BCF). Then, simple spectral parameters, such as band intensities and band ratios, were tested to determine which one best correlates to characteristics of spermatozoa. The region of the amide I band, between 1700 cm(-1) and 1590 cm(-1), was defined as a specific spectral region that correlates to the concentration of spermatozoa. The parameter that gave the linear dependence to the concentration of spermatozoa was the intensity of the amide I band. For VSL, the bands between 1119 cm(-1) and 943 cm(-1) were defined as the specific spectral region. The relative amount of nucleic acids with respect to proteins showed linear dependence on the straight-line velocity of spermatozoa. BCF showed the best correlation to the bands between 3678 cm(-1) and 2749 cm(-1), which largely represent lipids and proteins. These results suggest that FT-IR spectroscopy can serve as an adjunct to conventional histopathology studies.

Disorders of male rat reproductive tract under the influence of atrazine.

The effects of atrazine exposure on testicular sperm number, epididymal sperm number and motility and alpha-glucosidase activity in the epididymis were studied in Fischer rats. Histological changes in the testicular tissue were followed by light and electron microscopy. Groups of adult animals were treated i.p. with 60 and 120 mg atrazine kg(-1) body wt. twice a week over 60 days. The results indicate a decrease in the body weight and relative weights of pituitary and ventral prostate vs control, measured on the last day of treatment in both treated groups. Testicular sperm number (expressed as number of sperm per 500 Sertoli cells) in atrazine-treated groups increased with the treatment time due to the reduced sperm motility. Therefore atrazine treatment provoked a significant decrease in sperm number and motility in epididymis, measured after the last day of treatment. alpha-Glucosidase activity in the epididymis, after the last day of treatment, showed a decrease in both treated groups vs control values. Histological analysis of testicular tissue from treated rats showed the cell disorganization and cell clusters together with spermatocytes. Electron microscopy presented differently vacuolated cytoplasm, collagen fibre was reduced, Leydig cells were of irregular shape with unequal form and cisternae of rough endoplasmic reticulum were accentuated and softly widened. In Sertoli cell cytoplasm, atrazine treatment provoked degenerative changes. According to the results obtained, it is evident that atrazine exerted morphological changes and a toxic effect on sperm and their motility.

Sperm motility and kinetics of dynein ATPase in astheno- and normozoospermic samples after stimulation with adenosine and its analogues.

We tested the effects of adenosine and 2-deoxyadenosine on the activation of human spermatozoa. In the asthenozoospermic group of patients adenosine produces an increase in sperm motility from 33.3 +/- 2.1% to 42.1 +/- 3.4%, progressive motility from 22.5 +/- 1.3% to 28.6 +/- 1.7% and forward progression rating from 2.1 +/- 0.2% to 2.8 +/- 0.1%. 2-Deoxyadenosine stimulated asthenozoospermic samples to a greater degree than adenosine. Sperm motility rose to 48.9 +/- 3.4%, progressive motility to 32.1 +/- 3.4% and forward progression rating to 3.0 +/- 0.1% following stimulation with 2-deoxy-adenosine. The kinetic parameters and basic characteristics of dynein ATPase were determined. The maximum activity of dynein ATPase, Vmax, was significantly different (P < 0.001) for asthenozoospermic and normozoospermic samples: 6.46 +/- 2.1 nmol Pi/mg/min and 16.99 +/- 3.7 nmol Pi/mg/min respectively. However, the enzyme affinity for ATP was not different. Stimulation of asthenozoospermic samples with adenosine and 2-deoxyadenosine caused an increase of Vmax (70-90% and 90-110% respectively) and no significant change in KM was observed. In order to block the nucleoside transporter and to eliminate the action of adenosine inside the cell, dipyridamole was used but the effects of adenosine were not neutralized. 5'-(N-ethylcarboxy-amido)-adenosine showed effects similar to those of adenosine, even when applied in 1 microM concentration. These results indicate that adenosine and its analogues stimulate sperm motility and activity of dynein ATPase, most probably via A2 receptors.