Iain S. Young

Isometric and isotonic muscle properties as determinants of work loop power output

The power output of rabbit latissimus dorsi muscle was calculated under isotonic conditions and during oscillatory work. Isotonic shortening studies yielded a maximum power output of 120 W·kg−1 at a P/P0 of 0.4 compared to a maximum power output of 32 W·kg−1 obtained using the work loop technique. This difference can largely be explained by comparing actual work loops with those constructed using force velocity (P/V) and isometric data. At low cycle frequencies, work loop power output is quite close to that predicted from P/V and isometric data. However, at higher frequencies other dynamic muscle properties appear to exert a more marked effect.

Distribution of the AQP4 Water Channel in Normal Human Tissues: Protein and Tissue Microarrays Reveal Expression in Several New Anatomical Locations, including the Prostate Gland and Seminal Vesicles

Aquaporins facilitate osmotically driven water movement across cell membranes. Aquaporin 4 (AQP4) is a major water channel in the central nervous system where it participates in cerebral water balance. AQP4 is also present in basolateral membranes of lower respiratory tract airway and renal collecting duct epithelial cells, gastric parietal cells and skeletal muscle cells. However, the distribution of AQP4 in many other tissues is still unknown. The aim of this study was to determine the expression and relative abundance of AQP4 in human Tissue MicroArrays (TMAs) and human protein microarrays by immunohistochemistry and chemiluminescence. In the central nervous system AQP4 was abundantly expressed in the cerebral cortex, cerebellar cortex (purkinje/granular layer), ependymal cell layer, hippocampus and spinal cord. Lower levels were detected in choroid plexus, white matter and meninges. In the musculoskeletal system AQP4 was highly expressed in the sarcolemma of skeletal muscle from the chest and neck. In the male genital system AQP4 was moderately expressed in seminiferous tubules, seminal vesicles, prostate and epidiymis. In the respiratory system AQP4 was moderately expressed in lung and bronchus. AQP expression was abundant in the kidney. In the gastrointestinal system AQP4 was moderately present in basolateral membranes of parietal cells at the base of gastric glands. AQP4 was also detected in salivary glands, adrenals, anterior pituitary, prostate and seminal vesicles. Human protein microarrays verified the TMA data. Our findings suggest that AQP4 is expressed more widely than previously thought in human organs and may be involved in prostatic and seminal fluid formation.

Mutually exclusive muscle designs: the power output of the locomotory and sonic muscles of the oyster toadfish (Opsanus tau).

Animals perform a vast array of motor activities. Although it has generally been accepted that muscles are well suited to the function that they must perform, specialization for performing one function may compromise their ability for carrying out another. We examined this principle in the toadfish muscular system: slow–twitch red and fast–twitch white myotomal muscles are used for powering swimming at relatively low frequencies, while the superfast swimbladder muscle powers mating calls by contracting at 100 Hz. We measured muscle power output over a wide range of frequencies. The red and white locomotory muscles could not generate power over ca. 2.2 and 12 Hz, respectively and, hence, could not power sound production. In contrast, the swimbladder muscle has many specializations that permit it to generate power at frequencies in excess of 100 Hz. However, these specializations drastically reduce its power output at low frequencies: the swimbladder muscle generated only one–twentieth of the power of the red muscle and one–seventh of the power of the white muscle at the frequencies used during swimming. To generate the same total power needed for swimming would require unfeasibly large amounts of swimbladder muscle that could not fit into the fish. Hence, the designs of the swimbladder and locomotory muscles are mutually exclusive.

What Do Zebrafish Want? Impact of Social Grouping, Dominance and Gender on Preference for Enrichment

Although environmental enrichment is known to improve laboratory rodent wellbeing and enhance scientific data collection, relatively little is known with regards to the type of enrichment that might be useful for zebrafish (Danio rerio). Therefore, this study explored if zebrafish displayed preferences for a range of enrichments, including substrates, artificial plants, combinations thereof and airstones. Tanks divided into two compartments containing different enrichment cues were used to determine the preferences of zebrafish housed in pairs and groups of eight. When comparing time spent in enriched versus barren compartments, dominant individuals in a pair displayed a preference for substrate and behaviourally excluded the subordinate (p < 0.05). In groups there was a preference for all substrate (p < 0.01) and plant (p < 0.05) enrichments over barren conditions. The strongest preference was for gravel substrate and images of gravel attached to the bottom of the tank. When preferences were compared for different enrichments, gravel (both sexes, p < 0.01) again emerged as the cue attracting the most significant preferences, with any combination featuring gravel substrate preferred over any combination featuring sand (p < 0.05). The study has demonstrated that zebrafish reared in barren conditions preferred structural enrichment over standard conditions; however, when fish were held in pairs this was influenced by dominance status and in groups this was influenced by gender.

Regional variation in parvalbumin isoform expression correlates with muscle performance in common carp (Cyprinus carpio)

SUMMARY The mechanical properties of the axial muscles vary along the length of a fishs body. This variation in performance correlates with the expression of certain muscle proteins. Parvalbumin (PARV) is an important calcium binding protein that helps modulate intracellular calcium levels which set the size and shape of the muscle calcium transient. It therefore has a central role in determining the functional properties of the muscle. Transcript data revealed eight specific isoforms of PARV in common carp (Cyprinus carpio) skeletal muscle which we classified as α1 and β1–7. This study is the first to show expression of all eight skeletal muscle PARV isoforms in carp at the protein level and relate regional differences in expression to performance. All of the PARV isoforms were characterised at the protein level using 2D-PAGE and tandem mass spectrometry. Comparison of carp muscle from different regions of the fish revealed a higher level of expression of PARV isoforms β4 and β5 in the anterior region, which was accompanied by an increase in the rate of relaxation. We postulate that changes in specific PARV isoform expression are an important part of the adaptive change in muscle mechanical properties in response to varying functional demands and environmental change.

Growth Induced by Incremental Static Stretch in Adult Rabbit Latissimus Dorsi Muscle

Incrementally applied static stretch over 3 weeks resulted in a 72% increase in the weight of the in situ latissimus dorsi muscle in rabbits. True growth rather than tissue oedema was confirmed by increases in the protein content (130%), the cross‐sectional area of the type I fibres (30%) and the muscle length (i.e. number of sarcomeres in series increased 25%). Despite an increase in the proportion of fibres staining positive for the enzyme succinate dehydrogenase (SDH), the myosin ATPase stain showed no appreciable fibre type transformation. While total power output in the stretched muscle was unchanged, its maximum mass specific power output, as determined by oscillatory work loops, was decreased by 50%. The cross‐sectional area that was occupied by connective tissue increased from 15 to 19% in the stretched muscles, with a concomitant increase in passive energy dissipation. Some incrementally stretched muscles were then allowed an additional 3 weeks of maintained stretch to determine whether the adaptive changes would be preserved or reversed. Previous gains in muscle weight, length and area of type I fibres all remained. In contrast, the connective tissue content and the passive properties returned to control values during this period.

Direct Analysis and Quantification of Metaldehyde in Water using Reactive Paper Spray Mass Spectrometry

Metaldehyde is extensively used worldwide as a contact and systemic molluscicide for controlling slugs and snails in a wide range of agricultural and horticultural crops. Contamination of surface waters due to run-off, coupled with its moderate solubility in water, has led to increased concentration of the pesticide in the environment. In this study, for the first time, rapid analysis (<~1 minute) of metaldehyde residues in water is demonstrated using paper spray mass spectrometry (PS-MS). The observed precursor molecular ions of metaldehyde were confirmed from tandem mass spectrometry (MS/MS) experiments by studying the fragmentation patterns produced via collision-induced dissociation. The signal intensity ratios of the most abundant MS/MS transitions for metaldehyde (177 → 149 for protonated ion) and atrazine (221 → 179) were found to be linear in the range 0.01 to 5 ng/mL. Metaldehyde residues were detectable in environmental water samples at low concentration (LOD < 0.1 ng/mL using reactive PS-MS), with a relative standard deviation <10% and an R2 value >0.99, without any pre-concentration/separation steps. This result is of particular importance for environmental monitoring and water quality analysis providing a potential means of rapid screening to ensure safe drinking water.

A proteomics strategy for determining the synthesis and degradation rates of individual proteins in fish

In order to study the protein dynamics in the tissues of fish we have developed a proteomics-based strategy to determine the rates of synthesis and degradation of individual proteins. We have demonstrated the feasibility of this approach by measuring the turnover of multiple isoforms of parvalbumin (β1-7) in the skeletal muscle of common carp (Cyprinus carpio). A stable isotope-labelled amino acid ([(2)H(7)] l-leucine) was administered to the carp via the diet and its incorporation into the isoforms of parvalbumin in muscle over time was monitored by LC-MS analysis of signature peptides. The relative isotope abundance was calculated and used to deconvolute the data. The β7 parvalbumin isoform had a rate of synthesis that was greater than the rate of degradation. In contrast the rate of degradation of the β5 isoform exceeded its rate of synthesis, whilst the analysis revealed that the other parvalbumin β-isoforms (β1, β2, β3, β4 and β6) had a rate of synthesis that was equal to the rate of degradation. This work has addressed a number of technical challenges and represents the first study to use proteomic approaches to measure the turnover of individual proteins in fish.

Observer variation in the evaluation and classification of severe central tarsal bone fractures in racing Greyhounds

OBJECTIVES To determine observer agreement on radiographic evaluation of central tarsal bone (CTB) fractures and compare this with evaluation of the same fractures using computed tomography (CT). METHODS Radiographs and CT scans were obtained of the right tarsi from limbs of Greyhounds euthanatized after sustaining severe CTB fracture during racing. Four observers described and classified each fracture. Inter- and intra-observer agreements were calculated. RESULTS Inter-observer agreement was higher for assessment of fractures using CT. Several fractures assessed by radiography were misclassified as a less severe type. Intra-observer agreement for assessment and classification of CTB fractures via radiography versus CT was variable. Overall agreement among all four observers was higher for CT than radiography. Additionally, when identifying fractures of the adjacent tarsal bones, observer agreement was higher for CT than radiography. CLINICAL SIGNIFICANCE Computed tomography improved observer ability to correctly evaluate CTB fracture and detect the degree of displacement and extent of any comminution. Identification of fractures of adjacent tarsal bones was also improved when tarsi were assessed using CT. These data suggest that treatment decisions based solely on radiographic assessment of CTB fractures may not produce the expected outcome.

Immunohistochemical evidence for expression of fast-twitch type sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA1) in German shepherd dogs with dilated cardiomyopathy myocardium

OBJECTIVES Dilated cardiomyopathy (DCM) is one of the most common acquired canine heart diseases. It is particularly common in large and giant breed dogs. Although a great deal is known about the clinical progression and manifestations of the disease, the underlying cellular and molecular mechanisms remain poorly understood. One widely held belief is that calcium-handling abnormalities are critically involved in the disease process. This study investigates the changes in expression of the sarco(endo)plasmic reticulum calcium ATPase (SERCA) isoforms in DCM myocardium from German shepherd dogs. ANIMALS, MATERIALS AND METHODS Affected tissue samples were obtained from German shepherd dogs with DCM, euthanized for intractable congestive heart failure while normal myocardial tissue samples were obtained from German shepherd dogs, euthanized for non-cardiovascular reasons. Tissue microarrays containing normal and DCM myocardium samples were prepared, immunostained with SERCA1 and SERCA2 antibodies and analyzed. RESULTS We were able to demonstrate, for the first time, that while there is little change in the expression of the cardiac isoform (SERCA2), there is clear expression of the fast-twitch skeletal muscle isoform SERCA1 in the myocardium of dogs diagnosed with DCM. CONCLUSION We propose that SERCA1 expression is evidence of a natural adaptive response to the impaired Ca2+ handling thought to occur in German shepherd dogs with DCM and heart failure.