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To investigate some possible effects of low doses of nonsteroidal anti-inflammatory drugs upon functional activity of mouse sperm, the authors injected lysine acetyl salicylate (im 14.3 mg/kg day(-1), ibuprofen (ip 5.6 mg/kg day(-1)), or piroxicam (ip 0.28 mg/kg day(-1) to pregnant females (the male cohort was sacrificed at adulthood) (A) or to adult males during 35 (B) or 60 (C) days. Parameters measured were motility, viability, acrosomal integrity, responses to hypoosmotic shock, in vitro fertilization index, and testosterone plasma levels. Salicylate evoked a slight reduction in the percentage of swollen gametes in A, and ibuprofen diminished testosterone plasma levels in B. The other parameters remained unchanged in all groups. Results are well supported by the low doses assayed, which are equivalent to the content of one tablet commercially available for each compound.

In epididymal mouse spermatozoa, the effects of dibutyryl cyclic adenosine monophosphate 1 mmol/L (dbcAMP), pentoxifylline 5 mmol/L (PX), and/or mastoparan 50 mumol/L (MT) were evaluated for the following parameters: percentage of motile cells and response to hypoosmotic shock (HOS). The gametes were incubated during 80 min (A) or 200 min (B) in Tyrode's medium, and the drugs were added during the last 20 min. In A, dbcAMP + PX (61.5 +/- 5.4%; n = 10) enhanced and MT decreased significantly the population of motile cells (13.4 +/- 5.4%; n = 6) (control: 47.6 +/- 3.9%; n = 11). In B, PX significantly increased this parameter and MT plus PX also exerted a significant detrimental effect. Responses to HOS dropped significantly in the presence of PX + MT in A or in B; in this latter condition a similar decrease was evoked by MT alone. A positive correlation between percentages of swollen and motile spermatozoa was detected in A or in B in samples incubated with PX (r = .58, n = 11 and r = .76, n = 10; p < .05, respectively). These results that support that, in mouse sperm tail, PX would preserve functional membrane integrity, a relevant condition for adequate motility.

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.

Chinchilla laniger is an endangered species and improved cryopreservation of spermatozoa would constitute a significant advance in the development of assisted reproductive techniques in this species. The functional activity of epididymal spermatozoa from adult males was studied immediately after extrusion and after 24 hours incubation, and the ability of five extenders to protect these gametes during cryopreservation was determined. A decrease in sperm motility, viability, acrosome intact cells and response to hypo-osmotic swelling test was detected 24 hours later. The extenders here assayed showed well-defined differences in their cryoprotective ability; however such differences could not be attributed to any one of their individual components. The presence of TES plus Tris, the proportion of the individual constituents and/or the differences in metabolic substrate content could explain the above-mentioned finding. The results indicate that cryo-buffer II (TES-Tris-egg yolk-fructose-glycerol) is the most powerful protector of sperm functional activity in this species.

Several studies indicate that reactive oxygen species (ROS) are involved in defective sperm function pathophysiology. In this study we attempted to determine differentially the effects of xanthine (0.12 mM) plus xanthine oxidase (0.035 U/mL) (X+XO, a ROS promoter system), ROS scavengers (Tiron (TIR, 15 mM); catalase (CAT, 10 micrograms/mL); dimethylsulfoxide (DMSO, 140 mM)), and X+XO plus scavengers on several epididymal mouse spermatozoa functional parameters, incubated in NTPC medium, for 29 min. In the presence of X+XO, progressive gametes significantly diminished. TIR or CAT attenuated this effect, but DMSO did not. Inversely, X+XO increased the bending-forms population; only TIR reversed this phenomenon. The ROS promoter system diminished the viable cell population; all scavengers assayed maintained sperm viability at levels similar to control ones. When exposed to hypoosmotic shock after 29 min incubation with X+XO, the percentage of swollen cells decreased; TIR, CAT, or DMSO did not prevent this effect. Our experiments demonstrate that it is possible to differentiate the deleterious ROS effects upon sperm functional activity. O-2. and H2O2 preferentially seem to modify sperm motility, O-2. exhibiting the greatest ability for generating bending-form gametes, OH-being the most lethal ROS. In addition, sperm membrane clearly appears as the most damaged structure.

The kinetics of functional modifications during in vivo or in vitro incubation was studied in spermatozoa from Albino Swiss mice, obtained from cauda epididymis (E) or from uterus (U) or oviduct (O) post copulation (p.c.). Results clearly suggest that functional activity as reflected by measurements of motility, hypoosmotic swelling test and acrosome reaction differs according to the segments of the reproductive tract from which they were collected. The comparison of results obtained after equivalent periods of in vitro or in vivo incubation, shows that the time course of changes in sperm motility and percentage of swollen sperm is similar in both conditions. Since the percentage of acrosome-reacted gametes was higher in O, at all times scored, oviductal environment appeared to be capable of inducing this phenomenon. Our experimental conditions seemed to be adequate for completion of spermatozoa maturation and successful interaction between male and female gametes as reflected by the results of fertilization rate and embryo cleavage.

The evaluation of spermatic morphology is of importance due to its prognostic value in potential male fertility, both in natural and in assisted reproduction. In the present work, the morphologic profile of samples of semen from normal and dispermic patients was determined, in order to establish a correlation between structure and function. Samples were obtained from male partners of infertile couples who attended andrologic consultation. The smears were stained by the Papanicolaou technique, and the criteria of the OMS and the recent modification of Kruger were followed to evaluate spermatic morphology. Of all the patients studied (n = 46), 35% were normospermics (A), 39% asthenospermics, 7% oligospermics, 2% terastospermics and 17% presented combined alterations. The average percentage of normal spermatozoa was of 23.7 +/- 1.23 in the whole population. When group A patients were compared with asthenospermics, significant differences were found in the percentage of spermatozoa with middle piece and tail alterations, in the last group (p < 0.02). Similarly, when samples were analysed according to the presence of alterations: none (A), one (B) two or more (C), it was observed that the percentage of normal forms decreased in the groups in that order (A < B < C); (A or B vs C p < 0.05). The percentage of tapering spermatozoa was significantly higher in group C (p < 0.01 vs A or B). From the above results, it appears that 1) the larger number of functional alterations is related to a higher percentage of structural anomalies, and 2) the deficient motility in the asthenospermic group, is associated to the middle piece and tail alterations.(ABSTRACT TRUNCATED AT 250 WORDS)