A novel mutation of the CLCN1 gene associated with myotonia hereditaria in an Australian cattle dog

BACKGROUND: Heritable myotonia is a genetic muscle disorder characterized by slow relaxation of skeletal muscles. The main clinical signs are skeletal muscle stiffness, especially after vigorous contraction, and muscle hypertrophy. Muscle stiffness may be enhanced by inactivity, and often is relieved by exercise. Myotonia can be inherited in an autosomal dominant or recessive manner (Thomsen- or Becker-type myotonia, respectively). In mice, goats, Miniature Schnauzer dogs, and most affected humans, the disorder is caused by mutations in CLCN1, which encodes the skeletal muscle voltage-gated chloride channel, Cl1C-1. HYPOTHESIS: We hypothesized that an Australian Cattle Dog with generalized muscle stiffness and hypertrophy examined at the Ontario Veterinary College would have a mutation in the CLCN1 gene. ANIMALS: A pure-bred Australian Cattle Dog from Ontario, Canada, was used. METHODS: Based on clinical signs and electromyographic test results, a diagnosis of myotonia hereditaria was made, and a muscle biopsy was collected for genetic analysis. RESULTS: Sequence data obtained from the affected dog confirmed that it was homozygous for a single base insertion in the CLCN1 coding sequence. This mutation would result in a truncated ClC-1 protein being expressed, which, based on molecular evidence from other studies, would result in functionally compromised chloride conduction in the skeletal muscles of the animal. CONCLUSIONS AND CLINICAL IMPORTANCE: To the authors' knowledge, this report describes the Ist case of myotonia in an Australian Cattle Dog and represents the 1st non-Schnauzer canine myotonia to be genetically characterized. In addition, we developed a polymerase chain reaction-based genetic screen to detect heterozygotes with this mutation in the at-large Australian Cattle Dog population.     

Muscular dystrophy in female dogs

The most common form of muscular dystrophy in dogs and humans is caused by mutations in the dystrophin gene. The dystrophin gene is located on the X chromosome, and, therefore, disease-causing mutations in dystrophin occur most often in males. Therefore, females with dystrophin deficiency or other forms of muscular dystrophy may be undiagnosed or misdiagnosed. Immunohistochemistry was used to analyze dystrophin and a number of other muscle proteins associated with muscular dystrophy in humans, including sarcoglycans and laminin alpha2, in muscle biopsy specimens from 5 female dogs with pathologic changes consistent with muscular dystrophy. The female dogs were presented with a variety of clinical signs including generalized weakness, muscle wasting, tremors, exercise intolerance, gait abnormalities, and limb deformity. Serum creatine kinase activity was variably high. One dog had no detectable dystrophin in the muscle; another was mosaic, with some fibers normal and others partly dystrophin-deficient. A 3rd dog had normal dystrophin but no detectable laminin alpha2. Two dogs could not be classified. This study demonstrates the occurrence of dystrophin- and laminin alpha2-associated muscular dystrophy and the difficulty in clinical diagnosis of these disorders in female dogs.     

骨骼肌纤维烟酰胺腺嘌呤二核苷酸四唑氧化还原酶组化染色的检测

本文应用烟酰胺腺嘌呤二核苷酸四唑氧化还原酶（NADH-TR）组化染色，将大鼠与家兔小腿胫前肌与腓肠肌的肌纤维型分成染色最深的快缩氧化FO型、染色较深的慢缩氧化SO型、染色浅的快缩氧化酵解FOG型及染色最浅的快缩酵解FG型纤维。这与定量分析结果相吻合。并对低温液氮保存骨骼肌标本进行观察，发现小块肌组织＋生理盐水和小块肌组织＋5％二甲基亚砜（DMSO）冷冻保护剂的玻璃化冻存一起浸入液氮制成切片，都未见冰晶形成；而在用5％二甲基亚砜玻璃化冻存大块肌组织切片见有少量冰晶出现；未使用5％二甲基亚砜玻璃化冻存大块肌组织切片见有大量冰晶出现。提示小块标本超快速浸入液氮内能减少冰晶形成，保证酶组化染色切片的质量。并进一步对烟酰胺腺嘌呤二核苷酸四唑氧化还原酶组化作用机理与孵育液的科学配制进行了探讨。     

Evaluation of jitter by stimulated single-fiber electromyography in normal dogs

Single-fiber electromyography (SFEMG), a technique used to investigate neuromuscular transmission, has been described previously in the pelvic limb of dogs. Because preferential involvement of isolated muscle groups can occur in disorders of neuromuscular transmission, SFEMG was done in the peroneus longus (PL), extensor carpi radialis (ECR), and orbicularis oculi (OO) muscles of 10 adult, clinically normal dogs. Jitter was calculated as the mean absolute value of the consecutive differences in latency of 50 single muscle fiber action potentials after stimulation of intramuscular nerve bundles at the level of the motor point in at least 20 muscle fibers per muscle. Bilateral recordings were performed in 3 dogs. Mean jitter values were determined for each muscle, and differences among muscle groups and among dogs were compared. The upper limits of mean consecutive difference (mean plus 3 standard deviations) for the PL, ECR, and OO muscles were 21.94, 22.53, and 23.39 micros, respectively, and the upper limit of mean consecutive difference for individual muscle fibers in the respective fiber pools was 28.62, 36.39, and 35.68 micros. Jitter values for the ECR and OO were significantly higher than the jitter value for the PL muscle (P < .05). Significant differences among muscles or dogs or between sides were not observed for the ECR. Significant differences among dogs were observed for OO jitter values and were attributed to extremely low jitter values in 1 dog. Significant differences were demonstrated between sides for the PL and were attributed to small sample size. Results of this study provide normative data that can be used in the application of the stimulated SFEMG technique to dogs with suspected disorders of neuromuscular transmission.     

莱芜猪肌肉组织学特性与肉质关系的研究

本研究采用二因子多水平有重复的试验方法,对１４头育肥莱芜猪不同屠宰体重及不同解剖部位肌肉的组织结构和肌纤维的组织学特性及肉质性状进行研究。结果表明：随屠宰体重的提高,肌肉纤维直径（ＤＭＦ）,密度（ＭＦＤ）和宰后僵直肌工降度（ＳＬ）没有发生显著变化（Ｐ〉０．０５）,组织学显示的肌肉３种构成分在不同体重组间差异也不显著,但除ｐＨ外,其他各测定性状在不同部位肌肉都存在极显著的差异（Ｐ〈０．０１）,研究下     

稻田和池塘养殖模式对中华绒螯蟹肠道菌群、免疫酶活及肌肉游离氨基酸的影响

为了探究稻田和池塘养殖模式对中华绒螯蟹(Eriocheir sinensis)肠道菌群、免疫力和肌肉风味影响,选取同一批次稻田和池塘的中华绒螯蟹在5月和10月对其肠道菌群、免疫酶活和肌肉游离氨基酸进行测定。结果表明：中华绒螯蟹肠道优势菌群为软壁菌门、拟杆菌门、变形菌门和厚壁菌门。在季节和养殖模式均对蟹的肠道菌群造成影响的情况下,季节变化对菌群变化的影响更大。从5月到10月,稻田养殖模式下中华绒螯蟹肠道菌群的丰富度和多样性均出现上升趋势,而池塘养殖模式下肠道菌群的丰富度和多样性则无显著变化。两种养殖模式下中华绒螯蟹肠道中的SOD (超氧化物歧化酶)在5月的活性均高于10月(P <0.05);在10月,池塘模式下的免疫酶活性LZM（溶菌酶）和ACP（酸性磷酸酶）显著高于稻田模式(P< 0.05)。各组肌肉中均检测出17种游离氨基酸,呈味氨基酸中甜味氨基酸含量(TSAA)>总苦味氨基酸(TBAA)>总鲜味氨基酸(TUAA),且池塘养殖模式下中华绒螯蟹肌肉中TSAA和总游离氨基酸TFAA高于稻田养殖模式下含量(P< 0.05)。因此,池塘养殖模式较稻田养殖模式下的中华绒螯蟹肠道菌群时间上更稳定,免疫酶活更强,肌肉风味更优。     

Muscle fibre type distribution of the thoracolumbar and hindlimb regions of horses: relating fibre type and functional role

Background

Although the majority of equine muscles have a mixed fibre type distribution indicative of diverse functional roles, the predominance of a fibre type can indicate the primary function of a muscle. The deep epaxial musculature has an important role in core spinal stability in humans, reflected as a predominantly muscle fibre type (MFT) I or postural fibre type. The fibre type of the deep epaxial musculature has not been determined in horses. The objective of the study was to determine the MFT distribution in selected muscles of thoracolumbar and hindlimb region of horses. This included deep epaxial and hypaxial muscles that were hypothesised to have a postural stabilising role. A second objective was to examine differences in MFT distribution between horses bred for endurance (Arabian) and sprinting (Quarter horse). Muscle biopsy samples were obtained from selected thoracolumbar and hind limb muscles of 5 Quarter horses, 4 Arabians, and 2 Thoroughbreds. The myosin heavy chain distribution was determined by gel electrophoresis. Mann–Whitney rank test was used to compare the proportional MFT and differences between breeds.

Results

Mm. sacrocaudalis dorsalis medialis and diaphragm had the highest proportion of MFT-I. The remaining deep epaxial muscles and the hypaxial muscle m. psoas minor had approximately equal MFT I and II proportions. Mm. psoas major, iliocostalis, longissimus dorsi and the hind limb muscles contained mostly MFT-IIX. The fibre type distribution was similar between Arabians and Quarter horses, although Quarter horses had more MFT-IIX fibres in psoas major (P = 0.02) while Arabians had more MFT-I fibres in m. longissimus dorsi (P = 0.03).

Conclusions

The fibre type distribution of the deep epaxial muscles, mm psoas minor and diaphragm varied from approximately equal MFT-I and II proportions to predominantly MFT-I suggesting a postural stabilising role possibly important in core spinal stability. In contrast the fibre type proportions of mm psoas major, iliocostalis, longissimus dorsi and the hind limb muscles were mainly MFT-II suggesting a locomotory role. Knowledge of fibre type distribution in such a clinically important area can direct diagnosis, prevention and treatment of muscular or neuromotor dysfunction.     

Sexual dimorphism of fast muscle fibre recruitment in farmed Atlantic halibut (Hippoglossus hippoglossus L.)

Commercially farmed Atlantic halibut (Hippoglossus hippoglossus) were reared at the Aga Marin's facility located on Dønna (Norway) under ambient environmental conditions in duplicate 15 × 15 × 8 m netpens (May 2004 to May 2005). Twenty fish were sampled five times over a twelve month production period during which time the average body weight increased from 1.26 to 2.08 kg (n = 100 fish). Body mass, fork length (L_F), and the number and size distributions of fast muscle fibres were determined in male and female fish. All males matured during the autumn whereas no maturation was observed in females. From the point of maturation females had superior growth performance to males and body mass and the total cross-sectional area of muscle were 1.4-fold (P < 0.05) and 1.3-fold higher (P < 0.01) respectively by May 2005. The total number of fast muscle fibres per trunk cross-section at 0.55 L_F was 24.5% higher in females (7.58 × 10⁵) than males (5.80 × 10⁵) prior to sexual maturation. In females, muscle fibre recruitment slowed with short days and low water temperatures (< 6.5 °C), but had increased to 1.01 × 10⁶ by May 2005 (P < 0.001). In contrast, there was no growth and no increase in muscle fibre number in males following the onset of maturation. The distribution of muscle fibre diameters prior to maturation in males was also significantly different between male and female fish matched for L_F. The results illustrate a sexual dimorphism of muscle fibre recruitment patterns in Atlantic halibut and highlight the adverse affects of sexual maturation in males on muscle growth.