Biomedical subjects

R J McCormick

Publications and source records attributed to R J McCormick.

At least 19 recordsLinked to original sources

PubMed2001

Exercise training attenuates aging-associated increases in collagen and collagen crosslinking of the left but not the right ventricle in the rat.

The purpose of this study was to compare the independent and interactive effects of age and exercise training on selected parameters of the right ventricle (RV), left ventricular septum (LVS) and left ventricular free-wall (LVFW) extracellular matrix. Specifically, we evaluated collagen and collagen crosslinking (hydroxylysylpyridinoline, HP) concentrations in the myocardial extracellular matrix in young adult, Y (5.5 months) and old, O (25.5 months) male Fischer 344 rats. Rats were assigned to either a sedentary control (YC, OC) or an exercise training group (YT, OT). Rats were trained for 45 min/day, 5 days/week, for 10 weeks at approximately equals 70% of maximal oxygen consumption. Following the training regimen, rats were sacrificed and their hearts were dissected into the RV, LVS and LVFW. Training resulted in a significant hypertrophy of the left ventricle (LV) but not RV relative to body weight. In young rats, collagen concentration was significantly higher (P<0.01) in RV compared to LVS, but not LVFW. With aging, collagen concentration increased significantly (P<0.05) in both ventricles, but more so in LV than RV so that differences in percent collagen observed between chambers in both YC and YT rats no longer existed in OC and OT animals. This aging effect was attenuated by training in the LVS but not the LVFW, so that collagen concentration, while higher in this region of the LV in OT compared to YT rats, was also lower than that seen in the LVS of OC rats. HP crosslink concentration in the LVS and LVFW, but not the RV of OC rats was significantly elevated above corresponding values seen in YC rats (P < 0.05 or greater). In YT rats, training had no effect on HP crosslinking concentration in any of the three regions of the heart. However, in OT rats, training completely prevented the age-associated increase in crosslinking seen in both the LVS and LVFW of OC animals. These findings illustrate the different responses of the RV, LVS and LVFW extracellular matrix components, collagen and HP crosslinking, to both aging and training in the rodent.

Aging↗

PubMed2001

Time course of collagen and decorin changes in rat cardiac and skeletal muscle post-MI.

We examined the temporal relationship between messages (type I and type III mRNAs) for the principal fibrillar procollagens and subsequent collagen accretion, cross-linking, and decorin expression in the left ventricle (LV) postmyocardial infarction (post-MI). We sought to determine 1) what role the proteoglycan decorin plays in extracellular matrix (ECM) remodeling known to take place as a consequence of MI and 2) the extent skeletal muscle ECM is altered early post-MI. Therefore, after surgically induced production of small- to moderate-sized infarcts (approximately 20% of LV mass), extent and time course of ECM remodeling was evaluated in remaining viable LV free wall and in slow- [soleus (SOL)] and fast-twitch [gastrocnemius (GAST)] skeletal muscles. Decorin, collagen, and hydroxylysylpyridinium cross-link concentrations and alpha1(I) (type I) and alpha1(III) (type III) procollagen mRNAs were measured in LVs from noninfarcted controls and at 72 h, 1, 2, 5, and 13 wk post-MI. These same data were collected in SOL and GAST muscles at all time points except 13 wk. Type I procollagen mRNA increased at both 72-h and 1-wk time points in LVs. Type III procollagen mRNA was elevated at 1 wk, returning to baseline by 2 wk post-MI. Collagen concentration was significantly increased by 1 wk, more than doubled by 5 wk, and was elevated 129% by 13 wk in the remaining viable LV. LV decorin expression was unaltered at early time points, but increased 38% at 5 wk post-MI and doubled by 13 wk post-MI. In skeletal muscle, procollagen mRNAs were transiently altered in SOL and GAST muscles without any demonstrable effect on the measured ECM parameters. This study reports, for the first time, the upregulation time course of decorin and its relationship to increased HP cross-linking and accumulation of collagen in viable myocardium post-MI.

Animals↗

PubMed2001

Collagen characteristics and organization during the progression of cholesterol-induced atherosclerosis in Japanese quail.

This study reports the concentration of collagen and its hydroxypyridinoline crosslinks, collagen fibril organization in the dorsal aortas, and systolic blood pressure during the progression of atherosclerosis in Japanese quail selected for cholesterol-induced atherosclerosis. The quail were placed on either a control or 0.5% cholesterol-added diet at approximately 16 weeks of age. The concentration of total collagen did not change in the control arteries during the course of the study, whereas at 5 and 10 weeks of cholesterol feeding, collagen levels decreased in the cholesterol-fed birds. Hydroxypyridinoline concentration increased during the duration of the study in the cholesterol-fed birds and by 15 and 20 weeks of cholesterol feeding, levels were significantly increased over those observed in the control arteries. Transmission electron microscopy showed changes in the organization of collagen fibrils. Increased systolic blood pressure was noted beginning at 10 weeks of cholesterol feeding, which is suggestive of other systemic changes induced by hypercholesterolemia. These results demonstrated remodeling of the collagen component of the dorsal aorta extracellular matrix during the progression of atherosclerosis and are suggestive of other systemic cardiovascular system changes.

Animals↗

PubMed2000

Collagen gene expression in rat left ventricle: interactive effect of age and exercise training.

Whether or not exercise training of sufficient intensity and duration to produce left ventricle (LV) hypertrophy also regulates deposition of interstitial collagen and cross-linking at the pretranslational level is unknown. Therefore, the effects of exercise training on gene expression for the two principal fibrillar collagens in LV, types I and III, were assessed in young adult (5 mo), middle-aged (15 mo), and old (26 mo) rats. We also evaluated the potential interaction of changes in mRNA for these procollagens with alterations in LV extracellular matrix characteristics by simultaneously measuring collagen concentration (hydroxyproline) and extent of mature collagen cross-linking (hydroxylysylpyridinoline, HP). Ten weeks of treadmill running resulted in LV hypertrophy and an increased maximal oxygen uptake in all three age groups of trained rats compared with sedentary controls. Percent collagen in rat LV almost doubled (P < 0.0001) from 5 to 26 mo of age, an increase unaffected by exercise training. With aging, a significant decline in expression of mRNAs for both collagen type I (P < 0.005) and type III (P < 0.001) was observed in LV free wall (LVF) but not septum (LVS). Training prevented this decline in LVF mRNAs for the two principal fibrillar collagens in middle-aged rats whereas it attenuated the decline in senescent animals. HP concentration increased significantly with aging in both LVF (P < 0.005) and LVS (P < 0.01). Training modulated this effect, but again only in LVF, so that HP was significantly lower (P < 0.05) in this region of the LV in old trained rats compared with sedentary counterparts. We conclude that exercise training modulates the effects of aging on collagen gene mRNAs and HP cross-linking regionally within the LV.

Aging↗

PubMed2000

Bovine placental protease specificity toward muscle connective tissue proteins.

Enzymes currently used to tenderize meat are not substrate-specific, resulting in extensive myofibrillar protein degradation that often produces an undesirable texture. Bovine placental metalloproteases, which selectively hydrolyze connective tissue proteins while leaving myofibrillar proteins intact, may tenderize meat without causing texture problems. Therefore, our objective was to extract and crudely purify bovine metalloproteases from bovine placenta for possible use as tenderizers in meat systems. Enzymes were extracted from homogenized tissue and purified by ammonium sulfate precipitation. Samples were collected before (crude enzyme) and after gel filtration on a Sephadex G-100 column. Spectrophotometric analysis identified one major peak (filtered enzyme). Gelatin, casein, and type I acid-soluble collagen zymography were used to determine substrate specificity. Beef myofibrillar proteins were incubated with crude and filtered enzyme fractions, enzymes quenched, and substrate degradation visualized using SDS-PAGE. Active gelatinases and collagenases exhibiting molecular weights of 57 to 65 kDa were detected on zymograms. Banding patterns from crude enzyme indicated two enzymes with both gelatinase and collagenase activity and a third enzyme with gelatinase activity only. Banding patterns from filtered enzyme indicated two enzymes with both gelatinase and collagenase activity. Proteolytic activity was not detected with casein, actin, or myosin heavy-chain substrates. Due to specificity for collagen and gelatin, these enzymes may be capable of improving the tenderness of certain cuts relatively high in connective tissue, while avoiding myofibrillar protein hydrolysis.

Animals↗

PubMed1999

Extracellular modifications to muscle collagen: implications for meat quality.

The extracellular matrix (EMC) of muscle is composed mostly of the protein collagen with lesser quantities of other constituents such as proteoglycans also present. The focus of this brief review is the extracellular modification of collagen, critical to forming a stable matrix, called crosslinking. Enzyme-mediated covalent collagen crosslinks are largely lysine-derived. Their formation is absolutely essential for stabilization of the EMC and a functional muscle. In cooked meat, the presence of crosslinks contribute to the shrinkage and tension development of collagen as it denatures with a subsequent increase in the toughness of meat. Both crosslink and collagen concentrations vary with differing muscle type, producing a wide range of textural differences among muscles. Furthermore, within a given muscle type, a wide range of conditions, often dependent on management choices, influence crosslinking patterns. Although information regarding the chemical structure, specific location, and quantity of collagen crosslinks is available, mechanisms that control and regulate their formation remain elusive. Recent studies, however, suggest a potential role for the proteoglycan decorin in regulating collagen fibrillogenesis, ordering the spatial arrangement of collagen molecules and, thus, influencing crosslinking patterns.

Animals↗

PubMed1998

Fatty acid composition and cholesterol concentration in tissues of white-tailed deer (Odocoileus virginianus) as influenced by lactation, age, and season of the year.

The purpose of this study was to determine effects of lactation, season, and age on fatty acid compositions of adipose tissue (subcutaneous and perirenal), liver, and muscle (m. longissimus dorsi), and on cholesterol concentration of liver and muscle, of female white-tailed deer (Odocoileus virginianus). Lactation did not affect fatty acid composition in adipose tissue or muscle, but in liver, weight percentages of 18:2 were lower, and of 20:4 higher in non-lactating does. Increased age (fawns, yearlings, 2 and 3+(-)years old) decreased 14:0, 15:0, and 16:0 in subcutaneous adipose tissue; decreased 14:0, 15:0, 16:1, 18:2, and increased 18:0 in perirenal adipose tissue; increased 18:1 and decreased 18:2 in liver; and increased 18:1, 18:3, and 20:4, and decreased 18:2 in muscle. Season of the year had little effect on adipose tissue and muscle fatty acids. Liver of fall season does had greater concentrations of most fatty acids than winter does, and cholesterol concentration was greatest in liver of winter does. It was concluded that season and lactation minimally affect fatty acid composition in adipose and muscle, whereas changes in saturated and unsaturated fatty acids occur with increased age. Also, liver fatty acids are influenced by each parameter studied, but cholesterol varies only with season.

Adipose Tissue↗

PubMed1998

Effect of exercise training on passive stiffness in locomotor skeletal muscle: role of extracellular matrix.

The purpose of this study was to evaluate the effect of endurance exercise training on both locomotor skeletal muscle collagen characteristics and passive stiffness properties in the young adult and old rat. Young (3-mo-old) and senescent (23-mo-old) male Fischer 344 rats were randomly assigned to either a control or exercise training group [young control (YC), old control (OC), young trained (YT), old trained (OT)]. Exercise training consisted of treadmill running at approximately 70% of maximal oxygen consumption (45 min/day, 5 days/wk, for 10 wk). Passive stiffness (stress/strain) of the soleus (Sol) muscle from all four groups was subsequently measured in vitro at 26 degreesC. Stiffness was significantly greater for Sol muscles in OC rats compared with YC rats, but in OT rats exercise training resulted in muscles with stiffness characteristics not different from those in YC rats. Sol muscle collagen concentration and the level of the nonreducible collagen cross-link hydroxylysylpyridinoline (HP) significantly increased from young adulthood to senescence. Although training had no effect on Sol muscle collagen concentration in either age group, it resulted in a significant reduction in the level of Sol muscle HP in OT rats. In contrast, exercise had no effect on HP in the YT animals. These findings indicate that 10 wk of endurance exercise significantly alter the passive viscoelastic properties of Sol muscle in old but not in young adult rats. The coincidental reduction in the principal collagen cross-link HP also observed in response to training in OT muscle highlights the potential role of collagen in influencing passive muscle viscoelastic properties.

Aging↗

PubMed1996

The leucine zipper may induce electrophoretic mobility anomalies without DNA bending.

Numerous proteins bend DNA upon binding, a phenomenon of potential significance for regulation of gene expression and chromatin. DNA bending is commonly predicted from the presence of electrophoretic mobility anomalies in protein-DNA complexes. However, as compared with electrophoretic methods, several DNA binding oncoprotein families do not display comparable evidence of DNA bends in x-ray structural studies. Herein, circularization kinetics and affinity measurements with prebent DNA templates were employed to assess bending and DNA structural preferences for Max and other basic helix-loop-helix/leucine zipper proteins. In this way, proteins in the Myc/Max basic helix-loop-helix/ leucine zipper family were found not to bend DNA in solution but to actually stabilize DNA in an unbent configuration that resists circularization. The mobility anomaly was found to be induced by the leucine zipper protein motif, rather than structural distortions of DNA. Thus rigid protein domain structures may induce anomalous electrophoretic mobility. Moreover, the energetic preference of non-DNA bending proteins for unbent templates suggests mechanisms whereby chromatin structure may regulate transcription.

Binding Sites↗

PubMed1996

The semidominant Mi(b) mutation identifies a role for the HLH domain in DNA binding in addition to its role in protein dimerization.

The mouse microphthalmia (mi) locus encodes a basic helix-loop-helix-leucine zipper (bHLH-Zip) transcription factor called MITF (microphthalmia transcription factor). Mutations at mi affect the development of several different cell types, including melanocytes, mast cells, osteoclasts and pigmented epithelial cells of the eye. Here we describe the phenotypic and molecular characterization of the semidominant Microphthalmia(brwnish) (Mi(b)) mutation. We show that this mutation primarily affects melanocytes and produces retinal degeneration. The mutation is a G to A transition leading to a Gly244Glu substitution in helix 2 of the HLH dimerization domain. This location is surprising since other semidominant mi mutations characterized to date have been shown to affect DNA binding or transcriptional activation domains of MITF and act as dominant negatives, while mutations that affect MITF dimerization are inherited recessively. Gel retardation assays showed that while the mutant MITF(Mi-b) protein retains its dimerization potential, it is defective in its ability to bind DNA. Computer modeling suggested that the Gly244Glu mutation might disrupt DNA binding by interfering with productive docking of the protein dimer onto DNA. The Mi(b) mutation therefore appears to dissociate a DNA recognition function of the HLH domain from its role in protein dimerization.

Animals↗

PubMed1996

Collagen crosslinks in longissimus muscle from lambs expressing the callipyge gene.

The objective of this study was to determine whether increased amounts of collagen and (or) hydroxylysylpyridinoline crosslinks in longissimus muscle are partially responsible for decreased tenderness when callipyge lambs are compared to normal lambs. The longissimus muscle was used because we believe tenderness is a greater problem in loin chops of callipyge lambs than it is in other cuts. Ten normal and 10 half-sibling lambs expressing the callipyge gene were compared. Lambs were slaughtered at approximately 8 mo of age when they reached a target weight of 59 kg. Dressing percentages were higher for the leaner callipyge lambs that exhibited larger longissimus muscle areas and heavier leg weights (P < .01). Warner-Bratzler shear values for longissimus muscle from callipyge lambs were higher (P < .01) than those from normal lambs. The higher shear values were not explained by amount of muscle collagen or by hydroxylysylpyridinoline crosslink concentration because both collagen percentage (P < .09) and crosslink concentration (P < .04) were lower in callipyge lambs. Therefore, attempts to modify shear values and make loin chops from callipyge lambs more acceptable to consumers should focus on the myofibrillar fraction of muscle.

Amino Acids↗

PubMed1996

Partial characterization of ovine skeletal muscle proteoglycans and collagen.

Ovine longissimus dorsi and biceps femoris muscles were analyzed for proteoglycan content, collagen and lysine aldehyde-derived collagen crosslinking concentrations at 2-4 days, six-month-old, and six-year-old stages of development. Tissue extracted proteoglycan molecular sieve distribution on a Sephacryl S-200HR column revealed two proteoglycan populations with estimated relative molecular weight ranges of 200,000 to 250,000 daltons and 23,000 to 70,000 daltons. The molecular sieve distribution was similar between the two muscles within a developmental age, but changed as a function of developmental age. Primary culture from both the longissimus dorsi and biceps femoris muscle liberated proteoglycans into the culture medium. In contrast to the tissue extracted proteoglycans, at the six-year-old stage of development, culture medium liberated proteoglycan Sephacryl S-200HR molecular sieve distribution differed between the two muscles. In both the tissue extracted and medium liberated proteoglycans at all developmental stages, nitrous acid deamination demonstrated the presence of heparan sulfate. Immunoblot analysis of the tissue extracted proteoglycans indicated the presence of decorin at each developmental stage. Longissimus dorsi and biceps femoris collagen concentrations (5.13 +/- 0.9 vs. 5.53 +/- 1.5%, respectively) and crosslink concentrations (0.07 +/- 0.01 moles HP/mole collagen) were initially similar between the two muscles; however, by six-months the muscles differed in both collagen concentration (1.72 +/- 0.5 and 2.53 +/- 0.7%, respectively) and crosslinking (0.24 +/- 0.02 and 0.27 +/- 0.03 moles HP/mole collagen, respectively). At six years of age, both the longissimus dorsi and biceps femoris exhibited slightly elevated collagen concentrations (2.49 and 3.05%, respectively) while crosslinking values were decreased relative to values at six-months of age (0.11 +/- 0.01 and 0.18 +/- 0.01 moles HP/mole of collagen, respectively). The results from this study indicate that skeletal muscle proteoglycans and collagen show developmental changes, which suggests that they are subject to developmental regulation.

Animals↗

PubMed1996

The avian low score normal muscle weakness alters decorin expression and collagen crosslinking.

Extracellular matrix development of chicken pectoral muscle was examined in the Low Score Normal (LSN) genetic muscle weakness and compared to both normal and avian muscular dystrophy (MD). At 20 days of embryonic development significant elevations were noted in LSN total glycosaminoglycan concentration and decorin, while at 14 days, LSN glycosaminoglycan and decorin levels were indistinguishable from the controls. Levels of a large skeletal muscle chondroitin sulfate proteoglycan (M-CSPG) appear to be unaffected. Morphologically, at 20 days, the extracellular matrix space between muscle fibers increased to a level characteristic to that observed in avian muscular dystrophy. At six weeks posthatch a marked increase in LSN collagen crosslinking relative to MD or control tissues was observed, while collagen concentration was not altered. By one year posthatch LSN collagen crosslink levels did not significantly differ from normal tissue. These data support the concept that the LSN muscle weakness is associated with changes in both proteoglycan and collagen characteristics.

Animals↗

PubMed1995

Pre-bending of a promoter sequence enhances affinity for the TATA-binding factor.

TATA-binding protein (TBP) binds the minor groove of the TATA element with the DNA bent 80 degrees towards the major groove. A constrained minicircle strategy has been used to test the effect of DNA topology on the affinity of TBP for the TATA element. We report here that TBP bound to DNA which was slightly pre-bent towards the major groove with 100-fold higher affinity than unbent (linear) DNA of identical sequence and 300-fold higher affinity than DNA pre-bent towards the minor groove. Similar discrimination was observed with the holo-TFIID transcription complex. DNA topology, particularly bending, is determined by many factors including chromatin in cells and may, through changes in the affinity of the TATA factor, be important in the control of transcription.

DNA↗

PubMed1995

Collagen characteristics of skin, fell, and epimysium from rams, wethers, and zeranol-implanted ram lambs.

Eighteen spring-born Columbia ram, wether, and zeranol-implanted ram lambs were examined to determine the influence of castration or zeranol implantation on collagen characteristics of skin, fell, and epimysium and possible relationships between collagen properties of each tissue and difficulty of pelt removal. Pelt removal force was lower in wethers than in rams (P < .05) and intermediate for zeranol-implanted rams. Collagen concentration in skin of rams was greater (P < .05) than that in wethers or implanted rams, but percentage of heat-soluble collagen in skin was higher in implanted rams. Percentage of type III collagen in skin was highest in rams and lowest in wethers (P < .05); that in the skin of implanted rams was intermediate. The fell of wethers contained a higher (P < .05) collagen concentration, higher insoluble collagen amount, and lower percentage of soluble collagen than that of rams or implanted rams. The fell had a lower percentage of type III collagen than that of rams (P < .05). Epimysium collagen concentration of rams and wethers was higher (P < .05) than that of implanted rams; however, percentage of soluble collagen was higher for the implanted rams than for the other classes. Type III collagen percentage in the epimysium did not differ by animal class (P < .05). Zeranol-implanted ram lambs had a higher percentage of soluble collagen in all tissues examined than did non-implanted rams and force required for pelt removal was reduced in implanted rams.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Fibronectin self-association is mediated by complementary sites within the amino-terminal one-third of the molecule.

The formation of a fibrillar fibronectin (FN) extracellular matrix requires self-association of FN dimers. In this report, we show that the major sites for self-association are the amino-terminal repeats I1-5 and the first type III repeats. Recombinant FNs and fragments were generated by baculovirus expression of cysteine-rich domains and by bacterial expression of type III repeats as fusion proteins with maltose binding protein. When recombinant polypeptides were immobilized on microtiter wells, FN bound to 70-kDa amino-terminal fragment and to fusion proteins containing repeats III1-2 and III1-6 but not to other type III repeats. Similar results were obtained with a gel overlay assay. Binding was concentration-dependent and saturable. The amino-terminal binding site for III1-2 was further localized to repeats I1-5. Therefore, at least two different sites for FN-FN interaction reside near the amino terminus of the molecule. A model for the regulation of FN matrix assembly is proposed based on intramolecular interactions between these amino-terminal sites.

Amino Acid Sequence↗

PubMed1994

Regional differences in LV collagen accumulation and mature cross-linking after myocardial infarction in rats.

To determine the extent of and any regional differences in remodeling response of the extracellular matrix (ECM) to myocardial infarction (MI), moderate-to-large transmural infarcts were surgically produced in left ventricular (LV) free wall of rats. Animals were killed 13 wk after surgery. In comparison to age-matched controls, infarction was associated with an overall increase in heart weight, which included hypertrophy of both the right ventricle and LV. Although the remaining viable myocardium in LV free wall was significantly reduced, the interventricular septum was hypertrophied some 30% compared with control tissues (247 +/- 9 vs. 189 +/- 8 mg). Collagen concentration more than doubled in remaining viable free wall (8.92 +/- 0.59 vs. 3.95 +/- 0.25 mg/100 mg, P < 0.0001), and a smaller but still highly significant 27% increase occurred (P < 0.01) in the more remote septum. Degree of covalent cross-linking of collagen fibrils as assessed by hydroxylysylpyridinoline (HP) concentration also revealed regional differences in response of the ECM to infarction. Although HP concentration was increased 60% in viable free wall (P < 0.05) post-MI, it was unchanged in the septum. With respect to collagen characteristics of the transmural infarct per se, the scar exhibited still further increases in both collagen and HP concentrations compared with the already elevated values for these two parameters in viable free wall. The results indicate that any evaluation of the remodeling response of viable myocardium post-MI must include not only the myocyte but also the ECM, the principal component of which is collagen.

Amino Acids↗

PubMed1993

Copper deficiency alters collagen types and covalent cross-linking in swine myocardium and cardiac valves.

Dietary copper deficiency induces alterations of connective tissue metabolism that are associated with lesions in cardiovascular and other organ systems. To determine the impact of copper deficiency on characteristics of collagen in porcine myocardium and cardiac valves, weaned pigs were fed diets with adequate or deficient levels of copper. Although dietary copper did not affect the concentration of collagen in either myocardium or bicuspid valves, the degree of collagen cross-linking, as assessed by the level of hydroxylysylpyridinoline, was lower in both tissues of copper-deficient pigs. Proportions of type III collagen were increased in the left ventricle and bicuspid valves of copper-deficient pigs. Copper deficiency induced extensive remodeling, however, of the collagen fraction of cardiac interstitium. Reduction in left ventricular collagen cross-linking may provide the stimulus for the development of cardiac hypertrophy, which characterizes severe copper deficiency, by increasing the compliance of the ventricular wall. The shift in the phenotypic profile of collagen that is associated with this cardiac hypertrophy indicates synthesis of new collagen, which could affect collagen cross-linking irrespective of copper status.

Animals↗