Maria Lonnett Burgess

Integrin-mediated collagen gel contraction by cardiac fibroblasts. Effects of angiotensin II.

Angiotensin II (Ang II), a vasoactive octapeptide, has been implicated in cardiac growth and the development of hypertrophy and fibrosis secondary in hypertensive disease. These consequences of Ang II imply an effect on the function and morphology of cardiac interstitial cells (fibroblasts). The present investigation was designed to (1) determine whether neonatal heart fibroblasts (NHFs) possess functional Ang II receptors on their plasma membrane and (2) examine the effects of Ang II on NHFs in vitro using three- and two-dimensional (3D and 2D, respectively) cultures. Several analytic techniques were used to test the specific questions of the present study. Since cardiac fibroblast phenotype can be influenced by culture conditions, both 2D and 3D cultures were used in the present investigations. Reverse-transcriptase polymerase chain reaction and radioligand binding analysis were used to test for the presence of Ang II receptors on NHFs. Both revealed that NHFs in 2D culture possess Ang II receptor mRNA and Ang II receptors. When isolated NHFs were cultured in 3D collagen gels and treated with Ang II, gel contraction was stimulated by NHFs. This effect was attenuated by the specific Ang II receptor antagonist [Sar1,Ala8]Ang II. Ang II-stimulated gel contraction was completely inhibited by extracellular matrix receptor (beta 1-integrin) antibodies (P < .05), supporting previous studies indicating that collagen gel contraction is mediated via the integrins. Immunofluorescent staining was used to test the localization of cell-surface integrins. A more intense staining pattern for beta 1-integrin in Ang II-treated versus control cells was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Age-associated changes in cardiac matrix and integrins

The progressive shift from young age to senescence is characterized by structural and functional changes in the cardiac extracellular matrix (ECM), which supports and aligns myocytes and blood vessels, and maintains myocardial mass, structure and function. As cardiac function declines with advancing age, ECM collagen and fibronectin influence diastolic stiffness. ECM binding to membrane-bound receptors, or integrins, directly links ECM to cardiac muscle and fibroblast cells, affording it the permissive role to modulate heart function. To better understand the ECM structure-function relationship in the old heart, we studied the relative protein content of these ECM proteins and integrins across three age groups. Old Balb-c mice (20 months) exhibit biventricular, cardiac hypertrophy, and greater left ventricular (LV) collagen, fibronectin, alpha 1 and alpha 5 integrin protein than middle-aged (12 months) or young (2 months) LV (P<0.05). beta1 integrin protein content is lower in old LV (P<0.05). These data show that advancing age is associated with greater collagen, fibronectin, alpha 1 and alpha 5 integrin content, suggesting that these matrix proteins undergo coordinated regulation in the aging heart. The differential integrin and ECM protein content suggests that there is regulatory signaling to the fibroblasts, which maintain the cardiac ECM.

Differential integrin expression by cardiac fibroblasts from hypertensive and exercise-trained rat hearts

The cardiac fibroblast is the principal cell type responsible for extracellular matrix (ECM) synthesis in the heart during growth and pathophysiological conditions. A dynamic interaction exists between the cardiac ECM and fibroblasts that is sensitive to the local mechanical and chemical tissue environment. We propose here that cardiac fibroblasts structurally and functionally adapt to changing local environments by altering their expression of receptor integrins. Changes in the extracellular environment are communicated in part by integrins, which link the ECM to the cell and regulate phenotype and function. In this report, we analyze integrin protein expression, migration and gel contraction by cardiac fibroblasts from rats subjected to 10 weeks of treadmill exercise (XTR), experimental hypertension (HYP) or controls (CONT). Immunoprecipitation shows that beta1 protein increases in XTR and HYP. Also, alpha1 and alpha2 integrins are lower in XTR and HYP, and alpha5 integrin is higher in XTR and lower in HYP. Functional assays show that XTR and HYP migrate slower on collagen, while XTR migrate faster and HYP slower on fibronectin. Cell isolation procedure, population expansion number or a general adaptation to culture conditions does not explain the differences observed. No significant differences in collagen gel contraction are detected. These results indicate that cardiac fibroblasts retain their in vivo patterns in vitro for a limited number of population expansions. This tissue-specific phenotype is exhibited in early passage (< or =6). However, by late passage (>8), cells begin to show adaptation to the in vitro conditions. These results show that cardiac fibroblasts respond to changing environments in pathophysiological conditions by modulating integrin expression, which is associated with changes in cell migration. They also suggest a pragmatic use for primary cardiac fibroblasts as a model to study the cardiac matrix remodeled by physiological (exercise) and pathological (hypertension) stressors.

Validity of Field Tests of Upper Body Muscular Strength

The purpose of this study was to determine the validity of five field tests (FTs) of upper body muscular strength and endurance (UBMSE) in 9-10-year-old children. Ninety-four children (38 boys, 56 girls) performed five FTs of UBMSE: pull-ups, flexed arm hang, push-ups, Vermont modified pull-ups (VMPU), and New York modified pull-ups. They also performed three criterion tests (CTs) of strength and three CTs of muscular endurance using a supported weight, set-resistance device. Zero-order correlations between the sum of the standard scores on the three CTs of strength (SUM1RM) and the FTs were nonsignificant. However, when SUM1RM was expressed relative to body weight (SUM1RM.kg-1), significant (p < .01) correlation coefficients were obtained for each FT. Highest correlations with SUM1RM.kg-1 were observed for the VMPU, and this same test yielded the smallest percentage of zero scores. Principal components analysis of the CTs, normalized for body weight, and FTs yielded a factor on which both the FTs and CTs of strength loaded significantly. These data indicate that the five FTs, though invalid as measures of absolute strength and muscular endurance, manifest concurrent and construct validity as measures of weight-relative muscular strength.

Extracellular Matrix Expression, Organization, and Interaction with Heart Myocytes During Development and Disease

The extracellular matrix (ECM) of the heart is intimately associated with cardiac function in development and disease. The heart ECM is composed of the interstitial collagens, including predominantly type I and type III collagen, various proteoglycans and noncollagenous glycoproteins such as laminin and fibronectin [2, 7, 22]. These ECM components are organized into an elaborate three-dimensional network which is associated with the structure and physiology of the heart.

Effects of angiotensin II on cardiac fibroblast contractility

Angiotensin II is known to mediate cardiac remodeling during hypertension by promoting fibrosis and myocyte hypertrophy. This study investigates the effects of angiotensin II on the contractility of the cardiac fibroblast. Using a version of the elastic substratum method, angiotensin II at 1 /spl mu/M was shown to increase the average traction stress exerted by the cardiac fibroblast up to 75% after one hour of stimulation. The enhanced contractility caused by angiotensin II was eliminated by 1 /spl mu/M of irbesartan which is a competitive blocker of the AT/sub 1/-receptor. Irbesartan not only inhibited the effects of angiotensin II, it actually decreased traction compared to untreated controls. This indicates that the effects of angiotensin II on contractility are mediated at least in part by the AT/sub 1/-receptor. Although cardiac fibroblasts are a minor component of the myocardium, a chronic increase in their contractility could be sufficient to increase resistance to ventricular relaxation during diastole, and thereby negatively affect the end-diastolic filling capacity.