PMID- 36362173 OWN - NLM STAT- MEDLINE DCOM- 20221114 LR - 20230701 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 23 IP - 21 DP - 2022 Nov 2 TI - Ligand-Dependent Downregulation of Guanylyl Cyclase/Natriuretic Peptide Receptor-A: Role of miR-128 and miR-195. LID - 10.3390/ijms232113381 [doi] LID - 13381 AB - Cardiac hormones act on the regulation of blood pressure (BP) and cardiovascular homeostasis. These hormones include atrial and brain natriuretic peptides (ANP, BNP) and activate natriuretic peptide receptor-A (NPRA), which enhance natriuresis, diuresis, and vasorelaxation. In this study, we established the ANP-dependent homologous downregulation of NPRA using human embryonic kidney-293 (HEK-293) cells expressing recombinant receptor and MA-10 cells harboring native endogenous NPRA. The prolonged pretreatment of cells with ANP caused a time- and dose-dependent decrease in (125)I-ANP binding, Guanylyl cyclase (GC) activity of receptor, and intracellular accumulation of cGMP leading to downregulation of NPRA. Treatment with ANP (100 nM) for 12 h led to an 80% decrease in (125)I-ANP binding to its receptor, and BNP decreased it by 62%. Neither 100 nM c-ANF (truncated ANF) nor C-type natriuretic peptide (CNP) had any effect. ANP (100 nM) treatment also decreased GC activity by 68% and intracellular accumulation cGMP levels by 45%, while the NPRA antagonist A71915 (1 microM) almost completely blocked ANP-dependent downregulation of NPRA. Treatment with the protein kinase G (PKG) stimulator 8-(4-chlorophenylthio)-cGMP (CPT-cGMP) (1 microM) caused a significant increase in (125)I-ANP binding, whereas the PKG inhibitor KT 5823 (1 microM) potentiated the effect of ANP on the downregulation of NPRA. The transfection of miR-128 significantly reduced NPRA protein levels by threefold compared to control cells. These results suggest that ligand-dependent mechanisms play important roles in the downregulation of NPRA in target cells. FAU - Khurana, Madan L AU - Khurana ML AD - Department of Physiology, School of Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA. FAU - Mani, Indra AU - Mani I AUID- ORCID: 0000-0001-7690-5517 AD - Department of Physiology, School of Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA. FAU - Kumar, Prerna AU - Kumar P AD - Department of Physiology, School of Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA. FAU - Ramasamy, Chandramohan AU - Ramasamy C AD - Department of Physiology, School of Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA. FAU - Pandey, Kailash N AU - Pandey KN AD - Department of Physiology, School of Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA. LA - eng GR - R01 DK133833/DK/NIDDK NIH HHS/United States GR - R01 HL057531/HL/NHLBI NIH HHS/United States GR - R01 HL062147/HL/NHLBI NIH HHS/United States GR - HD062147 DK133833/NH/NIH HHS/United States PT - Journal Article DEP - 20221102 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - EC 4.6.1.2 (Guanylate Cyclase) RN - 85637-73-6 (Atrial Natriuretic Factor) RN - GVO776611R (Iodine-125) RN - 0 (Ligands) RN - H2D2X058MU (Cyclic GMP) RN - 0 (MicroRNAs) RN - 114471-18-0 (Natriuretic Peptide, Brain) RN - 0 (MIRN128 microRNA, human) RN - 0 (MIRN195 microRNA, human) SB - IM MH - Humans MH - *Guanylate Cyclase/genetics/metabolism MH - Atrial Natriuretic Factor/genetics/pharmacology/metabolism MH - Ligands MH - Down-Regulation MH - HEK293 Cells MH - Cyclic GMP/metabolism MH - *MicroRNAs/genetics MH - Natriuretic Peptide, Brain/metabolism PMC - PMC9657974 OTO - NOTNLM OT - ANP-NPRA/cGMP signaling OT - GC/NPRA OT - HEK-293 cells OT - MA-10 cells OT - internalization COIS- The authors declare no conflict of interest. EDAT- 2022/11/12 06:00 MHDA- 2022/11/15 06:00 PMCR- 2022/11/02 CRDT- 2022/11/11 01:28 PHST- 2022/07/29 00:00 [received] PHST- 2022/10/23 00:00 [revised] PHST- 2022/10/25 00:00 [accepted] PHST- 2022/11/11 01:28 [entrez] PHST- 2022/11/12 06:00 [pubmed] PHST- 2022/11/15 06:00 [medline] PHST- 2022/11/02 00:00 [pmc-release] AID - ijms232113381 [pii] AID - ijms-23-13381 [pii] AID - 10.3390/ijms232113381 [doi] PST - epublish SO - Int J Mol Sci. 2022 Nov 2;23(21):13381. doi: 10.3390/ijms232113381.