#Braina pro alternative Activator

A consistent passage (P3вЂ“P6 from the initially cryopreserved cells) of Neuro-2A cells was used throughout. Mouse Neuro-2A cells (American Type Culture Collection, Manassas, VA) were plated at a density of 1 Г— 10 5 cells/ml in DMEM (Invitrogen, Gaithersburg, MD) containing 10% FBS ( 25). We also demonstrate that cAMP response element-binding protein (CREB) acts downstream of ANG II/AT 1R signaling to mediate the increase in PRR expression in response to DOCA-salt by binding to sites identified in the promoter region of the PRR gene ( Atp6ip2). Here, we report that ANG II acts through the type 1 angiotensin receptor (AT 1R) to increase PRR expression in the CNS during DOCA-salt hypertension. However, it is still not clear how the PRR is upregulated in hypertension.ДЄ common mechanism among these hypertensive animal models is activation of the brain RAS and increased ANG II formation. Taken together, these reports suggest that the PRR plays a pivotal role in the central regulation of blood pressure. We also recently reported that PRR expression levels are elevated throughout the brain during DOCA salt-induced hypertension ( 17), and neuron-specific PRR deletion prevents the development of DOCA-salt hypertension. Knocking down PRR in the brain through virus-mediated introduction of small hairpin (inhibitory) RNA against the PRR (PRR-shRNA) attenuates hypertension in both SHR and RA mice. We and others have reported that the PRR is upregulated in the central nervous system (CNS) in different hypertensive models, including spontaneously hypertensive rats (SHR) ( 27) and human renin-angiotensinogen (RA) double-transgenic hypertensive mice ( 16).

#Braina pro alternative Activator

Binding of renin or prorenin to the PRR also directly initiates an intracellular signaling pathway independent of ANG II that increases the synthesis of profibrotic molecules, including plasminogen activator inhibitor-1, fibronectin, collagen, and transforming growth factor-ОІ ( 10, 25, 26). Prorenin also gains enzymatic activity by binding to the PRR ( 24). Binding of renin to the PRR increases the catalytic activity of renin by approximately fourfold to fivefold. The (pro)renin receptor (PRR) is a newly discovered component of the RAS that is highly expressed in the brain ( 16, 26). The brain renin-angiotensin system (RAS) plays an essential role in neurogenic hypertension ( 4, 21). Collectively, these findings indicate that ANG II acts via AT 1R to upregulate PRR expression both in cultured cells and in DOCA-salt hypertensive mice by increasing CREB binding to the PRR promoter. This increase in CREB activity was reversed by AT 1R blockade. Chromatin immunoprecipitation assays revealed that ANG II treatment increased CREB binding to the endogenous PRR promoter in both cultured neurons and hypothalamic tissues of DOCA-salt hypertensive mice.

ANG II also induced PRR upregulation in primary cultured neurons. CREB and AP-1 inhibitors, as well as CREB knockdown abolished ANG II-induced increases in PRR levels.

To test which ANG II/AT 1R downstream transcription factors were involved in PRR regulation, we treated Neuro-2A cells with ANG II with or without CREB (cAMP response element-binding protein) or AP-1 (activator protein-1) inhibitors, or CREB siRNA. Intracerebroventricular infusion of losartan or captopril attenuated DOCA-salt-induced PRR mRNA elevation in the paraventricular nucleus of the hypothalamus, suggesting a role for ANG II/AT 1R signaling in regulating PRR expression during DOCA-salt hypertension.

Concurrently, mice were intracerebroventricularly infused with the AT 1R blocker losartan, angiotensin-converting-enzyme inhibitor captopril, or artificial cerebrospinal fluid for 3 wk. Sham pellet-implanted mice that were provided regular drinking water served as controls. To address whether ANG II type 1 receptor (AT 1R) signaling is involved in this regulation, we implanted a DOCA pellet and supplied 0.9% saline as the drinking solution to C57BL/6J mice. We reported that brain (pro)renin receptor (PRR) expression levels are elevated in DOCA-salt-induced hypertension however, the underlying mechanism remained unknown.