Seasonal changes in the activity of the adrenal medulla of Viscacha (Lagostomus maximus maximus).

Animals living in nontropical climates modify their physiology and behavior to adapt to seasonal environmental changes. Part of this adaptation involves the release of catecholamine from sympathetic nerve endings and the adrenal medulla, which play a major role in regulating energy balance. The aim of this work was to investigate whether adult male viscachas in their natural habitat exhibits structural changes in the adrenal medulla during the annual seasonal cycle. In August-September, chromaffin granules revealed ultrastructural changes suggestive of piecemeal degranulation. Quantitative morphometric analysis by transmission electron microscopy showed a significantly lower percentage of resting chromaffin granules and a higher percentage of altered granules and empty containers in August-September (late winter) compared to February-March (late summer), suggesting an increased secretory process of catecholamines in August-September. The mechanism of piecemeal degranulation might amplify this process, encouraging the adaptive response to winter environmental conditions. Tissue levels of epinephrine, norepinephrine, and dopamine (analyzed by high-performance liquid chromatography) changed throughout the year, reaching maximum values in February-March and minimum values in August-September. These results demonstrate morphological and biochemical seasonal variations of the adrenal medulla, suggesting that epinephrine might promote energy mobilization, which allow the Lagostomus to cope with adverse environmental conditions and thus to survive during winter season.

Seasonal morphological variations and age-related changes of the seminal vesicle of viscacha (Lagostomus maximus maximus): an ultrastructural and immunohistochemical study.

The viscacha is a seasonal rodent that exhibit an annual reproductive cycle with periods of maximum reproductive activity and gonadal regression. We studied seasonal variations in the morphology and cellular population of the seminal vesicles (SVs) during both periods and in impuber animals. Seminal vesicles were studied by light and electronic microscopy. Measurements of epithelial height, nuclear diameter, luminal diameter, and muscular layer were performed. Also, we studied the distribution of androgen receptors (AR) in this gland during the reproductive cycle and in impuber animal. During gonadal regression, principal and clear cells showed signs of reduced functional activity. These were characterized by an epithelium of smaller height, irregular nuclei, and cytoplasm with few organelles, dilated cisterns, and glycogen granules. In impuber animals, the principal cells showed large nuclei with chromatin lax and cytoplasm with small mitochondria, poorly developed Golgi apparatus, and granules of glycogen. On the other hand, the cells exhibited seasonal variations in the distribution and percentage of immunolabeled cells to AR throughout the annual reproductive cycle. During the gonadal regression period, glandular mucosa exhibited numerous epithelial cells with intense nuclear staining. However, fibromuscular stromal cells were weakly positive for AR in contrast to what was observed during the activity period. Considering that testosterone values are lower in adult animals during the period of gonadal regression and in impuber animals, our immunohistochemical results show a significant correlation with the percentage of AR-immunopositive cells. In conclusion, these results demonstrate that the structure of the SVs changes in the activity period of viscacha, probably because of elevated levels of testosterone leading to an increase in the secretory activity of epithelial cells.

Effect of the photoperiod and administration of melatonin on the pars tuberalis of viscacha (Lagostomus maximus maximus): an ultrastructural study.

The pituitary pars tuberalis (PT) is a glandular zone exhibiting well-defined structural characteristics. Morphologically, it is formed by specific secretory cells, folliculostellate cells, and migratory cells coming from the pars distalis. The purpose of this work was to investigate differences in specific cellular characteristics in the PT of viscachas captured in summer (long photoperiod) and winter (short photoperiod), as well as the effects of chronic melatonin administration in viscachas captured in summer and kept under long photoperiod. In summer, the PT-specific cells exhibited cell-like characteristics with an important secretory activity and a moderate amount of glycogen. In winter, the PT-specific granulated cells showed ultrastructural variations with signs of a reduced synthesis activity. Also, PT showed a high amount of glycogen and a great number of cells in degeneration. After melatonin administration, the ultrastructural characteristics were similar to those observed in winter, but the amount of glycogen was higher. These results suggest possible functional implications as a result of morphological differences between long and short photoperiods, and are in agreement with the variations of the pituitary-gonadal axis, probably in response to the natural photoperiod changes through the pineal melatonin. The ultrastructural differences observed in PT, after melatonin administration, were similar to those observed in the short photoperiod, thus supporting the hypothesis that these cytological changes are induced by melatonin.

Ultrastructural and biochemical seasonal changes in epididymal corpus and cauda of viscacha (Lagostomus maximus maximus).

The reproductive and adaptative behavior of wild rodents is synchronized primarily by the photoperiod. The viscacha, a South American rodent of nocturnal habits and seasonal reproduction is photoperiod-dependent and its reproductive behavior is regulated by the retinohypothalamic-pituitary pineal axis. Adult males exhibit an annual reproductive cycle with periods of maximum gonadal activity (summer-early autumn) and gonadal regression (winter). The corpus and the cauda, the most sensitive segments of the epididymis to changes induced by the photoperiod, were analyzed using electron microscopy and enzymatic biochemistry. During gonadal regression, principal and clear cells showed signs of involution with respect to the activity period. These were characterized by more irregular nuclei, smaller cytoplasms, large vacuoles, altered mitochondria, and glycogen deposits. All cellular populations of the epididymal epithelium in regression presented abundant lysosome-like dense bodies during the active period. In addition, we measured the activity of four acid glycosidases in the cauda epididymis along the reproductive cycle. N-acetyl-beta-D-glucosaminidase (NAG), an enzyme that degrades endocytosed substances from the epididymal lumen, increased significantly during gonadal regression relative to the active period. These results demonstrate that the viscacha epididymis exhibits significant ultrastructural and biochemical changes during the reproductive cycle. We demonstrate that during regression, melatonin secretion in viscacha increases. This study shows that the epididymal epithelium is reduced. Thus, we postulate that the changes observed in the epididymis are modulated by pineal melatonin. Despite these changes, the epididymis might maintain a microenvironment suitable for the survival of stored spermatozoa.

Morphological and endocrine study of the ovarian interstitial tissue of viscacha (Lagostomus maximus maximus).

The morphological and endocrine aspects of the ovarian interstitial tissue of adult female viscachas were investigated to establish the probable function and the biological significance of this compartment in this rodent. Pregnant and nonpregnant adult female viscachas were used. The histological characteristics, histochemical properties, and ultrastructural features of the interstitial tissue were studied. A morphometric study was carried out to measure the relative area of lipid droplets. The progesterone and androstenedione levels in ovarian tissue as well as in serum were determined by radioimmunoassay. In this species, the histological observations showed an abundant interstitial tissue that contained a large amount of lipids. The cholesterol and its esters were present in nonpregnant females and were scarce in pregnant animals. The most ultrastructural differences were observed at mid-pregnancy. At this stage, the interstitial cells showed features that suggested higher steroidogenic activity. Furthermore, during mid-pregnancy, the relative area of lipid droplets was smaller. Both progesterone and androstenedione levels in ovarian tissue and serum were higher during pregnancy. Our results suggest that the interstitial tissue may be storage of precursor substances for the steroidogenesis via. These precursors are probably used when the endocrine requirements are high, that is, during the pregnancy. Thus, this compartment may contribute to the normal gestation of Lagostomus. However, the relation between the interstitial tissue and the pregnancy is complex, and further studies are needed to clearly establish it.

Effects of chronic exposure to cadmium on prostate lipids and morphology.

Cadmium is an environmental toxic metal implicated in human prostate carcinogenesis. The mechanism of its toxicity is not fully understood. Previously, we showed that cadmium exposure induces oxidative stress, especially lipid peroxidation. This study evaluates the effect of chronic exposure to 0.886 mM of cadmium (Cd) per liter in the drinking water on prostate lipid content and metabolism in Wistar rats. We determined the lipid profile and measured the expression of lipogenic enzymes: FAS, GPAT, LPL, DGAT-1, DGAT-2, ACO, CPT-1 and CT, and of certain factors involved in lipid regulation and fatty acid transporters: FAT/CD36, E-FABP, SREBP-2, PPAR-gamma and PPAR-alpha by RT-PCR. Ultrastructure was analyzed by electron microscopy and, as prostate is an androgen controlled gland, AR expression was measured by RT-PCR and Western blot. Cd altered the prostatic lipid profile. Triglycerides (TG) and esterified cholesterol (EC) decreased, free cholesterol (FC) and phospholipids (PL) increased and total cholesterol (TC) did not change. FAS, MDH and IDH activities did not vary but G6PDH decreased significantly in Cd group. Regarding TG synthesis, DGAT-1 decreased while GPAT increased and FAS, LPL and DGAT-2 remained unchanged. Regarding beta oxidation, CPT-1 increased while ACO expression decreased in Cd group. In the PL pathway, CT expression was increased. All these results would justify the decrease of TG in Cd group when compared to control. In the cholesterol metabolic pathway, HMGCoAR and SREBP-2 increased. PPAR-alpha increased but PPAR-gamma did not change. Regarding fatty acid transporters, FAT/CD36 decreased, while E-FABP increased. AR mRNA and protein expression decreased. Ultrastructural analysis showed a decrease in lipid droplets and signs of cellular damage in the Cd group. Cadmium exposure induces important changes in prostatic lipid profile and metabolism, confirmed by the morphology analyses, which also showed signs of cellular damage. These results could be important to further understanding the complex mechanism of cadmium toxicity in prostate and in the development of better treatments for people and animals exposed to the heavy metal.

Seasonal variations in the heterologous binding of viscacha spermatozoa: A scanning electron microscope study

Seasonal changes in the reproductive activity of the adult male viscacha (Lagostomus maximus maximus) were investigated during the annual reproductive cycle. Assays of heterologous in vitro binding between compatible gametes were used to evaluate the ability of viscacha spermatozoa to achieve primary binding during its annual reproductive cycle. Sperm were collected by mincing cauda epididymis in HECM-3 medium and the sperm concentration and motility were evaluated. Cumulus-free and zona-free oocytes obtained from superovulated hamsters were inseminated in vitro with capacitated sperm suspensions, incubated at 37ºC, 5% CO2 for 3 h, and then processed for studies by scanning electronic microscopy. Statistical analysis was used to compare the quantitative differences. The number of spermatozoa significantly decreases during the regression period, while sperm motility was progressive speed in both periods. During the active period elevated sperm binding to cumulus-free and zona-free oocytes was observed, while the binding during the regression period decreased drastically. In both periods, oocyte microvilli covered sperm heads and tails. These results suggest that the ability of viscacha spermatozoa to participate in gamete recognition is profoundly affected. This would likely be related to different functional stages of the spermatozoa and their epididymal microenvironment during the annual reproductive cycle of viscacha.

Morphological and biochemical study of the pineal gland of pregnant and non-pregnant female vizcachas (Lagostomus maximus maximus).

OBJECTIVES: The morphological and biochemical aspects of the pineal gland of pregnant and non-pregnant female vizcachas were investigated to examine whether there is a relationship between the pregnancy and this gland. METHODS: Pregnant and non-pregnant adult female vizcachas were used. The ultrastructure of type I cell (dark and light pinealocytes) was studied. Pineal and serum melatonin were investigated by radioimmunoassay. Radiometric methods were applied to investigate the arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O-methyltransferase (HIOMT) activities. RESULTS: The morphological study showed the existence of dark and light pinealocytes. The differences between both cells are slight. However, the appearance of dark pinealocytes suggests that they have a high activity. These cells were more abundant in the pineal gland of pregnant females. The pineal melatonin levels and the AA-NAT activity were higher in the pregnant females. Serum melatonin and HIOMT activity did not show significant differences. CONCLUSION: Our results suggest that the morphological and biochemical changes in the pineal gland of pregnant vizcachas are associated with the gestation. However, the relation between the pineal gland and the pregnancy is complex and further studies are necessaries to clearly establish it.

Ultrastructural and immunocytochemical studies of the viscacha (Lagostomus maximus maximus) pituitary pars tuberalis.

The hypophyseal pars tuberalis (PT) has been the focus of numerous studies attempting to understand its physiological role in the reproductive regulation and modulation by the neuroendocrine system. Ultrastructural studies of the PT in a number of species have shown that it consists of a well-developed hypophyseal area with important secretory activity, demonstrated by the abundance of secretory granules in the cytoplasm and the marked blood irrigation. This article describes ultrastructural and immunocytochemical aspects of the PT in viscachas captured in their habitat. The cell types identified were PT-specific cells, agranulated cells, and Folliculostellate cells. PT-specific cells are divided into type I and II. Type I cells have cytoplasms with secretory granules of 150-500 nm diameter. The secretory granules of type II PT-specific cells are 65-200 nm in diameter. Both cellular types exhibit numerous nerve endings on the plasmatic membranes. Agranulated cells exhibit nuclei with lax chromatin, mitochondria, phagosomes, scarce Golgi complex, and rough endoplasmic reticulum. Folliculostellate cells exhibit an irregularly shaped and moderately condensed nucleus. All the described cellular types exhibit deposits of cytoplasmic glycogen. The immunocytochemical study revealed the presence of cells immunostained for LH-beta and FSH-beta in the PT caudal zone. ACTH was only detected in the zona tuberalis. No staining was observed with antiprolactin, anti-TSH-beta, and anti-GH sera. Folliculostellate cells exhibited staining with anti-S-100. The results demonstrate that the viscacha PT is a hypophyseal zone with specific cellular types, which exhibits evident secretory activity. The presence of nerve endings suggests neural control of the function of PT cells.

Seasonal variations in the heterologous binding of viscacha spermatozoa. A scanning electron microscope study.

Seasonal changes in the reproductive activity of the adult male viscacha (Lagostomus maximus maximus) were investigated during the annual reproductive cycle. Assays of heterologous in vitro binding between compatible gametes were used to evaluate the ability of viscacha spermatozoa to achieve primary binding during its annual reproductive cycle. Sperm were collected by mincing cauda epididymis in HECM-3 medium and the sperm concentration and motility were evaluated. Cumulus-free and zona-free oocytes obtained from superovulated hamsters were inseminated in vitro with capacitated sperm suspensions, incubated at 37 degrees C, 5% CO2 for 3 h, and then processed for studies by scanning electronic microscopy. Statistical analysis was used to compare the quantitative differences. The number of spermatozoa significantly decreases during the regression period, while sperm motility was progressive speed in both periods. During the active period elevated sperm binding to cumulus-free and zona-free oocytes was observed, while the binding during the regression period decreased drastically. In both periods, oocyte microvilli covered sperm heads and tails. These results suggest that the ability of viscacha spermatozoa to participate in gamete recognition is profoundly affected. This would likely be related to different functional stages of the spermatozoa and their epididymal microenvironment during the annual reproductive cycle of viscacha.

Epididymis of viscacha (Lagostomus maximus maximus): morphological changes during the annual reproductive cycle.

Little is known about morphological changes in the epididymis in relation to the natural photoperiod or their influence on sperm maturation. The viscacha is a seasonal rodent living in the Southern Hemisphere. The adult males exhibit an annual reproductive cycle with periods of maximum gonadal activity and gonadal regression. In this work, we studied seasonal variations in the morphology and cellular population of the epididymis during both periods, and we compared these results with those recorded at the testicular level. Epididymides were removed and studied by light microscopy. Measurements of luminal diameter, epithelial height, thickness of the lamina propria, and relative cellular distribution were performed. Analysis of variance (ANOVA) or nonparametric ANOVA was used to compare the results. Striking quantitative and qualitative changes were observed. Epididymides in periods of gonadal regression showed a significant decrease in luminal diameter and epithelial height in cauda, while the thickness of the lamina propria increased. In the epididymal corpus, the number of clear cells increased, and the cytoplasm of principal cells showed numerous giant vacuoles. During the active period, the number of halo cells increased and the cytoplasm of these cells was filled with dense bodies. In conclusion, the epididymis of the viscacha exhibits important seasonal morphological changes throughout annual reproductive cycle. The epididymal corpus and cauda segments appeared to be the segments most sensitive to seasonal cyclical variations of the external environment. We therefore postulate that the epididymal morphology of the viscacha probably could be regulated by the natural photoperiod.

Seasonal variations in the heterologous binding of viscacha spermatozoa. A scanning electron microscope study.

Seasonal changes in the reproductive activity of the adult male viscacha (Lagostomus maximus maximus) were investigated during the annual reproductive cycle. Assays of heterologous in vitro binding between compatible gametes were used to evaluate the ability of viscacha spermatozoa to achieve primary binding during its annual reproductive cycle. Sperm were collected by mincing cauda epididymis in HECM-3 medium and the sperm concentration and motility were evaluated. Cumulus-free and zona-free oocytes obtained from superovulated hamsters were inseminated in vitro with capacitated sperm suspensions, incubated at 37 degrees C, 5

CO2 for 3 h, and then processed for studies by scanning electronic microscopy. Statistical analysis was used to compare the quantitative differences. The number of spermatozoa significantly decreases during the regression period, while sperm motility was progressive speed in both periods. During the active period elevated sperm binding to cumulus-free and zona-free oocytes was observed, while the binding during the regression period decreased drastically. In both periods, oocyte microvilli covered sperm heads and tails. These results suggest that the ability of viscacha spermatozoa to participate in gamete recognition is profoundly affected. This would likely be related to different functional stages of the spermatozoa and their epididymal microenvironment during the annual reproductive cycle of viscacha.

Morphological changes and oxidative stress in rat prostate exposed to a non-carcinogenic dose of cadmium.

Cadmium chloride is an environmental toxicant implicated in human prostate carcinogenesis. The mechanism of its toxicity is far from fully understood. This study evaluates the effect of exposure to an oral non-carcinogenic dose of cadmium (15 ppm in drinking water for three months) on different parameters of the ventral prostatic lobe of normal and exposed rats. We analyzed the histology by optic light microscopy, activities of antioxidant enzymes (CAT, SOD, GPx and G-6-PDH), expression of iNOS and COX-2 by Western blot, expression of MT-I, MT-II, IGF-I, IGF-BP5 and rtert by RT-PCR. Histological changes were found: the height of the cells decreased, acinar lumen were enlarged and they lost the typical invaginations. Lipoperoxidation was increased in the Cd group and the antioxidant enzymes changed their activities: SOD increased, CAT and G-6-PDH decreased and GPx did not show variations. iNOS and COX-2 did not change their expressions. MT-I and IGF-BP5 mRNA increased while MT-II, IGF-I and rtert did not show variations. Cd exposure induces important morphological changes in the prostate, which could be a consequence of lipoperoxidation and oxidative stress, which are not related to iNOS and COX-2. The histology suggests an involution state of the gland, confirmed by the expression of IGF-I, IGF-BP5 and rtert.

Bilateral enucleation and captivity influence the reproductive cycle of male Viscacha (Lagostomus maximus maximus).

The viscacha (Lagostomus maximus maximus) is a seasonal rodent living in the Southern Hemisphere. The adult males exhibit an annual reproductive cycle characterized by a gonadal regression period during winter. In this study, we investigated the effects of bilateral enucleation and captivity on their annual reproductive cycle. Testicular volume relative to body weight was recorded monthly in intact and bilaterally enucleated animals placed under natural photoperiod, water, and food ad lib. and constant temperature. Testes and accessory organs were evaluated by qualitative and quantitative light microscopic studies. The intact animals showed an annual reproductive cycle with complete gonadal atrophy in the first year. In the second year, testicular regression was observed but attenuated in regard to that recorded in the first winter period, indicating that adaptive changes might be involved. Bilateral enucleation in the viscacha dampened and extended the period of its annual reproductive cycle. The results suggest that both conditions, constant captivity and enucleation, produced stimulatory effects on the reproductive system of this rodent. Furthermore, local control mechanisms might be responsible for the morphological differences observed in testes, epididymis, and seminal vesicles from both groups, which exhibited similar levels of serum testosterone. Finally, an intact retinohypothalamic-pineal axis and/or photoperiodic input would be necessary to maintain the reproductive cycle amplitude and timing in viscacha.

Chronic zinc deficiency induces an antioxidant adaptive response in rat lung.

Few studies are available about the role of dietary zinc (Zn) in respiratory diseases. Adult male rats were divided into 2 groups and fed respectively a moderate Zn-deficient diet and a Zn-adequate control diet. In lung tissue at 2 months, thiobarbituric acid-reactive species (TBARS), total glutathione, glutathione disulfide, protein carbonyls, metallothionein, and the activities of glutathione peroxidase (GPx), catalase, CuZn-superoxide dismutase (CuZnSOD) and glucose-6-phosphate dehydrogenase (G-6-PDH) were increased, but protein thiols decreased. In lung tissue at 4 months, TBARS, metallothionein, and the activities of CuZnSOD, Mn-superoxide dismutase (MnSOD) increased. The activities GPx, catalase, G-6-PDH were lower than control group. The changes were accompanied by histological alterations in Zn-deficient lung. The results provide evidence of the pro-oxidative effects of Zn-deficiency in lung, and suggest that the time of treatment play a key role in determining lung susceptibility to oxidative stress.