Ramón S. Piezzi

Ultrastructural and morphometric study of the sertoli cell of the viscacha (Lagostomus maximus maximus) during the annual reproductive cycle

Changes in the morphology of viscacha Sertoli cells were studied during the annual reproductive cycle. Sertoli cells exhibited marked nuclear and cytoplasmic changes. Seasonal variation in nuclear size and shape, chromatin texture, and nucleolus characteristics was observed. The seasonal patterns of the volume densities of the endoplasmic reticulum (ER), mitochondria, Golgi complex, dense bodies and lipid inclusions were distinct. Morphometric analysis revealed that the Golgi complex is the organelle most sensitive to seasonal change. It declined drastically in the regressed testes and its recovery was slow. The ER and mitochondria exhibited seasonal variations in their pattern and content, that was minimal during winter. In contrast, an accumulation of lipid and dense bodies, such as primary and secondary lysosomes, accompanied the spermatogenic arrest. The volume densities of both organelles were maximum during the restoration of spermatogenesis. The length and organization of the inter‐Sertoli junctions also changed with the reproductive cycle. The Sertoli cell number per tubular cross section decreased significantly during the testicular regression, coincident with the presence of Sertoli cells with marked signs of involution. The degree of regression and recovery exhibited by the viscacha Sertoli cells was closely related to that shown by the associated germ cells. Therefore, seasonal endocrine fluctuations and local factors could be involved in the regulation of the morphological and functional characteristics of the viscacha Sertoli cells. These hormonal fluctuations are synchronized by the photoperiod through the pineal gland and its hormone, melatonin. Anat Rec 262:176–185, 2001.

A Light and Electron Microscopic Study of the Pineal Gland of the Viscacha (Lagostomus maximus maximus)

The pineal of viscacha (Lagostomus maximus maximus) is formed by two main cellular populations of pinealocytes and interstitial cells. A connective capsule surrounds the gland sending numerous septa containing blood vessels and nerves. In the specimens collected during summer the pinealocyte displays a defined contour and a clear cytoplasm with abundant ribosomes and numerous cisternae of endoplasmic reticulum. The pinealocytes observed during winter have cytological characteristics of active cells with dense and elongated mitocondria, dense bodies, and a developed Golgi complex. These seasonal glandular changes could be related to the environmental condition and mainly to the summer photoperiod. The presence of abundant nerve terminals in the perivascular spaces suggests a neural control on the blood vessels.

SEASONAL CHANGES OF THE LEYDIG CELLS OF VISCACHA (LAGOSTOMUS MAXIMUS MAXIMUS). A LIGHT AND ELECTRON MICROSCOPY STUDY

The Leydig cells of viscacha (seasonal rodent) show cytoplasmic hypertrophy and regional distribution during the breeding period (summer-autumn). The dominant organelles are smooth endoplasmic reticulum (SER) and mitochondria. A moderately well-developed Golgi, abundant lipid inclusions, dense bodies like lysosomes in different stages, and centrioles are observed. Extensive or focal desmosome and gap-like junctions between neighbouring Leydig cells are present. These cells exhibit an evident hypotrophy and an increase in the number of dense bodies during the gonadal regression in winter (July and August). Cells in different stages of involution are observed in this period. Their nuclei are irregular and heterochromatic. The cytoplasm contains few mitochondria. The vesicular SER is scarse. Irregular and large intercellular spaces with microvilli and amorphous material are present. The junctional complexes are absent. The nuclear and cytoplasmic volume and development of SER and mitochondria increase during the recovery period (spring). The lipid inclusions decrease. Dilatations of the intercellular space with microvilli and limited by focal desmosome-like junctions are observed. In conclusion, the Leydig cells of Lagostomus maximus maximus show deep changes alongside the reproductive cycle. The photoperiod variations, through pineal hypothalamus pituitary axis and the hormone melatonin, are probably responsible for them. Moreover, the fall of serum and tubular testosterone would be one of the factors responsible for gonadal regression.

Stages of the cycle of the seminiferous epithelium of the Viscacha (Lagostomus maximus maximus)

The adult male viscacha (Lagostomus maximus maximus) is a seasonal rodent. It exhibits a short period of testicular regression with partial arrest of spermatogenesis during winter (July‐August). The present study provides the first description of the viscacha spermatogenic cycle during the period of maximum gonadal activity (summer‐autumn).

Colloid in the pituitary pars distalis of viscacha (Lagostomus maximus maximus): ultrastructure and occurrence in relation to season, sex, and growth.

Randomly distributed extracellular colloidal accumulations were observed in the pars distalis of viscacha (Lagostomus maximus maximus). They were preferentially located in the peripheral zone of the gland and showed variability in shape and size. Two different types of colloidal accumulations were found by electron microscopy: 1) those surrounded by nongranulated follicular cells that correspond to characteristic follicles, and 2) those surrounded by granulated cells. In the follicles lined by nongranulated follicular cells, long, prominent microvilli and cytoplasmic processes protruded into the lumen. The frequency of these accumulations varies during the year in adult male animals, showing an increase in number during summer and a decrease during winter. The lowest value was registered in August (winter). The mean follicular diameter did not vary seasonally. The number of colloidal accumulations did not vary seasonally in adult female viscachas, but a significant difference in the mean follicular diameter between pregnant and non‐pregnant females was observed. Pituitaries of immature animals contain fewer colloidal accumulations than those of adults. In fetuses, these accumulations were absent. The administration of melatonin provoked a decrease in the number of these structures. The numeric changes of the colloidal accumulations observed in this study are associated with: 1) the seasonal reproductive activity in adult males, and 2) the reproductive condition, body weight and sexual maturity in males and females. The fact that melatonin administration decreases the population of colloidal accumulations in males suggests participation of the pineal gland in these changes. Anat Rec 258:252–261, 2000.

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. Anat Rec, 2007.

Probable effect of photoperiod on seasonal variation in the nuclear volume of the adrenal cortex of viscacha (Lagostomus maximus maximus)

The neuroendocrine system regulates several organic functions such as reproduction, metabolism and adaptation to the environment. This system shows seasonal changes linked to the environment. The experimental model used in the present study was Lagostomus maximus maximus (viscacha). The reproduction of males of this species is photoperiod dependent. Twenty-four adult male viscachas were captured in their habitat at different times during one year. The adrenal glands were processed for light microscopy. Serial cuts were stained with hematoxylin-eosin for the morphometric study, and 100 nuclei of each zone of the adrenal cortex were counted per animal. Data were analyzed statistically by ANOVA and the Tukey test. The cells of the glomerulosa zone are arranged in a tube-shaped structure. The fasciculata zone has large cells with central nuclei and clearly visible nucleoli and with a vacuolar cytoplasm. In the reticularis zone there are two of types of cells, one with a nucleus of fine chromatin and a clearly visible nucleolus and the other with nuclear pycnosis. Morphometric analysis showed maximum nuclear volumes during the February-March period with values of 133 +/- 7.3 microm3 for the glomerulosa, 286.4 +/- 14.72 microm3 for the fasciculata, and 126.3 +/- 9.49 microm3 for the reticularis. Minimum nuclear volumes were observed in August with values of 88.24 +/- 9.9 microm3 for the glomerulosa, 163.7 +/- 7.78 microm3 for the fasciculata and 64.58 +/- 4.53 microm3 for the reticularis. The short winter photoperiod to which viscacha is subjected could inhibit the adrenal cortex through a melatonin increase which reduces the nuclear volume as well as the cellular activity.

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. Anat Rec, 293:871–878, 2010.

Permeability of the Pineal Gland of the Viscacha (Lagostomus maximus maximus) to the Lanthanum Hydroxide

Lanthanum hydroxide together with a fixative solution was injected into the third ventricle of the viscacha. The tracer penetrated freely into the pineal gland demonstrating an extensive network of intercommunicating intercellular spaces. Numerous gap junctions were observed indicating that they are the predominant interparenchymal contacts. The capillaries were characterized by non‐fenestrated endothelial cells lining a moderate‐sized perivascular space. Precipitates of lanthanum hydroxide were found in the basal membranes of the perivascular and interendothelial spaces, but not in the vascular lumina. The tracer stopped at tight junctions; these constitute a barrier between the pinealocytes and the blood compartment. Active micropinocytosis observed in the capillary walls suggests a transendothelial transport.