Reptilian skeletal muscle: contractile properties of identified, single fast-twitch and slow fibers from the lizard Dipsosaurus dorsalis

Contractile properties and innervation patterns were determined in identified single fibers from the iliofibularis muscle of the desert iguana, Dipsosaurus dorsalis. Single fibers from both the red and white regions of the iliofibularis muscle were dissected along their length under oil and a portion was mounted on transducers for determination of maximum isometric tension (Po) and unloaded shortening velocity (Vmax) using the slack test method. Fibers were chemically skinned and activated by high Ca++. The remaining portion of the muscle fiber was mounted on a glass slide and histochemically treated to demonstrate myosin ATPase activity. Fibers studied functionally could therefore be classified as fast or slow according to their myosin ATPase activity, and they could also be classified metabolically according to the region of the muscle from which they were dissected. Fast-twitch glycolytic (FG) fibers from the white region and fast-twitch oxidative, glycolytic (FOG) and slow fibers from the red region had shortening velocities at 25 degrees C of 7.5, 4.4, and 1.5 l X s-1, respectively. Po did not differ in the three fiber types, averaging 279 kN X m-2. In a second experiment, 10 microns sections were examined every 30 microns through the proximal-most 7.5 mm of the iliofibularis muscle for motor endplates. Sections were stained to demonstrate regions of acetylcholinesterase activity. Fibers with visible endplates were classified in serial sections by histochemical treatment for myosin ATPase and succinic dehydrogenase. All slow fibers examined (n = 22) exhibited multiple endplates, averaging one every 725 microns.(ABSTRACT TRUNCATED AT 250 WORDS)     

Histochemical determination of the fiber composition of locomotory muscles in a lizard, Dipsosaurus dorsalis

A histochemical survey was done on the fiber composition of 12 different locomotory muscles in the lizard Dipsosaurus dorsalis. Three types of fibers were found in all muscles: (1) fast-twitch-glycolytic (FG); (2) fast-twitch-oxidative-glycolytic (FOG); and (3) tonic fibers. Virtually all locomotory muscles contain some tonic fibers. Most muscles have bulk white regions (containing mostly FG fibers) and distinct red, oxidative regions (with FOG and tonic fibers). These red regions are predominantly located around the joints in the hind limb muscles, and probably serve a postural and joint-stabilizing function. The predominance of FG fibers in the bulk white regions is well-correlated with the rapid, anaerobically supported predator escape behavior of D. dosalis.     

Lactate: a substrate for reptilian muscle gluconeogenesis following exhaustive exercise

Summary The pattern of lactate and glycogen metabolism in red and white muscle fibers was examined in fasted, cannulated lizards (Dipsosaurus dorsalis) run on a treadmill to exhaustion. The white and red portions of the iliofibularis (wIF, rIF) muscle of the hindlimb were analyzed post-exercise and at intervals over 120 min of recovery for lactate and glycogen changes. Five min of exercise resulted in lactate concentrations of from 35 mM (rIF) to 48 mM (wIF) while blood lactate concentrations were elevated to 21 mM from resting levels of 1.8 mM. Glycogen depletion was significant (p<0.05) in whole hindlimb (–30%) and in wIF (–42%) but not in rIF (–25%). Metabolite changes were consistent with a pattern of fiber type recruitment favoring fast-twitch glycolytic (FG) fibers during high intensity locomotion. Glycogen replenishment during recovery was fiber typespecific. After 2 h recovery, whole hindlimb glycogen concentration had increased 24% above pre-exercise levels (p<0.05). Rates of glycogen resynthesis during recovery were significant only in oxidative fibers of the red iliofibularis. Animals were infused with either [U-¹⁴C]-lactate or [U-¹⁴C]-glucose at the point of exhaustion, and label incorporation into muscle glycogen was used to estimate the substrate preference for glycogenesis during recovery. Lactate uptake and incorporation occurred in both wIF and rIF. Glucose uptake and incorporation into glycogen was greatest in the rIF, where it equalled 9% of the rate of lactate incorporation. The rate of lactate incorporation could account for 67% of the rate of glycogen synthesis that occurred in oxidative fibers of the rIF. The data indicate that in contrast to mammalian muscle, reptilian muscle replenishes glycogen while it removes lactate, utilizing lactate directly as a gluconeogenic substrate. The data also suggest that lactate produced by FG fibers during exercise is utilized by oxidative fiber types post-exercise to synthesize glycogen in excess of pre-exercise levels.Abbreviations wIF, rIF white, red portions of iliofibularis muscle - FG fast-twitch, glycolytic muscle fiber - FOG fast-twitch, oxidative, glycolytic muscle fiber - HPLC high performance liquid chromatography - SA specific activity - [LA] lactate concentration - GLU glucose - ANOVA analysis of variance - C.I. confidence interval     

Beta-adrenergic control of trans-cutaneous evaporative cooling mechanisms in birds

Summary The decreasing effect of -adrenergic blockade on skin resistance to vapor diffusion and the onset of cutaneous water evaporation in the pigeon (Columba livia) was investigated. Oral administration of 1, 2.3 and 5 mg propranolol to pigeons (268±53 g) initiated intensive trans-cutaneous water evaporation (CWE) up to 29.1 mg H₂O·cm^–2·h^–1 in resting birds at 30°C air temperature (Ta), but had only a slight effect on CWE of birds exposed to 50 °C Ta.After 7 h of effective -adrenergic blockade (oral administration of 5 mg propranolol), skin and body temperature stabilized at 39.0±0.5 °C and 41.0±0.7 °C, compared to 40.2±0.8 °C and 41.9±0.6 °C in the control group, respectively. A slight hypothermia was accompanied by feather fluffing.Intradermal injection of 0.001, 0.01 and 0.12 mg propranolol also caused intensive CWE. Local -adrenergic blockade in relatively low blocker doses (0.001 and 0.01 mg propranolol) decreased skin resistance from a high value of 44.5 s·cm^–1 to about 6.0 s·cm^–1, and caused a sharp increase in CWE from a control value of about 4 to a high of 26.4 mg H₂O·cm^–2·h^–1 during the first two hours of exposure to 30°C Ta.The possible role of -adrenergic blockade in regulation of trans-cutaneous water evaporation of latent heat dissipation is discussed.     

On the relationship of ultrastructural and cytochemical features to color in mammalian skeletal muscle

Summary The semitendinosus muscle of the albino rat is divided grossly into two clearly distinguishable parallel longitudinal bands, one red (anterior) and the other white (posterior). By using mitochondrial content as a criterion for distinguishing fiber types, it is demonstrated that the red portion of the muscle is composed predominantly of red (52%) and intermediate (40%) fibers, while the white portion consists primarily of white fibers (82%). Red fibers have the smallest and white fibers have the largest average diameter. Ultrastructural characteristics of the three fiber types resemble closely those previously described for the rat diaphragm. Red fibers are rich in large mitochondria with abundant cristae, and possess the widest Z lines. In red fibers, the H-band region of the sarcoplasmic reticulum consists of an elaborate network of narrow tubules. In white fibers, mitochondria are smaller, less numerous, and have fewer cristae; Z lines are about half as wide as in red fibers. In the H-band region of the sarcoplasmic reticulum there is a more compact arrangement of broad more or less parallel tubules. Intermediate fibers are similar to red fibers except that their diameters are larger; mitochondria are somewhat smaller and cristae are less abundant; the width of the Z lines is close to that of white fibers. The consistent difference in Z line width establishes this dimension as an important criterion for distinguishing fiber types and facilitates ultrastructural identification, especially of the intermediate fiber.The clear relationship between color of the semitendinosus and cytological features of its component fibers supports the use of the terms red, white, and intermediate as simple and valid designations for fiber types in mammalian skeletal muscle. Measurement of the cross-sectional area contributed by each fiber type to the total area indicates that both red and intermediate fibers may contribute to redness in mammalian skeletal muscle.An early portion of this work was carried out with MissSharon Whelan (Mrs.Bernard Weiss). The author acknowledges the important contribution of Mr.Richard Stearns through his skillful work on the photographic illustrations and the technical assistance of MissAnn Campbell and Mrs.Joan Normington. — This study was supported by Grant No. HD 01026-04 from the United States Public Health Service.     

Glycogen synthesis from lactate in skeletal muscle of the lizardDipsosaurus dorsalis

Summary The capacity of skeletal muscle to synthesize glycogen from lactate was tested in the iliofibularis muscle of the desert iguana,Dipsosaurus dorsalis. Like other reptiles,Dipsosaurus accumulates significant lactic acid concentrations following vigorous exercise. After 5 min of progressively faster treadmill running at 35°C (final speed=2.2 km/h), blood lactate concentration increased over 14 mM, which decreased 11 mM after 2 h of recovery. Blood glucose concentration remained unchanged throughout at 8.6±0.46 mM. The role that muscle gluconeogenesis might play in the removal of post-exercise lactate was evaluated. Animals were run to exhaustion at 1.5 km/h on a treadmill thermostatted at 35°C. Animals (n=43) ran 6.9±0.75 min prior to exhaustion. Animals were sacrificed and iliofibularis muscles of both hindlimbs removed and stimulated at 2 Hz for 5 min, reducing twitch tension to 6% of prestimulus tension. Fatigued muscles were then split into red and white fiber bundles and incubated 2 h or 5 h at 35°C in Ringer solution or in Ringer plus 20 mM lactate. In muscles tested in August, red fiber bundles incubated in lactate demonstrated a rate of glycogen synthesis of approximately 1 mg/(g muscle·h). In muscles tested in December, red fiber bundles synthesized glycogen at a reduced rate that was not statistically different than in fiber bundles incubated in Ringer solution without lactate. Glycogen synthesis from lactate was not evident in white fiber bundles in either August or December. The period of peak gluconeogenic capacity coincides with the field active season ofDipsosaurus. In vivo rates of lactate removal and in vitro rates of glycogen synthesis suggest that muscle gluconeogenesis may potentially account for 20% of the lactate removed during recovery from exhaustive activity.Abbreviations IF iliofibularis - FG fast twitch, glycolytic - FOG fast twitch, oxidative-glycolytic     

Quantitative analysis of muscle breakdown during starvation in the marine flatfish Pleuronectes platessa

Summary The present study describes the effects of starvation for a duration of four months on the ultrastructure of skeletal muscles from the marine flatfish (Pleuronectes platessa L.). Starvation is associated with a decrease in resting metabolic rate from 20.1±2.2 to 11.6±1.5mg-O₂/kg/h (P<0.05) and muscle wasting. Median fibre size fell from 700 m² to 500 m² in intermediate (fast oxidative) and from 1,800 m² to 600 m² in starved, white (fast-glycolytic) muscle fibres. In contrast, median fibre size in red (slow oxidative) muscle remained within the range 300–400 m². The fraction of red fibre volume occupied by myofibrils (58.6%) and mitochondria (24.5%) did not change significantly with starvation. There was, however, a decrease in stored lipid (10.7% to 3.2%) and an alteration in the structure of the cristae in mitochondria from red muscle.Atrophy of white muscle fibres is associated with a decrease in both the diameter and fractional volume occupied by myofibrils (85.7% to 61.9% P < 0.01). In a high proportion of white fibres peripheral degeneration of Z-discs is evident causing an unravelling of the thin filament lattice. It is suggested that this allows a partial decrease in myofibril diameter and hence the maintenance of contractile function in muscle from starved fish. In severely degenerating white fibres, disorganised thick and thin filaments and numerous multimembrane lysosome-like vesicles are observed.Starvation results in an increase in the average content of mitochondria in white fibres from 2.2 to 6.7% (P<0.01). In fed plaice mitochondria constitute less than 1% of the volume of the white fibre in 43.5% of the fibres. The proportion of white fibres containing more than 6% mitochondria increases from 6.5% to 58% with starvation.     

Tumor necrosis factor-α in macrophages of heart,liver, kidney,and in the pituitary gland

The relationship between myonuclear number, cellular size, succinate dehydrogenase activity, and myosin type was examined in single fiber segments (n=54; 9±3 mm long) mechanically dissected from soleus and plantaris muscles of adult rats. One end of each fiber segment was stained for DNA before quantitative photometric analysis of succinate dehydrogenase activity; the other end was double immunolabelled with fast and slow myosin heavy chain monoclonal antibodies. Mean±S.D. cytoplasmic volume/myonucleus ratio was higher in fast and slow plantaris fibers (112±69 vs. 34±21x10³ m³) than fast and slow soleus fibers (40±20 vs. 30±14x10³ m³), respectively. Slow fibers always had small volumes/myonucleus, regardless of fiber diameter, succinate dehydrogenase activity, or muscle of origin. In contrast, smaller diameter (<70 m) fast soleus and plantaris fibers with high succinate dehydrogenase activity appeared to have low volumes/myonucleus while larger diameter (>70 m) fast fibers with low succinate dehydrogenase activity always had large volume/myonucleus. Slow soleus fibers had significantly greater numbers of myonuclei/mm than did either fast soleus or fast plantaris fibers (116±51 vs. 55±22 and 44±23), respectively. These data suggest that the myonuclear domain is more limited in slow than fast fibers and in the fibers with a high, compared to a low, oxidative metabolic capability.     

Sexual dimorphism in histological characteristics and contractility of the iliofibularis muscle in the lizard Sceloporus torquatus

The iliofibularis is a hindlimb muscle used in lizard locomotion that is composed of at least three types of fibres: fast‐twitch‐glycolytic (FG), fast‐twitch‐oxidative‐glycolytic fibre (FOG) and slow‐twitch‐oxidative (SO). The striated skeletal muscle is a highly plastic tissue undergoing phenotypic change in response to activity. The lizard Sceloporus torquatus has sexual differences associated with microhabitat use, which can be reflected in the physiology and anatomy of the muscle, and thus, in our study, we analysed the morphological and contractile characteristics of the iliofibularis muscle (IF) of S. torquatus males and females. We found a larger prevalence of FOG compared with FG and SO fibres in the muscle of both sexes. We also found that males show larger areas of the three types of fibres, develop greater strength but also faster fatigue than females, suggesting that strength is a key functional feature that enables males to perform faster movements (but for shorter periods), associated with the demands of territoriality.     

Intermitochondrial junctions in the heart of the frog,Rana esculenta

Summary In muscle fibers of the frog heart, junctions between outer membranes of adjacent mitochondrial profiles are occasionally found. In thin sections of embedded tissue and of mitochondrial pellets, the intermitochondrial junctional space is 5.4±0.15 nm; the external leaflets of the membranes are joined by periodic structures separated from each other by 16.3±0.29 nm. There are 65.3±2 periodic structures per m of membrane measured on a section perpendicular to the junction. After cryofracture, the outer membrane is cleaved into two parts. Closely packed, parallel rows of large particles and furrows are found either on the P-, or on the E-faces. The rows of particles are 11±0.3 nm thick and are separated from each other by 16.5±0.46 nm, their density being 65±2.28 per m of the membrane. In junctional areas, rows of particles on one membrane correspond with the furrows on the other membrane. Intermitochondrial junctions appear to be real structures and not artifacts due to preparation procedures. The conditions of their occurrence are discussed.     

Mitochondrial DNA mutations and oxidative damage in skeletal muscle of patients with chronic uremia

Abundant evidence has been gathered to suggest that mitochondrial DNA (mtDNA) sustains many more mutations and greater oxidative damage than does nuclear DNA in human tissues. Uremic patients are subject to a state of enhanced oxidative stress due to excess production of oxidants and a defective antioxidant defense system. This study was conducted to investigate mtDNA mutations and oxidative damage in skeletal muscle of patients with chronic uremia. Results showed that large-scale deletions between nucleotide position (np) 7,900 and 16,300 of mtDNA occurred at a high frequency in muscle of uremic patients. Among them, the 4,977-bp deletion (mtDNA⁴⁹⁷⁷) was the most frequent and most abundant large-scale mtDNA deletion in uremic skeletal muscle. The proportion of mtDNA⁴⁹⁷⁷ was found to correlate positively with the level of 8-hydroxy 2-deoxyguanosine (8-OHdG) in the total DNA of skeletal muscle (r=0.62, p<0.05). Using long-range PCR and DNA sequencing, we identified and characterized multiple deletions of mtDNA in skeletal muscle of 16 of 19 uremic patients examined. The 8,041-bp deletion, which occurred between np 8035 and 16,075, was flanked by a 5-bp direct repeat of 5-CCCAT-3. Some of the deletions were found in more than 1 patient. On the other hand, we found that the mean 8-OHdG/10⁵ dG ratio in the total cellular DNA of muscle of uremic patients was significantly higher than that of the controls (182.7 ± 63.6 vs. 50.9 ± 21.5, p=0.05). In addition, the mean 8-OHdG/10⁵ dG ratio in muscle mtDNA of uremic patients was significantly higher than that in nuclear DNA (344.0 ± 56.9 vs. 146.3 ± 95.8, p=0.001). Moreover, we found that the average content of lipid peroxides in mitochondrial membranes of skeletal muscle of uremic patients was significantly higher than that of age-matched healthy subjects (23.76 ± 6.06 vs. 7.67 ± 0.95 nmol/mg protein; p<0.05). The average content of protein carbonyls in the mitochondrial membranes prepared from uremic skeletal muscles was significantly higher than that in normal controls (24.90 ± 4.00 vs. 14.48 ± 1.13 nmol/mg protein; p<0.05). Taken together, these findings suggest that chronic uremia leads to mtDNA mutations together with enhanced oxidative damage to DNA, lipids, and proteins of mitochondria in skeletal muscle, which may contribute to the impairment of mitochondrial bioenergetic function and to skeletal myopathy commonly seen in uremic patients.     

Histochemical and biochemical properties of flight muscle fibers in the little brown bat,Myotis lucifugus

Summary The purpose of this investigation was (1) to determine the fiber composition of pectoralis muscle of the little brown bat,Myotis lucifugus; (2) to compare the fiber composition of this muscle with two of the animal's accessory flight muscles; and (3) to study the effect of hibernation on pectoralis muscle fiber composition. Bat skeletal muscle fibers were also compared with those of white laboratory rats (Rattus norvegicus). Bat pectoralis muscles possessed exceptionally high oxidative capacities as indicated by their succinate dehydrogenase activities, but relatively low glycolytic potentials (phosphofructokinase activities). Muscle histochemistry demonstrated that fiber composition of bat pectorlis muscle was homogeneous; all fibers possessed high aerobic and low glycolytic potentials, and high myofibrillar ATPase activities indicating fast contractile properties. In contrast, accessory flight muscles possessed three distinguishable fiber types. During hibernation there was a significant decline in oxidative potential, no change in glycolytic potential, and no alteration in basic fiber composition of bat pectoralis muscle. The findings of this study suggest that pectoralis muscles ofM. lucifugus may approach the ultimate adaptation of a mammalian locomotory muscle for aerobic generation of muscular power.Abbreviations FG fast-twich glycolytic - FOG fast-twitch-oxydative-glycolytic - -GPDH -glycerophosphate dehydrogenase - LDH lactate dehydrogenase - NADH-D reduced nicotinamide adenine dinucleotide diaphorase - PFK phosphofructokinase - SDH succinate dehydrogenase - SO slowtwich-oxidative     

Freeze-fracture study of the receptor membranes in the olfactory organ of Alburnus alburnus (Teleostei)

Summary Olfactory receptor molecules are assumed to be integral membrane proteins which may be visualized on fracture faces of the membrane as intramembrane particles (IMPs). In the present study, the plasma membrane of the receptor dendrites and ciliated epithelial cells in the teleost fish Alburnus alburnus were studied by freeze-fracture electron microscopy. The IMP diameters on the membrane P-faces of both receptor dendrites and ciliated epithelial cells ranged from 5 nm to 11 nm. The average IMP densities on membrane fracture faces of the ciliated and microvillous sensory dendrites were 3130±780 for the cilia, 2070±550 for the microvilli, 2390±1190 on the knob regions and 3050±1130/m on the lateral dendrite membranes. The IMP densities on the P fracture faces of the cilia and knob regions were compared with the densities found on the lateral membranes of each individual dendrite. The ratios ranged from 0.5 to 0.96 in the case of the cilia/lateral membrane and from 0.5 to 0.90 in that of the knob/lateral membrane, indicating that, in contrast to the average densities, it is the lateral membrane which has the higher IMP densities and not the cilia. The great variations in the average IMP densities, as well as the considerable variety of the ratios, may be explained by the maturation and turnover of the olfactory sensory neurons.     

Efficiency in thin-film liquid system for <Emphasis Type="Italic">Hosta</Emphasis> micropropagation

Three varieties of Hosta (Striptease, Minuteman and Stiletto) at four densities (40, 80, 120 and 200 explants per litre) were micropropagated on semi-solid agar and a thin-film liquid system with intermittent wetting of plant tissue. The mechanics of wetting by a small wave front required a larger rectangular vessel (11 × 27=297 cm ²) compared to the common cylindrical baby food jar (18 cm ²). Plants multiplied more rapidly in the agitated thin-film system than on agar. Lower plant densities increased rates of multiplication in liquid, but had little or no effect on multiplication rate on agar. Increasing plant density lowered the overall multiplication rate, but yielded greater numbers of plants per vessel. Yield, tabulated for utilization of shelf-space in growth room, was greater at all densities in rectangular vessels of liquid than conventional jars of agar media. Increased plant density lowered the sugar residual in media following the culture cycle and liquid media had less residual sugar than agar media. A liquid medium with 50 g l^–1 sucrose was concentrated enough so that sugar depletion did not limit growth, even at the highest densities. The liquid system allows the technician to skip the step of manually spacing and orienting the freshly cut bud tissue at the transfer station. Harvesting 75–100 plants per vessel from the large rectangular vessels resulted in most efficient use of technician time. Plants from liquid and agar acclimatized to greenhouse. Increased multiplication, space utilization, sugar availability and worker efficiency was demonstrated to be greater in thin-film liquid than more conventional agar-based system.     

Fine structure and metabolic adaptation of red and white muscles in tuna

Synopsis An electron microscopic study of the red and white muscle fibres in the trunk musculature of the Kawakawa tuna (Euthynnus affinis) was carried out with a view to correlating their structure with metabolic adaptation. The red fibres which are considerably smaller in diameter (34.58 m ± 6.16 S.D.) are characterized by their high content of myoglobin, mitochondria, lipid droplets and glycogen granules. The white fibres which are relatively larger in diameter (66.03 m ± 11.59 S.D.) are characterized by their lack of myoglobin, low mitochondria) density, high content of glycogen granules and the conspicuous absence of lipid droplets. The characteristics in fine structure of the two fibre types are discussed in the light of their metabolic adaptation, the red fibres as being adapted for long term cruising movement utilizing lipid as the main source of energy and the white fibres for short bursts of activity metabolizing glycogen as the chief fuel.The tuna, with the acquisition of the counter-current heat exchange system which provides for the retention of the heat generated from high substrate oxidation in the red muscle and an efficient respiratory system, it is postulated, is well adapted for high speed sustained swimming.     

Mitochondrial regular arrangement in muscle cells: a "crystal-like" pattern

The aim of this work was to characterize quantitatively the arrangement of mitochondria in heart and skeletal muscles. We studied confocal images of mitochondria in nonfixed cardiomyocytes and fibers from soleus and white gastrocnemius muscles of adult rats. The arrangement of intermyofibrillar mitochondria was analyzed by estimating the densities of distribution of mitochondrial centers relative to each other (probability density function). In cardiomyocytes (1,820 mitochondrial centers marked), neighboring mitochondria are aligned along a rectangle, with distance between the centers equal to 1.97 ± 0.43 and 1.43 ± 0.43 µm in the longitudinal and transverse directions, respectively. In soleus (1,659 mitochondrial centers marked) and white gastrocnemius (621 pairs of mitochondria marked), mitochondria are mainly organized in pairs at the I-band level. Because of this organization, there are two distances characterizing mitochondrial distribution in the longitudinal direction in these muscles. The distance between mitochondrial centers in the longitudinal direction within the same I band is 0.91 ± 0.11 and 0.61 ± 0.07 µm in soleus and white gastrocnemius, respectively. The distance between mitochondrial centers in different I bands is 3.7 and 3.3 µm in soleus and gastrocnemius, respectively. In the transverse direction, the mitochondria are packed considerably closer to each other in soleus than in white gastrocnemius, with the distance equal to 0.75 ± 0.22 µm in soleus and 1.09 ± 0.41 µm in gastrocnemius. Our results show that intermyofibrillar mitochondria are arranged in a highly ordered crystal-like pattern in a muscle-specific manner with relatively small deviation in the distances between neighboring mitochondria. This is consistent with the concept of the unitary nature of the organization of the muscle energy metabolism. confocal microscopy; quantitative analysis; cardiac and skeletal muscles; probability density function; unitary structure of cells     

8-Oxoguanosine and uracil repair of nuclear and mitochondrial DNA in red and white skeletal muscle of exercise-trained old rats.

Oxoguanine DNA glycosylase (OGG1) and uracil DNA glycosylase (UDG) are two of the most important repair enzymes that are involved in the base excision repair processes to eliminate oxidative damage from mammalian DNA, which accumulates with aging. Red and white skeletal muscle fibers have very different antioxidant enzyme activities and resistance to oxidative stress. In this paper, we demonstrate that the activity of OGG1 is significantly higher in the red type of skeletal muscle compared with white fibers from old rats. Exercise training resulted in increased OGG1 activity in the nuclei of red fibers and decreased activity in nuclei of white fibers and in the mitochondria of both red and white fibers. The activities of UDG were similar in both red and white muscle fibers. Exercise training appears to increase the activity of UDG in the nuclei and mitochondria. However, exercise training affects the activity of OGG1 in nuclei and mitochondria differently, suggesting different regulation of the enzymes. In contrast, UDG showed similar activities in nuclei and mitochondrial extracts of exercise-trained animals. These data provide evidence for differential regulation of UDG and OGG1 in maintaining fidelity of DNA in oxidatively stressed cells.     

Excitation-calcium release uncoupling in aged single human skeletal muscle fibers

The biological mechanisms underlying decline in muscle power and fatigue with age are not completely understood. The contribution of alterations in the excitation-calcium release coupling in single muscle fibers was explored in this work. Single muscle fibers were voltage-clamped using the double Vaseline gap technique. The samples were obtained by needle biopsy of the vastus lateralis (quadriceps) from 9 young (25–35 years; 25.9 ± 9.1; 5 female and 4 male) and 11 old subjects (65–75 years; 70.5 ± 2.3; 6 f, 5 m). Data were obtained from 36 and 39 fibers from young and old subjects, respectively. Subjects included in this study had similar physical activity. Denervated and slow-twitch muscle fibers were excluded from this study. A significant reduction of maximum charge movement (Q_max) and DHP-sensitive Ca current were recorded in muscle fibers from the 65–75 group. Q_max values were 7.6 ± 0.9 and 3.2 ± 0.3 nC/F for young and old muscle fibers, respectively (P < 0.01). No evidences of charge inactivation or interconversion (charge 1 to charge 2) were found. The peak Ca current was (–)4.7 ± 0.08 and (–)2.15 ± 0.11 A/F for young and old fibers, respectively (P < 0.01). The peak calcium transient studied with mag-fura-2 (400 m) was 6.3 ± 0.4 m and 4.2 ± 0.3 m for young and old muscle fibers, respectively. Caffeine (0.5 mm) induced potentiation of the peak calcium transient in both groups. The decrease in the voltage-/ Ca-dependent Ca release ratio in old fibers (0.18 ± 0.02) compared to young fibers (0.47 ± 0.03) (P < 0.01), was recorded in the absence of sarcoplasmic reticulum calcium depletion. These data support a significant reduction of the amount of Ca available for triggering mechanical responses in aged skeletal muscle and, the reduction of Ca release is due to DHPR-ryanodine receptor uncoupling in fast-twitch fibers. These alterations can account, at least partially for the skeletal muscle function impairment associated with aging.This work was supported by Grant-in-Aid from the American Heart Association (National) and Muscular Dystrophy Association, and National Institutes of Health (2-P60AG18484-06)     

Qualitative and quantitative freeze-fracture studies on olfactory and respiratory epithelial surfaces of frog,ox, rat,and dog

Summary A comparison of the necklaces of sensory olfactory, and non-sensory nasal respiratory cilia of four vertebrate species (frog, ox, rat and dog) shows that the olfactory cilia have 7±1 (mean±standard deviation) strands in the three mammalian species and 6±1 strands in the frog; for the respiratory cilia these values are 5±1 and 4±1. This function- and species-dependency of ciliary necklace strand numbers is supported by a review of the literature. Necklaces show no other structural differences. Necklace strand densities range from 25–33 strands/m. In both sensory and non-sensory cilia ciliogenesis is preceded by the formation of necklace strands. Sometimes cilia do not develop properly, as demonstrated by the presence of necklace-like structures in the membranes of olfactory dendritic endings and respiratory axonemal aggregates.     

DNA buoyant densities ofTrypanosoma (Schizotrypanum) species from bats in Ontario,Canada

Summary Two species of trypanosomes,Trypanosoma hedricki andT. myoti, isolated respectively fromEptesicus fuscus andMyotis lucifugus, were cloned.T. hedricki produced long trypomastigotesin vitro (mean length 22.1±0.3 m); the buoyant densities of its kinetoplastic and nuclear deoxyribonucleic acid (kDNA and nDNA) were respectively 1.700 and 1.710 g ml^–1 and it possessed a small amount of a satellite component (C-DNA) of density 1.716 g ml^–1. This species is in this respect closely related toT. dionisii. T. myoti produced long trypomastigotesin vitro (mean length 20.1±0.2 m); its DNA buoyant densities (g ml^–1) were: kDNA 1.700 and nDNA 1.710 (no C-DNA was detected). In this respect, therefore,T. myoti is closer toT. cruzi than the other bat trypanosomes. ac]19830129