Browsing by Subject "heart"

Now showing 1 - 4 of 4
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    Development of imaging methods to quantify the laminar microstructure in rat hearts
    (Texas A&M University, 2004-11-15) Hudson, Kristen Kay
    The way in which the myocardium responds to its mechanical environment must be understood in order to develop reasonable treatments for congestive heart failure. The first step toward this understanding is to characterize and quantify the cardiac microstructure in healthy and diseased hearts. Myocardium has a laminar architecture made up of myolaminae, which are sheets of myocytes surrounded by a collagen weave. By enhancing the contrast between the myocytes and the surrounding collagen, the myocardium can be investigated and its laminar structure can be quantified. Many of the techniques that have been used to view the microstructure of the heart require the use of toxic or caustic chemicals for fixation or staining. An efficient imaging method that uses polarization microscopy and enhances the contrast between the collagen and myocytes while minimizing the use of harmful chemicals was developed in this research. Collagen is birefringent; therefore its visibility should be enhanced through polarization microscopy and image processing. The sheet angles were viewed directly by cutting slices of a rat septum perpendicular to the fiber angle. Images of different polarization combinations were taken and a region of interest was selected on the sample. Image processing techniques were used to reduce the intensity variation on the images and account for the variable gain of the camera. The contrast between the collagen and myocytes was enhanced by comparing adjusted images to the background and looking at a single image this comparison produced. Although the contrast was enhanced, the embedding media reduced the collagen signal and the enhancement was not as striking as expected.
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    Exercise training modulates apoptotic signaling in the aging rat heart
    (Texas A&M University, 2005-11-01) Kwak, Hyo Bum
    Aging is characterized by a progressive decline in cardiac function. A critical contributor to the age-related impairment in heart function is the loss of cardiac myocytes through ??apoptosis??, or programmed cell death. A dramatic increase in the rate of apoptosis has been reported with aging in the rat left ventricle. In contrast, exercise training not only improves cardiac function, but also reduces the risk of heart disease. However, the ability of exercise training to modulate apoptotic signaling and apoptosis in the aging heart remains unknown. Therefore, the purpose of this study was to determine the effects of exercise training on apoptotic signaling and apoptosis in the aging heart. We hypothesized that (1) aging would increase pro-apoptotic signaling and apoptosis in the rat left ventricle, and (2) exercise training would ameliorate upregulation of Bcl-2 family-driven apoptosis in the heart. Four and 25 month old Fischer-344 rats were assigned to four groups: young control (YC), young trained (YT), old control (OC), and old trained (OT). Exercise training groups ran on a treadmill for 60 min/day at 15 m/min (15Ėš incline), 5 d/wk for 12 wk. Protein expression of Bax, Bcl-2, caspase-9, and cleaved caspase-3 was measured using Western immunoblot analysis. Apoptosis (DNA fragmentation) was assessed using a cell death detection ELISA. Bax levels in OC were dramatically higher (+176.0%) compared to YC. In contrast, exercise training resulted in a significant decrease (-53.4%) in Bax in OT compared to OC. Bcl-2 levels in OC were lower (-26.3%) compared to YC. Conversely, exercise training significantly increased Bcl-2 levels by 117.8% in OT compared to OC. Caspase-9 levels were higher (+98.7%) in OC than YC, while exercise training significantly reduced caspase-9 levels in YT (-52.6%) and OT (-76.9%), respectively. Aging resulted in a dramatic increase (+122.8%) in cleaved caspase-3 levels and a significant decrease (-32.9%) with exercise training. Finally, apoptosis (DNA fragmentation) significantly increased (+163.8%) with aging and decreased (-43.9%) with exercise training. These novel data indicate that aging increases pro-apoptotic signaling and apoptosis in the left ventricle, while exercise training is effective in diminishing pro-apoptotic signaling and apoptosis in the aging heart.
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    Exercise training regulation of extracellular matrix and remodeling in the aging rat heart
    (2009-05-15) Kwak, Hyo Bum
    Aging is characterized by a progressive impairment of cardiac structure and function. The cardiac remodeling involves loss of cardiac myocytes, reactive hypertrophy of the remaining cells, and increased extracellular matrix (ECM) and fibrosis in the aging heart. In contrast, exercise training not only improves cardiac function, but also reduces the risk of heart disease. However, the ability of exercise training to modulate ECM and remodeling in the aging heart remains unknown. Therefore, the purpose of this study was to determine the effects of exercise training on ECM remodeling in the aging heart. We hypothesized that (1) exercise training would attenuate age-related changes in left ventricle morphology including extramyocyte space and collagen contents, and (2) exercise training would ameliorate age-induced changes in ECM-related factors including MMPs, TIMPs, TNF-?, TGF-?1, and ?-SMA in the heart. Three and 31 month old Fischer 344 ? Brown Norway F1 hybrid rats were assigned to four groups: young sedentary (YS), young exercise-trained (YE), old sedentary (OS), and old exercise-trained (OE). Exercise training groups walked briskly on a treadmill for 45 min/day (12? incline) at 20m/min (young) or 10 m/min (old), 5 d/wk for 12 wk. We found that endurance exercise training might ameliorate the ageinduced increase in extramyocyte space and collagen contents of the left ventricle. Exercise training might protect against age-induced fibrosis by increasing MMP-2, MMP-14 in the soluble fraction and MMP-1, MMP-3, MMP-14 in the insoluble fraction of old rat hearts. Conversely, exercise training might reduce the fibrosis by decreasing TIMP-1 in the soluble fraction of old rat hearts. Further, exercise training reduced potential upstream pro-fibrotic mediators including TNF-? and TGF-?1 in the aging rat hearts. These results are the first to demonstrate that exercise training has a protective effect against age-induced extracellular collagen matrix remodeling in the aging heart, associated with increased MMP-1, -2, -3, -14 and decreased TIMP-1, TNF-?, and TGF- ?1.
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    The development of processing methods for a quantitative histological investigation of rat hearts
    (Texas A&M University, 2004-11-15) Jetton, Emily Hope
    In order to understand the mechanical functions of the cardiac muscle it is important to first understand the microstructure of the tissue. Young et al. (1998) realized that quantitative three-dimensional information about the ventricular myocardium is necessary to analyze myocardial mechanics. They developed a technique using confocal fluorescence laser scanning microscopy to obtain three-dimensional images. While this method worked well in rebuilding the myocardial tissue image by image, it was quite extensive and costly. Costa et al. (1999) developed a method that was used to perform three-dimensional reconstruction as well. Their method, while less expensive and much less time consuming, required sheet assumptions and did not look directly at the cross-fiber plane. From Dr. Criscione's previous work on canines (Ashikaga et al., 2004), we found that the sheet structure can be accurately determined from cross-fiber sections without making any sheet assumptions. We have now expanded on those ideas and created a method to perform the quantitative histological investigation of the rat hearts in a way that is both timely and cost effective. We developed a processing method that preserves the orientation of the fiber and sheet angles. This method was carried out using plastic embedding since the dehydration process used in paraffin embedding has a tendency to grossly distort tissue. Once the heart was fixed in formalin, we then removed the septum and sliced it several times vertically. This allowed us to image the tissue at several depths and find an average fiber angle for each slice. Next, the specimen was hardened, and the sheet orientation was evaluated using polarized light. Once both fiber and sheet angles were obtained from several depths within the septum, we then constructed a three-dimension model of the wall. This method was both cost effective and less time consuming than previous ones and will be a method that can be used in the future to compare the myocardial tissue of diseased and healthy rat hearts so that we may better understand the mechanical functions of the heart as it remodels due to disease.