Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Morphometric and ultrastructure features of the ventral prostate of rats (Rattus norvegicus) submitted to long-term nicotine treatment

The harmful effects of nicotine on male genital system fertility have been reported in experimental and clinical studies. However, its effects on prostatic cells and glandular pathogenesis remain unclear. The aim of the present study was to analyse the histological, histochemical and ultrastructural alterations, in addition to stereology, of the ventral lobe of the prostate of rats, submitted to chronic nicotine administration, as well as to establish the relationship between these changes and prostate diseases. Twelve male Wistar rats (Rattus norvegicus) were divided into two experimental groups: group I (nicotine) and group II (control). Samples of the ventral prostate were collected, processed and submitted to histological analysis, acid phosphatase histochemistry and ultrastructural analysis by transmission and scanning electron microscopies. The results showed that in the nicotine group, the secretory epithelial cells of the ventral lobe of the prostate were atrophied, and prostatic intraepithelial neoplasia occurred and reduced the expression of acid phosphatase. The disorganisation of organelles involved in the glandular secretory process, accompanied by biomembrane destructuring, was also observed. In conclusion, nicotine causes drastic alterations in the secretory epithelium of the ventral prostate, compromising its function. Furthermore, nicotine also induces premalignant lesions in the prostate gland, thus representing a risk factor in the development of prostate diseases.

THE HYPOPHYSEAL-TESTICULAR AXIS AND SEX ACCESSORY-GLANDS FOLLOWING CHEMICAL SYMPATHECTOMY WITH GUANETHIDINE OF PRE-PUBERTAL TO MATURE RATS

Selective chemical sympathectomy of the internal sex organs of prepubertal to mature male Wistar rats was performed by chronic treatment with low doses of guanethidine. Plasma testosterone and luteinizing hormone and the intratesticular level of testosterone were determined. The weight and fructose content of seminal vesicle and ventral prostate were also investigated. The results showed that sympathetic innervation is related to the control of the hypophyseal-testicular axis as well as to the growth and potential secretory activity of the male sex accessory glands.

Effects of guanethidine-induced sympathectomy on the spermatogenic and steroidogenic testicular functions of prepubertal to mature rats

Selective chemical sympathectomy of the internal genital organs of prepubertal to mature male Wistar rats was performed by chronic treatment with low doses of guanethidine. Sympathetic denervation caused an increase in intratesticular progesterone levels in prepubertal and early pubertal rats in addition to a decrease in androstenedione and testosterone levels in prepubertal animals, thus indicating a decrease in the conversion of progesterone into androgen, probably by blocking the steroidogenic enzymatic pathway at the 17 alpha-hydroxylase/17,20 desmolase level. A lower degree of testicular maturation, probably related to reduced androgen activity, was observed in prepubertal and early pubertal sympathectomized rats. Concentration of spermatozoa, on the other hand, was increased in the enlarged cauda epididymidis of late pubertal and mature denervated animals. This result is discussed in terms of the impairment of epididymal mechanisms of seminal emission, fluid resorption and spermatozoal disposal.

Estudo do perfil proteico nas membranas de espermatozóides congelados de caprinos

Pós-graduação em Medicina Veterinária - FMVZ

Características seminais e resfriamento de sêmen de tamanduá (Myrmecophaga tridactyla) de vida livre

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)