Functional Aspects of the Juxtaglomerular Apparatus: Control
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ill Johanna och Isak List of Papers This thesis is based on the following papers, which are referred to in the text by their Roman numerals. I Neuronal nitric oxide synthase-deficient mice have impaired renin release but normal blood pressure. Sällström J, Carlström M, Jensen BL, Skøtt O, Brown RD, Persson AEG. Am J Hypertens, 21: 111-6, 2008. II Neuronal nitric oxide synthase supports renin release during sodium restriction through inhibition of phosphodiesterase 3. Sällström J, Jensen BL, Skøtt O, Xiang G, Persson AEG. Manuscript. III Diabetes-induced hyperfiltration in adenosine A1-receptor deficient mice lacking the tubuloglomerular feedback mechanism. Sällström J, Carlsson PO, Fredholm BB, Larsson E, Persson AEG, Palm F. Acta Physiol (Oxf), 190: 253-9, 2007. IV Inhibition of sodium-linked glucose reabsorption in the kidney normalizes diabetes-induced glomerular hyper- filtration in conscious adenosine A1-receptor-deficient mice. Sällström J, Engström T, Fredholm BB, Persson AEG, Palm F. Manuscript. Reprints were made with permission from The Nature Publishing Group and Wiley-Blackwell, respectively. Contents Introduction...................................................................................................11 Regulation of glomerular filtration...........................................................12 The myogenic response .......................................................................13 The TGF mechanism ...........................................................................14 Proximal reabsorption..........................................................................14 Nitric oxide...............................................................................................15 Nitric oxide synthases..........................................................................15 Modulation of TGF response by nitric oxide.......................................15 Nitric oxide and hypertension..............................................................16 The renin angiotensin aldosterone system................................................16 Renin release........................................................................................17 Intracellular regulation of renin release...............................................19 Nitric oxide and renin release..............................................................20 Glomerular hyperfiltration and diabetic nephropathy ..............................20 Renal glucose handling........................................................................21 Mechanisms causing diabetes-induced hyperfiltration........................21 Aims of the investigation..............................................................................23 Methods ........................................................................................................24 Animals ....................................................................................................24 nNOS knockout mice...........................................................................24 A1AR knockout mice...........................................................................24 Experimental protocols ............................................................................25 Protocol for study I ..............................................................................25 Protocol for study II.............................................................................26 Protocol for study III ...........................................................................27 Protocol for study IV ...........................................................................27 Telemetric blood pressure measurements ................................................27 Implantation of transmitters (Study I&II)............................................27 Data processing of chronic data (Study I) ...........................................28 Acute blood pressure response (Study II)............................................29 Measurements of renin and aldosterone...................................................29 Plasma renin concentration (Study I&II).............................................29 Plasma aldosterone concentration (Study I) ........................................30 Renin mRNA levels in the renal cortex (Study I)................................30 Renin genes (Study I) ..........................................................................30 Renal excretion and plasma potassium concentration..............................31 Urine collection in metabolic cages (Study I&III) ..............................31 Measurement of plasma potassium concentration (Study I)................31 Measurements of cAMP (Study II) ..........................................................31 cAMP concentration in the renal cortex ..............................................32 cAMP concentration in cultured As4.1 cells .......................................32 Acid dissection of renal tissue (Study II) .................................................32 Measurements of GFR and renal plasma flow .........................................33 GFR during anaesthesia (Study III).....................................................33 GFR in conscious mice (Study IV)......................................................34 GFR and renal plasma flow in conscious mice (Study II)...................34 Histology (Study III) ................................................................................35 Statistics ...................................................................................................36 Results...........................................................................................................37 nNOS in renin and blood pressure regulation (Study I&II) ....................37 Blood pressure and heart rate regulation .............................................37 Renin and aldosterone regulation ........................................................38 Renal excretion ....................................................................................38 PDE3 and renin regulation...................................................................39 cAMP measurements...........................................................................41 PDE5 and renin regulation...................................................................42 Effect of nNOS deletion on JG cell index ...........................................42 Diabetes-induced hyperfiltration (Study III&IV).....................................43 TGF and diabetes-induced hyperfiltration...........................................43 Glucose reabsorption and diabetes-induced hyperfiltration ................44 Discussion.....................................................................................................45 The role of nNOS in renin regulation.......................................................45 Aldosterone regulation in nNOS-/- with impaired renin regulation ..........47 Cardiovascular consequences of nNOS deficiency..................................47 TGF and diabetes-induced hyperfiltration ...............................................49 Glucose reabsorption and diabetes-induced hyperfiltration.....................50 Summary and conclusions ............................................................................52 Future perspectives .......................................................................................53 Sammanfattning på svenska..........................................................................54 Kväveoxid, hypertension och renin (Arbete I&II) ...................................55 Diabetes, hyperfiltration och nefropati (Arbete III&IV)..........................56 Acknowledgements.......................................................................................54 References.....................................................................................................61 Abbreviations 7-NI 7-nitroindazole -/- A1AR adenosine A1-receptor knockout mice AC adenylate cyclase cAMP cyclic adenosine monophosphate bw body weight cGMP cyclic guanosine monophosphate eNOS endothelial nitric oxide synthase GFR glomerular filtration rate GU Goldblatt units JG juxtaglomerular JGA juxtaglomerular apparatus Kf filtration coefficient nNOS-/- neuronal nitric oxide synthase knockout mice PAH para-amino hippuric acid PBS hydrostatic pressure in Bowman's space PGC hydrostatic pressure in the glomerular capillaries Pprox hydrostatic pressure in the proximal tubule PDE phosphodiesterase