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      Adrenal glands of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831): morphological and morphometric aspects Translated title: Glândulas adrenais de preás (Galea spixii, Wagler, 1831): aspectos morfológicos e morfométricos

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          Abstract

          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.

          Translated abstract

          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.

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          Adrenal glands of mouse and rat do not synthesize androgens

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            Androgen synthesis in adrenarche.

            The enzymes and pathways of steroidogenesis are central to an understanding of adrenarche. The quantitative regulation of steroidogenesis occurs at the first step, the conversion of cholesterol to pregnenolone. Chronic quantitative regulation is principally at the level of transcription of the CYP11A1 gene encoding P450scc, which is the enzymatically rate-limiting step. Acute regulation is mediated by the steroidogenic acute regulatory protein (StAR), which facilitates the rapid influx of cholesterol into mitochondria, where P450scc resides. Qualitative regulation, which determines the type of steroid produced in a cell, is principally at the level of P450c17 (CYP17). In the absence of P450c17 in the zona glomerulosa, C21 deoxy steroids are produced, leading to the mineralocorticoid, aldosterone. In the presence of the 17alpha-hydroxylase but not the 17,20 lyase activity of P450c17 in the zona fasciculata, C21, 17-hydroxy steroids are produced, leading to the glucocorticoid, cortisol. When both the 17alpha-hydroxylase and 17,20 lyase activities of P450c17 are present in the zona reticularis, the androgen precursor DHEA is produced. The discrimination between 17alpha-hydroxylase and 17,20 lyase activities is regulated by two post-translational events, the serine phosphorylation of P450c17 and the allosteric action of cytochrome b5, both of which act to optimize the interaction of P450c17 with its obligatory electron donor, P450 oxidoreductase. In the adrenal zona reticularis, the abundant expression of P450 oxidoreductase and cytochrome b5, and the low expression of 3beta-hydroxysteroid dehydrogenase (HSD3B2) result in the production of the large amounts of DHEA that characterize adrenarche.
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              Comparison of the 17 alpha-hydroxylase/C17,20-lyase activities of porcine, guinea pig and bovine P450c17 using purified recombinant fusion proteins containing P450c17 linked to NADPH-P450 reductase.

              The cDNAs for cytochrome P450c17 (P450c17) of three species, pig, guinea pig, and cow, representing three families of mammals (suidae, procaviidae, and bovidae, respectively) were each engineered into an expression plasmid (pCWori+). The P450c17 domain of the coding sequence was connected to a truncated form of rat NADPH-P450 reductase by a linker sequence encoding two amino acids (SerThr). These fusion proteins were expressed in E. coli and purified for use in enzymatic assays to determine similarities and differences in 17 alpha-hydroxylase and lyase activities. The fusion proteins were found to catalyze both the 17 alpha-hydroxylation of progesterone (P4) and pregnenolone (P5) to 17 alpha-hydroxylated P4 and P5 (17 alpha-OH P4 and 17 alpha-OH P5) followed by the C17,20-lyase reaction for the conversion of these C(21)-17 alpha-hydroxylated steroids to C(19)-steroids (the C17,20-lyase reaction). These in vitro studies show that (a) porcine P450c17 possesses cytochrome b(5) (b(5))-stimulated C17,20-lyase activity that converts 17 alpha OH-P4 to androstenedione (AD) but also converts 17 alpha-OHP5 to dehydroepiandrosterone (DHEA); (b) guinea pig P450c17 possesses a b(5)-stimulated C17,20-lyase activity that converts 17 alpha-OH P4 to AD but does not convert 17 alpha-OH P5 to DHEA., and (c) bovine P450c17 possesses a b(5)-stimulated C17,20-lyase activity that converts 17 alpha-OH P5 to DHEA but does not convert 17 alpha-OH P4 to AD. Thus, the P450c17 of each species differs in its ability to catalyze in vitro the conversion of C(21)-steroids to C(19)-steroids. In addition, each P450c17 is capable of catalyzing additional hydroxylation reactions leading to low levels of 2 alpha-, 6 beta-, 16- and 21-hydroxy-metabolites. Porcine P450c17 also catalyzes the b(5)-dependent synthesis of andien-beta (androsta-5,16-dien-3beta-ol) from P5. When the amino acid sequences of the three P450c17s were aligned there was an approximate 50% variation in the alignment identity (227 differences in the sequences of 509 amino acids). Alignment did not permit the assignment of specific amino acids or domains to the observed differences in enzymatic activities.
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                Author and article information

                Contributors
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                Journal
                bjb
                Brazilian Journal of Biology
                Braz. J. Biol.
                Instituto Internacional de Ecologia
                1678-4375
                September 2016
                : 76
                : 3
                : 645-655
                Affiliations
                [1 ] Universidade de São Paulo Brazil
                [2 ] Universidade Federal de Minas Gerais Brazil
                [3 ] Universidade Federal do Mato Grosso Brazil
                [4 ] Universidade Federal Rural do Semi-Árido Brazil
                Article
                S1519-69842016000300645
                10.1590/1519-6984.23514
                257f24af-4c2c-4ad0-980f-5243dc5208c9

                This work is licensed under a Creative Commons Attribution 4.0 International License.

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1519-6984&lng=en
                Categories
                BIOLOGY

                General life sciences
                adrenal cortex,adrenal medulla,CYP,steroidogenic enzymes,wild rodents,córtex adrenal,medula adrenal,CYP17,enzimas esteroidogênicas,roedores silvestres

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