Loss-of-function mutations in HINT1 cause axonal neuropathy with neuromyotonia.

Inherited peripheral neuropathies are frequent neuromuscular disorders known for their clinical and genetic heterogeneity. In 33 families, we identified 8 mutations in HINT1 (encoding histidine triad nucleotide-binding protein 1) by combining linkage analyses with next-generation sequencing and subsequent cohort screening of affected individuals. Our study provides evidence that loss of functional HINT1 protein results in a distinct phenotype of autosomal recessive axonal neuropathy with neuromyotonia.

Calcium, calmodulin and 3',5'-cyclic nucleotide phosphodiesterase activity in human muscular disorders.

3',5'-Cyclic nucleotide phosphodiesterase (PDE) is known to play an important role in the regulation of cyclic nucleotide levels in various tissues including the muscle. Previous studies have estimated the level of this enzyme in several neuromuscular disorders but the results have been variable. Moreover, there was no attempt made to correlate the enzyme levels with the levels of calcium and calmodulin, both of which regulate diverse biological processes including muscle contraction. In the present study we have estimated phosphodiesterase in the muscle of normal controls as well as patients with myotonic (MyD) and Duchenne muscular dystrophy (DMD) and amyotrophic lateral sclerosis (ALS). PDE was found to be increased significantly in all of the diseased muscles as compared to controls (P less than 0.01). But the increase could be coupled with an increase in calcium and calmodulin only in Duchenne dystrophic muscle.

Effect of myotonia induced by anthracene-9-carboxylic acid on mitochondrial calcium, plasma creatinine-phosphokinase and aldolase activity in the rat.

The frequent association of myotonia with dystrophy and the knowledge that calcium is increased in injured skeletal muscle cells suggest a possible relationship between cell calcium and myotonic alterations. This investigation has been performed to study the role of calcium in experimental myotonia induced by anthracene-9-carboxylic acid (9-AC) in rats treated with several regimens of food and exercise. Thirty-two rats were divided into 4 groups of 8 rats each, one control and 3 experimental groups. The treatments included caffeine plus exercise (group 2), and a calcium-rich diet (group 3); these procedures were designed to increase intracellular calcium; another group was treated with 9-AC as a myotonia-inducer (group 4). The treatment for all groups lasted 60 days. No significant differences in plasma sodium, potassium, chloride and calcium between control and experimental groups were observed. Whole muscle calcium in wet tissue samples did no change with any treatment. On the contrary, mitochondrial calcium showed a significantly higher concentration in group 3 and 4. CPK and aldolase activities in groups 1, 2 and 3 were similar; but in group 4 these enzyme activities were significantly higher (p less than 0.05). The electrical and mechanical responses were not altered in any rat with any experimental treatment. Our data suggest that myotonia is a predisposing factor for an altered mitochondrial calcium homeostasis in this model; in addition, the enzyme activities of CPK and aldolase were increased in the rats of group 4 implicating that myotonia is a crucial factor in the development of enzymatic abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS)

Glyconeogenic and glycogenic enzymes in chronically active and normal skeletal muscle.

The chronically active (pseudomyotonic) gastrocnemius muscle in the C57B16J dy2J/dy2J mouse contains both elevated lactate and glycogen as well as fibers that have high amounts of glycogen and enhanced glyconeogenic activity. In the present study we analyze the activities of some key glyconeogenic enzymes to assess the causes of elevated muscle glycogen and to determine the pathway for glycogen synthesis from lactate. Glycogen synthase, malate dehydrogenase, phosphoenolpyruvate carboxykinase, and malic enzyme were all elevated in homogenates of the chronically active muscle. Activities of glycogen phosphorylase and fructose 1,6-bisphosphatase were decreased in whole muscle homogenates. Histochemistry demonstrated that the high-glycogen fibers were typically fast-twitch glycolytic fibers that had high glycogen synthase, glycogen phosphorylase, and malic enzyme activities. Malate dehydrogenase activity followed succinate dehydrogenase activity and did not correlate to high-glycogen fibers. Thus the high-glycogen fibers have an elevated enzymatic capacity for glycogen synthesis from lactate, and the pathway may involve use of the pyruvate kinase bypass enzymes.

The myotonic mouse mutant ADR: physiological and histochemical properties of muscle.

The muscle physiology and histochemistry of a hereditary neuromuscular syndrome of the mouse, "arrested development of righting response" (ADR), was studied. The speed of single twitches of fast ADR limb muscles was normal up to an age of about 60 days but decreased at later ages. At any age between 10 and 120 days postnatal, fast and slow muscles of the mutant displayed after-contractions of 1-3 (5) seconds duration. These coincided with electrical after-activity of muscle, as demonstrated by electromyography. After-contractions and EMG signals were suppressed by the membrane-stabilizing drug tocainide. These physiological data suggest that ADR is a myotonia. With a few exceptions, limb and trunk muscles of ADR animals showed a uniform oxidative phenotype with a lack of large diameter glycolytic fibers. The histochemical muscle phenotype of the ADR mouse was partially reversed by a long-term treatment with tocainide.

Clofibrate-induced myotonia in rats.

Determination of enzymatic activity, protein profile and phospholipid composition of muscle plasma membranes and sarcoplasmic reticulum in rats were carried out after clofibrate injections in a dose of 0.4 g/kg body weight. In the plasma membranes, the activity of Na+ + K+, Mg2+ ATPase was insignificantly decreased, and that of 5'-nucleotidase significantly diminished. A non-significant change was observed in the total amount of phopholipids. The amount of phosphoethanolamine appeared to be lower. Changes in the protein profile were seen. In the sarcoplasmic reticulum, the major abberation was the decrease of Mg2+ ATPase activity. No evident changes were observed in the phospholipid behaviour. Abnormalities in the protein profile appeared. In the myofibrillar proteins, increases of alpha-actinin and troponin at the expense of myosin were observed. In the clofibrate model of myotonia in rats, the changes in the biochemical parameters were less pronounced as compared to the previously tested 20,25-diazacholesterol model.

Increase of muscle peroxisomal enzymes and myotonia induced by nafenopin, a hypolipidemic drug.

The chronic administration of nafenopin, a hypolipidemic drug, induced an increase in catalase and acyl-CoA oxidase activities in various skeletal muscles, including the gracilis, diaphragm, soleus, and extensor digitorum longus. The magnitude of the increase was around 100% for both enzymes in each of the muscles studied in spite of the different basal level. These changes seem to be specific of the peroxisomal enzymes because acetylcholinesterase, which is not peroxisomal, did not follow the same pattern in all the muscles. Concomitant with the increase in muscle peroxisomal enzymes, the skeletal muscles presented an altered electromyogram with prolonged insertional activity, repetitive firing of action potentials, and myotonic runs characteristic of myotonia. Our results suggest a role for peroxisomes in the myotonic disorder.

Erythrocyte membrane abnormalities in human myotonic dystrophy.

Membrane-bound enzyme activities and cardiac glycoside binding were determined in red blood cell membrane preparations from patients with myotonic dystrophy and in age matched controls. Na+-K+-activated ATPase activity was significantly increased in myotonic patients. [3H]Ouabain binding to erythrocyte membranes was also significantly increased in myotonic dystrophy patients. The Mg2+-ATPase (ouabain-insensitive) was, however, unchanged. The K+-stimulated paranitrophenyl phosphatase (KPNPPase) activity was markedly enhanced in myotonic patients as compared to controls. The kinetic analysis showed a marked change in Vmax of Na+-K+ ATPase with respect to the activation by Na+, K+ and ATP. However, the Km values were the same in control as well as in myotonic groups. The increased erythrocyte membrane Na+-K+-ATPase activity, KPNPPase and [3H]ouabain binding in myotonic patients supports the hypothesis that generalized membrane abnormality may be involved in pathogenesis of the human myotonic dystrophy.

Erythrocyte ghosts (Na+ + K+) ATPase activity in Duchenne's dystrophy and myotonia.

In Duchenne muscular dystrophy the activity of (Na+ + K+)ATPase in erythrocyte ghosts is reduced and its reaction to ouabain is paradoxical both in low sodium and high sodium systems. No such changes were seen in a case of Becker dystrophy, in limb-girdle dystrophy, and in neurogenic atrophy of muscles. In myotonic dystrophy and congenital myotonia the activity of ATPase and its inhibition by ouabain were depressed.

The metabolism of cyclic-3'-5'-adenosine monophosphate (cAMP) in diseased muscle.

Adenyl cyclase (AC) and cyclic nucleotide phosphodiesterase (PDE) have been studied in muscle from patients affected with muscular and neuromuscular diseases as well as in muscle of rats after experimental denervation. A diminution of AC was observed in Duchenne's dystrophy, in neurogenic atrophies, in inflammatory forms, in myasthenia and myotonia. PDE was diminished in Duchenne's dystrophy and increased in all other diseases considered. Both enzymes were increased in rat muscle after nerve transection. The possible mechanism underlying the biochemical alterations observed are discussed including unmasking of extra-activity due to changed metabolic environment, tissue destruction and activation of lysosomal activity.