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Biomedical subjects

J Kucera

Publications and source records attributed to J Kucera.

At least 91 records · Page 5Linked to original sources
PubMed1993

Differential effects of neonatal denervation on intrafusal muscle fibers in the rat.

The response of developing muscle spindles to denervation was studied by sectioning the nerve to the medial gastrocnemius muscle of rats at birth. The denervated spindles were examined daily throughout the first postnatal week for changes in ultrastructure and expression of several isoforms of myosin heavy chain (MHC). Each of the three different types of intrafusal muscle fiber exhibited a different response to denervation. Within 5 days after the nerve section nuclear bag2 fibers degenerated completely; nuclear bag1 fibers persisted, but ceased to express the 'spindle-specific' slow-tonic MHC isoform and thereby could not be differentiated from extrafusal fibers; nuclear chain fibers did not form. The capsules of spindles disassembled, hence spindles or their remnants could no longer be identified 1 week after denervation. Neonatal deefferentation has little effect on these features of developing spindles, so removal of afferent innervation is presumably the factor that induces the loss of spindles in denervated muscles. Degeneration of the bag2 fiber, but not bag1 or extrafusal fibers, reflects a greater dependence of the bag2 fiber than the bag1 fiber on afferent innervation for maintenance of its structural integrity. This difference in response of the two types of immature bag fiber to denervation might reflect an origin of the bag2 fibers from a lineage of myogenic cells distinct from that giving rise to bag1 or extrafusal fibers, or a difference in the length of contact with afferents between the two types of bag fiber prior to nerve section.

Animals↗
PubMed1993

Transient expression of a slow-tonic MHC isoform by extrafusal fibers in the developing rat.

ALD 19, a monoclonal antibody that recognizes the slow-tonic myosin heavy chain (MHC) isoform, has been used extensively as a marker for nuclear bag intrafusal fibers of muscle spindles in developing and adult rats. Extrafusal fibers of adult rat hindlimb muscles do not express slow-tonic MHC. However, while using ALD 19 to trace the fate of intrafusal fibers following neonatal denervation, we noted that some extrafusal fibers of neonates also bound this antibody. The immunolabeled extrafusal fibers were a subset of slow fibers located in the deep axial regions of crural muscles. The same fiber subset transiently displayed a weak affinity for ALD 19 during the first postnatal week in normal muscles. Denervation at birth increased the intensity of ALD 19 immunolabelling by these extrafusal fibers and extended the duration of the slow-tonic immunoreactivity into the 2nd postnatal week, after which expression diminished or ceased. Demonstration that some developing extrafusal fibers have a nerve-independent capacity for transiently expressing slow-tonic MHC, an MHC previously though to be expressed only by intrafusal fibers, raises the possibility that both types of fiber originate from a subset of bipotential slow primary myotubes in rat hindlimbs.

Animals↗
PubMed1993

Fusimotor-free spindles in reinnervated muscles of neonatal rats treated with nerve growth factor.

Crushing the nerve to the medial gastrocnemius muscle in newborn rats and administering nerve growth factor afterwards results in a reinnervated muscle containing supernumerary muscle spindles. The structure and innervation of 88 spindles in the reinnervated muscles were reconstructed from serial thick and thin transverse sections at 30-35 days after the nerve crush, and compared to those of five control spindles. The spindles consisted of one to four small-diameter encapsulated fibers with features of nuclear chain intrafusal fibers, or infrequently a nuclear bag intrafusal fiber. Some of the spindles were located within a capsule that also contained an extrafusal fiber. Each spindle was innervated by an afferent with features of the primary afferent. The density of secondary afferents was lower in reinnervated muscles than in controls. Endplates were observed on extrafusal fibers in the experimental muscles, attesting to restoration of skeletomotor (alpha) innervation after the nerve crush. However, 78% of the experimental spindles were entirely devoid of efferent innervation. The remainder received either one or two fusimotor (gamma) axons or a skeletofusimotor (beta) axon, compared to the six to eight motor axons that innervated control spindles. The presence of supernumerary spindles composed of fibers that resemble normal intrafusal fibers in the absence of motor innervation suggests that afferents alone can induce the formation and subsequent differentiation of intrafusal fibers in nerve-crushed muscles of neonatal rats. In addition, the paucity of gamma innervation in nerve-crushed muscles suggests that immature gamma neurons are more susceptible than spindle afferents or alpha efferents to cell death after axotomy at birth.

Animals↗
PubMed1993

Influence of muscle cell substrates on differentiation of intrafusal fiber types in neonatal rats.

Rat muscle spindles contain one nuclear bag2, one nuclear bag1, and two nuclear chain fibers. The three different types of intrafusal fiber in spindles may be a reflection of concomitant changes in proportions of slow primary, slow/fast secondary, and fast secondary myotubes during the period of spindle development. We examined whether experimentally altering the available muscle substrates would impact the intrafusal fiber type composition of spindles. De novo formation of spindles in muscles devoid of primary myotubes was induced by crushing the nerve to the medial gastrocnemius muscle in newborn rats and administering nerve growth factor for ten days afterwards. Encapsulated fibers of the reinnervated muscles examined one month after nerve crush had myofibrillar adenosine triphosphatase and myosin heavy chain profiles similar to normal bag2, bag1, or chain intrafusal fibers. However, spindles in reinnervated muscles contained fewer fibers than controls. Most experimental spindles contained chain and/or bag1 fibers, the two fiber types which ordinarily arise during secondary myogenesis. In contrast, bag2 fibers, fibers that normally form concomitant with primary myogenesis, were absent from nearly 90% of spindles in reinnervated muscles. The paucity of bag2 fibers may reflect the absence of primary myotubes, whereas the prevalence of chain and/or bag1 fibers may reflect that secondary myotubes or myofibers that descended from the secondary myotubes were the principal muscle substrates available for spindle formation in the nerve-crushed muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗
PubMed1993

Rapid response of large, drought-stressed beech trees to irrigation.

Large, declining beech (Fagus sylvatica L.) trees (diameter at breast height = 50 cm), growing on heavy clay soils in the highlands near Zurich, Switzerland, were amply irrigated in late summer. During irrigation, the xylem sap flow rate, Q(wt), was measured by the stem-tissue heat balance method with internal heating and sensing. Only a gradual and slight increase in Q(wt) in response to irrigation was observed in the control trees, whereas Q(wt) in the declining trees, whose transpiration rates were only 2-20% those of the control trees, increased 2-5 times within minutes. This suggests, that severe local drought was the major factor limiting tree growth at the site. The extent of the response permits estimation of the supply-limited (soil water) and demand-limited (tree structure) components of stress. Drought caused a decline in Q(wt) in the trees with short crowns and limited root systems that had originally been growing in dense canopies and had become suddenly exposed to full illumination as a result of a severe wind storm and thinning. Trees with deep, narrow, dense crowns, growing in more open places and adapted over a long period to high irradiance remained healthy during drought. Prolonged, periodic water shortage reduced the amount of foliage up to 90% but during drought stimulated the growth of fine roots in the surface and upper soil layers. The stem conductive systems of the declining trees were still partially functional.

Journal Article↗
PubMed1992

Formation of muscle spindles in the absence of motor innervation.

Whether muscle spindles can form in muscles innervated only by afferents was investigated by removing the lumbosacral segment of the spinal cord immediately after crushing the nerve to the medial gastrocnemius (MG) muscle in newborn rats, and administering nerve growth factor for 10 days afterwards. The nerve-crushed MG muscles reinnervated by afferents in the absence of motor innervation were examined at postnatal (P) days 7, 9 and 30 for the presence of spindles by light and electron microscope. Reinnervated MG muscles contained spindle-like encapsulations of 1-4 fibers at 7, 9 and 30 days after the nerve crush. The number of spindles exceeded that of normal MG muscles, suggestive of de novo formation of spindles. All nerve-muscle contacts in the spindles had features of sensory endings, and intrafusal fibers expressed the spindle-specific slow-tonic myosin heavy chain (MHC) isoform at P30. No motor endplates were visible on any muscle fibers and extrafusal fibers were atrophied, as would be predicted in the absence of motor innervation. Thus, efferents are not essential for the formation and differentiation of muscle spindles in reinnervated muscles of neonatal rats.

Animals↗
PubMed1992

Vanadium levels in hair and blood of normal and exposed persons.

Vanadium was determined by both instrumental neutron activation analysis (INAA) and NAA with radiochemical separation (RNAA) in hair of normal children and of children potentially exposed by accidental drinking of vanadium contaminated water (long-term, low-dose exposure). Vanadium hair levels in the two groups did not differ significantly and were in the range 46-313 micrograms/kg (median 98 micrograms/kg) and 24-235 micrograms/kg (median 88 micrograms/kg for the normal and exposed groups, respectively. Using RNAA with proven reliability at the ultratrace level, vanadium was determined in whole blood of the exposed and normal children, normal adults and workers professionally exposed to vanadium in a factory producing vanadium pentoxide. Significantly increased vanadium concentrations were found in blood of exposed children (range 0.018-0.239 micrograms/l, median 0.078 micrograms/l) compared to normal children (range 0.024-0.226 micrograms/l, median 0.042 micrograms/l), while no differences could be detected between blood vanadium levels of normal children and normal adults (range 0.032-0.095 micrograms/l, median 0.056 micrograms/l). Preliminary results for vanadium in blood of occupationally highly exposed persons showed values 2-4 orders of magnitude higher than in normal adults.

Adult↗
PubMed1992

Axotomy induces fusimotor-free muscle spindles in neonatal rats.

Crushing the nerve to the medial gastrocnemius (MG) muscle at birth and administering nerve growth factor to rats afterwards results in a reinnervated muscle with supernumerary muscle spindles, some of which must have formed de novo. Structure and innervation of spindles in the reinnervated MG muscles were studied in serial 1 micron transverse sections. Two types of spindle-like encapsulations were observed. The prevalent type consisted of one to three small diameter intrafusal fibers with features of nuclear chain fibers or infrequently a nuclear bag fiber. The second type of encapsulation consisted of the small-diameter fibers located in a compartment which abutted a compartment containing a large diameter extrafusal fiber. All intrafusal fibers in spindles of the experimental muscles were innervated by afferents, but most of them (85%) were devoid of efferent innervation. Thus, immature fusimotor neurons may be more susceptible than spindle afferents to cell death after axotomy at birth.

Animals↗
PubMed1992

Superfluousness of motor innervation for the formation of muscle spindles in neonatal rats.

Muscle spindles form de novo in reinnervated muscles of neonatal rats treated with nerve growth factor. Whether the spindles can also form in muscle reinnervated only by afferents was investigated by removing the lumbosacral segment of the spinal cord immediately after crushing the nerve to the medial gastrocnemius muscle at birth, and administering nerve growth factor for 10 days afterwards. As predicted, the medial gastrocnemius muscles were reinnervated by afferents, but not efferents. No motor endplates were visible on any muscle fibers, and extrafusal fibers were atrophied. The reinnervated muscles contained spindle-like encapsulations of one to four fibers at 5, 7, 9 and 30 days after the nerve crush. The number of spindles as well as encapsulated fibers exceeded that of normal medial gastrocnemius muscles. The encapsulated fibers resembled typical intrafusal fibers. They had normal sensory-muscle contacts, but no motor endings. The fibers displayed equatorial clusters of myonuclei and expressed the spindle-specific slow-tonic myosin heavy chain isoform at postnatal day 30. Thus, efferents are not essential for the formation and differentiation of muscle spindles in reinnervated muscles of neonatal rats.

Animals↗
PubMed1992

Radial velocity profiles of water flow in trunks of Norway spruce and oak and the response of spruce to severing.

Trunk-tissue heat balance, volumetric and staining methods were used to study xylem water flow rates and pathways in mature Norway spruce (Picea abies (L.) Karst.) and pedunculate oak (Quercus robur L.) trees. The radial profile of flow velocity was confirmed to be symmetrical in spruce, i.e., maximum flow velocity was in the center of the conducting xylem and tailed with low amplitude (about 30 cm h(-1)) in the direction of the cambium and heartwood. Variability around the trunk was high. In contrast, in oak, the radial profile of flow velocity was highly asymmetrical, reaching a peak of about 45 m h(-1) in the youngest growth ring and tailing centripetally for about 10 rings, but variability around the trunk was less, under non-limiting soil water conditions, than in spruce. In spruce, the flow rate increased abruptly within seconds when the tree was severed while immersed in water, and then decreased gradually, showing significant root resistance. We conclude that water flow through an absorbing cut surface differs from the flow higher in a tree trunk because of the presence of hydraulic capacitances in the conductive pathways. The staining technique always yielded higher estimates of flow velocity than the non-destructive tree-trunk heat balance method.

Journal Article↗
PubMed1992

Expression of type-specific MHC isoforms in rat intrafusal muscle fibers.

Myosin heavy chain (MHC) expression by intrafusal fibers was studied by immunocytochemistry to determine how closely it parallels MHC expression by extrafusal fibers in the soleus and tibialis anterior muscles of the rat. Among the MHC isoforms expressed in extrafusal fibers, only the slow-twitch MHC of Type 1 extrafusal fibers was expressed along much of the fibers. Monoclonal antibodies (MAb) specific for this MHC bound to the entire length of bag2 fibers and the extracapsular region of bag1 fibers. The fast-twitch MHC isoform strongly expressed by bag2 and chain fibers had an epitope not recognized by MAb to the MHC isoforms characteristic of developing muscle fibers or the three subtypes (2A, 2B, 2X) of Type 2 extrafusal fibers. Therefore, intrafusal fibers may express a fast-twitch MHC that is not expressed by extrafusal fibers. Unlike extrafusal fibers, all three intrafusal fiber types bound MAb generated against mammalian heart and chicken limb muscles. The similarity of the fast-twitch MHC of bag2 and chain fibers and the slow-tonic MHC of bag1 and bag2 fibers to the MHC isoforms expressed in avian extrafusal fibers suggests that phylogenetically primitive MHCs might persist in intrafusal fibers. Data are discussed relative to the origin and regional regulation of MHC isoforms in intrafusal and extrafusal fibers of rat hindlimb muscles.

Aging↗
PubMed1992

Morphometric analysis of coated vesicles in developing rat muscle spindles.

The incidence of coated vesicles under sarcolemmal surfaces of equatorial, juxta-equatorial and polar regions in developing and adult spindles of the rat soleus muscle was examined by quantitative morphometry of transverse ultrathin sections. Coated vesicles were more numerous: 1) under primary sensory endings than under other types of neuromuscular contacts; 2) under the appositional sarcolemma between neighbouring intrafusal fibres than under free surfaces of the sarcolemma; and 3) in developing than in mature spindles. Factors such as location and age of the animal often interacted to produce an additive effect on the incidence of coated vesicles. Although there was a high incidence of coated vesicles at the postsynaptic surface under sensory terminals of bag2 fibres in 18 and 19 day gestational embryonic rats, it peaked in 4 day postnatal animals. The high incidence of coated vesicles at sensory endings supports the view that coated vesicles mediate neurotrophic interactions between afferents and intrafusal fibres during the critical late gestation and early postnatal time period, as sensory axons first contact their target fibres and exert a maximal directing influence on the differentiation of intrafusal fibre types. In addition, the preferential localization of coated vesicles under appositional rather than free surfaces of developing intrafusal fibres in 0-4 day rats suggests that they play a role in the transport of active substances among intrafusal fibres exhibiting different stages of maturity.

Aging↗
PubMed1991

Non-neural and neural expression of myosin heavy chains by regenerated intrafusal fibers of rats.

Expression of myosin heavy chain (MHC) isoforms was studied in rat soleus (SOL) and extensor digitorum longus (EDL) muscles which regenerated in the presence or absence of innervation. Frozen sections of two 5 day denervated SOL and EDL grafts, two 40 day denervated SOL and EDL grafts, and two reinnervated 40 day SOL and EDL grafts were processed for demonstration of motor endplates, sensory endings, myosin adenosine triphosphatase (mATPase) and for expression of 4 MHCs. No qualitative differences in MHC expression were noted between 5 day or 40 day denervated grafts of the SOL and EDL muscles. All regenerated intrafusal and extrafusal myotubes or myofibers reacted to antibodies against neonatal and fast-twitch MHCs, but not to antibodies against slow-twitch and slow-tonic MHCs in these grafts. These data indicate that MHCs expressed by regenerated intrafusal myotubes do not parallel those expressed by myotubes which give rise to the three types of intrafusal fibers during development and that MHC expression by regenerated intrafusal myotubes parallels that of regenerated extrafusal myotubes prior to innervation. However, some regenerated intrafusal fibers in 40 day nerve-intact grafts bound antibodies to slow-twitch and slow-tonic MHCs, thus expressions of these two MHCs are nerve-dependent in regenerated muscle spindles.

Animals↗
PubMed1991

Neural organization of spindles in three hindlimb muscles of the rat.

The neuroanatomical organization of the dynamic (bag1) and static (bag2 and chain) intrafusal systems was compared by light and electron microscopy of serial sections among 71 poles of muscle spindle in soleus (SOL), extensor digitorum longus (EDL), and lumbrical (LUM) muscles in the rat. Eighty-four percent of 195 fusimotor (gamma) axons to the spindles innervated either the dynamic bag1 fiber or the static bag2 and/or chain fibers. Sixteen percent of the gamma axons coinnervated the dynamic and static intrafusal fibers. Some of these nonselective axons were branches of effernts that also gave rise to axons selective to either the dynamic or static types of intrafusal fibers in one or more spindles. Thus activation of individual stem gamma efferents might not have a purely dynamic or purely static effect on the integrated afferent outflow from spindles of a hindlimb muscles in the rat. In addition, primary afferents in all muscles had terminations that cross-innervated the dynamic bag1 and static bag1 and/or chain intrafusal fibers in individual spindles, an arrangement that may enhance the mixed dynamic/static behavior of afferents when different intrafusal fibers are activated concurrent. Spindles of the slow SOL and fast EDL muscles had similar features, whereas differences were observed in the organization of the proximal (SOL and EDL) and distal (LUM) muscles. Spindles in LUM muscles had fewer static intrafusal fibers, a higher ratio of dynamic to static gamma axons, and a higher incidence of skeletofusimotor (beta) innervation to intrafusal fibers than spindles in the SOL or EDL muscles. Thus, the relative contribution of dynamic and static systems to muscle afferent outflow may differ among spindles located in different segments of the rat hindlimb. However, the dynamic and static intrafusal systems of spindle were less sharply demarcated in each of the three hindlimb rat muscles than in the cat tenuissimus muscle.

Animals↗
PubMed1991

Slow-tonic MHC expression in paralyzed hindlimbs of fetal rats.

Whether nerve activity and active contraction of myotubes are essential for the assembly and initial differentiation of muscle spindles was investigated by paralyzing fetal rats with tetrodotoxin (TTX) from embryonic day 16 (E16) to E21, prior to and during the period when spindles typically form. TTX-treated soleus muscles were examined by light and electron microscopy for the presence of spindles and expression of myosin heavy chain (MHC) isoforms by the intrafusal fibers. Treatment with TTX did not inhibit the formation of a spindle capsule or the expression of a slow-tonic MHC isoform characteristic of intrafusal fibers, but did retard development of spindles. Spindles of TTX-treated E21 muscles usually consisted of one intrafusal fiber (bag2) only rather than two fibers (bag1 and bag2) typically present in untreated (control) E21 spindles. Intrafusal fibers of TTX-treated spindles also had only one sensory region supplied by multiple afferents, and were devoid of motor innervation. These features are characteristic of spindles in normal E18-E19 muscles. Thus, nerve and/or muscle activity is not essential for the assembly of muscle spindles, formation of a spindle capsule, and transformation of undifferentiated myotubes into the intrafusal fibers containing spindle-specific myosin isoforms. However, activity may promote the maturation of intrafusal bundles, as well as the maturation of afferent and efferent nerve supplies to intrafusal fibers.

Animals↗
PubMed1991

Aggregation of myonuclei and the spread of slow-tonic myosin immunoreactivity in developing muscle spindles.

The pattern of regional expression of a slow-tonic myosin heavy chain (MHC) isoform was studied in developing rat soleus intrafusal muscle fibers. Binding of the slow-tonic antibody (ATO) began at the equator of prenatal intrafusal fibers where sensory nerve endings are located, and spread into the polar regions of nuclear bag2 and bag1 fibers but not nuclear chain fibers during ontogeny. The onset of the ATO reactivity coincided with the appearance of equatorial clusters of myonuclei (nuclear bag formations) in bag1 and bag2 fibers. Moreover, the intensity of the ATO reaction was strongest in the region of equatorial myonuclei and decreased with increasing distance from the equator of bag1 and bag2 fibers at all stages of prenatal and postnatal development. The polar expansion of ATO reactivity continued throughout the postnatal development of bag1 fibers, but ceased shortly after birth in bag2 fiber coincident with innervation by motor axons. Thus, afferents that innervate the equator might induce the slow-tonic MHC isoform in bag2 and bag1 fibers by regulating the myosin gene expression by equatorial myonuclei, and efferents or twitch contractile activity might inhibit the spread of the slow-tonic MHC isoform into the poles of bag2 but not bag1 fibers. Absence of ATO binding in chain fibers suggests that chain myotubes may not be as susceptible to the effect of afferents as are myotubes that develop into bag2 and bag1 fibers. The different patterns of slow-tonic MHC expression in the three types of intrafusal fiber may therefore result from the interaction of three elements: sensory neurons, motor neurons, and intrafusal myotubes.

Aging↗
PubMed1991

Enzyme variants of Eimeria parasitizing the domestic fowl and possibilities of species diagnostics.

Electrophoretic variation of the enzymes lactate dehydrogenase (LDH) and glucosephosphate isomerase (GPI) of Eimeria parasitizing the domestic fowl in Czechoslovakia is summarized and the differentiation of species of poultry coccidia is discussed. A new method for evaluation of zymograms of coccidial enzymes is presented. This method enables the results of different experiments to be compared by calculating standardized rates of mobility of each enzyme band relative to the positions of reference variants coded LDH-8 or GPI-9.

Animals↗