Caracterização morfológica e frequência dos estádios do ciclo do epitélio seminífero em preás (Galea spixii Wagler, 1831) criados em cativeiro / Morphological characterization and frenquency of stages of the seminiferous epithelium cycle in captive bred Spix's yellow-toothed (Galea spixii Wagler, 1831)

Estudos baseados nas características testiculares estão altamente relacionados com a eficiência reprodutiva de varias espécies. Assim, o projeto desenvolvido teve como objetivo identificar as células do epitélio seminífero, caracterizar histologicamente suas associações, que formam os estádios, e determinar a frequência destes. Os fragmentos de testículos, com 30, 45, 60, 75, 90, 105, 120, 150 dias foram coletados no Centro de Multiplicação da Universidade Federal Rural do Semi-árido, Mossoró/ RN. Passando pelos processos de fixação, lavagens em soluções de concentrações crescentes de álcoois (70-100 por cento), desidratação em xilol, inclusão em Histosec®, preparação das lâminas histológicas, colorações em Hematoxilina e Eosina (HE) e suas fotomicrografias para a caracterização dos núcleos celulares do epitélio germinativo e a definição dos oitos estágios do ciclo do epitélio seminífero (CES) baseados no Método da Morfologia Tubular. Das faixas etárias analisadas todos os animais de 90-150 dias de idade apresentaram todos os estádios do CES. Os estádios I e III foram os que apresentaram maior e menor freqüência, respectivamente. Os animais caracterizados como pré-púberes (30 dias), púberes (45-90 dias de idade) e pós-púberes (105150 dias de idade) apresentaram os estádios I, VIII e IV com uma maior freqüência, respectivamente.

Studies based on the testicular characteristics are strongly associated with the reproductive efficiency of various species. Thus, the developed project aimed to identify the cells of the seminiferous epithelium, histologically characterized their associations, which form stages, and determine the frequency of these. The fragments of testes, 30, 45, 60, 75, 90, 105, 120, 150 days were collected Multiplication Center of Universidade Federal Rural do Semi-Árido, Mossoró, RN. Through the process of fixing, washing in solutions of increasing concentrations of alcohols (70-100 percent), dehydration in xylene, inclusion in Histosec ®, preparation of histological slides, stained with hematoxylin and eosin (HE) and their photomicrographs for the characterization of cell nuclei of the germinal epithelium and the definition of the eight stages of the seminiferous epithelium cycle (CES) based on the tubular morphology method. The different age groups all animals at 90 to 150 days of age showed all stages of the CES. Stages I and III showed the highest and lowest frequency, respectively. Animals categorized as prepubertal (30 days), pubertal (45 to 90 days old) and postpubertal (105 to 150 days of age) had stage I, IV and VIII with a higher frequency, respectively.

Nuclear calcium signaling: a cell within a cell

Calcium (Ca2+) is a versatile second messenger that regulates a wide range of cellular functions. Although it is not established how a single second messenger coordinates diverse effects within a cell, there is increasing evidence that the spatial patterns of Ca2+ signals may determine their specificity. Ca2+ signaling patterns can vary in different regions of the cell and Ca2+ signals in nuclear and cytoplasmic compartments have been reported to occur independently. No general paradigm has been established yet to explain whether, how, or when Ca2+ signals are initiated within the nucleus or their function. Here we highlight that receptor tyrosine kinases rapidly translocate to the nucleus. Ca2+ signals that are induced by growth factors result from phosphatidylinositol 4,5-bisphosphate hydrolysis and inositol 1,4,5-trisphosphate formation within the nucleus rather than within the cytoplasm. This novel signaling mechanism may be responsible for growth factor effects on cell proliferation.

Maturation of pig oocytes in vitro in a medium with pyruvate

The aim of in vitro maturation oocyte systems is to produce oocytes of comparable quality to those derived in vivo. The present study was designed to examine the surface morphological changes of the cumulus-oocyte complex (COC) and nuclear maturation in a culture system containing pyruvate. Ovaries were obtained from a slaughterhouseand transported to the laboratory within 2 h at 35-39°C,and rinsed three times in 0.9 percent NaCl. The COCs were harvested from the ovaries and in vitro maturation was evaluated in San Marcos (SM) medium, a chemically defined culture system containing 22.3 mM sodium pyruvate. Oocytes were cultured in SM, SM + porcine follicular fluid (pFF) and in SM + pFF + gonadotropins (eCG and hCG) for 20-22 h and then without hormonal supplements for an additional 20-22 h. After culture, the degree of cumulus expansion and frequency of nuclear maturation were determined. Oocytes matured in SM (40.9 percent) and SM + pFF (42.9 percent) showed moderate cumulus expansion, whereas oocytes matured in SM + pFF + gonadotropins (54.6 percent) showed high cumulus expansion. The maturation rate of cultured oocytes, measured in function of the presence of the polar corpuscle, did not differ significantly between SM (40.9 ± 3.6 percent) and SM + pFF (42.9 ± 3.7 percent). These results indicate that pig oocytes can be successfully matured in a chemically definedmedium and suggest a possible bifunctional role of pyruvate as an energy substrate and as an antioxidant protecting oocytes against the stress of the in vitro environment.

Gametogenesis and nucleotypic effects in the tetraploid red vizcacha rat, Tympanoctomys barrerae (Rodentia, Octodontidae)

Nucleotypic effects link DNA content with nuclear size and cell dimensions of reproductive cells in polyploid organisms. We studied the gametogenesis of the allotetraploid rodent Tympanoctomys barrerae, aiming to determine these effects in reproductive cells. The species' cofamily members, Octodon degus and Spalacopus cyanus were used as control. Spermatogenesis and oogenesis in T. barrerae follows the pattern of differentiation and sequence of events of the control species, but varied nucleotypic effects were observed. Exceedingly large, spatulated spermatozoa with a submedially attached flagellum are characteristic of male T. barrerae. The diameter of the nuclei of primordial and growing follicles as well as those of the Graaff follicles, of the granulose, and of luteal cells are significantly larger and heavily heterochromatic. Moreover, the width of the pellucid zone is 108 percent thicker in T. barrerae than in S. cyanus. Binucleation was recorded in 26 percent of luteal bodies examined whereas no binucleated cells are detected in the diploid control. Likewise, large heterochromatic nucleoli were observed in the follicle cells but not in S. cyanus. This finding and the high heterochromatin content of reproductive cells in the red vizcacha rat is probably associated with its genome complexity so that redundant genetic information is silenced through heterochromatinization.

Sperm ultrastructure and spermatogenesis in the lizard, Tropidurus itambere

Spermatogenesis, with emphasis on spermiogenesis, is described for the lizard, Tropidurus itambere, using light microscopy, phase contrast and epifluorescence, as well as scanning and transmission electron microscopy. Cellular differentiation involves events of chromatin condensation, nuclear elongation and the formation of structural complexes, such as the acrosomal and axonemal ones. Other new characteristics, exclusive for this species, include various aspects of the subacrosomal granule, the insertion of the pro-acrosomal vesicle and the development of these structures to participate in the acrosomal complex. Radial projections occurjust above the nuclear shoulders, which have been recognized already from the beginning of cellular elongation. The development of the midpiece, the dense bodies, formation of the flagellum and elimination of residual cytoplasm result in the final characterization of the mature spermatozoon. Comparisons between Tropiduridae and other lizard families are made.

A unique mechanism of nuclear division in Giardia lamblia involves components of the ventral disk and the nuclear envelope

The fine structure of the binucleate, parasitic protist Giardia lamblia during interphase and divisional stages was studied by serial thin sectioning and three-dimensional reconstructions. The earlier sign of nuclear division is the development of a few peripheral areas of densely packed chromatin directly attached to the inner nuclear envelope. An intracytoplasmic sheet of ventral disk components grows from the cell periphery towards one of the nuclei, apparently constricting this nucleus, which becomes located at a ventral bulge. After the basal bodies become duplicated, a full nuclear division occurs in trophozoites, giving two pairs of parent-daughter nuclei. This full division occurs in a dorsal-ventral direction, with the resulting nuclear pairs located at the sides of the two sets of basal bodies. A new ventral disk is formed from the disk-derived sheets in the cell harboring the four nuclei. Cytokinesis is polymorphic, but at early stages is dorsal-to-dorsal. Encysting trophozoites show the development of Golgi cisternae stacks and dense, specific secretory granules. 3-D reconstructions show that cysts contain a single pair of incompletely strangled nuclei. The dividing Giardia lacks a typical, microtubular spindle either inside or outside the nuclei. The nuclear envelope seems to be the only structure involved in the final division of the parent-daughter nuclei.

Study of evaluation criteria used for in vitro bovine oocyte selection and maturation

Production of bovine preimplantation embryos in vitro requires beneficial maturation conditions and high quality oocytes at the germinal vesicle stage. The current classification of oocytes is based on character of the cumulus cell investment around the oocyte. We wished to study the nuclear stage of immature oocytes selected for in vitro maturation according to cumulus cell character and, in the other hand, to compare the relationship among 3 parameters utilized to evaluate in vitro maturation of bovine oocytes (degree of cumulus expansion, meiotic maturation rate and in vitro fertilization rate) when fetal calf serum, steer serum and bovine follicular fluid supplementation were used. Ovaries were collected at an abattoir and the oocytes harvested. As regards selection criteria, immature oocytes were classified as Class A, B, C and D according to the character of the cumulus cells. A high percentage of Class A oocytes (87.7) were in the germinal vesicle stage with respect to the other classes (p < 0.05). Significant differences were found in the meiotic maturation rate in Class A oocytes (76.5) versus those of the other classes (p < 0.05). The meiotic maturation rate diminished to 47.5 when Class A oocytes were denuded and then matured in vitro (p < 0.05). As regards maturation criteria, there was no cumulus expansion when oocytes were matured in TCM-199 without supplementation, partial expansion with the addition of fetal calf serum and full expansion when supplemented with steer serum or bovine follicular fluid. No significant differences were found in the meiotic maturation rate for the various treatments. In vitro fertilization rate was significantly lower in media without supplementation versus supplemented media (p < 0.05), but no significant differences were found between the supplemented media inter se. There is no direct relationship between the three studied parameters to evaluate in vitro maturation. Class A oocytes are the most likely to mature in vitro as they not only have a close association with their surrounding cumulus cells, but are also very numerous in the germinal vesicle stage. The degree of cumulus expansion and the meiotic maturation rate have a relative importance in evaluating in vitro maturation, as oocyte maturation implies not only nuclear events but also at other cellular levels, as evaluated by in vitro fertilization

Nuclear maturation inhibitors in Bufo arenarum oocytes

In Bufo arenarum, as in other amphibian species, nuclear maturation--characterized by germinal vesicle breakdown (GVBD) and meiosis resumption--is induced by progesterone. Seasonal variations in the sensitivity to progesterone were observed in Bufo arenarum oocytes. The gonadotropin FSH and drugs such as dbcAMP and xanthine did not affect nuclear maturation per se, although they acted as negative modulators of progesterone-induced nuclear maturation. Similar results to FSH in nuclear maturation were obtained with 17 beta-estradiol

Effect of nuclear localization signal-receptor interaction on nuclear envelope-associated ATPase activity.

The possibility that interaction of the nuclear localization signal (NLS) with its pore receptor may directly stimulate nuclear envelope-associated ATPase activity and consequently provide energy for protein translocation across the pore has been studied. ATPase activity was assayed after cross-linking of the prototype NLS peptide with its pore receptor, or after preincubation of envelopes with NLS-albumin conjugates. Neither treatment enhanced enzyme activity. A more complex series of events may be required for energy-generation at the nuclear pore.

Estudo morfométrico da hiperplasia gengival dilantínica em comparaçäo à gengiva normal e gengivite / Morphometric study of dilantin gingival hyperplasia in relation to normal gingiva and gingivitis

A presente pesquisa teve como objetivo, realizar em indivíduos do sexo masculino, um estudo quantitativo do epitélio oral e lâmina própria de gengiva marginal afetada pelo uso do difenil-hidantoinato de sódio e compará-la à gengiva normal e inflamada, a fim de estabelecer, a nível de microscópio de luz, as possíveis diferenças entre os grupos analisados. Em relaçäo ao epitélio, tanto na camada basal como na espinhosa, a análise estatística evidenciou diferenças significantes entre os três grupos nos seguintes parâmetros: relaçäo núcleo/citoplasma; densidade de volume nuclear e citoplasmático; volume nuclear, citoplasmático e celular; número de celular por mmü e espessura do epitélio. Quanto à avaliaçäo morfométrica das estruturas presentes na lâmina própria, a densidade de volume de fibras colágenas e substância intercelular; densidade de volume de núcleos de fibroblastos e células imigrantes; densidade de volume de vasos sanguíneos; número de fibroblastos e número de células imigrantes mostraram diferenças significantes entre si