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N L Stephens

Publications and source records attributed to N L Stephens.

At least 55 records · Page 3Linked to original sources
PubMed1992

The biophysics and biochemistry of smooth muscle contraction.

In this review the biophysics and biochemistry of smooth muscle contraction are dealt with. We describe a new model for the study of bronchial smooth muscle, which facilitates study of cellular contractile mechanisms. A new concept emerging is that study of steady-state mechanical parameters such as maximal isometric force (Po) velocity is inadequate because two types of crossbridges (normally cycling (NBR) and latch) seem to be sequentially active during smooth muscle contraction. Thus quick-release techniques are required to characterize the force-velocity properties of the two types of bridges. Pathophysiological processes that affect the muscle's shortening ability seem to affect the early NBRs only. With respect to maximal shortening capacity of the smooth muscle, the role of loading is very important. The differences between isotonic, elastic, and viscous loading are considerable. Ultimately, the time course and magnitude of loading should exactly resemble that operative in vivo. Once again, it is the characteristic of loading in the early phase of contraction that is crucial, as most of the shortening in smooth muscle occurs early in the contraction. While the maximum force developed by smooth muscle per unit cross-sectional area is the same as for striated muscle, the velocity is 50 times less. The properties of the series and parallel elastic elements of smooth muscle are described. The latter, when in compression mode, acts as an internal resistance to shortening and probably limits it. Isotonic relaxation has therefore not been studied in smooth muscle. We have developed a shortening parameter that is independent of the load on the muscle and of the initial length of the muscle's contractile element. We report the novel observation that isotonically relaxing smooth muscle reactivates itself, resulting in terminal slowing of the relaxation process. With respect to the biochemistry of smooth muscle contraction, contractile (actin isoforms, myosin heavy and light chains and their isoforms), regulatory (calmodulin-4 Ca2+, myosin light chain kinase, myosin light chain and its phosphorylation, tropomyosin, caldesmon, and calponin), and cytoskeletal (chiefly desmin and vimentin) proteins are discussed. While the kinase activates the contractile system, caldesmon and calponin modulate the activity downward. The cytoskeletal proteins desmin, vimentin, and alpha-actinin could constitute the muscle cell's internal resistor.

Animals↗
PubMed1992

Normalization of contractile parameters in canine airway smooth muscle: morphological and biochemical.

Asthma research has recently highlighted the importance of correctly normalizing force development for purposes of comparing stiffness properties of smooth muscle between different airways, between airways at different stages of maturity, and between airways from different animal species. This problem does not exist in striated muscle where the entire tissue consists almost entirely of muscle and where cross bridges cycle at the same rate throughout a contraction when load correlation is made. In the bronchus, cross-sectional area of true muscle may constitute only 20-30% of the total tissue cross section, and load-independent cycling rate varies fourfold during the course of a contraction because of the occurrence of normally cycling and latch bridges. These features are responsible for the difficulty in force normalization in smooth muscle. Our studies indicate that normalization with respect to true muscle cell cross-sectional area (derived by quantitative morphometry of appropriate tissue transverse sections) is the most valid. This is only so, however, when it has been proved that the actomyosin content per unit weight of the different muscle tissues is the same.

Animals↗
PubMed1992

Pulmonary and systemic vascular smooth muscle mechanical characteristics in newborn sheep.

To investigate the hypothesis that the higher pulmonary vascular resistance in newborn sheep is the result of developmental differences in the vascular muscle mechanical properties, we evaluated pulmonary arteries from newborn and adult sheep and compared them with their respective systemic counterparts (common carotid arteries). The newborn pulmonary artery mechanical stress (13.0 +/- 1.4 mN/mm2) and shortening capacity (11.4 +/- 1.1% of optimal length) were lower (P less than 0.01) than in the adult (20.4 +/- 2.5 and 15.6 +/- 1.3, respectively). The adult carotid artery muscle developed a greater stress (97.6 +/- 18.5 mN/mm2) than the newborn (40.7 +/- 5.0; P less than 0.01), whereas no age differences in shortening capacity were observed (newborn = 19.4 +/- 1.7; adult = 18.4 +/- 1.5% of optimal length). The contraction half-time was similar for the pulmonary and carotid arteries and was not affected by age, whereas the relaxation half-times of the newborn pulmonary (30.7 +/- 2.9 s) and carotid artery (23.3 +/- 1.5) were greater than in the adult (24.9 +/- 2.9 and 14.6 +/- 1.4, respectively; P less than 0.01). The myosin contents of the pulmonary and carotid arteries, as an indicator of the tissue muscle mass, were similar and did not change with age. In conclusion, while the lower stress and shortening capacity of the newborn pulmonary arteries limit their maximum capacity to vasoconstrict, the significantly greater relaxation time of their vascular muscle, a new observation, may account for the higher resistance to blood flow after birth.

Aging↗
PubMed1992

Isotonic relaxation of sensitized bronchial smooth muscle.

Although the mechanisms underlying asthma are not clear, we have demonstrated that in dogs antigen sensitization results in alterations of contractile properties, such as greater early maximum shortening velocity and capacity of bronchial smooth muscle. These changes could account for the hyperresponsiveness reported in asthma. The failure of the muscle to relax could be another important factor responsible for maintaining high airway resistance. This, a relatively less-studied area, requires study. In the current study, we therefore developed an index of isotonic relaxation, T1/2,CE (half-time for relaxation which is independent of muscle load and initial contractile element length) for evaluation of the relaxation process. Because the maximum shortening velocity at 2 s but not at 10 s was greater in sensitized bronchial smooth muscle (SBSM) than that in controls studies of relaxation were also undertaken at these two times. The mean half-relaxation time indicated by T1/2,CE showed no difference between sensitized and control muscles after 10 s of stimulation (8.38 +/- 0.92 s vs. 7.78 +/- 0.93, means +/- SE); however, it was prolonged significantly in SBSM stimulated only for 1 s (12.74 +/- 2.5 s, mean +/- SE) compared with the control (6.98 +/- 1.01). Unexpectedly, we found that during the late phase of isotonic relaxation, both groups showed spontaneous increase in zero load shortening velocity, which is an index of cross-bridge cycling rate.2

Animals↗
PubMed1992

Bronchial smooth muscle mechanics of a canine model of allergic airway hyperresponsiveness.

Although we have reported that tracheal smooth muscle from sensitized dogs shows altered mechanical properties, we did not know, because of technical difficulties with the preparation, whether similar changes occur in the properties of sensitized central bronchial smooth muscle (BSM), the site at which the acute asthmatic response is believed to develop. We have now succeeded in developing a cartilage-free BSM preparation that retains optimal mechanical properties. Such strips were obtained from mongrel dogs that had been sensitized to ragweed pollen. Controls were littermates injected with adjuvant alone. Length-tension relationships were obtained for both control and sensitized BSM strips (CBSM and SBSM, respectively). The maximal active stresses were the same (P greater than 0.05) when normalized to muscle fraction in total tissue cross-sectional area [6.2 +/- 0.6 x 10(4) and 5.9 +/- 0.6 x 10(4) (SE) for SBSM and CBSM, respectively]. This suggests that optimal tension is an insensitive indicator of bronchial hyperresponsiveness and that isotonic studies might be more revealing. The maximal shortening velocity (Vo) for SBSM at 2 s [0.35 +/- 0.017 (SE) lo/s, where lo signifies optimal muscle length], in the course of a 10-s contraction, was significantly greater (P less than 0.05) than Vo measured for CBSM (0.27 +/- 0.015 lo/s). However, Vo did not differ at the 8-s point of contraction. The sensitized group demonstrated a statistically significantly greater maximal shortening capacity (0.67 +/- 0.04 lo) than the control group (0.51 +/- 0.04 lo). At 2 s of contraction, 80% of maximal SBSM shortening had been completed and was significantly greater than for CBSM.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Ragweed sensitization-induced increase of myosin light chain kinase content in canine airway smooth muscle.

Previous studies have identified changes of mechanical properties of airway smooth muscle (ASM) from a canine model of atopic airway hyperreactivity. These changes, including increased maximum shortening capacity (delta Lmax) and early shortening velocity (Vo), may be responsible for the airway hyperresponsiveness in asthma. We have suggested that these changes may be due to increased actomyosin ATPase activity, controlled via phosphorylation of the 20 kD myosin light chain (MLC20) by MLC kinase (MLCK). Therefore, ATPase activity, MLC20 phosphorylation, and MLCK content and activity were assessed in tracheal and bronchial smooth muscles (TSM and BSM) of ragweed pollen-sensitized dogs (S) and their littermate controls (C). Specific ATPase activities from STSM and SBSM were significantly higher than their control counterparts (CTSM, CBSM). Phosphorylation of MLC20 in STSM was greater both at rest and during electrical stimulation due to the increased amount of MLCK in STSM and SBSM by 30 and 25%, respectively. MLCK activity was also increased significantly in STSM and SBSM (from 46.99 +/- 8.33 and 42.85 +/- 5.92 to 91.9 +/- 6.43 and 64.12 +/- 7.88 32P mmol/mg fresh tissue weight/min respectively [mean +/- SEM]). When normalized to the amount of MLCK in the tissue, however, specific MLCK activity in STSM and SBSM was similar to that in controls. It is unlikely that myosin phosphatase plays any role in the changes of MLC20 phosphorylation in sensitized animals. Peptide mapping showed no visible change in primary structure of MLCK in STSM and SBSM compared with those of controls. We report that ASM actomyosin ATPase activity is increased in STSM and SBSM. The increased ATPase activity is the result of increased MLC20 phosphorylation, the latter likely resulting from the increased MLCK content, which may account for the hyperresponsiveness found in ASM from these animals.

Allergens↗
PubMed1992

Exercise blood pressure and echocardiographic abnormalities in genetically comparable populations.

OBJECTIVE: To determine the relationship between the systolic blood pressure response to exercise and the echocardiographic dimensions of the left atrium and ventricle in two geographically separate, but genetically comparable, populations. DESIGN AND SETTING: The study was cross-sectional. The settings were two semirural communities, one in Iceland and one in Manitoba. SUBJECTS: Individuals from families where there had been no intermarriage with non-Icelandic individuals were eligible. Of the 200 eligible men in Manitoba and the 150 men in Iceland, 157 and 121 men, respectively, agreed to participate. In the majority of cases, those who chose not to participate stated that work commitments prevented them from taking part. The subjects were aged 25 to 63 years and had blood pressure less than 160/95 mmHg at rest. INTERVENTIONS: Blood pressure was taken at rest and during standardized bicycle ergometry. Left ventricular and atrial echocardiography was performed on a selected number of subjects. MAIN RESULTS: The Canadians of 'pure' Icelandic descent had a higher prevalence of exaggerated exercise systolic blood pressure (ESBP), left atrial enlargement (LAE) and left ventricular hypertrophy (LVH) than native Icelanders. Given their genetic similarity, it is suggested that the difference between the two groups is due to environmental factors. Within each group, subjects with exaggerated ESBP had a significantly greater left atrial dimension index and left ventricular mass index than subjects without an exaggerated ESBP. The relationship of ESBP with left atrial dimension index and left ventricular mass index was independent of age, body mass index and resting blood pressure. LVH was of the eccentric type and was absent in the majority of cases with LAE; however, LAE was present in nearly all LVH cases. CONCLUSIONS: This study demonstrates that an exaggerated ESBP in individuals with resting blood pressure less than 160/95 mmHg is not an innocuous finding it is associated with demonstrable cardiac abnormalities (LAE and LVH). Furthermore, it is proposed that the presence of LAE, with or without LVH, may add to the value of an exaggerated ESBP in identifying individuals at increased risk of developing sustained resting essential hypertension.

Adult↗
PubMed1991

Myosin heavy chain isoform distribution in normal and hypertrophied rat aortic smooth muscle.

Causes of hypertension have been well scrutinized, whereas the secondary, disabling effects of high blood pressure are less well investigated. We have used a rat model of hypertension and developed a technique to study the secondary vascular smooth muscle component of the disorder. Banding patterns of myosin heavy chain isoforms from rat aortae were examined using denaturing electrophoresis, Western blotting, immunochemical identification, and degradation studies. Myofibrillar ATPase activities were also measured. Left ventricular hypertrophy and hypertension were induced in rats by aortic banding just proximal to the renal artery. Aortic banding increased the heart weight/body weight (mg/g) ratio from 2.8 to 3.8 and mean aortic weight by 53%. Two distinct myosin heavy chain isoforms, molecular masses of 204 and 200 kDa, were identified by 4% sodium dodecyl sulphate-polyacrylamide electrophoresis of crude aortic extracts from normal rats in a relative molar ratio of 1.54:1. The development of significant thickening of the aorta was marked by substantial increases in aortic wall smooth muscle content but was not associated with any changes in distribution of the isoforms. The band patterns obtained on gel electrophoresis were not the result of contamination by other proteins, as Western blotting studies with specific antibodies demonstrated that the isoforms were smooth muscle in origin and were not derived from nonmuscle myosin sources. Myofibrillar ATPase activity of aortic smooth muscle from hypertensive rats was increased. It is suggested that this increase may be the result of post-transcriptional alterations of one or more sarcomeric proteins involved in the regulation of smooth muscle contraction.

Adenosine Triphosphatases↗
PubMed1991

Effect of maturational changes in myosin content and morphometry on airway smooth muscle contraction.

We studied the relationship of airway morphometry, the content of myosin heavy-chain and isoform stoichiometry, and the distribution of bronchoconstrictor responses in the airways of maturing swine. Lungs were excised in 2-wk-old farm swine (2ws; n = 13) and 10-wk-old swine (10ws; n = 13), and tracheal smooth muscle strips and bronchial rings from generations 2-5 were fixed for in vitro isometric measurement of force generation. Split samples were placed in formaldehyde solution or glutaraldehyde for light- or electron-microscopic morphometry or frozen for analysis of tissue myosin content. The rank order of force generation elicited by both receptor- and nonreceptor-dependent mechanisms for both 2ws and 10ws was generation 4 greater than 3 greater than or equal to 2. For all matched airway generations, contractile force was 25-100% greater in 2ws than 10ws. Differences in force generation were not related to morphometric differences in smooth muscle mass content among airways. The relative cross-sectional area of smooth muscle derived by computerized morphometry was 5.5-7% for each airway generation and did not change with age. Electron-microscopic morphometry demonstrated comparable myocyte content within muscle bundles for all airways in both age groups. In generation 4 airways, myocyte size in 2ws (27.3 +/- 0.8 nuclei/2,500 microns2) hypertrophied approximately 15% in 10ws (20.4 +/- 0.6 nuclei/2,500 microns2; P less than 0.01). Tissue content of myosin measured by computerized laser densitometry of gel electrophoresis of homogenates was greater in trachea from 2ws than 10ws (135 +/- 10 vs. 90 +/- 4 micrograms/g tissue; P less than 0.01); homology of 200- and 205-kDa isoforms was confirmed by Western blot against polyclonal myosin antibody and Cleveland digest analysis of each band. Differences in contractile forces between generations in 2ws and 10ws were not correlated to functional myosin isoform content. We demonstrate a maturational downregulation of contractile forces in maturing swine. This response is independent of smooth muscle receptor distribution and is not related to morphological changes in airways muscle mass, cellularity, changes in content of nonmyocyte tissues, or tissue content of functional myosin isoform.

Aging↗
PubMed1991

Normalization of force generated by canine airway smooth muscles.

A variety of normalizations have been employed to compare maximal isometric force (Po) produced by smooth muscles at different locations and stages of maturation. Because these procedures have not always been based on rigorous principles, confusion has resulted. To obtain a less ambiguous index of force production, we measured in vitro Po from mongrel canine tracheal (TSM) and bronchial (BSM) smooth muscle with an electromagnetic lever and normalized it to force per unit cross-sectional area of whole tissue (tissue stress), to force per unit cross-sectional area of muscle in the cross section of total tissue (muscle stress), and to force per fractional unit of myosin in the tissue cross section (myosin stress). Proportion of myosin in cross-sectional area of tissue was deduced from data obtained by sodium dodecyl sulfate gel electrophoresis of crude muscle extracts. For TSM, tissue stress was 1.499 X 10(5) N/m2 +/- 0.1 (SE), whereas it was only 0.351 X 10(5) N/m2 +/- 0.05 (SE) for BSM, representing a 4.27-fold difference (P less than 0.01). There was a 1.60-fold difference (P less than 0.05) in muscle stress, which was correlated to the morphometric finding that 79 +/- 1.4% (SE) of the tracheal strip cross section was muscle, whereas only 30 +/- 1.0% (SE) of bronchial tissue was occupied by muscle. Average myosin content was the same in smooth muscle cells of TSM and BSM, indicating that total amount of myosin in tissue cross sections was essentially a function of proportional area of muscle cells in total tissue cross sections.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Changes in Schultz-Dale reaction in sensitized canine tracheal smooth muscle.

Tracheal smooth muscles from adult dogs 17 to 20 months of age sensitized with ragweed pollen demonstrated a Schultz-Dale phenomenon in response to specific antigen challenge. Seventy percent of the sensitized tracheal smooth muscles developed a Schultz-Dale reaction that consisted only of a phasic response, and the remaining 30% developed a Schultz-Dale reaction that consisted of a phasic component followed by a discrete tonic component. All the Schultz-Dale reactions were mediated only by histamine. The triggering of presynaptic acetylcholine release by histamine during the Schultz-Dale reaction from tracheal smooth muscles of ragweed-pollen-sensitized puppies 6 to 8 months of age was not detected in sensitized adult dogs. Hyperresponsiveness to acetylcholine was detected in tissues from sensitized puppies but not from sensitized adult dogs. Maximal active tension obtained from the sensitized adult canine trachealis during the Schultz-Dale reaction was lower than that obtained from trachealis from sensitized puppies. Dose-response studies showed that sensitized tissues used in the present studies were hyperresponsive to histamine when compared with their nonsensitized control littermates. These results suggest that the nature of the Schultz-Dale response and the identity of the transmitters is age-dependent.

Acetylcholine↗