Oviduct morphology and estrogen receptors ERα and ERß expression in captive Chinchilla lanigera (Hystricomorpha: Chinchillidae).

Chinchilla lanigera is a hystricomorph rodent from South America whose reproductive biology presents particular characteristics that distinguishes it from other Rodentia species, such as low reproductive rate, seasonal breeding pattern, and long estrous cycle. Nevertheless, reproductive features in female chinchillas are still poorly investigated, with a scarce knowledge concerning the estrous cycle and the histology of reproductive organs. In this study, we investigate the morphology, histomorphometry, secretory activity, and immunolocalization of estrogen receptors ERα and ERß in oviducts of nulliparous chinchillas, euthanized at fall season in Brazil. Follicular phase of estrous cycle of all studied animals was characterized by ovary and uterine morphology inspection, as well as vaginal cytology. Similar to other mammals, the oviduct wall of infundibulum, ampulla and isthmus was composed of mucosa, muscle, and serosa layers. Morphometric data of oviduct layers were used for identifying each oviduct segment. In the follicular phase, the oviduct was characterized by intense secretory activity, mainly in the ampulla, and expression of ERα and ERß throughout the oviduct epithelium. Both ERα and ERß were also detected in the connective tissue and smooth muscle cells. Our findings point out to the important role of estrogen in this female organ. Similar wide distribution of both ER proteins has been described for human Fallopian tube. Taken together, our data add to the understanding of the reproductive biology of female chinchillas, and may assist in the intensive breeding of this species and any eventual endeavor for conservation of chinchillas in the wild.

Effects of dantrolene on apoptosis and immunohistochemical expression of NeuN in the spinal cord after traumatic injury in rats.

Dantrolene has been shown to be neuroprotective by reducing neuronal apoptosis after brain injury in several animal models of neurological disorders. In this study, we investigated the effects of dantrolene on experimental spinal cord injury (SCI). Forty-six male Wistar rats were laminectomized at T13 and divided in six groups: GI (n = 7) underwent SCI with placebo and was euthanized after 32 h; GII (n = 7) underwent laminectomy alone with placebo and was euthanized after 32 h; GIII (n = 8) underwent SCI with dantrolene and was euthanized after 32 h; GIV (n = 8) underwent SCI with placebo and was euthanized after 8 days; GV (n = 8) underwent laminectomy alone with placebo and was euthanized after 8 days; and GVI (n = 8) underwent SCI with dantrolene and was euthanized after 8 days. A compressive trauma was performed to induce SCI. After euthanasia, the spinal cord was evaluated using light microscopy, TUNEL staining and immunochemistry with anti-Caspase-3 and anti-NeuN. Animals treated with dantrolene showed a smaller number of TUNEL-positive and caspase-3-positive cells and a larger number of NeuN-positive neurons, both at 32 h and 8 days (P ≤ 0.05). These results showed that dantrolene protects spinal cord tissue after traumatic SCI by decreasing apoptotic cell death.

[Proliferation, apoptosis and mammary gland histomorphometry of thyroxine-treated rats on lactation and after weaning and development of offspring]. / Proliferação, apoptose e histomorfometria da glândula mamária de ratas tratadas com tiroxina na lactação e ao desmame e desenvolvimento dos filhotes.

The purpose of this study was to evaluate mammary gland histomorphometry and proliferation rate and apoptosis of thyroxine-treated rats by CDC-47 and caspase-3 immunoexpression. The development of thyroxine-treated rats offspring was also evaluated. Thirty-six female rats were used, distributed in two groups, treated and non-treated with thyroxine. After 60 days of treatment, with thyroxine, rats were mated. Six animals/group were sacrificed on the 2nd and 21st days of lactation and on the 5th day after weaning. A significant difference was observed between groups only on the 5th day after weaning. Thyroxine treatment increased apoptosis rate, which was characterized by a higher caspase-3 expression in mammary epithelial cells. Thyroxine-treated mothers presented changed behavior, but there was no significant difference regarding taking care of offspring, as for cleaning offspring and keeping them warm. Taking into account sex and size of offspring, those from control and thyroxine-treated mothers presented no significant difference of weight and weaning. In conclusion, administering low doses of thyroxine increases apoptosis rate, which is characterized by the increased caspase-3 immunoexpression in mammary epithelial cells 5 days after weaning. But does not affect proliferation rate and development of thyroxine-treated rats offspring.

Proliferação, apoptose e histomorfometria da glândula mamária de ratas tratadas com tiroxina na lactação e ao desmame e desenvolvimento dos filhotes / Proliferation, apoptosis and mammary gland histomorphometry of thyroxine-treated rats on lactation and after weaning and development of offspring

O objetivo deste estudo foi avaliar a histomorfometria e a taxa de proliferação e apoptose da glândula mamária de ratas tratadas com tiroxina pela imuno-expressão de CDC-47 e caspase-3, respectivamente. Também foi avaliado o desenvolvimento dos filhotes de ratas tratadas com tiroxina. Foram utilizadas 36 ratas distribuídas em dois grupos, tratado com tiroxina e controle. Após 60 dias de tratamento com tiroxina, as ratas foram acasaladas. Seis animais/grupo foram sacrificados no 2° e 21° dias de lactação e no 5° dia após o desmame. Houve diferença significativa entre grupos apenas no quinto dia após o desmame. O tratamento com tiroxina aumentou a taxa de apoptose caracterizada pela maior expressão de caspase-3 nas células do epitélio mamário. As mães tratadas com tiroxina apresentaram comportamento alterado, mas não houve diferença significativa no que se refere aos cuidados com o filhote quanto a higienização e aquecimento. Levando-se em consideração o sexo e o tamanho da ninhada, os filhotes das ratas tratadas com tiroxina e controle não apresentaram diferença significativa de peso ao desmame. Conclui-se que a administração de baixas doses de tiroxina aumenta a taxa de apoptose, caracterizada pelo aumento da expressão de caspase-3 no epitélio mamário cinco dias após o desmame, mas não altera a taxa de proliferação celular e o comportamento materno.

The purpose of this study was to evaluate mammary gland histomorphometry and proliferation rate and apoptosis of thyroxine-treated rats by CDC-47 and caspase-3 immunoexpression. The development of thyroxine-treated rats offspring was also evaluated. Thirty-six female rats were used, distributed in two groups, treated and non-treated with thyroxine. After 60 days of treatment, with thyroxine, rats were mated. Six animals/group were sacrificed on the 2nd and 21st days of lactation and on the 5th day after weaning. A significant difference was observed between groups only on the 5th day after weaning. Thyroxine treatment increased apoptosis rate, which was characterized by a higher caspase-3 expression in mammary epithelial cells. Thyroxine-treated mothers presented changed behavior, but there was no significant difference regarding taking care of offspring, as for cleaning offspring and keeping them warm. Taking into account sex and size of offspring, those from control and thyroxine-treated mothers presented no significant difference of weight and weaning. In conclusion, administering low doses of thyroxine increases apoptosis rate, which is characterized by the increased caspase-3 immunoexpression in mammary epithelial cells 5 days after weaning. But does not affect proliferation rate and development of thyroxine-treated rats offspring.

Ehrlich tumor as model to study artificial hyperthyroidism influence on breast cancer.

This study used Ehrlich solid tumor as an experimental model for breast cancer to investigate the effects of thyroid hormones and castration on tumor development in adult female mice. Artificial hyperthyroidism was induced in animals, and after a 30-day-treatment, they received subcutaneous injection of neoplastic cells between left plantar cushions. We measured the growth of tumor inoculated in the paws for 10 days at necropsy. Hyperthyroidism induction led to significantly increased tumor size in non-castrated animals, and alterations were less intense in association with artificial hyperthyroidism and castration (p<0.05). Histomorphologic and histomorphometric analyses and neoplastic cell characterization were carried out by measuring nuclear diameter, by evaluating AgNORs, by mitotic count, and by measuring cell proliferation using immunohistochemical marker CDC47. At the end of the experiment, we noted metabolism and a decrease in cell proliferation in groups having received l-thyroxine, which were more evident in the non-castrated group (p<0.001).

Técnica histoquímica aplicada ao tecido ósseo desmineralizado e parafinado para o estudo do osteócito e suas conexões / Histochemical tecnique for osteocyte and its connections study in descalcified and paraffined bone tissue

O osteócito vem sendo alvo de pesquisas recentes. A avaliação in situ de sua morfologia, da atividade e das características de suas conexões é difícil, portanto é realizada apenas com técnicas avançadas. Devido à importância desse tipo celular na manutenção da matriz óssea, este estudo propõe uma técnica de coloração pela prata como alternativa para o estudo do osteócito e suas conexões em tecido ósseo desmineralizado e parafinado. Cortes de 4æm do fêmur de ratas foram desmineralizados, desparafinados em xilol e hidratados em concentrações decrescentes de álcool etílico (ETOH) e água miliQ. Para a impregnação foram utilizadas soluções de nitrato de prata a 50 por cento e de ácido fórmico a 1 por cento com 2 por cento de gelatina microbiológica em estufa a 40ºC. Essa técnica permite visualizar facilmente as bordas lacunares dos osteócitos e suas conexões, proporcionando uma alternativa simples e eficaz para o estudo da morfologia desse tipo celular até então ainda não proposta com essa finalidade.