Citations

Fredberg, J. J., K. A. Jones, M. Nathan, S. Raboudi, Y. S. Prakash, S. A. Shore, J. P. Butler, and G. C. Sieck. Friction in airway smooth muscle: mechanism, latch, and implications in asthma. J. Appl. Physiol. 81(6): 2703–2712, 1996.—In muscle, active force and stiffness reflect numbers of actin-myosin interactions and shortening velocity reflects their turnover rates, but the molecular basis of mechanical friction is somewhat less clear. To better characterize molecular mechanisms that govern mechanical friction, we measured the rate of mechanical energy dissipation and the rate of actomyosin ATP utilization simultaneously in activated canine airway smooth muscle subjected to small periodic stretches as occur in breathing. The amplitude of the frictional stress is proportional to ηE, where E is the tissue stiffness defined by the slope of the resulting force vs. displacement loop and η is the hysteresivity defined by the fatness of that loop. From contractile stimulus onset, the time course of frictional stress amplitude followed a biphasic pattern that tracked that of the rate of actomyosin ATP consumption. The time course of hysteresivity, however, followed a different biphasic pattern that tracked that of shortening velocity. Taken together with an analysis of mechanical energy storage and dissipation in the cross-bridge cycle, these results indicate, first, that like shortening velocity and the rate of actomyosin ATP utilization, mechanical friction in airway smooth muscle is also governed by the rate of cross-bridge cycling; second, that changes in cycling rate associated with conversion of rapidly cycling cross bridges to slowly cycling latch bridges can be assessed from changes of hysteresivity of the force vs. displacement loop; and third, that steady-state force maintenance (latch) is a low-friction contractile state. This last finding may account for the unique inability of asthmatic patients to reverse spontaneous airways obstruction with a deep inspiration.

Hypothyroidism (HYPO) often manifests as neuromuscular symptoms; however, little is known about its effects on the neuromuscular junction (NMJ). The present study examined changes in NMJ morphology and neuromuscular transmission failure (NTF) in the rat diaphragm muscle (Dimus) after 3 wk of HYPO. Three-color fluorescence immunocytochemistry and confocal microscopy were used to simultaneously visualize nerve terminals and axons, motor end plates, and myosin heavy chain isoform expression in Dimus fibers. NTF was assessed in vitro by comparing muscle fatigue induced by nerve with that induced by direct muscle stimulation. Diameters of axons innervating type I fibers were 30% smaller in the HYPO Dimus than in control (CTL). Planar areas of nerve terminals and end plates on type I and IIa fibers were 15-35% smaller in HYPO than in CTL. The extent of overlap between nerve terminals and end plates of type I fibers was 10% less in HYPO. Susceptibility to NTF during repetitive nerve stimulation was 20% greater in the CTL Dimus than in HYPO; however, changes in NMJ morphology could not fully account or the effect of HYPO on NTF.

We hypothesized that inactivity effects on diaphragm muscle contractile and morphometric properties are attenuated if phrenic motoneurons are also inactive. Three models of rat diaphragm inactivity were compared: 1) spinal isolation; 2) tetrodotoxin (TTX) nerve blockade; and 3) denervation (Dnv). Motoneuron and muscle fiber inactivities were matched only in spinal isolated animals. After 2 wk, maximum tetanic force decreased in all three groups compared with control group but to a greater extent in TTX and Dnv animals. Fatigue resistance improved, and maximum unloaded shortening velocity slowed only in TTX and Dnv groups. Type IIa fiber proportions decreased in all three groups, and type IIx fiber proportions increased in TTX and Dnv animals. Type I fiber cross-sectional area increased in all three groups but to a greater extent in TTX and Dnv animals. Type IIa fibers hypertrophied, whereas type IIx and IIb fibers atrophied only in TTX and Dnv groups. These results support the hypothesis that muscle adaptations to prolonged inactivity are attenuated when muscle fiber and motoneuron inactivities are matched.

In the rat diaphragm muscle, the histochemical classification of type I, IIa, IIb, or IIx fibers was correlated with myosin heavy chain (MHC) immunoreactivity. Expression of MHC isoforms in single dissected fibers was also assessed electrophoretically. Most fibers (approximately 86%) expressed a single MHC isoform, and when present, coexpression of MHC-2X and MHC-2B isoforms was most prevalent. Type I and IIa fibers were the smallest, type IIb fibers were the largest, and type IIx fibers were intermediate. Succinate dehydrogenase (SDH) and calcium-activated myosin adenosinetriphosphatase (actomyosin ATPase) activities were measured with quantitative histochemical procedures. Type I and IIa fibers had the highest SDH activities, followed in rank order by type IIx and IIb fibers. Type I fibers had the lowest actomyosin ATPase activity, followed in rank order by type IIa, IIx, and IIb fibers. Across all fibers, there was an inverse relationship between fiber SDH activity and cross-sectional area and a positive correlation between fiber actomyosin ATPase activity and cross-sectional area. The SDH and actomyosin ATPase activities of muscle fibers were also inversely correlated. These phenotypic differences in SDH and ATPase activities may be important in determining the contractile and fatigue properties of different fiber types in the rat diaphragm muscle.

The acute period of unilateral diaphragm denervation (DNV) is associated with increases in cell mitotic activity, protein synthesis, and muscle fiber hypertrophy. Our purpose was to determine whether acute unilateral diaphragm DNV is associated with changes in muscle isometric contractile properties, cross-sectional area of different muscle fiber types, mitotic activity of muscle fiber satellite cells, and muscle fiber ultrastructural properties indicative of injury. Adult male Fischer 344 rats underwent a right phrenicotomy, and DNV and intact (INT) hemidiaphragms were studied 72 h later. DNV hemidiaphragm displayed a significant decline in maximal isometric force (8.7 vs. 24.3 N/cm2) and a prolonged time to peak twitch force (47.8 vs. 37.5 ms) and time to half relaxation (72.3 vs. 44.3 ms) compared with INT contralateral hemidiaphragm (P < 0.05). DNV resulted in a significant increase in cross-sectional area of types I (33%), IIa (35%), and IIb (28%) fibers relative to INT hemidiaphragm (P < 0.05). Satellite cell mitotic activity (assessed by incorporation of bromodeoxyuridine) was approximately 5.5 times greater in DNV than in INT muscle (DNV 25.0 +/- 3.8, INT 4.5 +/- 1.4 labeled satellite cell nuclei/1,000 nuclei; P < 0.05). Ultrastructural examination of electron micrographs revealed alterations in Z-line and sarcomeric structure indicative of muscle injury. Cellular infiltration and segmental necrosis were also noted in some fibers. We conclude that acute unilateral diaphragm DNV results in muscle fiber injury that induces satellite cell activation. We also speculate that the specific force decrement associated with DNV is at least partially the result of muscle injury.

The effects of maternal nutritional deprivation (ND) on the proportions and cross-sectional areas (CSAs) of type I and II fibers in the developing rat diaphragm (Dia) were examined. In addition, the effects of ND on in vitro contractile and fatigue properties were also studied. Compared with in the control (CTL) group (from pups of well-nourished mothers), proportions of type II fibers were significantly lower in the ND group during the first 3 postnatal wk. However, this difference did not persist into adulthood. CSAs of type I and II fibers were larger in ND Dia than in CTL Dia during early postnatal development, but in adults type I and II fiber CSAs were larger in CTL than in ND Dia. Interstitial space comprised a smaller fraction of total muscle area in ND Dia throughout postnatal development and persisted in adults. Estimated fiber density (no. of fibers/CSA muscle) was lower in ND groups at all ages. In addition, the total number of fibers was significantly lower in ND Dia at all ages. Optimal muscle fiber length for peak twitch and maximum isometric force generation was shorter in ND Dia at all ages. At postnatal days 7 and 14, specific force (maximum tetanic force in N/cm2) of ND Dia was significantly lower than that of CTL Dia. At day 21, Dia specific force was comparable between the two groups but was significantly lower in adult ND compared with adult CTL Dia. Except for days 0 and 14, ND Dia was more fatigue resistant than CTL Dia.(ABSTRACT TRUNCATED AT 250 WORDS)

Emberger syndrome with underlying guanine-adenine-thymine-adenine 2 (GATA2) mutation is a rare disorder and very few successful nonmyeloablative allogeneic hematopoietic stem cell transplants (HSCTs) have been reported. We report a case of Emberger syndrome with GATA2 mutation in a 9-year-old girl who presented with congenital sensorineural deafness, warts, lymphedema, and Myelodysplastic syndrome. Her sister had died of a similar illness. She underwent a nonmyeloablative matched related donor peripheral blood HSCT with rabbit antithymoglobulin (5 mg/kg), fludarabine (160 mg/m2), cyclophophamide (29 mg/kg), and total body irradiation (2 Gray). Graft versus host disease prophylaxis consisted of tacrolimus and mycophenolate moefetil. She had neutrophil engraftment on day+15 and fully donor chimerism by day+30. She developed limited chronic skin graft versus host disease on tapering off immunosuppression. She is disease free on day+475. The review of literature showed a total of 28 patients with GATA2 mutation have undergone HSCT mostly nonmyeloablative and overall survival is 75%. Nonmyeloablatove HSCT is feasible and safe for the patients with GATA2 mutation.