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Am J Physiol Endocrinol Metab 292: E1418 –E1425, 2007.

First published January 23, 2007; doi:10.1152/ajpendo.00335.2006.

Growth hormone receptor deficiency in mice results in reduced systolic blood
pressure and plasma renin, increased aortic eNOS expression, and altered
cardiovascular structure and function
E. Egecioglu,1 I. J. Andersson,1 E. Bollano,2,3 V. Palsdottir,4 B. G. Gabrielsson,4 J. J. Kopchick,5
O. Skott,6 P. Bie,6 J. Isgaard,7 M. Bohlooly-Y,8 G. Bergstro¨m,1,7 and A. Wickman1
Departments of 1Physiology and 2Cardiology, 3Wallenberg Laboratory for Cardiovascular Research, 4Department of
Metabolism and Cardiovascular Research, Sahlgrenska Academy, Go¨teborg University, Gothenberg, Sweden; 5Edison
Biotechnology Institute and Department of Biomedical Sciences, Ohio University, Athens, Ohio; 6Department of Physiology
and Pharmacology, Institute of Medical Biology, Syddansk University, Odense, Denmark; 7Department of Clinical
Physiology, Sahlgrenska Academy, Go¨teborg University, Gothenberg, Sweden; and 8AstraZeneca R&D, Mo¨lndal, Sweden
Submitted 11 July 2006; accepted in final form 12 January 2007

Egecioglu E, Andersson IJ, Bollano E, Palsdottir V, Gabrielsson
BG, Kopchick JJ, Skott O, Bie P, Isgaard J, Bohlooly-Y M, Bergstro¨m G, Wickman A. Growth hormone receptor deficiency in mice
results in reduced systolic blood pressure and plasma renin, increased
aortic eNOS expression, and altered cardiovascular structure and function. Am J Physiol Endocrinol Metab 292: E1418 –E1425, 2007. First
published January 23, 2007; doi:10.1152/ajpendo.00335.2006.—To
study the role of the growth hormone receptor (GHR) in the development of cardiovascular structure and function, female GHR genedisrupted or knockout (KO) and wild-type (WT) mice at age 18 wk
were used. GHR KO mice had lower plasma renin levels (12 ⫾ 2 vs.
20 ⫾ 4 mGU/ml, P ⬍ 0.05) and increased aortic endothelial NO
synthase (eNOS) expression (146%, P ⬍ 0.05) accompanied by a
25% reduction in systolic blood pressure (BP, 110 ⫾ 4 vs. 147 ⫾ 3
mmHg, P ⬍ 0.001) compared with WT mice. Aldosterone levels were
unchanged, whereas the plasma potassium concentration was elevated
by 14% (P ⬍ 0.05) in GHR KO. Relative left ventricular weight was
14% lower in GHR KO mice (P ⬍ 0.05), and cardiac dimensions as
analyzed by echocardiography were similarly reduced. Myograph
studies revealed a reduced maximum contractile response in the aorta
to norepinephrine (NE) and K⫹ (P ⬍ 0.05), and aorta media thickness
was decreased in GHR KO (P ⬍ 0.05). However, contractile force
was normal in mesenteric arteries, whereas sensitivity to NE was
increased (P ⬍ 0.05). Maximal acetylcholine-mediated dilatation was
similar in WT and GHR KO mice, whereas the aorta of GHR KO mice
showed an increased sensitivity to acetylcholine (P ⬍ 0.05). In
conclusion, loss of GHR leads to low BP and decreased levels of renin
in plasma as well as increase in aortic eNOS expression. Furthermore,
GHR deficiency causes functional and morphological changes in both
heart and vasculature that are beyond the observed alterations in body
size. These data suggest an important role for an intact GH/IGF-I axis
in the maintenance of a normal cardiovascular system.
cardiac structure; cardiac function; vascular structure; vascular function; endothelial nitric oxide synthase; growth hormone receptor-null
mice

structural and functional changes of the cardiovascular system
are, however, unclear.
GH-deficient patients have a higher prevalence of cardiovascular morbidity and mortality than the general population
(16, 20, 26). However, patients with GH insensitivity/resistance (Laron syndrome), which is a much rarer condition, are
less well studied in relation to cardiovascular mortality and
morbidity. These patients have reduced cardiac dimension,
with reductions in left ventricular (LV) volume and mass, as
well as in stroke volume and cardiac output (CO) (11). However, systolic and diastolic functions appear intact, and the
cardiac response to exercise is normal. Thus, it appears that the
heart, despite its smaller size, can meet the metabolic need in
GH-resistant patients. To our knowledge, consequences of GH
insensitivity on vascular function and structure have not been
studied in patients with GH insensitivity/resistance, whereas
patients with GH deficiency, conversely, display endothelial
dysfunction (10) and decreased systemic formation of nitric
oxide (NO) (1).
An animal model resembling GH resistance is the GH
receptor (GHR)/GH-binding protein gene disrupted, or knockout (GHR KO), mouse (33). Although these mice secrete GH,
there are no biological effects because of the lack of the GHR.
It has previously been reported that the resultant mouse phenotype has very low levels of serum insulin-like growth factor
I (IGF-I) and insulin, loss of GHR and GH-binding protein, and
an ⬃50% weight reduction compared with littermates at 12 wk
of age (9, 33).
The aim of the study was to investigate the importance of an
intact GH/IGF-I axis in the development of normal cardiovascular function and morphology, in particular, the importance of
the GHR in cardiac and vascular function and structure.
MATERIALS AND METHODS

(GH) activity,
such as GH deficiency or GH insensitivity/resistance, or GH
excess as exemplified in acromegaly, are associated with an
increase in cardiovascular disease (5, 26). The mechanisms by
which abnormally low or high levels of GH signaling cause

Animals. The GHR KO mouse was generated as previously described (33). Homozygous GHR KO and wild-type (WT) littermate
females (Sv129Ola-Balb/c background) were identified by PCR analysis of DNA from tail biopsy specimens as previously described (33).
Animals were housed together with littermate controls during the
whole study. The environment of the animal rooms was controlled

Address for reprint requests and other correspondence: E. Egecioglu, Dept.
of Physiology, The Sahlgrenska Academy, Go¨teborg University, PO Box 434,
SE-405 30 Gothenberg, Sweden (e-mail: emil.egecioglu@medic.gu.se).

The costs of publication of this article were defrayed in part by the payment
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in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

CONDITIONS LINKED TO REDUCED GROWTH HORMONE

E1418

0193-1849/07 $8.00 Copyright © 2007 the American Physiological Society

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The Netherlands) following the manufacturer’s protocol. model ILS 943. Sweden) for a minimum of 20 min. The mice had free access to tap water and standard pellet chow. Aortic rings were stretched to 3 mN and equilibrated for 30 min. The internal circumferences of the mesenteric arteries (GHR KO n ⫽ 10. The endothelium-dependent relaxation response was studied by cumulative concentration response to acetylcholine (ACh. Chicago. Ring segments of thoracic aorta (⬃3 mm. pulsed. A heating pad was used to maintain body temperature. and the tail was gently heated before every recording session. A second concentration-response to ACh was performed in the presence of the NO synthase (NOS) inhibitor N␻-nitro-Larginine (L-NNA. mesenteric arteries. aldosterone. WT n ⫽ 13) were mounted in an organ bath (Department of Physiology. Systolic blood pressure (SBP) was measured using a computerized noninvasive tail-cuff system (RTBP Monitor. Offline measurements were done by one observer blinded to the type of mice. At 17 wk of age. and the plasma was frozen for later measurement of plasma renin concentration by radioimmunoassay of angiotensin I by use of the antibody-trapping technique (19).and 1. removing remaining hair. Bothell. Aortic sections (n ⫽ 6 in each group) were paraffin imbedded. RR interval and R amplitude were also calculated. Transcript levels of eNOS. twodimensional guided. CA) and performed on an ABI Prism 7900HT Sequence Detection System 292 • MAY 2007 • www. 10⫺1 mol/l) with 20-min washout between each procedure. LV fractional shortening (FS) was calculated as follows: (LVIDdLVIDs)/LVIDd ⫻ 100. COAT-A-COUNT.org . Relative quantification of caveolin-1 and ecSOD mRNA expression was performed using a LightCycler (Roche Diagnostics) and FastStart Master SYBR Green I (Roche Diagnostics). plasma renin concentration was measured (n ⫽ 17 in each group). n ⫽ 7) were anesthetized with isoflurane (Baxter Healthcare. MA). Ex vivo vascular function. GHR KO (n ⫽ 8) and WT (n ⫽ 8) mice were weighed and anesthetized with pentobarbital sodium (60 mg/kg. Time parameters were measured using Pharmlab 3. A graphic interface of the analysis program allowed visual reviewing and manual editing of the detected events.000-fold) were accepted. Roche Diagnostics. Louis.0 (AstraZeneca. Before the pharmacological protocol. Sodium nitroprusside (Sigma. The aorta was separated into three parts.asecho. and heart were dissected out. A standard curve. Renin values were standardized with renin standards obtained from the National Institutes for Biological Standards and AJP-Endocrinol Metab • VOL E1419 Control (Potters Bar. and end systole was taken at the peak inward of interventricular septum motion. Gothenberg. they were further stretched to 12 mN and allowed to stabilize.org). respectively. UK) and are expressed in milliGoldblatt units per milliliter (mGU/ml). Diagnostic Products. The chest was shaved and a peeling cream applied. Sweden) and a Multi myograph 610M (Danish Myo Technology. WT n ⫽ 13) were normalized according to the manufacturer’s protocol. Plasma renin. Go¨teborg University. All tracings were recorded at a sweep of 100 mm/s and were stored on magnetic optical discs for offline measurements. caveolin-1. and a mean QRS complex was generated as a mean of ⬃20 beats. Denmark). and extracellular superoxide dismutase (ecSOD) were designed using LightCycler Probe Design Software (version 1. Aortic wall thickness and number of muscle layers were measured on these sections. 10⫺5 and 10⫺4 mol/l) was applied after each ACh concentration-response to validate the endothelium-independent relaxation of the smooth muscle cells. St. The cumulative concentration-response relationship to NE was then studied (aorta 10⫺9 to 10⫺5 mol/l. and the level of anesthesia was adjusted to keep heart rate stable at ⬃400 beats/min. ECG was recorded at 2 kHz using Pharmlab 3. Harvard Apparatus. Following a very short anesthesia (isoflurane) blood was taken from the orbital plexa.ajpendo. WA). The ethics committee of Go¨teborg University gave prior approval of the animal procedures. respectively. MA). Only results with linearity in serial dilutions (between 50. mesenteric arteries 10⫺8 to 10⫺5 mol/l). sodium. IL). as described before (15). 10⫺9 to 10⫺5 mol/l) on vessels precontracted to 50 –100% of maximal recorded NE contraction. Sodium and potassium concentrations in plasma were determined using flame photometry (n ⫽ 12 in each group. Seattle. MO. and potassium measurements. The heart was separated into LV including septum and right ventricle (RV) and weighed. Germany).6 to 80 ng original RNA per well). and expression levels were normalized to GAPDH. CA). and prolactin receptor were analyzed using predesigned TaqMan assay-on-demands (Applied Biosystems. was included on each plate. Mannheim. and mounted on slides for hematoxylin staining. Hertfordshire. n ⫽ 8. This was done using minimal sample size and a pulsed frequency of 5 MHz. The vessels were placed into physiological salt solution (PSS). ECG. Echocardiography. one part was used for ex vivo functional tests. a warming pad was used to maintain body temperature. a relative humidity of 45–55%. sectioned in a microtome. South Natick. 10⫺5 mol/l) and KCl (K⫹.0. End diastole was taken at the onset of the QRS complex. GHR KO n ⫽ 8. Electrocardiography (ECG) leads were placed on the extremities. PCR primers for GAPDH. Real-time PCR analysis. The recordings were performed in the afternoon on four consecutive days with at least 10 measurements at each recording. The mice were anesthetized with isoflurane and ECG leads were placed on the extremities (GHR KO n ⫽ 7. one part was fixed in formalin for further morphological analysis. Doppler was used to record the estimated peak LV outflow tract velocity and the mitral inflow velocities (E and A wave and their ratio). WT. Final SBP was obtained from the three last days of recording by averaging the mean values. 10⫺4 mol/l). Lexington. Leiden. and the third part was trimmed free of surrounding tissues and frozen in liquid nitrogen and saved for further mRNA analysis. Primer sequences and thermal conditions for each gene are listed in Table 1. Relative wall thickness and velocity of circumferential shortening (Vcf) were calculated using formulas described previously (30). Standard cDNA from pooled aortic RNA was synthesized in parallel with the sample cDNAs. Aarhus. Eighteen-week-old mice were used in the study. WT n ⫽ 8) and mesenteric arteries (GHR KO n ⫽ 10. Morphological aorta analysis. the QRS time as the time from the start of the Q or R wave to the neutralization of the R wave. Instrumentation Laboratory. Plasma aldosterone was determined using a commercial kit (n ⫽ 12 in each group. and concentrations were determined by Ribo Green (Molecular Probes. Cardiac ultrasound studies were performed using a commercially available ultrasonograph (ATL HDI 5000 SonoCT. and the QT time as the start of the Q wave or R wave to the neutralization of the T wave. Briefly. This served as a guide for M-mode tracing. Sweden) before the aorta. Systolic blood pressure. Apoteksbolaget.0. a 15-MHz linear transducer was used to obtain two-dimensional parasternal short-axis images close to the papillary muscle. Foster City. Los Angeles. QT time was also correlated to the RR interval due to heart rate influence on the QT time. Mo¨lndal. Four or more beats were averaged for each measurement. Conscious animals (n ⫽ 8 in each group) were kept in a restraining cylinder with a standard acclimatization time of 10 min. Phillips Ultrasound. Gothenberg. Wall thickness was measured and muscle layers were counted by a blinded observer on five different sections of the aortic wall at ⫻40 magnification. Mice (GHR KO. M-mode measurements of LV internal diameters and wall thickness in diastole and systole were made using the convention of the American Society of Echocardiography (www. WT n ⫽ 7). All vessels were activated with norepinephrine (NE. PQ time was defined as the time from start of the P wave to the start of the Q or R wave. Total RNA from aorta (n ⫽ 8 in each group) was extracted with Tri Reagent (Sigma). obtained by serial dilution of the standard cDNA (range 0. where LVIDd and LVIDs are LV internal diameters in diastole (d) and systole (s). IGF-I.GH RECEPTOR AND CARDIOVASCULAR STRUCTURE AND FUNCTION with a 12:12-h light-dark cycle. and a temperature of 20°C.

08). The resultant relative wall thickness was similar in both groups. Statistics. 2A). WT 36 ⫾ 3 g.E1420 GH RECEPTOR AND CARDIOVASCULAR STRUCTURE AND FUNCTION Table 1. P values ⬍0. times. Table 2). 95°C 4 s.26 ⫾ 0.5 ⫾ 3. 28 g. WT 1. posterior wall thickness was decreased by 25% (P ⬍ 0.9 ⫾ 0.05.05 as analyzed with Student’s t-test. WT 7. no significant difference was detected between the groups (KO 2. WT 155 ⫾ 2 mmol/l.05. Reagents (TaqMan Universal PCR Mastermix. CO was decreased by 48% (P ⬍ 0.0 vs. 4B).3 vs. we apprehended that the smaller tail of the GHR KO animals possibly could affect our measurements.05) and LVDd was similarly decreased by 28% (P ⬍ 0.05). not significant (NS). heart rate was kept at ⬃400 beats/min in both groups (KO 412 ⫾ 17 vs. PPIA. ecSOD. CyberGene.05 as analyzed with Mann-Whitney U-test. PQ time and QRS time were similar in both groups (PQ: KO 37.05). If not stated otherwise. AJP-Endocrinol Metab • VOL The plasma renin concentrations were 40% lower in GHR KO compared with WT mice (KO 12 ⫾ 2 vs. 1). mGU.7 ⫾ 1. WT 26. Since the tail cuff technique is sensitive to the relation between the cuff and the size of the tail. Systolic blood pressure was reduced by 25% in growth hormone receptor (GHR) knockout (KO) vs.6 mg/10 g body wt). but no differences were detected between the groups.9.5 ⫾ 0. 95°C 4 s. 2D). Primer sequences. extracellular superoxide dismutase. Fig. The relative LV weights were also lower in the GHR KO mice compared with WT (KO 22. The plasma potassium concentrations were increased by 14% in GHR KO compared with WT mice (KO 7. unpublished observations]. Fig. P ⫽ 0.95 ⫾ 0. NS. GHR KO mice had 53% decreased body weight compared with WT (KO 17 ⫾ 1 vs. two-tailed t-tests for unpaired data were used for statistical comparisons between groups.3 vs. Fig. but when CO was related to body weight. caveolin-1.06.0 ⫾ 0. During ECG measurements.4 ms).7 ⫾ 1. 3). 1. no difference in maximum contractile response to K⫹ and NE could be detected between 292 • MAY 2007 • www. (Applied Biosystems). WT 20 ⫾ 4 mGU/ml.1 ⫾ 0. WT 22. WT 10. However. Fig. 4A) in GHR KO. glyceraldehyde-3-phosphate dehydrogenase. 95°C 60 s. 95°C 4 s. GHR KO mice showed longer QT time compared with WT (KO 27. suggesting decreased systolic cardiac function. WT 812 ⫾ 46 pg/ml.41 ⫾ 0.org . 56°C 9 s. 60°C Caveolin-1 15 s. Applied Biosystems) and reaction conditions were used according to the manufacturer’s instructions.05 vs.0. WT 147 ⫾ 3 mmHg. Fig.7 ⫾ 0. 72°C Forward primer Reverse primer Forward primer Reverse primer Forward primer Reverse primer Forward primer Reverse primer PPIA 15 s. Fig. when two different sizes of WT mice on a different background (C57/ BL6) were studied (18 vs. Fig. In the mesenteric arteries. 72°C ecSOD 15 s. Table 2).28 vs.51 ⫾ 0. WT 6. 67°C 3 s. Table 1) was used as a reference gene for eNOS and prolactin receptor and ␤-actin for IGF-I. WT 6. P ⬍ 0. and elongation for the genes GAPDH. Sodium concentrations were unchanged in GHR KO (KO 157 ⫾ 2 vs. The diastolic cardiac function was also studied by calculating the E/A quote. Cyclophilin A (PPIA. P ⬍ 0.05 were considered to be statistically significant. milliGoldblatt units. RESULTS Systolic blood pressure was reduced by 25% in GHR KO mice compared with WT (KO 110 ⫾ 4 vs. Huddinge.001. annealing. Variables without homogeneity in variance or normal distribution were analyzed with a Mann-Whitney U-test. wild-type (WT) mice (*P ⬍ 0. n ⫽ 5: 112 ⫾ 2 vs.5 ⫾ 0. 58°C 8 s.6 vs. similar systolic blood pressures were recorded in both groups [18 g. and temperatures for denaturizing. All standards and samples were analyzed in triplicate. When %FS and Vcf were calculated. and ecSOD used in real-time quantitative PCR Gene Denaturation Annealing Elongation Oligonucleotide GAPDH 0 s. P ⬍ 0. QRS: KO 12. no differences between the groups were detected (Table 2 and Fig.27 mmol/l.7 ⫾ 1.05) compared with WT (Table 2).8 ms). cyclophilin A.9 ⫾ 0. In GHR KO mice.6 vs. GHR KO mice displayed decreased %FS and Vcf by 11 and 24%.05. 28 g). The levels of aortic eNOS mRNA were increased by 146% in GHR KO compared with WT (KO 16. 2B). P ⬍ 0. Fig. 2C). There was no difference in the relative RV weight between GHR KO and WT mice (KO 5. WT 448 ⫾ 8 beats/min). No change in aldosterone levels was found in GHR KO compared with WT mice (KO 815 ⫾ 101 vs. WT 35.9 mg/10 g body wt. respectively (P ⬍ 0. P ⬍ 0. P ⬍ 0. Sweden. 72°C Sequence 5⬘-GTC 5⬘-TGC 5⬘-AGG 5⬘-TGC 5⬘-GAC 5⬘-AGG 5⬘-AGC 5⬘-GTC GTG CTG GTT CAT GAC AAG TAG CCC GAT CTT CCT TAT GTG GAG GAC GAA CTG CAC CCT GGC GTC AGA GAC CTC ACG CAC TTC GTG AAG ATG GAA ATG TGC C-3⬘ CTT CT ACA GAA TT-3⬘ TAA AGT C-3⬘ AT-3⬘ GC-3⬘ G-3⬘ C-3⬘ GAPDH.ajpendo.7 vs. when the QT time was corrected for the small difference in heart rate (QTc). n ⫽ 5: 110 ⫾ 2 g. Values are expressed as means ⫾ SE.001. However. PPIA.7 ⫾ 1.

4 ⫾ 0. 5C). WT ⫺7.83 ⫾ 4.18. P ⬍ 0. Fig. No difference in aldosterone (B) or sodium (C) was found between GHR KO and WT animals.1 vs. P ⬍ 0.05. GHR KO mice had increased mRNA expression of endothelial NO synthase (eNOS) normalized to cyclophilin A (PPIA) in the aorta (*P ⬍ 0.1 nM/mm.2 ⫾ 5. GHR KO showed an increased sensitivity to NE compared with WT in the mesenteric arteries (log EC50: KO ⫺5.2 log mol/l.58 ⫾ 0. P ⬍ 0. respectively.59 ⫾ 0. NE: KO 1. sensitivity to ACh was increased in aorta in GHR KO compared with WT (log EC50: KO ⫺7.1 vs. respectively.7 ⫾ 0.1 mN/mm. In the mesenteric arteries.2 ⫾ 0. Fig. WT 40. there was no difference in the maximal relaxation response induced by ACh relative to the maximal endothelium-independent relaxation induced by sodium nitroprusside (KO 49.5 ⫾ 0. WT ⫺6.6 ⫾ 0. respectively. WT 2.6 ⫾ 0.5 ⫾ 1.05 for both.9%.05). the maximum contractile response to K⫹ and NE was reduced in GHR KO compared with WT (K⫹ KO: 0. WT 44. 5F).5 ⫾ 4. respectively).05.2 ⫾ 6. WT mice (*P ⬍ 0. Fig. Blockade of NOS by L-NNA reduced ACh-induced dilations significantly more in the mesenteric arteries in GHR KO compared with WT (KO 11. WT 100 ⫾ 1. 5F). 3. Mann-Whitney U-test.5 vs. respectively. respectively.11 vs.6 vs.9 vs. WT 83. However.6%.44 ⫾ 0.5 vs.058 vs. Student’s t-test. Sodium nitroprusside was used to test the function of the smooth muscle cells and caused a less profound relaxation in the mesenteric arteries in GHR KO compared with WT (KO 72. D: potassium levels in plasma were increased by 14% in the GHR KO vs.1 ⫾ 0. Fig. 5E).07 vs. The presence of L-NNA totally abolished AChinduced relaxations (Fig. and sodium nitroprusside caused profound relaxation in aorta in both GHR KO and WT (KO 98.3 ⫾ 0. respectively. 5E). Fig.78 ␮m. WT (*P ⬍ 0. WT 27. AJP-Endocrinol Metab • VOL GHR KO and WT mice (K⫹: KO 1.58 ⫾ 4. respectively.35 ⫾ 0. A: plasma renin concentrations was decreased by 40% in GHR KO vs.05). n ⫽ 17 in each group).GH RECEPTOR AND CARDIOVASCULAR STRUCTURE AND FUNCTION E1421 Fig.7%) and no difference in the sensitivity to ACh (KO ⫺6. 2.4 ⫾ 6.73 ⫾ 0. WT 1. Fig.1 vs.4 ⫾ 2.1%. WT ⫺7. WT ⫺5. There were no differences in the sensitivity to NE (log EC50: KO ⫺7. P ⬍ 0.2 ⫾ 0.ajpendo. n ⫽ 12 in each group).11 vs.91 ⫾ 0.05.0 ⫾ 3. NE: KO 1.05).1 log mol/l.05. respectively.1 vs. 5D) or in ACh-induced maximal relaxation relative to the maximal endothelium-independent relaxation induced by sodium nitroprusside in aorta (KO 54. 5A). P ⬍ 0. 5B).9 ⫾ 0.9 ⫾ 6.7%: Fig. Fig.2 vs. WT 1.13 log mol/l. P ⬍ 0. The thickness of the GHR KO aortic media was significantly reduced by 29% compared with WT (KO 45.5 ⫾ 6.09 vs.05).1 mN/mm.org . WT 64. whereas no changes in the 292 • MAY 2007 • www. WT 1. In the aorta.068 nM/mm.

04 1. the reduction in plasma renin was not accompanied by a concomitant reduction in plasma aldosterone.52 vs. beats/min LVDd. A: cardiac output (CO) was reduced in GHR KO vs.12 ⫾ 122. we have shown that lack of GHR signaling causes a reduction in systolic blood pressure and plasma renin levels as well as an increase in aortic eNOS expression. Thus. AJP-Endocrinol Metab • VOL be high in some of the measurements.1). 24.001 404⫾55 332⫾31 NS 0.43 ⫾ 0. BW.75⫾0.6. but it has been speculated that a generally hypotrophic cardiovascular system could be causative (12). not significant. This.70. NS. the elevated potassium levels 292 • MAY 2007 • www.6 ⫾ 0. LVDs.05 0. WT 270. despite a reduced global systolic function. WT mice (*P ⬍ 0. posterior wall thickness.50 vs. number of muscle layers could be found between GHR KO and WT (average: KO 4. mm/10 g BW PWTh.05 NS 10. The aorta of the GHR KO shows a markedly hypotrophic phenotype with a reduction in media thickness and a reduced maximal contractile response. resulting in a reduction in total peripheral resistance. we conclude that functional GH signaling is important for the maintenance of normal cardiac and large vessel structure and function. cm/s E/A 36⫾3 17⫾1 0.28⫾0.ajpendo. mm/10 g BW LVDs.73 ⫾ 0. B: when CO was normalized to body weight. there was no difference between the groups.57 0.97 7. This effect could further be augmented by the altered endothelial function toward a more relaxed state with increased eNOS expression and increased NO release.39⫾0. might possibly be explained by the slightly increased plasma potassium levels in the GHR KO. Vcf.001 0. 4. % E Wave.org .8 vs. RWTh.04 61⫾2 55⫾2 0. growth hormone receptor knockout.38⫾0. Speculatively. body weight. Thus. heart rate.02 0.4 vs. The levels of aortic caveolin-1 mRNA tended to be lower in the GHR KO animals (KO 0. a possible altered regulation of plasma renin levels may be of importance for the lower blood pressure in the KO animals. NS) were unchanged as was that of the prolactin receptor (KO 217. velocity of circumfential shortening. in turn.03 NS 0. left ventricular diameter (systole). Expression levels of ecSOD (KO 1. Body weight and various variables from ECG measurements n WT GHR KO P value 7 8 BW. variances in the mRNA levels tended to Fig. WT 1. fractional shortening.60⫾0. cardiac weight and volume were reduced to a larger extent than could be explained by the lower body weight of the GHR KO mouse. However.62⫾0.57⫾0.15. Therefore.02 0. 31).03⫾0. Sensitivity to AChmediated dilatation is increased in the aorta with a matching increase in eNOS mRNA expression.14 NS WT.6. resulting in reduced systolic pressure.11 0. WT 0.15.26 1. reduced large-vessel compliance could be underlying the cardiovascular changes such that less contractile force would be needed for peak ejection.63⫾0. NS) and IGF-I mRNA (KO 1.47⫾0. wild type. The present study offers several explanations for the reduction in systolic blood pressure seen in GHR KO mice. DISCUSSION In the present study. PWTh.E1422 GH RECEPTOR AND CARDIOVASCULAR STRUCTURE AND FUNCTION Table 2.89 ⫾ 0. NS). However. P ⫽ 0. Furthermore. respectively).09 0.55 ⫾ 0.30⫾0.01 0. cm/s A Wave. FS. The observed reduction in systolic cardiac function may reduce systolic blood pressure. the GHR KO mice seem to be able to produce a CO that matches their reduction in body weight and metabolic rate but display a lower systolic blood pressure. Another important finding is the significant reduction in plasma renin activity. The mechanisms underlying this reduction remain unclear. The resistance vasculature appears normally developed but with an alteration in the factors responsible for ACh-mediated dilatation. However.01 0. left ventricular diameter (diastole).02 0. lack of significance in the statistical analysis of differences between groups does not necessarily indicate that there was no biological difference.04 NS 1. LVDd. which also has been found in hypophysectomized (Hx) rats (13.43 ⫾ 0. A general response to a reduction in blood pressure would be to increase plasma renin levels (14). Furthermore.98⫾0.44⫾0. circ/s FS.001 0. mm/10 g BW RWTh Vcf. g HR. relative wall thickness.05 0.08 vs. HR.64 ⫾ 128.51⫾0.001 0. GHR KO mice had reduced systolic blood pressure. WT 4.80⫾0. GHR KO.67⫾0. WT 0.05).

expressed as %maximal contractile force. diminishing the significance of size differences between the GHR KO and WT. ECG and posteuthanasia weights revealed a reduction in cardiac volume and mass that exceeded the observed reduction AJP-Endocrinol Metab • VOL in body weight in GHR KO mice. SNP. the lack of GHR signaling in GHR KO mice may result in a 292 • MAY 2007 • www. Thus. was also reduced in GHR KO mice compared with WT. 5.05. estimated by ECG (Bollano E. F: however. Maximal contraction in response to norepinephrine (NE) was unchanged in mesenteric arteries (A) and decreased in the aorta of GHR KO compared with WT mice (*P ⬍ 0. 28).05). E: response to ACh induced dilatation after N␻-nitro-L-arginine (L-NNA) treatment was impaired in mesenteric arteries of GHR KO (*P ⬍ 0. This is in agreement with data from Hx rats.ajpendo. which display reductions in relative LV weight (31) and cardiac dimensions.org .GH RECEPTOR AND CARDIOVASCULAR STRUCTURE AND FUNCTION E1423 Fig.05). the lack of statistical significance between the groups does not exclude a potential biological difference. personal communication). sensitivity to ACh was increased in aorta of GHR KO vs. even though we had a reasonable number of observations in the aldosterone (n ⫽ 12) measurements. B). However. Global systolic function. Relative response to NE. may contribute to the maintained aldosterone levels. C) but unchanged in the aorta of GHR KO compared with WT (D). WT mice (*P ⬍ 0. was increased in mesenteric arteries (*P ⬍ 0. sodium nitroprusside. measured as ejection fraction or fractional shortening. Systolic and diastolic blood pressure are essentially independent of mammal size (8).05. It has previously been shown that GH affects myocardial contractility via regulation of calcium handling during excitation/contraction coupling (4.

The cardiac function of GHR KO mice is also comparable with the situation in GH-deficient patients (22). Maria Sundling for excellent help with the aortic functional tests. AJP-Endocrinol Metab • VOL suggesting that vasodilatation is mediated by NO. Clark R. Vascular function and blood pressure in GH transgenic mice. it appears that intact GH signaling is of importance for development of both functional and structural aspects of large vessels. Gustafsson H. but important differences are observed relating to vascular function.ajpendo. A possible explanation to the increased eNOS expression and NO dependency is the observation of high levels of prolactin in GHR KO mice (18). 12 580). GHR KO mice showed an increased sensitivity to NE. Brabant G. agree with those in adults with primary GH resistance. Differential cardiac effects of growth hormone and insulinlike growth factor-1 in the rat. A combined in vivo and in vitro evaluation. Bergstrom G. Cittadini A. Cardiac changes in GHR KO mice have similarities with those observed in GH-resistant and GH-deficient patients. these data suggest that a functional GHR signaling is of importance for both cardiac structural development and cardiac contractile function. Wickman. 6. The GHR KO mice showed a 48% decrease in CO. The observed change in endothelial function in the direction of a more NO-dependent vasodilatation is hard to explain. SWEGENE supports the postdoctoral position of A. Wickman). Endocrinology 142: 3317–3323. and the Swedish National Heart and Lung Foundation (G. Tornell J. evidenced by a morphological reduction in the mean media thickness. increased activity of NO by prolactin might help improve NO-dependent endothelial function in GHR KO mice. It has been shown that prolactin activates the NO system (3). endothelial-derived relaxing factor. It is in line with our observation of increased expression of eNOS mRNA in the aorta of GHR KO mice. ACKNOWLEDGMENTS We thank Jing Jia for excellent help with blood pressure measurements. Bohlooly YM. Effects of 292 • MAY 2007 • www. 5. the Swedish Research Council (G. Frolich JC. which exhibit reduced cardiac dimensions but normal systolic and diastolic cardiac function at rest (11). the Novo Nordisk foundation (A. The mechanisms for the alterations in NE sensitivity caused by GH are not known. Mol Cell Endocrinol 174: 91–98. suggesting that it is principally mediated by NO. Growth hormone and the heart. in GHR KO mesenteric arteries. GH-deficient patients display impaired endothelial function (10). Olsson B. 13. Douglas PS. are responsible for the vasodilatation in this vascular segment. Endothelial function was intact in both aorta and mesenteric artery from GHR KO mice. Thus. The functional studies of GHR KO mesenteric vessels revealed a close to normal function with an intact contractile response. Carlson L. Moses AC. L-NNA effectively blocked vasodilatation. in part. and SWEGENE Centre for Mouse Physiology and Bio-Imaging. However.E1424 GH RECEPTOR AND CARDIOVASCULAR STRUCTURE AND FUNCTION defective cardiac calcium handling and resultant reduction of myocardial contractility. CO was reduced in GHR KO mice. Effect of GH on NE sensitivity is in line with findings from studies of Hx rats. 2001. In fact. Regulation of the postsynaptic alpha-adrenergic receptor in rat mesenteric artery. and its receptors are located in endothelial cells (21). further studies are needed. Interestingly. the GHR KO mice have an extended longevity (7). which may be linked to a decreased systemic production of NO (1). Gimbrone MA Jr. which is in accord with the situation in Hx rats (17. Bergstro¨m). Thus. suggesting that other factors. The role of nitric oxide in the biological activity of prolactin in the mouse mammary gland. Bode-Boger SM. Thus. L-NNA effectively blocked the ACh-mediated dilatory response in the aorta of both WT and GHR KO mice. in part. It is possible that GHR KO mice have developed compensatory systems that maintain NO production and endothelial function. or endothelialderived hyperpolarizing factor. Kopchick is supported. von zur Muhlen A. 32). Lombardi G. Loss of GHR causes functional and morphological changes in both heart and vasculature beyond the observed alterations in body size. In contrast to the GHR KO mice. 3. Taken together. by the State of Ohio Eminent Scholars Program. but the levels did not differ between the groups. Marzullo P. 4. All together. REFERENCES 1. L-NNA minimally affected the dilatory response to ACh in WT mesenteric arteries. despite systolic dysfunction. J Clin Invest 98: 2706 –2713. 2001. which is closely related to a similar reduction in body weight (⬃⫺43% in this part of the study). 25) and also with results from bovine GH transgenic mice.org . 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