Myocardial stretch produces an increase in developed force (DF) that occurs in two phases: the first (rapidly occurring) is generally attributed to an increase in myofilament calcium responsiveness and the second (gradually developing) to an increase in [Ca(2+)](i). Rat ventricular trabeculae were stretched from approximately 88% to approximately 98% of L(max), and the second force phase was analyzed. Intracellular pH, [Na(+)](i), and Ca(2+) transients were measured by epifluorescence with BCECF-AM, SBFI-AM, and fura-2, respectively. After stretch, DF increased by 1.94+/-0.2 g/mm(2) (P<0.01, n = 4), with the second phase accounting for 28+/-2% of the total increase (P<0.001, n = 4). During this phase, SBFI(340/380) ratio increased from 0.73+/-0.01 to 0.76+/-0.01 (P<0.05, n = 5) with an estimated [Na(+)](i) rise of approximately 6 mmol/L. [Ca(2+)](i) transient, expressed as fura-2(340/380) ratio, increased by 9.2+/-3.6% (P<0.05, n = 5). The increase in [Na(+)](i) was blocked by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA). The second phase in force and the increases in [Na(+)](i) and [Ca(2+)](i) transient were blunted by AT(1) or ET(A) blockade. Our data indicate that the second force phase and the increase in [Ca(2+)](i) transient after stretch result from activation of the Na(+)/H(+) exchanger (NHE) increasing [Na(+)](i) and leading to a secondary increase in [Ca(2+)](i) transient. This reflects an autocrine-paracrine mechanism whereby stretch triggers the release of angiotensin II, which in turn releases endothelin and activates the NHE through ET(A) receptors.

This study was designed to gain additional insight into the mechanism of the slow force response (SFR) to stretch of cardiac muscle. SFR and changes in intracellular Na(+) concentration ([Na(+)](i)) were assessed in cat papillary muscles stretched from 92% to approximately 98% of L(max). The SFR was 120+/-0.6% (n=5) of the rapid initial phase and coincided with an increase in [Na(+)](i). The SFR was markedly depressed by Na(+)-H(+) exchanger inhibition, AT(1) receptor blockade, nonselective endothelin-receptor blockade and selective ET(A)-receptor blockade, extracellular Na(+) removal, and inhibition of the reverse mode of the Na(+)-Ca(2+) exchange by KB-R7943. KB-R7943 prevented the SFR but not the increase in [Na(+)](i). Inhibition of endothelin-converting enzyme activity by phosphoramidon suppressed both the SFR and the increase in [Na(+)](i). The SFR and the increase in [Na(+)](i) after stretch were both present in muscles with their endothelium (vascular and endocardial) made functionally inactive by Triton X-100. In these muscles, phosphoramidon also suppressed the SFR and the increase in [Na(+)](i). The data provide evidence that the last step of the autocrine-paracrine mechanism leading to the SFR to stretch is Ca(2+) entry through the reverse mode of Na(+)-Ca(2+) exchange.

Abstract-Myocardial stretch is a well-known stimulus that leads to hypertrophy. Little is known, however, about the intracellular pathways involved in the transmission of myocardial stretch to the cytoplasm and nucleus. Studies in neonatal cardiomyocytes demonstrated stretch-induced release of angiotensin II (Ang II). Because intracellular alkalinization is a signal to cell growth and Ang II stimulates the Na ϩ /H ϩ exchanger (NHE), we studied the relationship between myocardial stretch and intracellular pH (pH i ). Experiments were performed in cat papillary muscles fixed by the ventricular end to a force transducer. Muscles were paced at 0.2 Hz and superfused with HEPES-buffered solution. pH i was measured by epifluorescence with the acetoxymethyl ester form of the pH-sensitive dye 2Ј,7Ј-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF-AM). Each muscle was progressively stretched to reach maximal developed force (L max ) and maintained in a length that was Ϸ92% L max (L i ). During the "stretch protocol," muscles were quickly stretched to L max for 10 minutes and then released to L i ; pH i significantly increased during stretch and came back to the previous value when the muscle was released to L i . The increase in pH i was eliminated by (1) Key Words: stretch, myocardial Ⅲ pH, intracellular Ⅲ Na ϩ /H ϩ exchange Ⅲ angiotensin Ⅲ endothelin A lthough it is well known that mechanical stimuli cause a variety of effects on the structure and function of the myocardial cells, little is known about how cells sense the mechanical stimuli, transmit the information to messenger systems, and finally regulate function and growth.1,2 Highly regarded experiments demonstrate that the release of angiotensin II (Ang II) contributes to stretch-induced hypertrophy in cultured neonatal cardiac myocytes, [2][3][4][5] and that the effect was suppressed by the AT 1 -receptor antagonist TCV-116. 4 The release of Ang II may involve an autocrine or paracrine mechanism because stretch-conditioned media mimicked the effect of stretch when transferred to nonstretched neonatal cardiomyocytes.2 An increase in PKC activity is an effect detected after stretching cultured neonatal cardiomyocytes 2 and the adult heart. 6 Because Ang II, by mechanisms still unresolved but probably linked to PKC, activates the Na ϩ /H ϩ exchanger (NHE), 7-8 the logical expectation is a rise in intracellular pH (pH i ) after myocardial stretch. Although we are unaware of measurements of myocardial pH i before and during stretch, pressure overload increased NHE-1 mRNA levels in hearts.9 Furthermore, the activity of mitogen-activated protein kinase (MAP kinase) was increased by stretch in cultured cardiomyocytes, and this increase was partially eliminated by NHE inhibition.9 Nevertheless, an unknown is whether stretch alters NHE activity in multicellular preparations from adult hearts. This point is critical, because Ang II released after stretch might increase NHE activity 10 -14 and promote the expression of endothelin (ET) as well as the upregulation of ET rec...