ABSTRACT
Stiff knee gait (SKG) occurrence after a stroke is associated with various abnormal muscle activities; however, the interactions among these muscles are unclear. This study aimed to elucidate the muscle synergy characteristics during walking in patients with SKG after a stroke. This cross-sectional study included 20 patients with poststroke SKG (SKG group), 16 patients without poststroke SKG (non-SKG group), and 15 healthy adults (control group). Participants walked a 10-m distance at a comfortable speed, and electromyographic data were recorded from six lower-limb muscles. Non-negative matrix factorization was employed to derive time-varying activity (C), muscle weights (W), and the percentage of total variance accounted for (tVAF) for muscle synergies. The SKG group showed a higher tVAF than the control group. The initial stance module (including knee extensors) showed increased activity during the swing phase. The initial swing module (including hip flexors and ankle dorsiflexors) exhibited a higher activity during the single-support phase but a lower activity during the swing phase. The synergy structure in patients with SKG after stroke was simplified, with specific abnormalities in synergy activities. SKG may arise from several synergy alterations involving multiple muscles, indicating that approaches focused on controlling individual muscle activities are unsuitable.
Subject(s)
Electromyography , Gait , Knee Joint , Muscle, Skeletal , Stroke , Humans , Male , Stroke/physiopathology , Stroke/complications , Female , Middle Aged , Muscle, Skeletal/physiopathology , Gait/physiology , Cross-Sectional Studies , Knee Joint/physiopathology , Aged , Walking/physiology , Adult , Biomechanical Phenomena , Gait Disorders, Neurologic/physiopathology , Gait Disorders, Neurologic/etiology , Knee/physiopathologyABSTRACT
OBJECTIVES: Few studies have examined motor function in determining the suitability of patients with stroke to resume driving a car. Patients with hemiplegia usually control car pedals with the unaffected lower limb. However, motor control on the unaffected side is also impaired in patients with stroke. This study aimed to clarify the neurophysiological characteristics of pedal switching control during emergency braking in patients with hemiplegia. METHODS: The study participants consisted of 10 drivers with left hemiplegia and 10 age-matched healthy drivers. An experimental pedal was used to measure muscle activity and kinematic data during braking, triggered by the light from a light-emitting diode placed in front of the drivers. RESULTS: The patient group took the same reaction time as the healthy group. However, from the visual stimulus to the release of the accelerator pedal, the patient group had higher muscle activity in the tibialis anterior and rectus femoris and had faster angular velocities of hip and knee flexion than the healthy group. In addition, the patient group had higher co-contraction activities between flexors and extensors. From the accelerator pedal release to brake contact, the patient group had slower angular velocities of hip adduction, internal rotation, ankle dorsiflexion, internal return, and internal rotation than the healthy group. CONCLUSIONS: Patients with hemiplegia exhibited poor control of pedal switching using their unaffected side throughout the pedal-switching task. These results indicate that the safety related to car-pedal control should be carefully evaluated while deciding whether a patient can resume driving a car after a stroke.
Subject(s)
Automobile Driving , Stroke , Humans , Hemiplegia/etiology , Automobiles , Accidents, Traffic , Muscle, Skeletal , Lower Extremity , Stroke/complicationsABSTRACT
BACKGROUND: The prevalence of low back pain (LBP) and its associated risk factors, including physical function, are not fully clarified among health, medical, and welfare facility professionals and workers. Furthermore, issues related to the prevention of LBP are not completely understood. OBJECTIVE: To investigate the prevalence of LBP and associated risk factors, including physical function, in professionals and workers in health, medical, and welfare facilities. DESIGN: Cross-sectional study. SETTING: Physical examinations for LBP were conducted, and data were collected at health, medical, and welfare facilities in Japan. PARTICIPANTS: A total of 1085 health, medical, and welfare facility professionals and workers participated in the study. MAIN OUTCOME MEASURES: Prevalence was calculated based on the presence or absence of symptoms of LBP at the time of survey. Then, bivariate analysis was conducted to examine the relationship between LBP and age, gender, occupation, history of treatment for LBP, passive straight leg raising (SLR) angle, and abdominal muscle strength. Furthermore, multivariate analysis was performed to examine the relationship between LBP and the six risk factors. RESULTS: The prevalence of LBP was high (74.8%). Bivariate analysis showed a significantly higher percentage of nursing professionals (46.2%) in the LBP group (p < .001). The association between LBP and the risk factors in multivariate analysis was as follows: history of treatment for LBP (odds ratio [OR] = 3.92, 95% confidence interval [CI] 2.63-5.84, p < .001), nursing professionals as occupation (OR = 2.12, 95% CI 1.55-2.90, p < .001), passive SLR angle (OR = 2.06, 95% CI 1.24-3.42, p = .005), abdominal muscle strength (OR = 1.61, 95% CI 1.07-2.41, p = .021), and gender (OR = 1.52, 95% CI 1.08-2.14, p = .015). CONCLUSION: In the future, it will be necessary to take preventive measures for risk factors and to follow up longitudinally on the prevalence of LBP and on workers with indicated risk factors.
Subject(s)
Low Back Pain , Occupational Diseases , Humans , Low Back Pain/epidemiology , Low Back Pain/etiology , Japan/epidemiology , Prevalence , Cross-Sectional Studies , Occupational Diseases/epidemiology , Occupational Diseases/etiology , Risk Factors , Surveys and Questionnaires , OccupationsABSTRACT
Age-related decline in lower limb motor control may cause errors in pedal operation when driving a car. This study aimed to clarify the kinematics and electrophysiological characteristics of the pedal-switching operation associated with emergency braking in the case of elderly drivers. The participants in this study consisted of 11 young drivers and 10 elderly drivers. An experimental pedal was used, and the muscle activity and kinematic data during braking action were analyzed using the light from a light-emitting diode installed in the front as a trigger. The results showed that elderly drivers took the same time from viewing the visual stimulus to releasing the accelerator pedal as younger drivers, but took longer to switch to the brake pedal. The elderly drivers had higher soleus muscle activity throughout the process, from accelerator release to brake contact; furthermore, the rectus femoris activity was delayed, and the simultaneous activity between the rectus femoris and biceps femoris was low. Furthermore, elderly drivers tended to have low hip adduction velocity and tended to switch pedals by hip internal rotation. Thus, the alteration in joint movements and muscle activity of elderly drivers can reduce their pedal operability and may be related to the occurrence of pedal errors.
ABSTRACT
OBJECTIVE: In patients with hemiplegia, botulinum toxin type A injection for ankle spasticity of the plantar flexors reportedly improves walking speed. This improvement may be affected by background factors and patient baseline physical performance. This study aimed to clarify the factors affecting gait velocity improvement after botulinum toxin type A injection. METHODS: Background and evaluation data were collected for 60 patients with stroke who received botulinum toxin type A injection for spasticity of the plantar flexors. The patients were divided into improvement (n=27) and non-improvement (n=33) groups based on the gait velocity change from before injection to 2 weeks after injection. Logistic regression analysis was performed with the improvement and non-improvement groups as response variables and background data and evaluation data at baseline as explanatory variables. RESULTS: The presence or absence of physical therapy following botulinum toxin type A injection (odds ratio: 7.82) was the only significant explanatory variable for gait velocity change. CONCLUSION: Background factors and physical performance at baseline did not affect gait velocity improvement after botulinum toxin type A injection. If botulinum treatment of the ankle plantar flexors in patients with stroke is targeted at walking performance improvement, then physical therapy following botulinum toxin type A injection should be an essential part of the treatment strategy.
ABSTRACT
BACKGROUND AND PURPOSE: Stiff-knee gait, which is a gait abnormality observed after stroke, is characterized by decreased knee flexion angles during the swing phase, and it contributes to a decline in gait ability. This study aimed to identify the immediate effects of pedaling exercises on stiff-knee gait from a kinesiophysiological perspective. METHODS: Twenty-one patients with chronic post-stroke hemiparesis and stiff-knee gait were randomly assigned to a pedaling group and a walking group. An ergometer was set at a load of 5 Nm and rotation speed of 40 rpm, and gait was performed at a comfortable speed; both the groups performed the intervention for 10 min. Kinematic and electromyographical data while walking on flat surfaces were immediately measured before and after the intervention. RESULTS: In the pedaling group, activity of the rectus femoris significantly decreased from the pre-swing phase to the early swing phase during gait after the intervention. Flexion angles and flexion angular velocities of the knee and hip joints significantly increased during the same period. The pedaling group showed increased step length on the paralyzed side and gait velocity. CONCLUSIONS: Pedaling increases knee flexion during the swing phase in hemiparetic patients with stiff-knee gait and improves gait ability.
Subject(s)
Accidental Falls/prevention & control , Bicycling , Exercise Therapy , Gait Disorders, Neurologic/rehabilitation , Gait , Knee Joint/physiopathology , Paresis/rehabilitation , Stroke Rehabilitation/methods , Stroke/therapy , Adult , Aged , Biomechanical Phenomena , Female , Gait Analysis , Gait Disorders, Neurologic/diagnosis , Gait Disorders, Neurologic/etiology , Gait Disorders, Neurologic/physiopathology , Humans , Japan , Male , Middle Aged , Paresis/diagnosis , Paresis/etiology , Paresis/physiopathology , Pilot Projects , Range of Motion, Articular , Recovery of Function , Single-Blind Method , Stroke/complications , Stroke/diagnosis , Stroke/physiopathology , Time Factors , Treatment OutcomeABSTRACT
BACKGROUND: Administration of botulinum neurotoxin A (BoNT-A) to the ankle plantar flexors in patients with hemiplegia reduces the strength of knee extension, which may decrease their walking ability. Studies have reported improvements in walking ability with physical therapy following BoNT-A administration. However, no previous studies have evaluated from an exercise physiology perspective the efficacy of physical therapy after BoNT-A administration for adult patients with hemiplegia. AIM: To investigate the effects of physical therapy following BoNT-A administration on gait electromyography for patients with hemiparesis secondary to stroke. DESIGN: Non-randomized controlled trial. SETTING: Single center. POPULATION: Thirty-five patients with chronic stroke with spasticity were assigned to BoNT-A monotherapy (N.=18) or BoNT-A plus physical therapy (PT) (N.=17). METHODS: On the paralyzed side of the body, 300 single doses of BoNT-A were administered intramuscularly to the ankle plantar flexors. Physical therapy was performed for 2 weeks, starting from the day after administration. Gait electromyography was performed and gait parameters were measured immediately before and 2 weeks after BoNT-A administration. Relative muscle activity, coactivation indices, and walking time/distance were calculated for each phase. RESULTS: For patients who received BoNT-A monotherapy, soleus activity during the loading response decreased 2 weeks after the intervention (P<0.01). For those who received BoNT-A+PT, biceps femoris activity and knee coactivation index during the loading response and tibialis anterior activity during the pre-swing phases increased, whereas soleus and rectus femoris activities during the swing phase decreased 2 weeks after the intervention (P<0.05). These rates of change were significantly greater than those for patients who received BoNT-A monotherapy (P<0.05). CONCLUSIONS: Following BoNT-A monotherapy, soleus activity during the stance phase decreased and walking ability either remained unchanged or deteriorated. Following BoNT-A+PT, muscle activity and knee joint stability increased during the stance phase, and abnormal muscle activity during the swing phase was suppressed. CLINICAL REHABILITATION IMPACT: If botulinum treatment of the ankle plantar flexors in stroke patients is targeted to those with low knee extension strength, or if it aims to improve leg swing on the paralyzed side of the body, then physical therapy following BoNT-A administration could be an essential part of the treatment strategy.
Subject(s)
Botulinum Toxins, Type A/therapeutic use , Gait/physiology , Muscle, Skeletal/physiopathology , Neuromuscular Agents/therapeutic use , Paresis/physiopathology , Physical Therapy Modalities , Aged , Female , Humans , Male , Middle Aged , Muscle Spasticity/drug therapy , Muscle Spasticity/etiology , Muscle Spasticity/physiopathology , Paresis/therapyABSTRACT
[Purpose] The aim of this study was to clarify the optimal timing for increasing muscle activity in the paralyzed lower limb of stroke survivors by evaluating the relationship between gait muscle activity patterns and gait parameters. [Participants and Methods] Electromyography of the tibialis anterior, soleus, rectus femoris, and biceps femoris on the paralyzed side and spatiotemporal gait parameters were evaluated in 40 chronic post-stroke patients as they walked at a comfortable speed. The normalized average amplitude and asymmetry indexes of each gait phase were calculated. The correlations between gait velocity or asymmetry indexes and the activity amplitudes of various muscles during each gait phase were analyzed. Multiple regression analysis was performed with gait velocity or asymmetry indexes as the response variable and the muscle activity amplitudes in the various gait phases as explanatory variables. [Results] The major determinants of gait velocity were the tibialis anterior activity (ß=-0.35) and biceps femoris activity (ß=0.45) during the swing phase. In addition, the biceps femoris activity during the swing phase was the major determinant of the asymmetry index for the swing phase duration (ß=-0.41). [Conclusion] For patients with hemiparesis, increasing the biceps femoris activity during the swing phase is considered optimal, which may lead to improvement in walking performance.
ABSTRACT
In hemiparetic stroke survivors, premature plantarflexor muscle activity (PPF) often appears as a gait abnormality from terminal swing to the loading response on the paretic side. This study aimed to discern factors giving rise to PPF. Lower extremity function, spasticity magnitude, and gait electromyograms were assessed in 31 hemiparetic stroke survivors. Mean amplitudes during gait phases were determined for the paretic soleus, tibialis anterior, rectus femoris, and biceps femoris. The subjects were classified into PPF and non-PPF groups based on their relative soleus amplitude at different phases of gait, and group differences in each measurement were calculated and subjected to logistic regression. The PPF group showed less activity of the tibialis anterior during the swing phase but greater activity of the rectus femoris during the swing phase and of the biceps femoris, both prematurely and during the loading response. Logistic regression revealed premature activity of the biceps femoris to be a significant variable related to presence of PPF (odds ratioâ¯=â¯1.054). PPF in hemiparetic gait may work with the biceps femoris to supplement compromised lower extremity extension strength. PPF might be reduced by attaining enhanced strength of the hip and knee extensors at the time of initial contact during gait.
Subject(s)
Gait Disorders, Neurologic/physiopathology , Muscle, Skeletal/physiopathology , Paresis/physiopathology , Stroke/physiopathology , Adult , Aged , Electromyography/methods , Female , Gait/physiology , Gait Disorders, Neurologic/diagnosis , Gait Disorders, Neurologic/etiology , Humans , Knee/physiopathology , Knee Joint/physiopathology , Lower Extremity/physiopathology , Male , Middle Aged , Muscle Spasticity/physiopathology , Paresis/diagnosis , Paresis/etiology , Quadriceps Muscle/physiopathology , Stroke/complications , Stroke/diagnosis , Stroke RehabilitationABSTRACT
[Purpose] The purpose of this study was to clarify the difference in gait parameters of at-home walking and the 10-meter walking test results of individuals with hemiparesis. [Subjects] A total of 14 hemiparetic stroke recovery patients participated in this study. Inclusion criteria were: living at home, the ability to walk independently, and demonstrated low extremity on recovery stages III-V on the Brunnstrom Approach. The average age of the subjects was 66â years. [Methods] We used video surveillance and the inked footprint technique to record usual walking speed and maximum speed patterns both in subjects' homes and during the 10-meter walking test. From these methods, walking speed, stride length, and step rate were calculated. [Results] While both usual and maximum walking speeds of the 10-meter walking test correlated with stride length and step rate, at-home walking speeds only significantly correlated with stride length. [Conclusion] Walking patterns of the 10-meter walking test are quantifiably distinct from those demonstrated in patients' homes, and this difference is mainly characterized by stride length. In order to enhance in-home walking ability, exercises that improve length of stride rather than step rate should be recommended.
ABSTRACT
BACKGROUND: Although fall predictions using motor ability have been well reported in elderly people, there are few reports on physical cognitive ability. OBJECTIVE: To examine the relationship of the results of motor function tests that include physical cognitive ability on the ability to predict falls and to determine which test is the most appropriate. METHODS: We studied 174 community-dwelling elderly adults (mean age 75.7 ± 5.7, 41 males and 133 females), and measured grip strength, one-leg standing time (OLS), timed up and go test (TUG), functional reach test, sit and reach test, and maximal step length (MSL). The estimation error (EE), which was defined as the difference between the predicted and actual values, was calculated in all motor ability tests. Other assessments included the number of falls in the previous year, BMI, frequency of going out, Mini-Mental State Examination score, and Falls Efficacy Scale. In the baseline study, we divided the subjects into a fall group (n = 33) and a nonfall group (n = 141) and compared motor ability and EE for the two groups. During a 1-year follow-up, the nonfall group (baseline study) was assessed for the same measurements by using the same methods. RESULTS: In the baseline study, the fall group had significantly lower values of OLS and MSL. Furthermore, the fall group significantly overestimated their OLS, TUG, and MSL. In logistic regression analysis, EE of TUG (OR = 1.27) and EE of MSL (OR = 1.08) were detected as risk factors for falls. During follow-up, 11 subjects (7.8%) experienced falls. In logistic regression analysis, TUG (OR = 1.89) and EE of MSL (OR = 1.06) were detected as significant risk factors for falls. Since EE of MSL had higher values of both the area under the receiver operating characteristic curve and the sum of sensitivity and specificity than EE of TUG, the nonfall group was divided into two groups with a cutoff value of 2 cm for EE of MSL. A significant distribution disparity in falls between the two groups was found during follow-up and showed a relative risk of 18.78 for EE of MSL. CONCLUSIONS: We suggest that EE of MSL is a potent predictor for falls among healthy elderly adults.
Subject(s)
Accidental Falls , Aging , Motor Activity/physiology , Motor Skills/physiology , Psychomotor Performance , Accidental Falls/prevention & control , Accidental Falls/statistics & numerical data , Aged , Aged, 80 and over , Aging/physiology , Aging/psychology , Exercise Test/methods , Female , Geriatric Assessment , Humans , Independent Living , Male , Predictive Value of Tests , Proportional Hazards Models , Risk Assessment/methodsABSTRACT
The rat controls the form of its tail, from straight to curved, by contraction and relaxation of its four tail muscles. The tendons of these muscles insert on any of the cranial articular, transverse, and hemal processes of each of 24 coccygeal vertebrae (Co5-Co28). In this study, we isolated for the four coccygeal muscles each muscular fascicle segment inserting on any process of the coccygeal vertebrae. We measured the length and weight of all muscular fascicles and tendons, and then divided all muscular fascicles into four groups based on their insertion: Co5-Co10, Co11-Co16, Co17-Co22, and Co23-Co28. Moreover, we used soft X-ray imaging to investigate the geometrical relationship between neighboring coccygeal vertebrae. Additionally we carried out serial sectioning at the sacral and caudal portions, and traced the course of the tendons of coccygeal muscles from their origin to the Co4 level. We discuss which muscles and tendons play important roles when coccygeal vertebrae bend along and rotate around the longitudinal axis.
Subject(s)
Movement/physiology , Muscle, Skeletal/anatomy & histology , Muscle, Skeletal/physiology , Tail/physiology , Tendons/anatomy & histology , Tendons/physiology , Animals , Body Weights and Measures , Radiography , Rats , Tail/diagnostic imagingABSTRACT
PGE, a potent vasodilator, plays a primary role in maintaining the patency of the ductus arteriosus (DA). Genetic disruption of the PGE-specific receptor EP4, however, paradoxically results in fatal patent DA (PDA) in mice. Here we demonstrate that EP4-mediated signals promote DA closure by hyaluronic acid-mediated (HA-mediated) intimal cushion formation (ICF). Chronic EP4 stimulation by ONO-AE1-329, a selective EP4 agonist, significantly enhanced migration and HA production in rat DA smooth muscle cells. When HA production was inhibited, EP4-mediated migration was negated. Activation of EP4, adenylyl cyclase, and PKA all increased HA production and the level of HA synthase 2 (HAS2) transcripts. In immature rat DA explants, ICF was promoted by EP4/PKA stimuli. Furthermore, adenovirus-mediated Has2 gene transfer was sufficient to induce ICF in EP4-disrupted DA explants in which the intimal cushion had not formed. Accordingly, signals through EP4 have 2 essential roles in DA development, namely, vascular dilation and ICF. The latter would lead to luminal narrowing, helping adhesive occlusion and permanent closure of the vascular lumen. Our results imply that HA induction serves as an alternative therapeutic strategy for the treatment of PDA to the current one, i.e., inhibition of PGE signaling by cyclooxygenase inhibitors, which might delay PGE-mediated ICF in immature infants.
Subject(s)
Ductus Arteriosus/embryology , Ductus Arteriosus/metabolism , Hyaluronic Acid/metabolism , Receptors, Prostaglandin E/metabolism , Animals , Cell Movement , Cyclic AMP-Dependent Protein Kinases/metabolism , Ductus Arteriosus/cytology , Gene Expression Regulation, Developmental , Glucuronosyltransferase/genetics , Hyaluronan Synthases , Mice , Mice, Knockout , Muscle, Smooth/cytology , Muscle, Smooth/embryology , Muscle, Smooth/metabolism , Rats , Receptors, Prostaglandin E/agonists , Receptors, Prostaglandin E/deficiency , Receptors, Prostaglandin E/genetics , Receptors, Prostaglandin E, EP4 Subtype , Signal Transduction , Tissue Culture Techniques , Transcription, Genetic/geneticsABSTRACT
Caveolin, a major protein component of caveolae, directly interacts with multiple signaling molecules, such as Ras and growth factor receptors, and inhibits their function. However, the role of the second messenger system in mediating this inhibition by caveolin remains poorly understood. We examined the role of Ca2+-dependent signal in caveolin- mediated growth inhibition using a rat cardiac myoblast cell line (H9C2), in which the expression of caveolin- 3, the muscle specific subtype, can be induced using the LacSwitch system. Upon induction with IPTG and serum-starvation, the expression of caveolin-3 was increased by 3.3-fold relative to that of mock-induced cells. The recombinant caveolin-3 was localized to the same subcellular fraction as endogenous caveolin-3 after sucrose gradient purification. Angiotensin II enhanced ERK phosphorylation, but this enhancement was significantly decreased in caveolin-3-induced cells in comparison to that in mock-induced cells. Similarly, when cells were stimulated with fetal calf serum, DNA synthesis, as determined by [3H]-thymidine incorporation, was significantly decreased in caveolin- 3-induced cells. When cells were treated with Ca2+ chelator (BAPTA and EGTA), however, this attenuation was blunted. Calphostin (PKC inhibitor), but not cyclosporine A treatment (calcineurin inhibitor), blunted this attenuation in caveolin-3 induced cells. Our findings suggest that caveolin exhibits growth inhibition in a Ca2+-dependent manner, most likely through PKC, in cardiac myoblasts.
Subject(s)
Calcium/metabolism , Caveolin 3/metabolism , Myoblasts, Cardiac/metabolism , Signal Transduction , Angiotensins/metabolism , Animals , Calcineurin Inhibitors , Caveolae/physiology , Cell Line , Cell Proliferation , Clone Cells , Extracellular Signal-Regulated MAP Kinases/metabolism , Protein Kinase C/antagonists & inhibitors , RatsABSTRACT
Thyroid hormone-mediated positive cardiotropic effects are differently regulated between the atria and ventricles. This regulation is, at least in part, dependent on sarcoplasmic reticulum (SR) proteins. Sarcolipin, a homologue of phospholamban, has been recently identified as an atrium-specific SR protein. The expression of sarcolipin mRNA was significantly decreased in the atria of mice with hyperthyroidism and in 3,5,3'-triiodo-l-thyronine-treated neonatal rat atrial myocytes. Promoter activity and mRNA stability analyses revealed that thyroid hormone post-transcriptionally down regulated the expression of sarcolipin mRNA. The atrium-specific effect of thyroid hormone may occur in part through the regulation of atrial sarcolipin gene expression.
Subject(s)
Down-Regulation/drug effects , Hyperthyroidism/metabolism , Muscle Proteins/biosynthesis , Myocardium/metabolism , Proteolipids/biosynthesis , Triiodothyronine/administration & dosage , Animals , Calcium-Binding Proteins/genetics , Cell Line , Heart Atria/metabolism , Hyperthyroidism/chemically induced , Hyperthyroidism/pathology , Mice , Muscle Proteins/genetics , Myocardium/pathology , Myocytes, Cardiac/metabolism , Promoter Regions, Genetic/genetics , Proteolipids/genetics , RNA Stability/drug effects , RNA, Messenger/metabolism , Rats , Sequence Homology, Amino Acid , Transcription, Genetic/drug effectsABSTRACT
Voltage-dependent Ca(2+) channels (VDCCs), which consist of multiple subtypes, regulate vascular tone in developing arterial smooth muscle, including the ductus arteriosus (DA). First, we examined the expression of VDCC subunits in the Wistar rat DA during development. Among alpha(1)-subunits, alpha(1C) and alpha(1G) were the most predominant isoforms. Maternal administration of vitamin A significantly increased alpha(1C)- and alpha(1G)-transcripts. Second, we examined the effect of VDCC subunits on proliferation of DA smooth muscle cells. We found that 1 microM nitrendipine (an L-type Ca(2+) channel blocker) and kurtoxin (a T-type Ca(2+) channel blocker) significantly decreased [(3)H]thymidine incorporation and that 3 microM efonidipine (an L- and T-type Ca(2+) channel blocker) further decreased [(3)H]thymidine incorporation, suggesting that L- and T-type Ca(2+) channels are involved in smooth muscle cell proliferation in the DA. Third, we found that a novel alternatively spliced variant of the alpha(1C)-isoform was highly expressed in the neointimal cushion of the DA, where proliferating and migrating smooth muscle cells are abundant. The basic channel properties of the spliced variant did not differ from those of the conventional alpha(1C)-subunit. We conclude that multiple VDCC subunits were identified in the DA, and, in particular, alpha(1C)- and alpha(1G)-subunits were predominant in the DA. A novel spliced variant of the alpha(1C)-subunit gene may play a distinct role in neointimal cushion formation in the DA.
Subject(s)
Alternative Splicing/genetics , Calcium Channels/genetics , Calcium Channels/metabolism , Ductus Arteriosus/embryology , Ductus Arteriosus/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Animals , Gene Expression Profiling , Protein Subunits , Rats , Rats, Wistar , Structure-Activity RelationshipABSTRACT
Upon agonist binding, beta-adrenergic receptors sequestrate from the cell surface plasma membrane to cytosol. In the present study, we examine the kinetics of sequestration of beta1-adrenergic receptor and beta3-adrenergic receptor subtypes by radioligand binding assays using whole cells ('whole cell binding assays'). We found that HEK293T cells, but not COS1 cells, were readily and uniformly detached from the culture dish upon exposure to ice-cold phosphate-buffered saline. Using this property of HEK293T cells, we conducted whole cell binding assays using a hydrophilic antagonist ([3H]CGP-12177) and HEK293T cells transiently overexpressing human beta1-adrenergic receptor or beta3-adrenergic receptor. The Bmax and Kd values were 5.96 +/- 0.97 pmol/mg protein and 1 +/- 0.23 nM for the beta1-adrenergic receptor, and were 1.84 +/- 0.13 pmol/mg protein and 44.7 +/- 2.5 nM for the beta3-adrenergic receptor, respectively. Isoproterenol treatment, but not 6-[3-(dimethylamino)propionyl]forskolin treatment, for 2 h resulted in a dose-dependent loss of the number of the cell surface beta1-adrenergic receptor. At 100 microM, 36.6 +/- 5.7% of the cell surface beta1-adrenergic receptor was lost. In contrast, the cell surface beta3-adrenergic receptor number remained unchanged with isoproterenol treatment. Thus, beta1-adrenergic receptor sequestrates upon agonist stimulation but the same agonist stimulation does not induce beta3-adrenergic receptor sequestration, as demonstrated by our whole cell binding assays.
Subject(s)
Radioligand Assay/methods , Receptors, Adrenergic, beta/metabolism , Animals , COS Cells , Cell Count/methods , Chlorocebus aethiops , Humans , Propanolamines/metabolismABSTRACT
Fatty acids are utilized as a cellular energy source. In the present study, we investigated whether fatty acids could affect axoplasmic transport. Cultured mouse superior cervical ganglion neurons were placed in the glucose-containing medium (145 mM NaCl, 5 mM KCl, 1 mM CaCl(2), 1 mM MgCl(2), 5 mM D-glucose, 10 mM Hepes, pH 7.3, 37 degrees C), and axoplasmic transport of particles in neurites was observed under video-enhanced contrast microscopy. A variety of fatty acids (acetate (C2), caproate (C6), caprylate (C8), caprate (C10), 2-decenoate (C10:1), arachidonate (C20:4); 0.1-1 mM) caused a transient increase in the amount of particles transported in both anterograde and retrograde directions. The increasing effects of fatty acids were dose-dependent. A half-maximum effective dose (ED(50)) for acetate was 0.8 mM, which is similar to the reported K(m) value of acetyl-CoA synthetase for acetate. The ED(50) for caprylate was 28 microM, which is near the K(m) value of acyl-CoA synthetase for medium- and long-chain fatty acids. Application of 5 mM malonate, an inhibitor of the citrate cycle, induced a steady-state decrease in axoplasmic transport, indicating that energy derived from the citrate cycle is required for the maintenance of axoplasmic transport. The increasing effect of acetate (1 mM) on axoplasmic transport was completely abolished by pretreatment with malonate (5 mM), suggesting that acetate produces ATP for axoplasmic transport via the citrate cycle. Alternatively, the effect of caprate (1 mM) was retained after treatment with malonate. Thus, fatty acids except acetate produce ATP probably through both the beta-oxidation pathway and the citrate cycle, increasing axoplasmic transport. Since the effect of fatty acids was transient, certain negative feedback mechanisms might be involved. The removal of glucose from the medium resulted in a low steady-state level of axoplasmic transport. Under such condition, the acetate (1 mM)-induced transient increase in axoplasmic transport remained. Since intracellular ATP must be low under glucose-free condition, intracellular ATP concentrations are unlikely to be involved in the feedback system. Instead, acetyl-CoA or its downstream products in the citrate cycle might lead to feedback inhibition. Application of citrate (5 mM) caused a strong decrease following a transient increase in axoplasmic transport, whereas no other acetyl-CoA product decreased axoplasmic transport. Thus, excessive citrate may be one of factors leading to feedback inhibition of metabolic pathways to arrest and reverse the increase in axoplasmic transport induced by fatty acids.
Subject(s)
Axonal Transport/physiology , Fatty Acids/metabolism , Superior Cervical Ganglion/physiology , Animals , Axonal Transport/drug effects , Cells, Cultured , Citrates/pharmacology , Dose-Response Relationship, Drug , Drug Interactions , Enzyme Inhibitors , Glucose/pharmacology , Iodoacetates/pharmacology , Malonates/pharmacology , Mice , Mice, Inbred C57BL , Radiation-Protective Agents/pharmacology , Succinic Acid/pharmacology , Time FactorsABSTRACT
We investigated whether polymorphism of the type 6 adenylyl cyclase gene influences the occurrence of left ventricular hypertrophy in a Japanese population. Type 6 adenylyl cyclase is a major cardiac adenylyl cyclase isoform and plays an important role in regulating cardiac function. We examined the type 6 adenylyl cyclase gene for single nucleotide polymorphism by heteroduplex analysis and found a mutation (T11215A) in intron 17. We genotyped the single nucleotide polymorphism (TT/TA/AA groups) by the mutagenically separated polymerase chain reaction method in 2068 subjects who underwent health screening for cardiovascular disease. Genetic variation was in the Hardy-Weinberg equilibrium. We found no significant association between the frequency of left ventricular hypertrophy and any of the genotype groups. In the TT and the TA genotype group, however, left ventricular hypertrophy was associated with increased blood pressure, while no association with increased blood pressure was found in the AA genotype group. It was concluded that the AA group may be at risk of developing left ventricular hypertrophy independent of increased blood pressure.