Abstract
Gender-related differences in cardiac function have been described in the literature,
but whether the presence of sex hormones is responsible for these differences remains
unclear. This study was designed to determine whether testosterone regulates the gene
expression of calcium regulatory proteins in rat heart, thus playing a role in gender-related
differences in cardiac performance. Ventricular myocytes were isolated from two-day-old
rats and treated with testosterone at varying duration; the levels of gene expression
for the androgen receptor (AR) and major calcium regulatory proteins were determined
by quantitative real-time PCR. Testosterone (1 µM) treatment induced a maximum increase
in β1-adrenergic receptor and L-type calcium channel mRNA levels following an eight hour
exposure. Six hours testosterone treatment stimulated a 300-fold increase in androgen
receptor message abundance, and Na/Ca exchanger mRNA levels reached a maximum level
following twenty-four hour testosterone treatment. Taken together, these data provide
the first evidence that testosterone regulates gene expression of the major calcium
regulatory proteins in isolated ventricular myocytes, and may thus play a role in
the gender-related differences observed in cardiac performance.
Key words
Testosterone - Calcium channel - Androgen receptor - Na/Ca exchanger - β1-adrenergic receptor
References
- 1
Kimmelstiel C D, Konstam M A.
Heart failure in women.
Cardiology.
1995;
86
304-309
- 2
Vaccarino V, Parsons L, Every N R, Barron H V, Krumholz H M.
Sex-based differences in early mortality after myocardial infarction.
N Engl J Med.
1999;
341
217-225
- 3
Makkar R R, Fromm B S, Steinman R T, Meissner M D, Lehmann M H.
Female gender as a risk factor for torsades de pointes associated with cardiovascular
drugs.
JAMA.
1993;
270
2590-2597
- 4
Rautaharju P M, Zhou S H, Wong S, Calhoun H P, Berenson G S, Prineas R, Davignon A.
Sex differences in the evolution of the electrocardiographic QT interval with age.
Can J Cardiol.
1992;
8
690-695
- 5
Rosenkranz-Weiss P, Tomek R J, Mathew J, Eghbali M.
Gender-specific differences in expression of mRNAs for functional and structural proteins
in rat ventricular myocardium.
J Mol Cell Cardiol.
1994;
26
261-270
- 6
Scheuer J, Malhotra A, Schaible T F, Capasso J.
Effects of gonadectomy and hormonal replacement on rat hearts.
Circ Res.
1987;
61
12-19
- 7
Marsh J D, Lehmann M H, Ritchie R H, Gwathmey J K, Green G E, Schiebinger R J.
Androgen receptors mediate hypertrophy in cardiac myocytes.
Circulation.
1998;
98
256-261
- 8
Bers D M.
Cardiac excitation-contraction coupling.
Nature.
2002;
415
198-205
- 9
Bassani R A, Bassani J W, Bers D M.
Relaxation in ferret ventricular myocytes: unusual interplay among calcium transport
systems.
J Physiol.
1994;
476
295-308
- 10
Bassani J W, Bassani R A, Bers D M.
Relaxation in rabbit and rat cardiac cells: species-dependent differences in cellular
mechanisms.
J Physiol.
1994;
476
279-293
- 11
Vizgirda V M, Wahler G M, Sondgeroth K L, Ziolo M T, Schwertz D W.
Mechanisms of sex differences in rat cardiac myocyte response to beta-adrenergic stimulation.
Am J Physiol Heart Circ Physiol.
2002;
282
H256-H263
- 12
Golden K L, Fan Q I, Chen B, Ren J, O’Connor J, Marsh J D.
Adrenergic stimulation regulates Na(+)/Ca(2+)Exchanger expression in rat cardiac myocytes.
J Mol Cell Cardiol.
2000;
32
611-620
- 13
Davidoff A J, Maki T M, Ellingsen O, Marsh J D.
Expression of calcium channels in adult cardiac myocytes is regulated by calcium.
J Mol Cell Cardiol.
1997;
29
1791-1803
- 14
Nakamura T, Sakaeda T, Ohmoto N, Tamura T, Aoyama N, Shirakawa T, Kamigaki T, Nakamura T,
Kim K I, Kim S R, Kuroda Y, Matsuo M, Kasuga M, Okumura K.
Real-time quantitative polymerase chain reaction for MDR1, MRP1, MRP2, and CYP3A-mRNA
levels in Caco-2 cell lines, human duodenal enterocytes, normal colorectal tissues,
and colorectal adenocarcinomas.
Drug Metab Dispos.
2002;
30
4-6
- 15
Wooley P H, Morren R, Andary J, Sud S, Yang S Y, Mayton L, Markel D, Sieving A, Nasser S.
Inflammatory responses to orthopaedic biomaterials in the murine air pouch.
Biomaterials.
2002;
23
517-526
- 16
Ritchie R H, Marsh J D, Lancaster W D, Diglio C A, Schiebinger R J.
Bradykinin blocks angiotensin II-induced hypertrophy in the presence of endothelial
cells.
Hypertension.
1998;
31
39-44
- 17
Vermeulen A.
Clinical review 24: Androgens in the aging male.
J Clin Endocrinol Metab.
1991;
73
221-224
- 18
Yan S M, Finato N, di Loreto C, Beltrami C A.
Nuclear size of myocardial cells in end-stage cardiomyopathies.
Anal Quant Cytol Histol.
1999;
21
174-180
- 19
Matturri L, Biondo B, Colombo B, Lavezzi A M, Rossi L.
Significance of the DNA synthesis in hypertrophic cardiomyopathies.
Basic Res Cardiol.
1997;
92
85-89
- 20
Lauer M S, Anderson K M, Larson M G, Levy D.
A new method for indexing left ventricular mass for differences in body size.
Am J Cardiol.
1994;
74
487-491
- 21
Vasan R S, Larson M G, Levy D, Evans J C, Benjamin E J.
Distribution and categorization of echocardiographic measurements in relation to reference
limits: the Framingham Heart Study: formulation of a height- and sex-specific classification
and its prospective validation.
Circulation.
1997;
96
1863-1873
- 22
Gardin J M, Henry W L, Savage D D, Ware J H, Burn C, Borer J S.
Echocardiographic measurements in normal subjects: evaluation of an adult population
without clinically apparent heart disease.
J Clin Ultrasound.
1979;
7
439-447
- 23
Hayward C S, Kalnins W V, Kelly R P.
Gender-related differences in left ventricular chamber function.
Cardiovasc Res.
2001;
49
340-350
- 24
Golden K L, Marsh J D, Jiang Y.
Castration reduces mRNA levels for calcium regulatory proteins in rat heart.
Endocrine.
2002;
19
339-344
- 25
Mora G R, Mahesh V B.
Autoregulation of the androgen receptor at the translational level: testosterone induces
accumulation of androgen receptor mRNA in the rat ventral prostate polyribosomes.
Steroids.
1999;
64
587-591
- 26
Wiren K M, Chapman E A, Zhang X W.
Osteoblast differentiation influences androgen and estrogen receptor-alpha and -beta
expression.
J Endocrinol.
2002;
175
683-694
- 27
Singh R, Artaza J N, Taylor W E, Gonzalez-Cadavid N F, Bhasin S.
Androgens Stimulate Myogenic Differentiation and Inhibit Adipogenesis in C3H 10T1/2
Pluripotent Cells Through an Androgen Receptor-Mediated Pathway.
Endocrinology.
2003;
144
5081-5088
- 28
Ubels J L, Wertz J T, Ingersoll K E, Jackson R S, Aupperlee M D.
Down-regulation of androgen receptor expression and inhibition of lacrimal gland cell
proliferation by retinoic acid.
Exp Eye Res.
2002;
75
561-571
- 29
Muller J G, Isomatsu Y, Koushik S V, O’Quinn M, Xu L, Kappler C S, Hapke E, Zile M R,
Conway S J, Menick D R.
Cardiac-specific expression and hypertrophic upregulation of the feline Na(+)-Ca(2+)
exchanger gene H1-promoter in a transgenic mouse model.
Circ Res.
2002;
90
158-164
- 30
Pogwizd S M, Schlotthauer K, Li L, Yuan W, Bers D M.
Arrhythmogenesis and contractile dysfunction in heart failure: Roles of sodium-calcium
exchange, inward rectifier potassium current, and residual beta-adrenergic responsiveness.
Circ Res.
2001;
88
1159-1167
- 31
Pogwizd S M.
Increased Na(+)-Ca(2+) exchanger in the failing heart.
Circ Res.
2000;
87
641-643
- 32
Hintz K K, Wold L E, Colligan P B, Scott G I, Lee K J, Sowers J R, Ren J.
Influence of ovariectomy on ventricular myocyte contraction in simulated diabetes.
J Biomed Sci.
2001;
8
307-313
- 33
Koenig H, Fan C C, Goldstone A D, Lu C Y, Trout J J.
Polyamines mediate androgenic stimulation of calcium fluxes and membrane transport
in rat heart myocytes.
Circ Res.
1989;
64
415-426
- 34
Gomez A M, Valdivia H H, Cheng H, Lederer M R, Santana L F, Cannell M B, McCune S A,
Altschuld R A, Lederer W J.
Defective excitation-contraction coupling in experimental cardiac hypertrophy and
heart failure.
Science.
1997;
276
800-806
- 35
Scamps F, Mayoux E, Charlemagne D, Vassort G.
Calcium current in single cells isolated from normal and hypertrophied rat heart.
Effects of beta-adrenergic stimulation.
Circ Res.
1990;
67
199-208
- 36
Richard S, Leclercq F, Lemaire S, Piot C, Nargeot J.
Ca2+ currents in compensated hypertrophy and heart failure.
Cardiovasc Res.
1998;
37
300-311
- 37
Keung E C.
Calcium current is increased in isolated adult myocytes from hypertrophied rat myocardium.
Circ Res.
1989;
64
753-763
- 38
Liu L, Fan Q I, El Zaru M R, Vanderpool K, Hines R N, Marsh J D.
Regulation of DHP receptor expression by elements in the 5'-flanking sequence.
Am J Physiol Heart Circ Physiol.
2000;
278
H1153-H1162
- 39
Engelhardt S, Boknik P, Keller U, Neumann J, Lohse M J, Hein L.
Early impairment of calcium handling and altered expression of junction in hearts
of mice overexpressing the beta1-adrenergic receptor.
FASEB J.
2001;
15
2718-2720
- 40
Evanko D S, Ellis C E, Venkatachalam V, Frielle T.
Preliminary analysis of the transcriptional regulation of the human beta 1-adrenergic
receptor gene.
Biochem Biophys Res Commun.
1998;
244
395-402
- 41
Tseng Y T, Stabila J P, Nguyen T T, McGonnigal B G, Waschek J A, Padbury J F.
A novel glucocorticoid regulatory unit mediates the hormone responsiveness of the
beta1-adrenergic receptor gene.
Mol Cell Endocrinol.
2001;
181
165-178
- 42
Jorgensen J S, Nilson J H.
AR suppresses transcription of the alpha glycoprotein hormone subunit gene through
protein-protein interactions with cJun and activation transcription factor 2.
Mol Endocrinol.
2001;
15
1496-1504
- 43
Golden K L, Ren J, O’Connor J, Dean A, DiCarlo S E, Marsh J D.
In vivo regulation of Na/Ca exchanger expression by adrenergic effectors.
Am J Physiol Heart Circ Physiol.
2001;
280
H1376-H1382
- 44
Golden K L, Ren J, Dean A, Marsh J D.
Norepinephrine regulates the in vivo expression of the L-type calcium channel.
Mol Cell Biochem.
2002;
236
107-114
- 45
Golden K L, Marsh J D, Jiang Y, Brown T, Moulden J.
Gonadectomy of adult male rats reduces contractility of isolated cardiac myocytes.
Am J Physiol Endocrinol Metab.
2003;
285
E449-E453
- 46
Maki T, Gruver E J, Davidoff A J, Izzo N, Toupin D, Colucci W, Marks A R, Marsh J D.
Regulation of calcium channel expression in neonatal myocytes by catecholamines.
J Clin Invest.
1996;
97
656-663
- 47
Wallukat G.
The beta-adrenergic receptors.
Herz.
2002;
27
683-690
K. L. Golden, Ph. D.
Wayne State University School of Medicine
421 E. Canfield Ave. · Detroit · MI 48201
Phone: +1(313)576-4480
Fax: +1(313)577-1112
Email: kgolden@med.wayne.edu