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1.
Domest Anim Endocrinol ; 74: 106572, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33039930

RESUMO

The aim of this study was to differentiate canine adipose-derived mesenchymal stem cells (ADMSCs) into insulin-producing cells by using culture media with different compositions to determine the most efficient media. Stem cells isolated from the fat tissues close to the bitch uterus were distributed into 6 groups: (1) Dulbecco's modified Eagle medium (DMEM)-high glucose (HG), ß-mercaptoethanol, and nicotinamide; (2) DMEM-HG, ß-mercaptoethanol, nicotinamide, and exendin-4; (3) DMEM-HG, ß-mercaptoethanol, nicotinamide, exendin-4, B27, nonessential amino acids, and l-glutamine; (4) DMEM-HG, ß-mercaptoethanol, and nicotinamide (for the initial 8-d period), and DMEM-HG, ß-mercaptoethanol, nicotinamide, exendin-4, B27, nonessential amino acids, l-glutamine, and basic fibroblast growth factor (for the remaining 8-d period); (5) DMEM-HG and fetal bovine serum; and (6) DMEM-low glucose and fetal bovine serum (standard control group). Adipose-derived mesenchymal stem cells from groups 1 to 5 gradually became round in shape and gathered in clusters. These changes differed between the groups. In group 3, the cell clusters were apparently more in numbers and gathered as bigger aggregates. Dithizone staining showed that groups 3 and 4 were similar in terms of the mean area of each aggregate stained for insulin. However, only in group 4, the number of insulin aggregates and the total area of aggregates stained were significantly bigger than in the other groups. The mRNA expression of PDX1, BETA2, MafA, and Insulin were also confirmed in all the groups. We conclude that by manipulating the composition of the culture medium it is possible to induce canine ADMSCs into insulin-producing cells, and the 2-staged protocol that was used promoted the best differentiation.


Assuntos
Diferenciação Celular , Meios de Cultura/farmacologia , Insulina/metabolismo , Células-Tronco Mesenquimais/fisiologia , Adipogenia/efeitos dos fármacos , Adipogenia/fisiologia , Animais , Carbazóis/química , Carbazóis/farmacologia , Condrogênese/efeitos dos fármacos , Condrogênese/fisiologia , Meios de Cultura/química , Cães , Imunofenotipagem , Mercaptoetanol/farmacologia , Niacinamida/química , Niacinamida/farmacologia , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia
2.
Braz. j. biol ; 76(3): 645-655, tab, graf
Artigo em Inglês | LILACS | ID: lil-785027

RESUMO

Abstract Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.


Resumo Considerando a importância fisiológica e a necessidade de melhor conhecimento morfofisiológico das glândulas adrenais, os objetivos do presente estudo foram comparar dados morfométricos entre adrenais direitas e esquerdas de machos e fêmeas; realizar estudos histológicos e microscopia eletrônica de varredura e transmissão revelando a constituição tecidual das glândulas; finalmente, foi realizado estudo imunohistoquímico com objetivo de definir a presença e correto sítio de expressão da citocromo P450c17 nas glândulas adrenais em 18 glândulas adrenais (direita n=9 e esquerda n=9) de nove Galea spixii adultos (quatro machos e cinco fêmeas). A glândula adrenal direita estava posicionada mais cranialmente que a adrenal esquerda; as dimensões (peso, comprimento e largura) da glândula adrenal direita foram maiores que as da esquerda; não houve diferença entre as medidas corpóreas e das adrenais entre machos e fêmeas; a morfologia das células e diferentes quantidades de gotículas lipídicas podem estar relacionadas a diferentes demandas na produção de hormônios esteroides relacionados a cada zona do córtex adrenal; e, a imunolocalização da citocromo P450 na zona fasciculada e reticular pode estar relacionado com a síntese de 17-hidroxi-pregnenolona, 17-hidroxi-progesterona, dehidroepiandrosterona ou androstenediona.


Assuntos
Animais , Masculino , Feminino , Roedores/anatomia & histologia , Glândulas Suprarrenais/anatomia & histologia , Cobaias
3.
Braz. j. biol ; 76(3): 645-655, July-Sept. 2016. ilus, tab
Artigo em Inglês | VETINDEX | ID: vti-25330

RESUMO

Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.(AU)


Considerando a importância fisiológica e a necessidade de melhor conhecimento morfofisiológico das glândulas adrenais, os objetivos do presente estudo foram comparar dados morfométricos entre adrenais direitas e esquerdas de machos e fêmeas; realizar estudos histológicos e microscopia eletrônica de varredura e transmissão revelando a constituição tecidual das glândulas; finalmente, foi realizado estudo imunohistoquímico com objetivo de definir a presença e correto sítio de expressão da citocromo P450c17 nas glândulas adrenais em 18 glândulas adrenais (direita n=9 e esquerda n=9) de nove Galea spixii adultos (quatro machos e cinco fêmeas). A glândula adrenal direita estava posicionada mais cranialmente que a adrenal esquerda; as dimensões (peso, comprimento e largura) da glândula adrenal direita foram maiores que as da esquerda; não houve diferença entre as medidas corpóreas e das adrenais entre machos e fêmeas; a morfologia das células e diferentes quantidades de gotículas lipídicas podem estar relacionadas a diferentes demandas na produção de hormônios esteroides relacionados a cada zona do córtex adrenal; e, a imunolocalização da citocromo P450 na zona fasciculada e reticular pode estar relacionado com a síntese de 17-hidroxi-pregnenolona, 17-hidroxi-progesterona, dehidroepiandrosterona ou androstenediona.(AU)


Assuntos
Animais , Roedores/anatomia & histologia , Roedores/classificação , Glândulas Suprarrenais/anatomia & histologia
4.
Braz J Biol ; 76(3): 645-55, 2016 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-27143060

RESUMO

Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.


Assuntos
Glândulas Suprarrenais/anatomia & histologia , Roedores/anatomia & histologia , Animais , Feminino , Cobaias , Masculino
5.
Braz. J. Biol. ; 76(3)2016.
Artigo em Inglês | VETINDEX | ID: vti-744769

RESUMO

Abstract Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.


Resumo Considerando a importância fisiológica e a necessidade de melhor conhecimento morfofisiológico das glândulas adrenais, os objetivos do presente estudo foram comparar dados morfométricos entre adrenais direitas e esquerdas de machos e fêmeas; realizar estudos histológicos e microscopia eletrônica de varredura e transmissão revelando a constituição tecidual das glândulas; finalmente, foi realizado estudo imunohistoquímico com objetivo de definir a presença e correto sítio de expressão da citocromo P450c17 nas glândulas adrenais em 18 glândulas adrenais (direita n=9 e esquerda n=9) de nove Galea spixii adultos (quatro machos e cinco fêmeas). A glândula adrenal direita estava posicionada mais cranialmente que a adrenal esquerda; as dimensões (peso, comprimento e largura) da glândula adrenal direita foram maiores que as da esquerda; não houve diferença entre as medidas corpóreas e das adrenais entre machos e fêmeas; a morfologia das células e diferentes quantidades de gotículas lipídicas podem estar relacionadas a diferentes demandas na produção de hormônios esteroides relacionados a cada zona do córtex adrenal; e, a imunolocalização da citocromo P450 na zona fasciculada e reticular pode estar relacionado com a síntese de 17-hidroxi-pregnenolona, 17-hidroxi-progesterona, dehidroepiandrosterona ou androstenediona.

6.
Braz. j. morphol. sci ; 30(1): 21-27, 2013. ilus, tab
Artigo em Inglês | LILACS | ID: lil-699326

RESUMO

The mdx mice model is widely used for Duchenne muscular dystrophy (DMD) studies, which is present in a high percentage from newborns human males. Therefore, the aim of this study was to evaluate possible morphological changes from spleens in these mice and to compare with normal mice (Mus musculus) in contribution to DMD understanding and its consequences on immune system by affected individuals. The study was performed by light and scanning electron microscopy (SEM) techniques beyond immunohistochemistry. Was found microscopically an increased number of lymph nodes and decreased in red pulp region by mdx, beyond a larger VEGF-C (vascular endothelial growth factor C) expression stimulates lymphangiogenesis in red pulp region from spleen. These findings suggest a spleen adaptation in order to supply immunological demand due upper respiratory infection, which are common in individuals affected by Duchenne muscular dystrophy.


Assuntos
Animais , Camundongos , Distrofia Muscular de Duchenne/imunologia , Sistema Imunitário , Infecções Respiratórias/imunologia , Linfócitos , Fator A de Crescimento do Endotélio Vascular , Eutanásia Animal , Camundongos Endogâmicos mdx , Microscopia Eletrônica de Varredura
7.
Braz. j. morphol. sci ; 30(3): 186-190, 2013. ilus, tab
Artigo em Inglês | LILACS | ID: lil-699347

RESUMO

The Duchenne Muscular Dystrophy (DMD) is a recessive genetic disease linked to chromosome X. Thisdisease is characterized by an absence or dysfunction in the expression of dystrophin. Experimental modelsmdxare widely used for the development of research addressing the DMD. The objective of this research is tocontribute to a detailed study of possible renal morphological changes resulting from DMD. We used five pairsof kidneys frommdxmice and five from normal mice, which were subjected to measurement, light microscopy,and scanning electron microscopy. The morphological findings of kidneys frommdxmice are within thepatterns described in animal studies with severe dehydration, which exhibit signs of diffuse hemorrhage inthe cortical and medullary area, while the glomeruli in the cortical region showed a decrease in urinary space,located between the Bowman’s capsule and the inner cell mass of the glomeruli. However, future experimentswith animals in different ages can assist in the proving of the morphological changes found here.


Assuntos
Animais , Masculino , Camundongos , Desidratação , Distrofia Muscular de Duchenne/complicações , Glomérulos Renais/anatomia & histologia , Rim/anatomia & histologia , Animais de Laboratório , Camundongos Endogâmicos mdx/anatomia & histologia , Eutanásia Animal , Microscopia Eletrônica de Varredura
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