Repression of vasopressin gene expression by glucocorticoids in transgenic mice: evidence of a direct mechanism mediated by proximal 5′ flanking sequence
Section snippets
Animals and treatments
The production and identification of transgenic mice that express a 3.5 kb bovine vasopressin gene construct has been described previously; in the present study the line designated VP-B1 was used, and bred on site.[3]Mice were maintained in a controlled environment and provided with freely available food and water. Animals that had been identified as either transgenic or non-transgenic by tail-tip genomic DNA screening were killed by cervical dislocation. In some experiments, mice were treated
Results
Northern analysis of total cellular RNA extracted from the adrenal glands of VP-B1 transgenic mice revealed easily detectable levels of transgene mRNA (Fig. 1) as observed previously.[3]Subsequent hybridization with an oligonucleotide probe specific for the endogenous mouse AVP mRNA did not reveal any detectable message in the adrenal gland, whereas mouse AVP mRNA was readily detectable in hypothalamic tissue (data not shown). Treatment with the adrenocortical 11 β-hydroxylase inhibitor
Discussion
In the present study we have shown that 3.5 kb of the BVP gene comprising 1.25 kb of 5′ flanking sequence, the coding region and 0.2 kb 3′ of the native poly (A) addition site, is able to mediate negative regulation by glucocorticoids in vivo. In a previous study it was shown that a similar region also mediates an osmotic response in magnocellular neurons, although additional sequence is required to restrict expression to these neurons.[3]Therefore, it would appear that proximal cis-acting
Conclusions
In conclusion, we have provided evidence which indicates that regulation of vasopressin gene expression by glucocorticoids may be mediated by a direct mechanism that is dependent, at least partially, upon regulatory sequence within the 5′ flanking region of the vasopressin gene. The use of a transgenic model has validated one experimental approach that may be used to define the molecular mechanisms of glucocorticoid repression in neuronally expressed genes. Similar studies are required to
Acknowledgements
Supported by grants to DC from the Medical Research Council (U.K.) and The Wellcome Trust. Mike Underwood (Animal Care), Bob Jones and Guy Pitt (Photography) are also gratefully acknowledged.
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2010, Journal of Biological ChemistryCitation Excerpt :When this balance is perturbed, as in glucocorticoid resistance, the decrease in the negative feedback control of cortisol in the hypothalamic-pituitary-adrenal axis leads to increased secretion of ACTH and cortisol and resistance to adrenal suppression by DEX (6, 7). The diverse pathway through which glucocorticoids regulate gene transcription is illustrated in reports where glucocorticoids directly affect expression of arginine vasopressin through a glucocorticoid-response element (GRE) (8) and negative glucocorticoid-response elements (9, 10) and is a receptor-mediated interaction with nonreceptor factors (11, 12). Glucocorticoids inhibit CRH secretion and in doing so inhibit pro-opiomelanocortin synthesis (13).
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2005, Endocrinology and Metabolism Clinics of North AmericaGlucocorticoid regulation of peptide genes in neuroendocrine CRH neurons: A complexity beyond negative feedback
2005, Frontiers in NeuroendocrinologyCitation Excerpt :Because glucocorticoids can control a wide range of cellular processes concerned with secretogogue physiology, it is likely that they can influence mechanisms in all six compartments of the CRH neuron illustrated in Fig. 3B. But their direct actions are probably confined to effects on receptors, signal transduction cascades, and genes that can affect a large number of processes [9,11,18,19,28,58,80,104]. But of course, the consequences of these actions can be quite wide-ranging.
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2005, Techniques in the Behavioral and Neural SciencesCitation Excerpt :Direct glucocorticoid negative feedback at the transcriptional level of the vasopressin promoter has been recently reported (Pearce et al., 1998; Kim et al., 2001). Glucocorticoid type II receptors are expressed in the parvocellular PVN neurons and, after coupling to these, the glucocorticoid–receptor complex inhibits vasopressin transcription by binding to a glucocorticoid regulatory element in the vasopressin promoter region (Uht et al., 1988; Burke et al., 1997). Additionally, evidence supports the binding of the glucocorticoid–receptor complex to Jun (co-transcription factor with Fos) that would repress AP-1 activity and thus attenuate vasopressin gene transcription responses (Diamond et al., 1990; Schule et al., 1990; Stauber et al., 1990; Yang-Yen et al., 1990; Unlap and Jope, 1994; Kovacs et al., 2000).
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