Sex differences in adrenal cortex beta-catenin immunolocalisation of the Saharan gerbil, Libyan jird (Meriones libycus, Lichtenstein, 1823).

BACKGROUND: The adrenal cortex provides adequate steroidogenic responses to environmental changes. However, in desert rodents, the adrenocortical activity varies according to several factors especially sex, age, and seasonal variations. Herein, we examined the sex differences in the adrenal cortex activity and explored the involvement of sex hormones in the regulation of this function in Libyan jird Meriones libycus. MATERIALS AND METHODS: Twenty-four adult male and female animals weighing 109-110 g were captured in the breeding season and equally assigned into control and gonadectomised groups. Animal euthanasia was performed 50 days after the gonadectomy. Adrenal gland was processed for structural and immunohistochemistry study of b-catenin, whereas plasma was used for cortisol assay. RESULTS: The results showed that female adrenal gland weight was heavier than male and gonadectomy reduced this dimorphism. The adrenal cortex thickness was greater in the female than in the male, mainly due to significant development of the zona fasciculata. Females presented higher cell density in fasciculata and reticularis zones. The plasma cortisol was higher in females than in males. The immunolocalisation of beta-catenin showed that the expression was particularly glomerular in both sexes. However, in the female, the immunostaining was present in the zona reticularis while it was absent in the control male. Orchiectomy reduced zona glomerulosa cell density and induced hypertrophy of zona reticularis characterised by strong beta-catenin immunoreactivity. CONCLUSIONS: Results indicated that sex hormones had a major role in the regulation of the Saharan gerbil's adrenal homeostasis by modulating beta-catenin signalling. Androgens seem to inhibit the Wnt-beta-catenin pathway and oestrogens are activators of the adrenal inner zones.

P53/Rb inhibition induces metastatic adrenocortical carcinomas in a preclinical transgenic model.

Adrenocortical carcinoma (ACC) is a rare cancer with poor prognosis. Pan-genomic analyses identified p53/Rb and WNT/ß-catenin signaling pathways as main contributors to the disease. However, isolated ß-catenin constitutive activation failed to induce malignant progression in mouse adrenocortical tumors. Therefore, there still was a need for a relevant animal model to study ACC pathogenesis and to test new therapeutic approaches. Here, we have developed a transgenic mice model with adrenocortical specific expression of SV40 large T-antigen (AdTAg mice), to test the oncogenic potential of p53/Rb inhibition in the adrenal gland. All AdTAg mice develop large adrenal carcinomas that eventually metastasize to the liver and lungs, resulting in decreased overall survival. Consistent with ACC in patients, adrenal tumors in AdTAg mice autonomously produce large amounts of glucocorticoids and spontaneously activate WNT/ß-catenin signaling pathway during malignant progression. We show that this activation is associated with downregulation of secreted frizzled related proteins (Sfrp) and Znrf3 that act as inhibitors of the WNT signaling. We also show that mTORC1 pathway activation is an early event during neoplasia expansion and further demonstrate that mTORC1 pathway is activated in ACC patients. Preclinical inhibition of mTORC1 activity induces a marked reduction in tumor size, associated with induction of apoptosis and inhibition of proliferation that results in normalization of corticosterone plasma levels in AdTAg mice. Altogether, these data establish AdTAg mice as the first preclinical model for metastatic ACC.

Adrenal tumorigenesis targeted by the corticotropin-regulated promoter of the aldo-keto reductase AKR1B7 gene in transgenic mice.

Studies of ACTH functions in adrenal steroidogenesis have been facilitated by the availability of immortalized mouse adrenocortical Y1 cells. In order to obtain alternative cell lines with a more differentiated zona fasciculata (ZF) phenotype we used targeted tumorigenesis strategy. We have generated transgenic mice expressing the SV40 T antigen under the control of the ACTH-dependent promoter for the AKR1B7/MVDP gene (aldo-keto reductase 1B7/mouse vas deferens protein), which encodes an enzyme responsible for detoxifying isocaproaldehyde, the product of side-chain cleavage of cholesterol generated by steroidogenesis. Our previous data indicated that in the mouse adrenal, AKR1B7 expression was restricted to the ZF and that a 0.5-kb promoter region was able to target specific adrenal expression in transgenic mice. In situ hybridization analyses indicate that AKR1B7 expression during fetal and post-natal periods paralleled the onset of glucocorticoid synthesis and the development of ZF. In transgenic mice, ACTH control and developmental programming of the CAT gene driven by the 0.5-kb promoter followed endogenous gene regulation. Then transgenic mice harboring the 0.5-kb/SV40 T antigen construct were generated and two founders out of three developed adrenal tumors. Cells derived from the tumor of founder 1 (ATC1) were grown in presence of forskolin to maintain ACTH receptor expression and were tested for ACTH responsiveness by immunocychemistry and northern blot analyses. Even after several passages, the ACTH induced AKR1B7 and P450c11beta mRNAs accumulations were similar to that observed in mouse primary adrenocortical cell cultures. Our findings suggest that ATC1 cells have conserved essential features of ZF cells. In order to achieve complete characterization of these cells further analyses are currently performed to investigate their steroidogenic activity.

The Drosophila gene stand still encodes a germline chromatin-associated protein that controls the transcription of the ovarian tumor gene.

The Drosophila gene stand still (stil) encodes a novel protein required for survival, sexual identity and differentiation of female germ cells. Using specific antibodies, we show that the Stil protein accumulates in the nucleus of all female germ cells throughout development, and is transiently expressed during early stages of male germline differentiation. Changes of Stil subnuclear localization during oogenesis suggest an association with chromatin. Several mutant alleles, which are point mutations in the Stil N-terminal domain, encode proteins that no longer co-localized with chromatin. We find that Stil binds to many sites on polytene chromosomes with strong preference for decondensed chromatin. This localization is very similar to that of RNA polymerase II. We show that Stil is required for high levels of transcription of the ovarian tumor gene in germ cells. Expression of ovarian tumor in somatic cells can be induced by ectopic expression of Stil. Finally, we find that transient ubiquitous somatic expression of Stil results in lethality of the fly at all stages of development.

Drosophila ovary morphogenesis: analysis of terminal filament formation and identification of a gene required for this process.

The adult ovary of Drosophila is composed of approximately 20 parallel repetitive structures called ovarioles. At the anterior tip of each ovariole is a stack of 8-9 disc-shaped cells, called the terminal filament. Ovariole morphogenesis starts with the formation of the terminal filaments. Using two enhancer trap markers for terminal filament cells, we show that terminal filaments form in a progressive manner from medial to lateral across the ovary and that the number of terminal filament cells in a developing stack increases gradually. This process occurs during the second half of the third larval instar. One of these enhancer trap mutations, which is in the bric à brac gene, demonstrates that this gene is necessary for terminal filament formation and that a terminal filament cell cluster is required for ovariole morphogenesis to take place.