J. L. Lacuara

Activity of Chinchilla laniger spermatozoa collected by electroejaculation and cryopreserved

Because reproductive studies and the application of assisted reproductive techniques are relevant issues for an endangered species such as Chinchila laniger, the availability of a source of viable spermatozoa becomes of utmost importance. In this paper, we evaluate several functional parameters (motility, viability, response to hypoosmotic swelling test and acrosomal integrity) of fresh or frozen-thawed spermatozoa. Electro-ejaculation trials (50-cyc/sec sinusoidal wave was applied for 5 of every 10 sec) were successful in all unanesthetized animals. After volume (108.3 +/- 12.0 microL, n = 15) and concentration (421.8 +/- 34.4 x 10(6) cells/mL, n = 15) measurements, the above mentioned parameters were determined. In frozen-thawed semen samples sperm motility, viability, hypoosmotic swelling test and acrosomal integrity were significantly lower than in fresh semen samples. The results clearly indicated that electro-ejaculation is a useful method for evaluating spermatozoa for genetic analysis or for used in Al in this species. In addition, the cryopreservation procedure in this study preserved adequate levels of functional sperm activity.

Contractility and pharmacological reactivity of isolated vascular smooth muscle from diabetic rats

Conflicting reports exist about the effects of diabetes on vascular function. In the present study we investigated (1) the influences of diverse stages of diabetes on mechanical activity and pharmacological reactivity of portal vein and tail ventral artery isolated from male rats 7, 21 or 30 days after alloxan injection (150-180 mg/kg) and (2) the effects of in vitro or in vivo insulin treatment. Various parameters were used to assess the diabetic state (serum glucose levels, body weight, percentage of glycosylated hemoglobin and glucosuria). Isometric developed tension of portal vein from control rats was 10.86 +/- 0.41 mN (n = 54), and was enhanced significantly in diabetics (+56% at 21 days and +45% at 30 days; p less than 0.001 vs. controls). When challenged with noradrenaline, portal veins from diabetics exhibited a greater contractility and lower reactivity (as reflected by EC50 values). The magnitude of responses to KCl remained similar to those obtained in controls, but nonetheless the reactivity seems to be higher. Tail ventral artery from diabetics also exhibits a greater contractility in response to noradrenaline with no significant changes in EC50 values. The results demonstrate that diabetes affects mechanical performance of the vascular smooth muscle in a differential manner depending on the stage of the endocrinopathy and on the types of vessel studied. These modifications were not avoided by insulin treatment.

In vitro parthenogenesis of mouse oocytes under several experimental conditions

Although the in vitro fertilisation index is a parameter commonly employed to investigate sperm functional activity, little attention has been given to the occurrence of parthenogenesis. The purpose of this study was to study at 6 h or 22 h incubation: (a) the cleavage-related events that occur in in vitro incubated mouse oocytes, in the absence (parthenogenesis) or presence of homologous spermatozoa; (b) the effect of mineral oil, commonly used in in vitro fertilisation assays; (c) the effect of piroxicam, a prostaglandin synthesis inhibitor, on the parthenogenetic rate; and (d) the influence on parthenogenesis of spontaneous loss of the cumulus oophorus coat during incubation. Under the experimental conditions employed, there was parthenogenetic activation and activation due to fertilisation. Both increased in a time-dependent manner. The mineral oil enhanced the parthenogenetic rate at 22 h incubation. However, it did not have any effect when the oocytes were inseminated. Since we can not discriminate how much of this activation was due to fertilisation and how much to parthenogenesis we must be very careful with this comparison. Piroxicam 10(-8) M did not show any effect on the mouse oocyte parthenogenetic rate at neither 6 h or 22 h incubation. Our results suggest that oocyte susceptibility to spontaneous parthenogenetic activation may be modified by the presence of the cumulus and corona radiata cells. In conclusion, we consider that further rigorous studies on these influences are necessary in order to confer more reliability on the results.

Modifications evoked by piretanide in the mechanical activity of cardiovascular tissues

The effects of piretanide upon mechanical activity and pharmacological reactivity of vascular and myocardial tissues from normotensive rats were investigated. Magnitude of phasic contractions of isolated rat portal vein was diminished by the drug in a dose-related manner; contractile depression induced by piretanide (10(-4)M) was less in the presence of insulin (0.1 U/mL), glucose (22 mM) or pyruvate (5 mM). Responses to KCl (90 mM), or norepinephrine (2.5 X 10(-5)M) were also reduced. Contractile activity of atria and ventricle strips was diminished only when piretanide reached 10(-4)M. Results support direct actions of piretanide upon cardiac and vascular tissues. Possible mechanisms of action are discussed.

Effects of vasoactive agents on oxygen uptake in portal venous smooth muscle.

The studies concerned the effects of epinephrine, norepinephrine, phenylephrine, acetylcholine and K+ ions, in doses inducing maximal contraction on oxygen consumption of isolated rat portal veins. All the agonists tested stimulated the rate of O2 uptake, whereas high K+ did not. After specific blockade of alpha-adrenoceptors, the effects of epinephrine and norepinephrine on O2 consumption were not affected, whilst those of phenylephrine were abolished, and beta-adrenoceptor blockade suppressed the metabolic effect of epinephrine and norepinephrine. Acetylcholine was the agonist that most elevated O2 consumption; atropine blocked both contractile and metabolic effects of this drug. It appears that there is a clear dissociation between contractile and metabolic actions of some pharmacological agents, and a question is posed concerning phenylephrine and acetylcholine effects. Results with K+ suggest that different metabolic pathways might be involved in the activation and maintenance of drug-induced contraction in smooth muscle.

Qualitative study of prostaglandins in pregnant uterine tissues: effects of estradiol- 17β and betamethasone

The presence of prostaglandins (PGs) E1 and F2 alpha in extracts of endometrial, myometrial and placental samples from 19 pregnant women at term, was investigated. The tissue samples were obtained from the following group of patients: untreated, A) with onset of labor, B) without onset of labor; pre-treated (without onset of labor); C) with betamethasone, D) with estradiol-17 beta. Thin-layer chromatography determinations demonstrated PGE1 presence in all tissue samples from the four groups. On the other hand, PGF2 alpha was consistently found in placenta and endometrial samples from groups A, C, and D, but it was absent in those from group B. The results suggest a physiological role of PGF2 alpha in human parturition, and a possible influence of estrogens and corticosteroids on the appearance of this prostaglandin.