Regulação hormonal e interações celulas-matriz extracelular na prostata ventral de ratos

Orientador: Hernandes Faustino de CarvalhoTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaDoutorad

Prostate telocytes change their phenotype in response to castration or testosterone replacement

Telocytes are CD34-positive cells with a fusiform cell body and long, thin cytoplasmic projections called telopodes. These cells were detected in the stroma of various organs, including the prostate. The prostate is a complex gland capable of undergoing involution due to low testosterone levels; and this condition can be reversed with testosterone replacement. Telocyte function in the mature prostate remains to be dermined, and it is not known whether telocytes can take place in tissue remodeling during prostate involution and regrowth. The present study employed structural, ultrastructural and immunohistochemical methods to investigate the telocyte's phenotypes in the ventral prostate (VP) from control (CT), castrated (CS) and testosterone replacement (TR) groups of adult male Wistar rats. Telocytes were found in the subepithelial, perimuscular and interstitical regions around glandular acini. Telocytes from CT animals have condensed chromatin and long and thin telopodes. In CS group, telocytes appeared quiescent and exhibited layers of folded up telopodes. After TR, telocytes presented loose chromatin, abundant rough endoplasmic reticulum and enlarged telopodes, closely associated with bundles of collagen fibrils. We called these cells "telocytes with a synthetic phenotype". As testosterone levels and glandular morphology returned toward to the CT group parameters, after 10 days ofTR, these telocytes progressively switched to the normal phenotype. Our results demonstrate that telocytes exhibit phenotypic plasticity upon androgen manipulation and interact with fibroblast and smooth muscle cells to maintain glandular architecture in control animals and during tissue remodeling after hormonal manipulation9CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP305391/2014-3; 306900/2016-5; 305840/2015-0; 310663/2018-02002/11102-4; 2014/26660-0; 2017/01063-

Estudo de alguns componentes macromoleculares do estroma da postata ventral de ratos : efeitos da orquiectomia

Orientador: Hernandes Faustino de CarvalhoDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A próstata é a glândula de maior volume e expressividade funcional anexa ao aparelho reprodutor masculino de mamíferos. Histologicamente é constituída de porções glandulares túbulo-alveolares. Em roedores, a glândula prostática está representada por pares de lobos ventrais, dorsais e laterais. O desenvolvimento embrionário e pós-natal, bem como a manutenção da estrutura e do funcionamento da próstata, são eventos dependentes de níveis constantes de andrógenos circulantes. A remoção destes andrógenos em animais adultos leva a próstata a um processo de involução que inclui uma redução de tamanho e diminuição ou perda da função secretora. O fato de a próstata humana ser o sítio de um grande número de doenças, nas quais os hormônios exercem um papel importante na etiologia, levou à necessidade de se conhecer as respostas deste órgão frente à ação hormonal em diferentes condições fisio-patológicas. O presente estudo teve como objetivo estudar os efeitos da remoção de andrógenos, por orquiectomia, sobre alguns componentes do estroma da próstata ventral de ratos adultos (Rattus rattus norvegicus), uma vez que as alterações decorrentes da castração no compartimento epitelial já são bem conhecidas e ainda pouco se sabe sobre as modificações no compartimento estromal. A identificação de alguns dos componentes da matriz extracelular foi efetuada utilizando-se métodos de rotina: hematoxilina-eosina e azul de toluidina, métodos histoquímicos, resorcina-fucsina de Weigert, para o sistema elástico; tricrômico de Masson e picrossíriushematoxilina, para fibras colágenas; impregnação pela prata para fibras reticulares; métodos imuno-histoquímicos, para detecção de glicosaminoglicanos(queratam sulfato e condroitim sulfato), e métodos ultra-estruturais. A análise dos efeitos da castração sobre o estroma foi efetuada em próstatas removidas aos 7, 14 e 21 dias após a castração. Os resultados demonstraram que após a castração ocorrem modificações no compartimento estromal, como o aumento da densidade dos componentes celulares e fibrilares tais como células musculares lisas e fibras do sistema elástico e colágenas, respectivamente. Observaram-se ainda um aumento da imunorreatividade do queratam sulfato e perda da imunorreatividade do condroitim sulfato na membrana apical das células epiteliais. Da análise dos componentes fibrilares foi possível observar um aumento na quantidade e espessura das fibras do sistema elástico e um aumento e uma maior agregação das fibras colágenas após a castração. Além da hiperplasia, observou-se que as células musculares lisas apresentaram fenótipos diferenciados, assumindo características ultra-estruturais de células sinteticamente ativas, nas condições de privação hormonal. Este fato indica que este tipo celular pode estar diretamente envolvido na síntese de alguns componentes macromoleculares do estroma prostático. As interações entre mesênquima e epitélio que ocorrem durante a morfogênese têm sua importância reconhecida, entretanto não se pode negligenciar a importância destas i nterações nos processos de manutenção e remodelação do estroma no órgão maduro. Neste sentido, o aprofundamento do conhecimento sobre a matriz extracelular, com base nos resultados obtidos neste trabalho, parece ser de grande importânciaAbstract: The prostate is an acessory gland of the mammalían reproductive system with great volume and high functional importance. Histologically, it is a tubuloalveolar gland. In rodents, the prostatic gland is composed of paired ventral, dorsal and lateral lobes. The embryonic and post.natal development, and the maintenance of the prostatic structure and phísiology are dependent on stable levels of circulanting androgens. The remova I of these androgens in adult animais leads to an involution process, characterized bya reduction in size and a decrease or loss of the secretory function. The fact that the human prostate is the site of a number of díseases, which are in many cases also dependent of hormonal regulation, stimulates the investigation on the response of this organ to different hormonal therapies. This study was developed to verify the effects of androgen withdrawal, by orchiectomy, on some components of the rat ventral prostate. The identification of some extracellular matrix components was performed using routine procedures such as haematoxylin_eosin and toluídine blue staining; hístochemical methods as Weigert's resorcin.fuchsin for elastic system fibers; Masson's trichrome and picrosirius.haematoxylin, for collagen fibers; silver impregnation for reticular fibers; immunohistochemical methods for keratan sulfate and chondroitin sulfate detection and ultrastructural methods. The castration effects on some of the stromal components were examined on prostates removed at the 7, 14 and 21 days after castration. The results showed that castration promotes modifications in the stromal compartment, such as an increased density of cellular (smoth muscle cells) and fibrilar (collagen and elastin) components. It was possible to identify an increase in the amount and thickness of elastic system fibers and an increase in the number and compactness of collagen fiber. The smooth muscle cells presented differentiated phenotypes, attaining ultrastructural features of synthetically active cells. This fact indicates that these cells may be directly involved in the synthesis of some macromolecular components of the prostatic stroma. There is also an increase in keratan sulfate immunoreactivity and loss of reactivity for chondroitin sulfate at the apical membrane of epithelial cells. The mesenchymal.epithelial interactions are important during morphogenesis and they cannot be neglected in the maintenance and remodelling processes of the adult organ. Thus, continuing studies on the extracellular matrix, based on our studies, seems to be of great importanceMestradoBiologia CelularMestre em Ciências Biológica

Modulation Of Smooth Muscle Cell Function: Morphological Evidence For A Contractile To Synthetic Transition In The Rat Ventral Prostate After Castration.

In this study, we evaluated the involvement of rat ventral prostate smooth muscle cells (SMC) in secretory activity and whether this function is modulated after castration. Cell morphology was examined at both light and electron microscopy levels and the organelles involved in secretory function were labeled by the zinc-iodide-osmium (ZIO) method at the ultrastructural level and their volume density was determined by stereology. Castration resulted in marked changes of the SMC, which adopted a spinous aspect and abandoned the layered arrangement observed in the prostates of non-castrated rats. The volume density of ZIO reactive organelles increased progressively after castration, reaching significantly higher levels 21 days after castration. Since previous studies have demonstrated that SMC express SMC markers (even 21 days after castration) and are able to respond to adrenergic stimulation, we concluded that differentiated SMC are able to shift from a predominantly contractile to a more synthetic phenotype without changing their differentiation status.29809-1

Modulação androgênica e estrogênica na próstata: uma abordagem em modelos experimentais de roedores com enfoque na biologia estrutural

O sistema endócrino é uma complexa rede de glândulas e hormônios que regulam muitas das funções do corpo, incluindo crescimento, desenvolvimento e maturação, como as vias de ação de muitos órgãos. A próstata é um importante alvo dos hormônios e sua maturidade funcional e seu desenvolvimento são influenciados pelos níveis de esteroides. O presente grupo de pesquisa tem estudado os potenciais efeitos dos agentes esteroides sobre a próstata masculina e feminina do gerbilo da Mongólia (Meriones unguiculatus), utilizando métodos morfológicos e imuno-histoquímicos. Os resultados têm revelado a próstata do gerbilo da Mongólia como uma importante ferramenta para estudos da ação dos hormônios esteroides e seus antagonistas.The endocrine system is a complex network of glands and hormones that regulates many of the body's functions; including growth, development and maturation, as well as the way several organs operate. The prostate is an important target of hormones and its functional maturity and development are influenced by steroids levels. Our research group has been evaluating the potential effects of the steroidal agents on the Mongolian gerbil (Meriones unguiculatus) male and female prostate using different morphological and immunohistochemical methods. Our results have revealed the Mongolian gerbil prostate as an important tool for the morphofunctional studies of steroid hormones and its antagonist actions.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Tissue Evidence Of The Testosterone Role On The Abnormal Growth And Aging Effects Reversion In The Gerbil (meriones Unguiculatus) Prostate.

Prostate differentiation during embryogenesis and its further homeostatic state maintenance during adult life depend on androgens. Abundant biological data suggest that androgens play an important role in the development of the prostate cancer and other prostatic diseases. The objective of this work was to evaluate the effects of the testosterone supplementation in gerbil (a new experimental model) at different ages. Tissues from experimental animals were studied by histological and histochemistry procedures, androgen receptor immunohistochemistry assay, morphometric-stereological analysis, and transmission electron microscopy (TEM). After the treatment were observed increase of prostate weight and epithelium height in all ages studied. In some adult and aged treated animals, hyperplasic and dysplastic process were observed, including prostatic intraepithelial neoplasias and adenocarcinomas. Increase of the thickness of the smooth muscle cell (SMC) layer was observed in pubescent and adult animals and TEM revealed apparent SMC hypertrophy. An apparent increase in the frequency of blood vessels distributed by the subepithelial stroma in the treated animals was noticed. Reversion of the natural effects of aging on the prostate was observed in the aged treated animals in some acini of the gland. These data demonstrate that the gerbil prostate is susceptible to androgenic action at the studied ages and it can serve, for example, as experimental model to studies of prostate neoplastic process induction and hormonal therapy in aged animals.2881190-20