Hyaluronic acid as an anti-angiogenic shield in the preovulatory rat follicle.

Angiogenesis in the preovulatory follicle is confined to the theca cell layers, and penetration of capillaries through the basement membrane into the granulosa cell layers does not occur until after ovulation. However, elevated expression of the angiogenic growth factor (VEGF) has been reported in the cumulus cells surrounding the oocyte, which are expelled from the follicle during ovulation. This spatial and temporal discrepancy between VEGF expression and angiogenesis was studied here in the rat ovarian follicle, and we showed that cumulus cells secrete to the follicular fluid, in addition to VEGF, material with antiangiogenic activity that blocks endothelial cell proliferation, migration, and capillary formation in vitro. Hyaluronic acid produced by the cumulus cells can account for this antiangiogenic activity. Degradation of hyaluronic acid by hyaluronidase restored proliferation and migration of endothelial cells directed toward the cumulus. Inhibition of hyaluronic acid synthesis with 6-diazo-5-oxo-1-norleucine restored endothelial proliferation and migration in vitro, and it also resulted in early penetration of capillaries across the follicular basement membrane in vivo. These results support the role of hyaluronic acid produced by the cumulus cells as a high-molecular-weight, antiangiogenic shield that prevents premature vascularization of the preovulatory follicle by blocking endothelial cell migration and proliferation.

Spatial and temporal modulation of perfusion in the rat ovary measured by arterial spin labeling MRI.

The hemodynamic changes triggered by luteinizing hormone/human chorionic gonadotropin (LH/hCG) in ovaries of immature pregnant mare serum gonadotropins (PMSG)-primed female Wistar rats were followed by pulsed arterial spin labeling magnetic resonance imaging. Decreased perfusion was monitored in the first 2 hours after administration of hCG followed by a transient significant rise in perfusion. Subsequently, constant ovarian perfusion of 10.9 +/- 4.3 mL min(-1) g(-1) was maintained during the exponential increase in ovarian volume. However, ovarian perfusion was not uniform, and prior to ovulation poorly perfused regions were detected that were assigned to the follicular fluid in preovulatory follicles. This result implied that in the time scale of seconds, corresponding to the T1 relaxation time of water in the follicular fluid, exchange of arterial water with water in the follicular fluid was negligible. Along with the drop in the levels of high-energy phosphate metabolites detected by 31P nuclear magnetic resonance spectroscopy and the shift to glycolytic metabolism, these results support the hypothesis that physiological hypoxia could play a role in large preovulatory follicles as part of the normal ovarian cycle.

Perfusion of the rat ovary: application of pulsed arterial spin labeling MRI.

Pulsed arterial spin labeling was used for mapping ovarian perfusion and measurement of blood velocity in the ovarian artery. Arterial blood was tagged upstream by pulsed slice selective saturation, and saturation transfer due to perfusion was monitored within the rat ovary. The velocity of arterial blood was determined from the dependence of the saturation transfer on the thickness of the saturation slice and the delay between successive saturation pulses. This method allows for determination of arterial velocity, even when the artery itself is not identified in the images. The arterial velocity of blood to the ovary was 3.6+/-0.6 cm x s(-1). The mean ovarian perfusion was 8.7+/-3.5 ml x min(-1) x g(-1) during the surge of luteinizing hormone and 5.9+/-3.0 ml min(-1) x g(-1) during the luteal phase. Arterial labeling can thus be used for following vascular remodeling and angiogenesis during the ovarian cycle by MRI.

Regulation of angiogenesis by hypoxic stress: from solid tumours to the ovarian follicle.

The preovulatory follicle provides a unique physiological example of rapid growth accompanied by neovascularization, two processes that are generally characteristic of pathologies such as wound repair or malignancy. During the hours preceding ovulation, follicular growth is accompanied by elevated levels of messenger RNA for vascular endothelial growth factor (VEGF). Angiogenic activity, mediated by VEGF, is manifested in the peripheral blood vessels surrounding the follicle, that show capillary sprouting and increased vascular permeability. Following ovulation, rapid infiltration of capillaries through the follicular wall is essential for the formation of the corpus luteum. In this review we compare the preovulatory follicle with a popular model of avascular solid tumour growth, namely the multicellular tumour spheroid, in particular the role of hypoxic stress in the regulation of angiogenesis in both systems.

Modulation of water diffusion during gonadotropin-induced ovulation: NMR microscopy of the ovarian follicle.

The preovulatory rat follicle reaches a diameter of 1 mm with no internal blood vessels. Nutrient supply to the enclosed oocyte depends solely on passive diffusion across the follicular wall and the follicular fluid. Spin-echo and stimulated-echo NMR microscopy experiments were applied here for studying modulations in water diffusion during gonadotropin-induced maturation of perfused rat ovarian follicles (32 degrees C). Two diffusion compartments were observed for the follicular wall. The intracellular water diffusion coefficient, measured at a short diffusion time (9 ms) was 0.28 x 10(-5) cm2/s. Diffusion at long diffusion times was restricted to 16 microns, the size of cells in the follicular wall, and did not change during maturation. In the follicular fluid a transient 26% decrease in the diffusion coefficient was observed 4-7 h after gonadotropin stimulation, a change that is bound to affect the metabolic balance of the oocyte before ovulation.

Cyclocreatine accumulation leads to cellular swelling in C6 glioma multicellular spheroids: diffusion and one-dimensional chemical shift nuclear magnetic resonance microscopy.

Cyclocreatine, an analogue of creatine, inhibits tumor cell proliferation in vitro and in vivo. The effects of cyclocreatine in large C6 glioma multicellular spheroids were mapped here by magnetic resonance microscopy. Diffusion-weighted images of C6 glioma spheroids resolved the bright viable rim and the dark necrotic center. Sequential sets of diffusion images, following cyclocreatine administration, showed increasing self-diffusion coefficients of the intracellular water in the viable rim (0.49 x 10(-5) cm2/s for untreated spheroids, 0.62 x 10(-5) cm2/s after 48 h perfusion with 20 mM cyclocreatine). This fact correlated with cellular swelling apparent in histological sections. The radial distribution of cyclocreatine and soluble lipids across perfused C6 spheroids was measured by one-dimensional chemical shift imaging. Cyclocreatine accumulation was prominent throughout the viable cell layer, with no cyclocreatine accumulation in the necrotic center. In both cyclocreatine-treated and control spheroids the lipid signal was highest in the necrotic center and lower in the inner viable cell layer.

Model of a new temperature-compensated optical current sensor using Bi(12)SiO(20).

A new optical bulk current sensor is presented with Bi(12)SiO(20) as a sensing crystal. Through the use of a mirror in the setup and the reciprocity of the optical activity of Bi(12)SiO(20), the sensor becomes insensitive to intrinsic linear birefringence and birefringence that is due to the enclosure of the crystal. Therefore the sensor is also insensitive to temperature, which affects the total linear birefringence. By making a computer model of the Bi(12)SiO(20) sensor, it was proved that the output signal of the sensor, which has a sinusoidal response, has a maximum relative error of 0.05%, apart from the variation of the Verdet constant, for a temperature change of approximately 100 °C.

Differences in the activation of m. biceps brachii in the control of slow isotonic movements and isometric contractions.

We have compared muscle activation in the control of slow isotonic movements and isometric contractions. Specific attention has been given to the contribution of the two force-grading mechanisms, the recruitment of motor units and the modulation of firing frequency in motor units that have already been recruited. The recruitment order of the m. biceps motor units under study was the same during isometric contractions and slow isotonic movements. However, the recruitment thresholds of the m. biceps units were considerably lower for both isotonic flexion and extension movements, even at velocities as low as 2 deg/s, than for isometric contractions. Furthermore, firing frequency at recruitment was found to depend on the motor task: at recruitment the motoneurone starts firing with a higher firing frequency during isotonic flexion movements and a lower firing frequency during isotonic extension movements than during isometric contractions. Two main conclusions can be drawn from these results. First of all, the concept of one single activation parameter (total synaptic drive?) cannot account for the motor-unit behaviour observed during our experiments: the relative contribution of the two force-grading mechanisms is different for different tasks. Secondly, the distribution of activity among flexor motoneurone pools is different for isometric contractions and isotonic movements.