RESUMO
INTRODUCTION: Worldwide, nanoparticles especially gold-nanoparticles (Au-NPs) are widely used in medicine, cancer treatment and cosmetic industry. They are easily conjugated with different biomedical and biological agents and effortlessly absorbed with few side effects. The pars distalis of the pituitary gland is considered as the maestro of the endocrine peripheral glands since it secrets trophic hormones that controls their functions. 5-10% of the non-granular pars distalis cells are folliculo-stellate cells (FSCs) that support the granular cells' functions. The aim of the study was to explore the histological and the biochemical effects of repeated exposure to Au-NPs on the pars distalis in adult male albino rats with highlighting the impact on FSCs. MATERIAL AND METHODS: Thirty-six adult male albino rats were divided equally into control group and Au-NPs group (received 40 µg/kg/day of 11 ± 2 nm spherical Au-NPs orally for 2 weeks). Then, rats were euthanized and deposition of Au-NPs in pars distalis was investigated. Biochemical investigations and histological studies including hematoxylin and eosin staining, periodic acid Schiff's reaction, immunohistochemistry (IHC) for S-100, connexin 43 (Cx43) and Cytochrome-C (Cyt-C) as well as electron-microscopic and morphometric studies were carried out. RESULTS: The Au-NPs group demonstrated structural disorganization in the pars distalis, inflammation, congestion and increased extracellular PAS-positive colloid deposition due to the accumulation of Au-NPs. A significant increase in the immunoreactivity of S-100, Cx43 and Cyt-c, along with a significant increase in TNF-a, MDA, and bFGF content in the pituitary homogenates, was noted as compared to the control group. Ultrastructurally, degenerative changes were observed in the secretory cells. FSCs showed proliferation and increased phagocytic activity. CONCLUSIONS: Repetitive exposure of adult male albino rats to Au-NPs prompted the accumulation of these nanoparticles in the pars distalis that was accompanied by cellular degeneration and dysfunction of the secretory cell and proliferation of FSCs. Thus, monitoring of the pars distalis hormonal levels might be useful for early detection of some hazardous effects possibly associated with the use of gold-nanoparticles.
Assuntos
Nanopartículas Metálicas/toxicidade , Adeno-Hipófise/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ouro/química , Ouro/toxicidade , Inflamação/patologia , Inflamação/fisiopatologia , Masculino , Nanopartículas Metálicas/química , Fagocitose/efeitos dos fármacos , Adeno-Hipófise/patologia , Adeno-Hipófise/ultraestrutura , Ratos WistarRESUMO
OBJECTIVE: Human patients with Duchenne muscular dystrophy (DMD) commonly exhibit a short stature, but the pathogenesis of this growth retardation is not completely understood. Due to the suspected involvement of the growth hormone/insulin-like growth factor 1 (GH/IGF1) system, controversial therapeutic approaches have been developed, including both GH- administration, as well as GH-inhibition. In the present study, we examined relevant histomorphological and ultrastructural features of adenohypophyseal GH-producing somatotroph cells in a porcine DMD model. METHODS: The numbers and volumes of immunohistochemically labelled somatotroph cells were determined in consecutive semi-thin sections of plastic resin embedded adenohypophyseal tissue samples using unbiased state-of-the-art quantitative stereological analysis methods. RESULTS: DMD pigs displayed a significant growth retardation, accounting for a 55% reduction of body weight, accompanied by a significant 50% reduction of the number of somatotroph cells, as compared to controls. However, the mean volumes of somatotroph cells and the volume of GH-granules per cell were not altered. Western blot analyses of the adenohypophyseal protein samples showed no differences in the relative adenohypophyseal GH-abundance between DMD pigs and controls. CONCLUSION: The findings of this study do not provide evidence for involvement of somatotroph cells in the pathogenesis of growth retardation of DMD pigs. These results are in contrast with previous findings in other dystrophin-deficient animal models, such as the golden retriever model of Duchenne muscular dystrophy, where increased mean somatotroph cell volumes and elevated volumes of intracellular GH-granules were reported and associated with DMD-related growth retardation. Possible reasons for the differences of somatotroph morphology observed in different DMD models are discussed.
Assuntos
Transtornos do Crescimento/patologia , Hormônio do Crescimento/metabolismo , Distrofia Muscular de Duchenne/patologia , Vesículas Secretórias/patologia , Somatotrofos/patologia , Animais , Animais Geneticamente Modificados , Contagem de Células , Modelos Animais de Doenças , Distrofina/genética , Transtornos do Crescimento/complicações , Transtornos do Crescimento/metabolismo , Microscopia Eletrônica , Distrofia Muscular de Duchenne/complicações , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/metabolismo , Tamanho do Órgão , Hipófise/patologia , Hipófise/ultraestrutura , Adeno-Hipófise/patologia , Adeno-Hipófise/ultraestrutura , Vesículas Secretórias/ultraestrutura , Somatotrofos/ultraestrutura , SuínosRESUMO
Folliculo-stellate (FS) cells are non-endocrine cells found in the adenohypophysis and are identified in many animals by the S100 protein marker. Although keratin is another FS marker in several animals, there is no information on localization of keratin in the avian adenohypophysis. In this study, localization of cytokeratin in chicken adenohypophyseal cells was investigated immunohistochemically. Basic cytokeratin (bCK)-positive cells were arranged radially in the cell cords with their cytoplasmic processes reaching the basal lamina. The cell bodies encircled a follicle in the center of the cell cord. Furthermore, the bCK-positive cells were also S100B-positive. Growth hormone, prolactin, adrenocorticotrophic hormone, and luteinizing hormone ß-subunit did not co-localize with the bCK-positive cells. In addition, the bCK-positive cells had a laminin-positive area in their cytoplasm. Transmission electron microscopy observed agranular cells equipped with several microvilli that encircled a follicle. These results indicate that bCK-positive cells in the chicken adenohypophysis may be a predominant FS cell population and produce laminin. It is suggested that they function as sustentacular cells to sustain the adjacent endocrine cells and the structure of the cell cords in the chicken adenohypophysis.
Assuntos
Queratinas/metabolismo , Adeno-Hipófise/citologia , Adeno-Hipófise/metabolismo , Animais , Galinhas , Citoplasma/metabolismo , Imuno-Histoquímica , Laminina/metabolismo , Microscopia Eletrônica de Transmissão , Microvilosidades/ultraestrutura , Adeno-Hipófise/ultraestruturaRESUMO
Macrophages are present throughout the anterior pituitary gland. However, the features and function of macrophages in the gland are poorly understood. Recent studies have indicated that there are two main macrophage classes: M1 (classically activated) and M2 (alternatively activated). In this study, we examine whether both M1 and M2 macrophages are present in the anterior pituitary gland of rats. Our findings indicate that macrophages that are positive for CD68 (a pan-macrophage marker) were localized near capillaries in rat anterior pituitary gland. These macrophages were positive for iNOS or mannose receptor (MR), which are markers of M1 and M2 macrophages, respectively. To determine the morphological characteristics of M2 macrophages under pathological conditions, diethylstilbestrol (DES)-treated rats were used as an animal model of prolactinoma. After 2 weeks of DES treatment, a number of MR-immunopositive cells were present in the gland. Immunoelectron microscopy revealed that MR-immunopositive M2 macrophages had many small vesicles and moderately large vacuoles in cytoplasm. Phagosomes were sometimes present in cytoplasm. Interestingly, M2 macrophages in prolactinoma tissues did not usually exhibit distinct changes or differences during the normal, hyperplasia and adenoma stages. This study is the first to confirm that both M1 and M2 macrophages are present in the anterior pituitary gland of rats. Moreover, the number of M2 macrophages was greatly increased in rats with DES-induced prolactinoma. Future studies should attempt to characterize the functional role of M2 macrophages in the gland.
Assuntos
Polaridade Celular , Estrogênios/efeitos adversos , Macrófagos/patologia , Adeno-Hipófise/patologia , Prolactinoma/patologia , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Dietilestilbestrol , Imuno-Histoquímica , Lectinas Tipo C/metabolismo , Macrófagos/metabolismo , Macrófagos/ultraestrutura , Masculino , Receptor de Manose , Lectinas de Ligação a Manose/metabolismo , Adeno-Hipófise/metabolismo , Adeno-Hipófise/ultraestrutura , Prolactinoma/metabolismo , Prolactinoma/ultraestrutura , Ratos Wistar , Receptores de Superfície Celular/metabolismoRESUMO
S100ß-positive cells exist in the marginal cell layer (MCL) of the adenohypophysis and follicle structure in the parenchyma of anterior lobe (ALFS) in pituitary. They have multiple functions as phagocytes or cells that regulate hormone secretion. Majority of S100ß-positive cells in the adenohypophysis express sex determining region Y-box 2 protein (SOX2), a stem cell marker; therefore, S100ß/SOX2 double positive cells are also considered as one type of stem/progenitor cells. MCL and ALFS are consisting of morphologically two types of cells, i.e., multiciliated cells and non-ciliated cells. However, the relationship between the S100ß-positive cells and multiciliated cells in the pituitary is largely unknown. In the present study, we first immunohistochemically verified the feature of multiciliated cells in MCL and ALFS. We then examined the expression patterns of FOXJ1, an essential expression factor for multiciliated cell-differentiation, and SOX2 in the S100ß-positive multiciliated cells by in situ hybridization and immunohistochemistry. We identified anew the S100ß/SOX2/FOXJ1 triple positive multiciliated cells, and revealed that they were dispersed throughout the MCL and ALFS. These results indicate that the MCL and ALFS are consisting of morphologically and functionally distinct two types of cells, i.e., S100ß/SOX2 double positive non-ciliated cells and S100ß/SOX2/FOXJ1 triple positive multiciliated cells.
Assuntos
Cílios/genética , Fatores de Transcrição Forkhead/genética , Adeno-Hipófise/metabolismo , Subunidade beta da Proteína Ligante de Cálcio S100/genética , Fatores de Transcrição SOXB1/genética , Células-Tronco/metabolismo , Animais , Diferenciação Celular , Cílios/metabolismo , Cílios/ultraestrutura , Fatores de Transcrição Forkhead/metabolismo , Expressão Gênica , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Masculino , Adeno-Hipófise/ultraestrutura , Ratos , Ratos Wistar , Subunidade beta da Proteína Ligante de Cálcio S100/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Células-Tronco/ultraestruturaRESUMO
Endocrine and endothelial cells of the anterior pituitary gland frequently make close appositions or contacts, and the secretory granules of each endocrine cell tend to accumulate at the perivascular regions, which is generally considered to facilitate secretory functions of these cells. However, three-dimensional relationships between the localization pattern of secretory granules and blood vessels are not fully understood. To define and characterize these spatial relationships, we used scanning electron microscopy (SEM) three-dimensional reconstruction method based on focused ion-beam slicing and scanning electron microscopy (FIB/SEM). Full three-dimensional cellular architectures of the anterior pituitary tissue at ultrastructural resolution revealed that about 70% of endocrine cells were in apposition to the endothelial cells, while almost 30% of endocrine cells were entirely isolated from perivascular space in the tissue. Our three-dimensional analyses also visualized the distribution pattern of secretory granules in individual endocrine cells, showing an accumulation of secretory granules in regions in close apposition to the blood vessels in many cases. However, secretory granules in cells isolated from the perivascular region tended to distribute uniformly in the cytoplasm of these cells. These data suggest that the cellular interactions between the endocrine and endothelial cells promote an uneven cytoplasmic distribution of the secretory granules.
Assuntos
Grânulos Citoplasmáticos/metabolismo , Células Endócrinas/ultraestrutura , Células Endoteliais/ultraestrutura , Adeno-Hipófise/ultraestrutura , Animais , Simulação por Computador , Células Endócrinas/classificação , Células Endócrinas/patologia , Células Endoteliais/classificação , Células Endoteliais/patologia , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Varredura , Adeno-Hipófise/anatomia & histologia , Ratos , Ratos WistarRESUMO
We investigated the effect of leptin on the postnatal development of gap junctions between folliculo-stellate cells by using Zucker fatty (fa/fa) rats that have defects of the functional leptin receptor. Male Zucker fatty rats (fa/fa) and male Zucker lean rats (+/+) were used at each of the following postnatal ages: 20, 30, 40, 50, 60, 70, 80, 90 days, and 1 year. On one of the aforementioned dates, the anterior pituitary glands were prepared for observation by transmission electron microscopy. We quantified the number of follicles and gap junctions, and calculated the rate of occurrence as the ratio of the number of gap junctions existing between folliculo-stellate cells per intersected follicular profile. In Zucker lean male rats, the number of gap junctions remained relatively constant from days 50 to 90 (0.44 ± 0.02 to 0.49 ± 0.03), and was similar in 1 year old rats (0.47 ± 0.03). These data were statistically higher compared to Zucker fatty male rats. In Zucker fatty male rats, very few gap junctions were observed in 30-day-old rats (0.04 ± 0.01: mean ± SE). This disruption of gap junction formation persisted, and the number of gap junctions remained constant and showed a low level from days 40 to 90 (0.11 ± 0.02 to 0.17 ± 0.02); this finding was similar in 1-year-old rats (0.17 ± 0.02). These observations indicate that the effect of leptin over the gap junction formation within the anterior pituitary glands was directly mediated by interaction with the functional leptin receptor present on the folliculo-stellate cells.
Assuntos
Junções Comunicantes/ultraestrutura , Adeno-Hipófise/ultraestrutura , Ratos/crescimento & desenvolvimento , Animais , Junções Comunicantes/metabolismo , Leptina/metabolismo , Masculino , Microscopia Eletrônica de Transmissão , Adeno-Hipófise/citologia , Adeno-Hipófise/crescimento & desenvolvimento , Adeno-Hipófise/metabolismo , Ratos/metabolismo , Ratos Zucker , Receptores para Leptina/metabolismoRESUMO
The adenohypophysis was studied by immunocytochemical and ultrastructural methods in juvenile grass carp (Ctenopharyngodon idella) from natural reproduction in Northern Italian rivers. The adenohypophysis included the rostral pars distalis (RPD), the proximal pars distalis (PPD) and the pars intermedia (PI), all deeply penetrated by branches of the neurohypophysis (Nh). The prolactin (PRL), adrenocorticotropic (ACTH), somatotropic (GH), thyrotropic (TSH), gonadotropic type I (GtH I) and type II (GtH II), somatolactin (SL), melanotropic (MSH) and endorphin (END) cells were identified with antisera raised against piscine and human pituitary hormones. In juveniles of 51-69 mm of total body length (TL) with undifferentiated gonads, the PRL cells, arranged in thick strands, occupied most of the RPD. The ACTH and GH cells organized in cords bordering Nh were, respectively, confined to RPD and PPD. The TSH cells were scattered among ACTH cells in RPD and among GH cells in PPD. Cells simultaneously immunoreactive to anti-follicle stimulating hormone and to anti-croaker gonadotropin were intermingled among GH and TSH cells, which were mostly in the dorsal PPD. The SL cells were detected in PI layers bordering the Nh. The MSH and END cells were intermingled in PI and, unlike what observed in other teleosts, their respective antisera did not cross-react. In individuals of 78-112 mm TL with gonads at the beginning of differentiation, the GtH II cells were detected in PPD; all other cell types increased in number. These results, supported by ultrastructural investigations, suggest that SL and GtH II cells are directly involved in gonadal differentiation in C. idella.
Assuntos
Carpas/crescimento & desenvolvimento , Gônadas/crescimento & desenvolvimento , Adeno-Hipófise/química , Adeno-Hipófise/ultraestrutura , Diferenciação Sexual/fisiologia , Animais , Imuno-Histoquímica , Itália , Microscopia Eletrônica de Transmissão , Hormônios Hipofisários/imunologia , Hormônios Hipofisários/metabolismo , RiosRESUMO
The aims of the present study were to determine whether castration results in quantitative immunohistochemical changes in androgen receptors (AR), LH-immunoreactive (IR) cells and FSH-IR cells, and to analyse the colocalisation of AR and gonadotropins in the pituitary pars distalis (PD) of viscachas. Pituitaries were processed for light and electron microscopy. AR-IR, LH-IR and FSH-IR cells were detected by immunohistochemistry. In morphometric studies, the percentage of AR-IR, LH-IR, FSH-IR, LH-IR/AR-IR and FSH-IR/AR-IR cells was determined. In intact viscachas, AR were distributed throughout the PD; they were numerous at the caudal end, with intense immunostaining. LH-IR cells and FSH-IR cells were found mainly in the ventral region and at the rostral end of the PD. Approximately 45%-66% of LH-IR cells and 49%-57% of FSH-IR cells expressed AR in the different zones of the PD. In castrated viscachas, there was a significant decrease in the percentage of AR-IR, LH-IR, FSH-IR, and FSH-IR/AR-IR cells. Some pituitary cells from castrated viscachas also exhibited ultrastructural changes. These results provide morphological evidence that gonadal androgens are directly related to the immunolabelling of AR, LH and FSH. Moreover, the colocalisation of AR and FSH is most affected by castration, suggesting the existence of a subpopulation of gonadotrophs with different regulatory mechanisms for hormonal synthesis, storage and secretion.
Assuntos
Gonadotropinas Hipofisárias/análise , Orquiectomia/veterinária , Adeno-Hipófise/química , Receptores Androgênicos/análise , Roedores/fisiologia , Animais , Núcleo Celular/química , Citoplasma/química , Hormônio Foliculoestimulante/análise , Imuno-Histoquímica/veterinária , Hormônio Luteinizante/análise , Masculino , Microscopia Eletrônica de Transmissão/veterinária , Adeno-Hipófise/ultraestruturaRESUMO
The architecture of luteinizing hormone-releasing hormone (LH-RH) nerve ends and the S-100 protein containing folliculo-stellate cells forming gap junctions in the pars tuberalis is basically important in understanding the regulation of the hormone producing mechanism of anterior pituitary glands. In this study, intact male rats 5-60 days old were prepared for immunohistochemistry and electron microscopy. From immunostained sections, the S-100 containing cells in pars tuberalis were first detected on day 30 and increased in number to day 60; this was parallel to the immunohistochemical staining of gap junction protein, connexin 43. LH-RH positive sites were clearly observed on just behind the optic chiasm and on the root of pituitary stalk on day 30. On day 60, the width of layer increased, while follicles and gap junctions were frequently observed between agranular cells in 10 or more layers of pars tuberalis. In the present study, we investigated the sexual maturation of the anterior pituitary glands through the postnatal development of S-100 positive cells, connexin 43 and LH-RH nerves. It is suggested that the folliculo-stellate cell system including the LH-RH neurons in the pars tuberalis participates in the control of LH secretion along with the portal vein system.
Assuntos
Comunicação Celular , Conexina 43/metabolismo , Hormônio Liberador de Gonadotropina/metabolismo , Adeno-Hipófise/metabolismo , Adeno-Hipófise/ultraestrutura , Proteínas S100/metabolismo , Animais , Comunicação Celular/fisiologia , Junções Comunicantes/metabolismo , Imuno-Histoquímica/métodos , Masculino , Microscopia Eletrônica/métodos , Neurônios/metabolismo , Neurônios/ultraestrutura , Ratos , Ratos WistarRESUMO
BACKGROUND: Gonadotropin cell is the main responsible for the secretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH), and immunocastration reduces the concentrations of serum FSH and LH. A few studies have reported the histological structure of gonadotropin cells obtained from immunocastration animals at the light microscopy level. However, the ultrastructure of gonadotropin cells remains largely unexplored. The aim of this study was to evaluate and to compare ultrastructure of gonadotropin cell in gonadally intact boars and immunologically castrated male animals. FINDINGS: In this study, serum and adenohypophysis tissue were collected from nine gonadally intact boars and nine male pigs treated with recombinant gonadotropin releasing hormone I (GnRH-I). Anti-GnRH-I antibodies in serum and the ultrastructure of gonadotropin cell in adenohypophysis were determined by enzymelinked immunosorbent assay and electron microscopy, respectively. The results demonstrated that active immunization against recombinant GnRH-I increased serum GnRH-I antibody levels (P<0.05). Ultramicroscopic analysis of gonadotropin cell revealed a decrease (P<0.05) in the number and size of the large granules and small granules in the recombinant GnRH-I immunized animals. CONCLUSIONS: We conclude that immunization against recombinant GnRH-I induces severe atrophy of granules in gonadotropin cell of boars, possibly reflecting GnRH-I regulation of gonadotropin cell.
Assuntos
Gonadotrofos/imunologia , Hormônio Liberador de Gonadotropina/imunologia , Imunização/métodos , Adeno-Hipófise/imunologia , Animais , Anticorpos/sangue , Ensaio de Imunoadsorção Enzimática , Hormônio Foliculoestimulante/sangue , Gonadotrofos/ultraestrutura , Hormônio Liberador de Gonadotropina/genética , Humanos , Hormônio Luteinizante/sangue , Masculino , Proteínas Ligantes de Maltose/genética , Proteínas Ligantes de Maltose/imunologia , Microscopia Eletrônica de Transmissão , Modelos Animais , Adeno-Hipófise/citologia , Adeno-Hipófise/ultraestrutura , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , SuínosRESUMO
Pericytes are perivascular cells associated with microcirculation. Typically, they are localized close to the capillary wall, underneath the basement membrane, and have sparse cytoplasm and poorly developed cell organelles. However, the specific properties of pericytes vary by organ and the conditions within organs. We recently demonstrated that pericytes in rat anterior pituitary gland produce type I and III collagens. The present study attempted to determine the morphological characteristics of these pituitary pericytes. Castrated rats were used as a model of hormonal and vascular changes in the gland. Pericytes, as determined by desmin immunohistochemistry, were more numerous and stained more intensely in castrated rats. Transmission electron microscopy revealed that pituitary pericytes displayed the typical characteristics of pericytes. In pituitary sections from castrated rats, the Golgi apparatus of pericytes was well developed and the rough endoplasmic reticulum was elongated. Additionally, scanning electron microscopy revealed four pericyte shapes: oval, elongate, triangular, and multiangular. As compared with normal rats, the proportion of oval pericytes was lower, and the proportions of the other three shapes were higher, in castrated rats. These results suggest that pericytes change their fine structure and cell shape in response to hormonal and vascular changes in the anterior pituitary gland. In addition, a novel type of perivascular cell was found by desmin immunoelectron microscopy. The morphological properties of these cells were dissimilar to those of pericytes. The cells were localized in the perivascular space, had no basement membrane, and contained dilated rough endoplasmic reticulum. This new cell type will require further study of its origin and characteristics.
Assuntos
Hormônios Gonadais/fisiologia , Pericitos/ultraestrutura , Adeno-Hipófise/ultraestrutura , Animais , Castração , Masculino , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Pericitos/fisiologia , Adeno-Hipófise/irrigação sanguínea , Adeno-Hipófise/fisiologia , Ratos , Ratos WistarRESUMO
The integrity of homeostasis can be affected by chronic stress, and hyposomatotropism is evident in chronic stress-associated illnesses. In the present study, we demonstrated that a continuous stress (CS) severely affected somatotrophs among hormone-secreting cells in the anterior lobe (AL) of the pituitary by using a rat CS model. Among AL cells, the proliferation of somatotrophs was almost entirely suppressed in rats that had 3-5 days of CS (5dCS), although other hormone-secreting cells continued to proliferate. The cell size of somatotrophs was reduced at 5dCS (P<0.01), the number of secretory granules was increased at 3dCS (P<0.01) and serum growth hormone (GH) was on declining trend during 1 to 5dCS, suggesting that GH release was inhibited. GH-releasing hormone (GHRH) mRNA level in the arcuate nucleus was transiently decreased, whereas its receptor expression in the AL was significantly increased in CS rats. When 5dCS rats were injected with GHRH, transient GH secretion was observed, whereas proliferation of somatotrophs did not occur. The GHRH administration failed to stimulate extracellular signal-regulated kinase (ERK) 1/2 phosphorylation and the nuclear translocation of ERK in somatotrophs. These results suggest that somatotrophs of 5dCS rats expressed sufficient GHRH receptor, which could transfer a signal for GH release. However, the GHRH-induced proliferation signal was blocked somewhere between the receptor and ERK1/2. Because significant increase of corticosterone in the initial stage (the 1-3dCS) was observed in this model, the corticosterone may affect the signalling. Although the mechanism underlying the blockage of the proliferation signal in somatotrophs under CS remains unclear, these somatotrophic disorder, suggesting that the present animal model may be useful for understanding the molecular mechanisms of chronic stress-associated illnesses.
Assuntos
Proteína Quinase 1 Ativada por Mitógeno/fisiologia , Proteína Quinase 3 Ativada por Mitógeno/fisiologia , Somatotrofos/fisiologia , Estresse Psicológico/metabolismo , Animais , Western Blotting , Proliferação de Células , Doença Crônica , Corticosterona/metabolismo , Ativação Enzimática/fisiologia , Hormônio do Crescimento/metabolismo , Hormônio Liberador de Hormônio do Crescimento/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Masculino , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosforilação , Adeno-Hipófise/fisiologia , Adeno-Hipófise/ultraestrutura , Transporte Proteico/fisiologia , Radioimunoensaio , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Somatotrofos/ultraestruturaAssuntos
Adenoma Oxífilo/cirurgia , Adeno-Hipófise/cirurgia , Neoplasias Hipofisárias/cirurgia , Adenoma Oxífilo/patologia , Adenoma Oxífilo/ultraestrutura , Idoso , Diagnóstico Diferencial , Evolução Fatal , Humanos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Procedimentos Neurocirúrgicos , Adeno-Hipófise/patologia , Adeno-Hipófise/ultraestrutura , Neoplasias Hipofisárias/patologia , Neoplasias Hipofisárias/ultraestrutura , Tomografia Computadorizada por Raios XRESUMO
Spindle cell oncocytoma (SCO) is a rare, non-adenomatous tumor originating from the anterior pituitary gland. Composed of fusiform, mitochondrion-rich cells sharing several immunophenotypic and ultrastructural properties with folliculo-stellate cells (FSC), SCO has been proposed to represent a neoplastic counterpart of the latter. To date, however, SCO has failed to meet one criterion commonly used in histological-based taxonomy and diagnostics; that of recapitulating any of FSCs' morphologically defined developmental or physiological states. We describe a unique example of SCO wherein a conventional fascicular texture was seen coexisting with and organically merging into follicle-like arrangements. The sellar tumor of 2.7 × 2.6 × 2.5 cm was transphenoidally resected from a 55-year old female. Preoperative magnetic resonance imaging indicated an isointense, contrast enhancing mass with suprasellar extension. Histology showed multiple rudimentary to well-formed, follicle-like cavities on a classical spindle cell background; while all the participating cells exhibited an SCO immunophenotype, including positivity for S100 protein, vimentin, EMA, Bcl-2, and TTF-1, as well as staining with the antimitochondrial antibody 113-1. Conversely no expression of GFAP, follicular-epithelial cytokeratin, carcinoembryonic antigen, or anterior pituitary hormones was detected. Ultrastructurally, tumor cells facing follicular lumina displayed organelles of epithelial specialization, in particular surface microvilli and apical tight junctions. This constellation is felt to be reminiscent of FSCs' metaplastic transition to follicular epithelium, as observed during embryonic development and physiological renewal of the hormone-secreting parenchyma. Such finding is apt to being read as a supporting argument for SCO's descent from the FSC lineage.
Assuntos
Adenoma Oxífilo/ultraestrutura , Adeno-Hipófise/ultraestrutura , Neoplasias Hipofisárias/ultraestrutura , Adenoma Oxífilo/complicações , Adenoma Oxífilo/metabolismo , Diabetes Mellitus Tipo 2 , Dislipidemias/complicações , Estrogênios/uso terapêutico , Feminino , Transtornos do Crescimento/complicações , Transtornos do Crescimento/tratamento farmacológico , Humanos , Hipertensão/complicações , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Adeno-Hipófise/metabolismo , Neoplasias Hipofisárias/complicações , Neoplasias Hipofisárias/metabolismoRESUMO
Folliculostellate (FS) cells in the anterior pituitary gland are believed to have multifunctional properties. Using transgenic rats that express green fluorescent protein (GFP) specifically in FS cells in the anterior pituitary gland (S100b-GFP rats), we recently revealed that FS cells in primary culture exhibited marked proliferation in the presence of laminin, an extracellular matrix (ECM) component of the basement membrane. In a process referred to as matricrine action, FS cells receive ECM as a signal through their receptors, which results in morphological and functional changes. In this study, we investigated matricrine signaling in FS cells and observed that the proliferation of FS cells is mediated by integrin ß1, which is involved in various signaling pathways for cell migration and proliferation in response to ECM. Then, we analyzed downstream events of the integrin ß1 signaling pathway in the proliferation of FS cells and identified caveolin 3 as a potential candidate molecule. Caveolin 3 is a membrane protein that binds cholesterol and a number of signaling molecules that interact with integrin ß1. Using specific small interfering RNA of caveolin 3, the proliferation of FS cells was inhibited. Furthermore, caveolin 3 drove activation of the mitogen-activated protein kinase (MAPK) signaling cascades, which resulted in upregulation of cyclin D1 in FS cells. These findings suggest that matricrine signaling in the proliferation of FS cells was transduced by a caveolin 3-mediated integrin ß1 signaling pathway and subsequent activation of the MAPK pathway.
Assuntos
Caveolina 3/metabolismo , Proliferação de Células , Matriz Extracelular/fisiologia , Integrina beta1/metabolismo , Sistema de Sinalização das MAP Quinases , Adeno-Hipófise/metabolismo , Animais , Cavéolas/metabolismo , Cavéolas/ultraestrutura , Caveolina 1/antagonistas & inibidores , Caveolina 1/genética , Caveolina 1/metabolismo , Caveolina 3/antagonistas & inibidores , Caveolina 3/genética , Polaridade Celular , Células Cultivadas , Técnicas de Cultura , Inativação Gênica , Genes Reporter , Laminina/fisiologia , Masculino , Microscopia de Vídeo , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Adeno-Hipófise/citologia , Adeno-Hipófise/ultraestrutura , RNA Mensageiro/metabolismo , RNA Interferente Pequeno , Ratos , Ratos Transgênicos , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/genética , Proteínas S100/metabolismoRESUMO
The primary cilium is now considered to function as a fundamental, not rudimentary, structure for mechanical and chemical sensing by individual cells. Primary cilia in neurons express type III adenylyl cyclase (ACIII) and GPCRs for somatostatin (somatostatin receptor 3, SSTR3), serotonin, and melanin-concentrating hormone. The present immunohistochemical and electron microscopic study revealed an abundant occurrence of SSTR3-expressing solitary cilia in insulin- and growth hormone-secreting cells of the mouse. The SSTR3 immunoreactivity was restricted to the plasma membrane of cilia in both cell types, differing from previously reported immunohistochemical localization of SSTRs to cell bodies. The primary cilia in the islet cells were longer than those in the pituitary cells and extended for a long distance in the intercellular canalicules endowed with microvilli. No other endocrine organs were provided with the SSTR3-expressing primary cilia, while the primary cilia in these organs were frequently immunolabeled with ACIII antibody. Since the somatostatin inhibition of both insulin and GH release is regulated mainly by SSTR1 and SSTR5, the primary cilia expressing SSTR3 may be involved in a signaling which differs from that via other SSTR subtypes expressing in cell bodies.
Assuntos
Ilhotas Pancreáticas/metabolismo , Adeno-Hipófise/metabolismo , Receptores de Somatostatina/biossíntese , Adenilil Ciclases/metabolismo , Animais , Linhagem Celular , Cílios/metabolismo , Cílios/ultraestrutura , Cricetinae , Glucagon/metabolismo , Hormônio do Crescimento/metabolismo , Cobaias , Insulina/metabolismo , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/ultraestrutura , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Adeno-Hipófise/citologia , Adeno-Hipófise/ultraestrutura , RatosRESUMO
OBJECTIVE: The aim of this work was to study the expression of NIS in the thyroid and anterior pituitary in rats after a single dose of iodide appropriate to the content of iodide in iodine-positive points in the thyroid and pituitary. METHODS: A total of 41 inbred rat females of local laboratory strain weighing 250-300 g at the stage of diestrus and/or metestrus were used. Pituitaries and thyroids were dissected from 15 control rats at the same time as these from four groups of 6-8 rats each which were given various doses of potassium iodide dissolved in 0.5 ml distilled water (6 rats - 1 µg/100 g body weight; 8 rats - 4 µg/100 g ; 6 rats - 8 µg/100 g ; 6 rats - 25 µg/100 g.) by gavage at 48 h before sacrifice. In 6 rats of control group the concentration of iodine in thyroids and pituitaries was estimated in terms of percent by weight in dry tissue (wt% I-2 dry tissue) using the wavelength dispersive spectrometry (WDS) quantitative analysis. The expression of NIS in thyroids and pituitaries in terms of the percentage of positive immunostained area (% PA) was measured by streptavidin-biotin method using specific polyclonal antibodies. RESULTS: In thyroids, the concentration of iodine in iodine-positive points ranged from 2.5 to 59.3 (mean of 16.7±3.0) in terms of wt% I-2 dry tissue (100 % iodine-positive points), while in pituitaries it ranged from 0.17 to 6.3 (mean of 1.4±0.3) in all points and 2.2±0.4 in iodine-positive points. Histochemical reaction for NIS in the pituitaries at 48 hours after iodide administration showed a dose related increase beginning from 4 µg/100 g (from 1.8±0.7 to 12.9±1.0 % PA, respectively to the dose of iodide), while such increase in the thyroids started from 8 µg/100g (from 3.7±1.2 to 9.1±2.0 % PA). It remained still increased in pituitaries after the dose of 8 µg/100g (11.4±1.0 % PA) and 25 µg/100g (13.9±1.5 % PA), while such increase in thyroids was found only after the dose of 25 µg/100g (11.9±2.8 % PA). CONCLUSION: It was found that in the pituitaries of rat females the expression of NIS started after the dose of 4 µg iodide/100g, while that in the thyroids started after 8 µg iodide/100g. Thus, it may be suggested that the pituitary appears more susceptible to the level of iodide in blood.
Assuntos
Adeno-Hipófise/metabolismo , Iodeto de Potássio/farmacocinética , Simportadores/metabolismo , Glândula Tireoide/metabolismo , Animais , Relação Dose-Resposta a Droga , Microanálise por Sonda Eletrônica , Feminino , Microscopia Eletrônica de Varredura , Especificidade de Órgãos , Adeno-Hipófise/efeitos dos fármacos , Adeno-Hipófise/ultraestrutura , Iodeto de Potássio/sangue , Ratos , Glândula Tireoide/efeitos dos fármacos , Glândula Tireoide/ultraestruturaRESUMO
Folliculo-stellate (FS) cells in the anterior pituitary gland are believed to have multifunctional properties. FS cells connect to each other not only by mechanical means, but also by gap junctional cell-to-cell communication. Using transgenic rats that express green fluorescent protein (GFP) specifically in FS cells in the anterior pituitary gland (S100b-GFP rats), we recently revealed that FS cells in primary culture markedly change their shape, and form numerous interconnections with neighboring FS cells in the presence of laminin, an extracellular matrix (ECM) component of the basement membrane. Morphological and functional changes in cells are believed to be partly modified by matricrine signaling, by which ECM components function as cellular signals. In the present study, we examined whether gap junction formation between FS cells is affected by matricrine cues. A cell sorter was used to isolate FS cells from male S100b-GFP rat anterior pituitary for primary culture. We observed that mRNA and protein levels of connexin 43 in gap junction channels were clearly higher in the presence of laminin. In addition, we confirmed the formation of gap junctions between FS cells in primary culture by electron microscopy. Interestingly, we also observed that FS cells in the presence of laminin displayed well-developed rough endoplasmic reticulum and Golgi apparatus. Our findings suggest that, in anterior pituitary gland, FS cells may facilitate functional roles such as gap junctional cell-to-cell communication by matricrine signaling.
Assuntos
Matriz Extracelular/metabolismo , Junções Comunicantes/metabolismo , Laminina/metabolismo , Adeno-Hipófise/metabolismo , Animais , Comunicação Celular , Células Cultivadas , Conexina 43/metabolismo , Retículo Endoplasmático Rugoso/metabolismo , Retículo Endoplasmático Rugoso/ultraestrutura , Complexo de Golgi/metabolismo , Complexo de Golgi/ultraestrutura , Proteínas de Fluorescência Verde/metabolismo , Masculino , Fatores de Crescimento Neural/metabolismo , Adeno-Hipófise/ultraestrutura , Ratos , Ratos Transgênicos , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/metabolismoRESUMO
Hypoplasia adrenal congenita is an extremely uncommon disease of early onset. This condition can be lethal in the absence of treatment. Some forms are due to the congenital adrenal hypoplasia of anencephalic type whose origin is even unknown. Here, we present two cases of congenital adrenal hypoplasia of anencephalic type with pituitary abnormalities. The two male newborns died because adrenal insufficiency in the neonatal period. The adrenal glands were hypoplastic with a histological structure of anencephalic type Immunocytochemical study of the pituitary revealed an absence of the gonadotrophs. No mutation of DAX 1 and SF-1 was found.