Construction of transgenic mice with specific Cre recombinase expression in the zona fasciculata in adrenal cortex / 生理学报

The adrenal gland is an important endocrine organ of human body. CYP11B1 gene was specifically expressed in the zona fasciculata in adrenal cortex. In order to better study the function of genes specifically expressed in the zona fasciculata in adrenal cortex, the mice with Cre recombinase specifically expressed in the zona fasciculata in adrenal cortex were constructed. It was then confirmed that CYP11B1 was specifically expressed in adrenal glands. Then, using CRISPR/Cas9 technique, CYP11B1-2A-GfpCre recombinant vector was constructed and subsequently injected into the fertilized eggs of mice. It was confirmed that the Cre gene was mainly expressed in the zona fasciculata in adrenal cortex of CYP11B1Cre mice by using mTmG and LacZ staining. The CYP11B1Cre mice were then mated with cystathionine γ-lyase (CTH) mice, thereby generating CTH/CYP11B1Cre mice. It was also confirmed that CTH gene in the zona fasciculata in adrenal cortex was specifically knocked out in these mice. These results suggest that transgenic mice with specific Cre recombinase expression in the zona fasciculata in adrenal cortex were constructed successfully. This animal model can be a powerful tool for the study of the function of genes expressed in the zona fasciculata in adrenal cortex.

POD-1/Tcf21 overexpression reduces endogenous SF-1 and StAR expression in rat adrenal cells

During gonad and adrenal development, the POD-1/capsulin/TCF21transcription factor negatively regulates SF-1/NR5A1expression, with higher SF-1 levels being associated with increased adrenal cell proliferation and tumorigenesis. In adrenocortical tumor cells, POD-1 binds to the SF-1 E-box promoter region, decreasing SF-1 expression. However, the modulation of SF-1 expression by POD-1 has not previously been described in normal adrenal cells. Here, we analyzed the basal expression of Pod-1 and Sf-1 in primary cultures of glomerulosa (G) and fasciculata/reticularis (F/R) cells isolated from male Sprague-Dawley rats, and investigated whether POD-1 overexpression modulates the expression of endogenous Sf-1 and its target genes in these cells. POD-1 overexpression, following the transfection of pCMVMycPod-1, significantly decreased the endogenous levels of Sf-1 mRNA and protein in F/R cells, but not in G cells, and also decreased the expression of the SF-1 target StAR in F/R cells. In G cells overexpressing POD-1, no modulation of the expression of SF-1 targets, StAR and CYP11B2, was observed. Our data showing that G and F/R cells respond differently to ectopic POD-1 expression emphasize the functional differences between the outer and inner zones of the adrenal cortex, and support the hypothesis that SF-1 is regulated by POD-1/Tcf21 in normal adrenocortical cells lacking the alterations in cellular physiology found in tumor cells.

Aldosterone secretion in patients with septic shock: a prospective study / Secreção de aldosterona em pacientes com choque séptico: estudo prospectivo

OBJECTIVE: To assess serum levels of the main factors that regulate the activation of the zona glomerulosa and aldosterone production in patients with septic shock, as well as their response to a high-dose (250 µg) adrenocorticotropic hormone (ACTH) stimulation test. SUBJECTS AND METHODS: In 27 patients with septic shock, baseline levels of aldosterone, cortisol, ACTH, renin, sodium, potassium, and lactate were measured, followed by a cortrosyn test. RESULTS: Renin correlated with baseline aldosterone and its variation after cortrosyn stimulation. Baseline cortisol and its variation did not correlate with ACTH. Only three patients had concomitant dysfunction of aldosterone and cortisol secretion. CONCLUSIONS: Activation of the zona glomerulosa and zona fasciculata are independent. Aldosterone secretion is dependent on the integrity of the renin-angiotensin-aldosterone system, whereas cortisol secretion does not appear to depend predominantly on the hypothalamic-pituitary-adrenal axis. These results suggest that activation of the adrenal gland in critically ill patients occurs by multiple mechanisms.

OBJETIVO: Avaliar os níveis séricos dos principais fatores que regulam a ativação da zona glomerulosa e a produção de aldosterona em pacientes com choque séptico, assim como sua resposta ao teste de cortrosina em alta dose (250 µg). SUJEITOS E MÉTODOS: Em 27 portadores de choque séptico, foram aferidos níveis basais de aldosterona, cortisol, ACTH, renina, sódio, potássio e lactato, bem como realizado teste de cortrosina. RESULTADOS: Renina se correlacionou com níveis basais de aldosterona e sua variação após teste de cortrosina. Cortisol basal e sua variação não se correlacionaram com ACTH. Apenas três pacientes apresentaram disfunção concomitante da secreção de aldosterona e cortisol. CONCLUSÕES: Ativação das zonas fasciculada e glomerulosa são independentes. Secreção de aldosterona é dependente da integridade do sistema renina-angiotensina-aldosterona, enquanto secreção de cortisol não parece predominantemente dependente do eixo hipotálamo-hipófise-adrenal. Esses resultados sugerem que a ativação da adrenal em pacientes críticos ocorre por múltiplos mecanismos.

Efectos del éstres crónico sobre la apoptosis y la proliferación celular en la corteza adrenal de ratas gestantes / Effects of chronic stress on apoptosis and cell proliferation in the adrenal cortex of pregnant rats

La homeostasis de los tejidos que constituyen a los seres vivos es regulada por los procesos de proliferación y apoptosis celular. El modelo de estrés crónico intermitente por inmovilización durante la gestación puede alterar diversos mecanismos que mantienen la homeostasis del organismo, los cuales son motivo de estudios actuales. En estudios previos se comprobó variaciones significativas en el peso de las glándulas adrenales y en los niveles plasmáticos de PRL, estrógenos y CORT. Bajo la hipótesis que el estrés crónico produce modificaciones en los procesos de proliferación y apoptosis en las glándulas adrenales de ratas gestantes, los objetivos fueron cuantificar las células en proliferación y apoptosis mediante la utilización de técnicas inmunocitoquímicas en la corteza adrenal de ratas en la segunda mitad de la gestación, y comprobar las características ultraestructurales del proceso apoptótico con microscopía electrónica de transmisión. Material y Métodos: En condiciones de bioterio controladas, se estudió la glándula adrenal de ratas de 12, 17 y 21 días de gestación, controles (RC) y experimentales (RE) sometidas a un estrés crónico, intermitente, homotípico e intenso. Se usaron técnicas combinadas de inmunomarcación, análisis estereológico y cuantificación de imágenes de cortes alternados para determinar las variaciones del fenómeno apoptótico y proliferativo que presentan las diferentes poblaciones celulares de la corteza adrenal. El tratamiento estadístico se realizó con los Softwares InfoStat y el SAS 9.1. Se aplicó ANOVA de una y tres vías y la distribución de Poisson de un modelo logarítmico lineal con efecto de grupo, tiempo e interacciones. Las diferencias fueron consideradas significativas si p<0.05. Resultados Apoptosis: a. En Ratas Gestantes Controles y Estresadas: 1. El índice apoptótico disminuyó a medida que progresó la gestación

SUMMARY: Homeostasis of tissues is regulated by cellular proliferation and apoptosis processes. Intermittent chronic stress by immobilization during gestation may alter several mechanisms that maintain the organism homeostasis. Those mechanisms are object of several studies. Previous studies using the immobilization model have shown significant variations in adrenal glands weight and plasmatic levels of PRL, estrogens and CORT. Under the hypothesis that chronic stress causes changes in proliferation and apoptosis processes in adrenal glands of gestant rats, the main objectives of this study were: 1) to identify and quantify cell nuclei in proliferation and apoptosis using immunocytochemical techniques in adrenal cortex samples of rats in the second half of gestation and, 2) to check the ultraestructural haracteristics of apoptotic process by means of transmission electronic microscopy. Materials and Methods: In controlled lab conditions, the adrenal glands of control (RC) and experimental (RE) rats at 12, 17 and 21 days of gestation were studied. Experimental rats were subject to an intermittent, chronic, homotypic and intense stress in order to avoid habituation. A combination of labeling techniques, stereological analysis and image quantification of alternate sections was used to determine the changes in proliferation and apoptosis in the different cell populations of the adrenal cortex. Statistical analyses were performed with InfoStat and SAS 9.1 softwares. Data was analyzed with one and three-way ANOVAs and, a linear logarithmic model with Poisson distribution including group, time and interactions effects

To observe and compare the histological features of fat induced adrenal cortex in albino rats

To observe and compare the histological features of fat induced adrenal cortex in albino rats. Prospective experimental study. Department of anatomy, Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre, Karachi from August 2008 to October 2008. Twenty, young, male albino rats were selected for this study and were divided into 2 groups. Group A served as control and received normal diet. Group B received high-fat diet with 20 gms of saturated fat in the form of unsalted dairy butter per 100 gms of diet. Both groups were further divided into two Al and A2, B1 and B2 according to period of study i.e. 4 and 6 weeks respectively. On completion of experimental period, animals were weighed then sacrificed. Adrenal glands were removed, weighed and processed for paraffin sectioning. Haematoxylin and eosin staining was done for histological study. Cortical zones examined for apparent zonal hypertrophy, and cellular and nuclear changes. Increase in body weight and weight of Adrenal glands were highly significant [P<0.001]. Apparent increase in zona fasciculata observed was consistent with increased activity of gland. Cells in this zone appeared vacuolated due to presence of fat. In some areas of zona glomerulosa pyknosis and distortion of cell out line seen. High-fat diet induces increase in body weight and weight of adrenal gland which were due to hypertrophied zona fasciculata mainly, suggesting that these findings lead to increased activity and secretion of glucocorticoids

Effects of infrasound on activities of 3beta hydroxysteroid dehydrogenase and acid phosphatase of polygonal cells in adrenal cortex zona fasciculate in mice / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the biological effects of infrasound on the polygonal cells in adrenal cortex zona fasciculation in mice.</p><p><b>METHODS</b>The biological effects of infrasound on the activities of 3beta hydroxysteroid dehydrogenase (3-betaHSDH) and acid phosphatase(ACP) of the polygonal cells in adrenal cortex zona fasciculate were observed when exposure to 8 and 16 Hz infrasound at 80, 90, 100, 110, 120 and 130 dB for 1 day, 7 days and 14 days or 14 days after the exposure.</p><p><b>RESULTS</b>When exposure to 8 Hz infrasound, the enzyme activities of 3-betaHSDH increase as the sound pressure levels increase. Only when the sound pressure levels reach 130 dB, the enzyme activities began to decrease exceptionally. When exposure to 16 Hz, 80 dB infrasound, no significant difference between the treatment and control group in the activities of 3-betaHSDH could be observed, but the injury of the polygonal cells had appeared. When exposure to 16 Hz, 100 dB infrasound, the activities of 3-betaHSDH started to increase. The cell injury still existed. When exposed to 16 Hz, 120 dB infrasound, the local tissue damage represented. Fourteen days after the mice exposure to 8 Hz, 90 dB and 130 dB infrasound for 14 days continuously, the local tissue injury of the adrenal cortex zona fasciculation began to recover at certain extent, but the higher the exposure sound pressure level, the poorer the tissue recovery.</p><p><b>CONCLUSION</b>The biological effects of infrasound on the polygonal cells in adrenal cortex zona fasciculation response to the frequency of the infrasound are found at certain action strength range, but this characteristic usually is covered by the severe tissue injury. When exposure to infrasound is stopped for a period of time, the local tissue injury of the adrenal cortex zona fasciculation could recovers at certain extent, but the higher the exposure sound pressure level, the more poorer the tissue recovery.</p>

effect of medroxyprogesterone acetate on the structure of adrenal cortex [zona fasciculata] of adult female albino rat. Light and electron microscopic studies

The injectable contraceptive medroxyprogesterone acetate [MPA] is an internationally established option of birth control. To evaluate the possible effects of MPA on the structure of zona fasciculata cells, the adrenals of forty adult female albino rats were used. The experimental animals were divided into four groups treated as follows: group I was kept as control, groups II and III were injected intramusculary with MPA for 2 and 4 months respectively. Animals of group IV were injected for 4 months and sacrificed one month after the last injection. Specimens from the adrenal gland were taken and processed for light and electron microscopic examination. It has been found that MPA caused an increase in the thickness of zona fasciculate and activation of the cells when administered for 2 months. The cells showed numerous lipid droplets, large number of mitochondria and prominent smooth endoplasmic reticulum. However, MPA administration for 4 months resulted in reduction in the thickness of zona fasciculata with degenerative changes in its cells. The cells appeared small and separated from each other and exhibited many lysosomes and intracellular vacuolar spaces. After withdrawal, most of the cells appeared more or less normal and few cells showed some structural changes. These findings indicated that MPA at first caused activation of fasciculata cells and increased steroidogenesis, but prolonged use of MPA induced inactivation and degenerative changes in these cells. However some of these changes were reversible on withdrawal of the drug. Therefore precaution and follow up must be considered with prescribing this drug for prolonged periods

Probable effect of photoperiod on seasonal variation in the nuclear volume of the adrenal cortex of viscacha (Lagostomus maximus maximus)

The neuroendocrine system regulates several organic functions such as reproduction, metabolism and adaptation to the environment. This system shows seasonal changes linked to the environment. The experimental model used in the present study was Lagostomus maximus maximus (viscacha). The reproduction of males of this species is photoperiod dependent. Twenty-four adult male viscachas were captured in their habitat at different times during one year. The adrenal glands were processed for light microscopy. Serial cuts were stained with hematoxylin-eosin for the morphometric study, and 100 nuclei of each zone of the adrenal cortex were counted per animal. Data were analyzed statistically by ANOVA and the Tukey test. The cells of the glomerulosa zone are arranged in a tube-shaped structure. The fasciculata zone has large cells with central nuclei and clearly visible nucleoli and with a vacuolar cytoplasm. In the reticularis zone there are two of types of cells, one with a nucleus of fine chromatin and a clearly visible nucleolus and the other with nuclear pycnosis. Morphometric analysis showed maximum nuclear volumes during the February-March period with values of 133 percet 7.3 µm3 for the glomerulosa, 286.4 percent 14.72 µm3 for the fasciculata, and 126.3 percent 9.49 µm3 for the reticularis. Minimum nuclear volumes were observed in August with values of 88.24 + or - 9.9 µm3 for the glomerulosa, 163.7 percent 7.78 µm3 for the fasciculata and 64.58 + or - 4.53 µm3 for the reticularis. The short winter photoperiod to which viscacha is subjected could inhibit the adrenal cortex through a melatonin increase which reduces the nuclear volume as well as the cellular activity

Adreno-Hepatic Fusion: A case report

Adreno-hepatic fusion is rare condition defined as adhesion of the liver and right adrenal cortex with close intermingling of the respective parenchyme. It is suggested to be an aging phenomenon, because its incidence is much higher in older age group. Clinically it may pose a problem of operability of the organ involved. We report a case of incidentally found adreno-hepatic fusion in a 49 year old female patient with adenocarcinoma of the sigmoid colon. The segementectomy of VIII segement of the liver was done due to a 6 4 cm sized metastatic nodule of adenocarcioma. Pathological examination of the liver revealed an ovoid shaped, 1 0.5 cm sized adrenal cortical tissue. It was subcapsularly located and about 1cm apart from the metastatic adenocarcinoma with an intervening normal hepatic tissue. The adrenal tissue was mainly composed of zona fasciculata without medullary tissue. In the interphase, the adrenal tissue and liver tissue were admixed closely and partially septated by thin fibrous tissue. There was no inflammatory response to the heterotropically located adrenal tissue and there was no symptom related to the adrenal gland.