USE OF COMBINATION OF FLUORESCENT PROBES TO IDENTIFY SPERM SUBPOPULATIONS FOR QUALITY ASSESSMENT OF FRESH AND CRYOPRESERVED CANINE SEMEN. PRELIMINARY RESULTS

The use of fluorescent markers in the evaluation of sperm morphophysiology allows a better accuracy, compared to the subjective nature of some routine tests in semen qualification. In this study was used the combination of fluorescence probes: propionate iodide, Hoechst 33342 and FITC-PSA in fresh and thawed dog semen, to the identification of the following morphological subpopulations: II (intact plasma and acrosomal membranes), IL (intact plasma membrane and lesioned acrosomal membrane), LI (lesioned plasma membrane and intact acrosomal membrane) and LL (both membranes lesioned). When comparing the results obtained with the results of the tests used conventionally in semen evaluation (sperm motility and vigor, hypoosmotic test and morphological alterations), little correlation was observed. The II population declined from fresh semen to thawed, while LL population increased (p <0.05). The IL population was composed of extremely small numbers of cells but increased (p <0.05) from fresh semen to thawed semen. In the thawed semen the major defects had a positive correlation with the LL population (p <0.01). For the thawed semen, the results of the hypoosmotic test (number of cells that reacted to the medium) correlated positively with population II (p <0.025), that is, different from that observed in fresh semen. Although all tests were able to detect decrease in sperm quality post-thawing (p <0.05). The use of this fluorescent probe association allowed qualification and more accurately quantification of plasma membrane and acrosomal insults mediated by cryopreservation. El uso de marcadores fluorescentes en la evaluación de la morfofisiología espermática permite una mayor precisión, comparada con la naturaleza subjetiva de algunas pruebas de rutina en la valoración del semen. En este estudio se usó la combinación de pruebas fluorescentes: yoduro de propidio; Hoechst 33342 y FITC-PSA en semen fresco y descongelado de perro, para la identificación de las siguientes subpoblaciones morfológicas: II (membranas plasmática y acrosomal intactas), IL (membrana plasmática intacta y membrana acrosomal dañada), LI (membrana plasmática dañada y membrana acrosomal intacta) y LL (ambas membranas dañadas). Cuando se comparan los resultados obtenidos con los resultados de las pruebas usadas convencionalmente en la evaluación seminal (motilidad y vigor espermáticos, prueba hipoosmótica y alteraciones morfológicas), se observó poca correlación. La población II disminuyó desde el semen fresco al descongelado, mientras que la población LL se incrementó (p<0.05). la población IL estuvo compuesta por un número extremadamente pequeño de células, pero incremento (p<0.05) desde el semen fresco al descongelado. En el semen descongelado los defectos mayores tuvieron una correlación positiva con la población LL (p<0.01). En el semen descongelado, los resultados de la prueba hipoosmótica (número de células que reaccionan al medio) se correlacionaron positivamente con la población II (p<0.05). El uso de esta asociación de pruebas fluorescentes permitió la valoración y la cuantificación más precisa de los daños a la membrana plasmática y acrosomal mediados por la criopreservación.

Neonatal treatment with naloxone increases the population of Sertoli cells and sperm production in adult rats

Endogenous opioid peptides play an important role in the ontogenesis of the functional and morphological parameters of the seminiferous epithelium. The aim of this study was to evaluate the effects of neonatal manipulations with naloxone, an opioid antagonist, on the population of Sertoli cells and on sperm production in adult rats. Rats were assigned to receive 8 $\mu$g per gram of body weight twice a day with interval of 8 h of naloxone and they were compared to a control group receiving saline. Naloxone groups presented the following findings when compared to the control group: increased body weight from the 2nd to the 27th day; a smaller seminiferous epithelium height, smaller seminiferous tubule diameter, increased number of Sertoli cells and daily sperm production per testis, increased daily sperm production per gram per testis and increased total length of the seminiferous tubule of the treated groups. According to our study, the neonatal treatment with naloxone during the critical period of testis development was able to change the proliferative dynamics of Sertoli cells by an intra and/or extra testicular blockage of opioid receptors, confirming the direct relation between the number of Sertoli cells and the number of spermatozoids

Effects of catuaba cristal® on the testis of wistar rats

The aim of this study was to evaluate the possible beneficial effects of Catuaba Cristal® (CC), an alcoholic drink made from wine and Erythroxylum catuaba Ar. Cam on testis. Wistar rats either received CC solution (n=8) or water (n=9). Results showed significant body weight reduction within the CC group, although, no weight changes were observed for liver, kidney, testis, epididymis, seminal vesicle and prostate. The volumetric proportion and volume of interstitial tissue and lymphatic space were reduced in the treated group. In the CC group, although the nuclear volume of Leydig cells (LC) decreased, the number of LC per testis increased. These results suggested that CC had no beneficial effect on spermatogenesis of Wistar rats