Lactate sensitive cells in newt kidneys

Summary The blood supply of duck muscle spindles from the extensor pollicis and the extensor digitorum comminis wing muscles has been studied by light and electron microscopy. Capillaries usually accompany the nerve bundle that innervates the spindle, approaching at an oblique angle around the midequatorial region. Capillaries may run for some distance along the surface of the outer capsule, or penetrate partially into the outer capsule and lie between layers of the capsule cell processes. Some capillaries also penetrate the outer capsule, running into the periaxial space and continuing further towards the polar region. They have been shown to be in close contact with intrafusal muscle fibres, from the juxta-equatorial to the polar region, but have not been encountered among sensory terminals in the mid-equatorial region.     

The affinity of concanavalin A and wheat germ agglutinin for components of the muscle spindle.

Sections through the soleus muscle of the rat were incubated with concanavalin A (Con A) or wheat germ agglutinin (WGA) conjugated to fluorescein isothiocyanate. Binding of these lectins to structures which comprise the muscle spindle was studied by fluorescence microscopy. The distribution of the lectins was heaviest in the outer capsule of the spindle and at the surface of intrafusal muscle fibres. The periaxial space in the equatorial region of spindles was unlabelled except in the immediate vicinity of the axial bundle. Binding by Con A was more extensive than by WGA, suggesting that more glucopyranoside units are accessible within the muscle spindle than are those of N-acetylglucosamine.     

The affinity of Concanavalin A and wheat germ agglutinin for components of the muscle spindle

Summary Sections through the soleus muscle of the rat were incubated with concanavalin A (Con A) or wheat germ agglutinin (WGA) conjugated to fluorescein isothiocyanate. Binding of these lectins to structures which comprise the muscle spindle was studied by fluorescence microscopy. The distribution of the lectins was heaviest in the outer capsule of the spindle and at the surface of intrafusal muscle fibres. The periaxial space in the equatorial region of spindles was unlabelled except in the immediate vicinity of the axial bundle. Binding by Con A was more extensive than by WGA, suggesting that more glucopyranoside units are accessible within the muscle spindle than are those of N-acetylglucosamine.     

Fine structure of the avian muscle spindle capsule

Summary The capsule of the muscle spindle from the anterior and posterior latissimus dorsi muscles of the adult domestic chicken has been studied with the electron microscope. As in other species, two distinct portions of the spindle capsule are distinguished: an outer capsule and an inner capsule.The outer capsule is structurally similar to and continuous with the perineural epithelium. Outer capsule cells are noted by the abundance of pinocytotic vesicles and a network of 6–7 nm microfilaments. The disposition of these microfilaments is circumferential with respect to the longitudinal axis of the spindle. It is proposed that they may provide a contractile mechanism for the capsule which may be related to the over-all functioning of the spindle during movements of the muscle.The inner capsule is composed of a contiguous network of cells possessing long cytoplasmic processes which envelop the intrafusal fibers and their nerve endings in sensory equatorial regions of the spindle. These cells may elaborate the fibrillar and amorphous extracellular material found in the periaxial spindle space. They also possess modified cilia with a 9+0 microtubular pattern. It is suggested that these cilia may behave as sensory transducers, relating fluid changes in the periaxial space to the intrafusal fiber nerve endings.Capillaries of the non-fenestrated variety commonly traverse the outer and inner portions of the capsule and are usually completely surrounded by tenuous overlapping processes of inner capsule cells. These findings suggest that the spindle capsule plays a role as a metabolically-active diffusion barrier to the entrance of substances from the external milieu.The excellent technical assistance of Ms. S.L. Shinn is gratefully acknowledged.Supported by grants from the Medical Research Council of Canada and the Muscular Dystrophy Association of Canada.     

The Fine Structure of Muscle Spindles in the Lumbrical Muscles of the Rat

Lumbrical muscles of young rats were fixed with OsO₄ and embedded in methacrylate for electron microscopy. The spindle capsule was found to be continuous with and similar in structure to the sheath of Henle surrounding the nerves supplying the spindle. The capsule consists of several closely applied concentric cytoplasmic sheets. Each sheet is about 1,000 A thick and has no fenestrations. Many caveolae and vesicles in the cytoplasm suggest active transport through the sheets. The periaxial space fluid contains much solid material. It is suggested that the capsule and periaxial space regulate internal chemical environment. The interfibrillar structures are less evident in the polar regions of intrafusal fibres than in extrafusal fibres. Simple motor end-plates occur on the polar regions of intrafusal fibres. In the myotube region of the intrafusal fibre a peripheral zone of myofibrils surrounds a cytoplasmic core containing nuclei, mitochondria, Golgi bodies, reticulum, and a few lipid-like granules. Naked sensory endings lie on the myotube "in parallel" with the underlying myofilaments. Naked processes of the primary sensory ending deeply indent the muscle plasma membrane and the underlying wisps of myofilament in the nuclear bag region. The plasma membranes of sensory nerve ending and intrafusal muscle fibre are about 200 A apart.     

Hyaluronan in human and rat muscle spindles

 Previous data on the composition of the periaxial fluid of muscle spindles have relied on indirect histochemical methods. We used a biotinylated hyaluronan-binding protein as a specific probe for the detection of hyaluronan in sections of human and rat limb muscles. Hyaluronan was identified in the axial and periaxial space of the muscle spindles as well as in the endoneurium and in the space in between individual axons. Hyaluronan was also present in the innermost layer of the spindle capsule in the A region and in all layers of the capsule in the B region. Accepted: 12 February 1998     

Immunohistochemical localization of alpha(1a)-adrenoreceptors in muscle spindles of rabbit masseter muscle

The expression of alpha(1a)-adrenoreceptors (alpha(1a)-ARs) within the muscle spindles of rabbit masseter muscle was investigated. The alpha(1a)-ARs were detected by immunohistochemical fluorescent method and examined along the entire length of 109 cross serially sectioned spindles. The sympathetic fibers were visualized by the immunofluorescent labeling of the noradrenaline synthesizing enzymes tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH). In order to recognize the intrafusal muscle fiber types, antibodies for different myosin heavy chain isoforms (MyHCI) were used. TH and DBH immunolabeled nerve fibers have been observed within the capsule lamellar layers, in the periaxial fluid space and close to intrafusal muscle fibers. The alpha(1a)-ARs were detected on the smooth muscle cells of the blood vessels coursing in the muscle and in the capsule lamellar layers or within the periaxial fluid space of the spindles. Moreover, at the polar regions of a high percentage (88.1%) of muscle spindles a strong alpha(1a)-ARs immunoreactivity was present on the intrafusal muscle fibers. In double immunostained sections for alpha(1a)-ARs and MyHCI it was evidenced that both bag, and nuclear chain fibers express alpha(1a)-ARs. The receptors that we have detected by immunofluorescence may support a direct control by adrenergic fibers on muscle spindle.     

Distribution of connective tissue proteins in chick muscle spindles as revealed by monoclonal antibodies: a unique distribution of brachionectin/tenascin

The distribution of chick muscle spindles of eight connective tissue proteins (collagen types I, IV, V, and VI, laminin, heparan sulfate, fibronectin, and brachionectin/tenascin) was examined by immunofluorescent histochemistry. Intrafusal fibers were surrounded by layers of collagen type VI and fibronectin, and by an external lamina containing collagen type IV, laminin, and heparan sulfate. Most of these layers displayed a different pattern of staining at the sensory region of the equator than at the polar region. The crescent-like sheath that caps each intrafusal fiber and sensory terminal at the equator was strongly positive for collagen type I and weakly positive for collagen type V. The outer spindle capsule contained laminin, heparan sulfate, collagen types IV and VI, brachionectin/tenascin, fibronectin, and to a lesser degree also collagen types I and V. Brachionectin/tenascin had the narrowest distribution of any of the connective tissue macromolecules studied. It was found only in the outer capsule and in the coverings of blood vessels and nerves associated with the outer capsule.     

Extracellular matrix and transmembrane linkages at the termination of intrafusal fibers and the outer capsule in chicken muscle spindles

Attachments of intrafusal fibers and of the outer spindle capsule at the far polar region were examined by immunohistochemistry in serially sectioned chicken leg muscles. Patterns of distribution of connective tissues and intracellular filaments suggest that, in this segment of the muscle spindle, intrafusal fibers bind laterally with the capsule. Contrary to extrafusal fibers at myotendinous junctions, folded plasmalemmas at the ends of intrafusal fibers were rare. Thus, there was little end-to-end interlocking between intrafusal fibers and the extracellular matrix. The tapered contours of terminating intrafusal fibers resembled those of extrafusal fibers which end in fascicles without tendinous connections. At points where the distal portions of intrafusal fibers closely adjoined and overlapped extrafusal fibers, α-actinin, vinculin, filamin, talin, β₁ integrin, spectrin, and dystrophin occurred with moderate to great frequency. It is generally accepted that these compounds are links in molecular chains that extend from the intracellular space across cell membranes to the extracellular matrix. Their location along substantial lengths of extrafusal fibers, distal capsule, and terminating intrafusal fibers suggests the presence of numerous transverse connections between elements of the terminal portion of the spindle and nonspindle tissues. Hence, it is likely that forces monitored by chicken spindles in muscles undergoing length changes are transferred from extrafusal fibers and extracellular matrix to the receptors in large part via lateral shear instead of by longitudinal tension. © 1996 Wiley-Liss, Inc.     

Histochemical study of an unusual cat intrafusal muscle fiber

A cat tenuissimus muscle spindle that contained two long chain intrafusal fibers in its distal pole is described. One of the fibers (lc1) had a histochemical profile (ATPase, NADH-TR, ChE reactions) of the kind which is characteristic for long chain fibers. The other fiber (lc2) consisted of two separate segments. The inner lc2 segment included the sensory equatorial region and was histochemically normal. The outer lc2 segment carried a motor plate, and did not stain for NADH-TR in the same way as the inner lc2 segment and the lc1 fiber. It is suggested that the unusual enzyme staining properties of the outer lc2 segment stemmed from its lack of sensory innervation, a situation which may have permitted the full expression of influences mediated by its motor nerve supply.     

The stroma of the thymus of the rat: morphology and antigen diffusion, a reconsideration

This work reconsiders aspects of the morphology of the capsule, of the blood vasculature, of the distribution of reticular fibers, and of the diffusion of intramediastinally injected antigens in the stroma of the thymus of the rat. This was done by an analysis of standard sections of normal thymuses, of sections of thymuses perfused with colloidal carbon, of silver-impregnated sections, and of sections of thymuses of rats injected intramediastinally with a fluorescent antigen or intravenously with Trypan blue, and by electron microscopy of the thymic capsule. The capsule consisted of two layers: an outer layer covering the entire periphery of a thymic lobe, and an inner layer which outlined the entire convoluted peripheral cortex of a lobe. Cortical vessels entered the capsule and septa in which they formed a capillary network. These capsular capillaries were fenestrated and leukocytes were often present near them. Adipocytes were also seen near these vessels in some areas of the capsule, and often at the bases of septa and trabeculae. Furthermore, much of the medulla had a dense network of coarse reticular fibers, whereas the remainder of the medulla and the cortex contained a loose network of fine fibers stretching out from the capsule, septa, and trabeculae. Intramediastinally injected fluorescent antigens were observed to spread in the capsule and septa and to diffuse in the fiber networks stretched across the cortex and the medulla. Fluorescence also highlighted cortical reticular cells but not the thymocytes. Intravenously injected Trypan blue stained the capsule, the septa, the cortical reticular cells, and the autofluorescent cells outlining the corticomedullary junction of each lobule. The unusual penetration of capillaries from the thymic parenchyma into the thymic capsule suggested that the capsular capillaries participate in peculiar thymic events, such as the recruitment of blood stem-cells. It is concluded that small amounts of blood antigens normally exude from capsular capillaries and diffuse into the fibers extending from the capsule across the cortex. The phenomenon would be increased under conditions causing thymic involution. An explanation is proposed to account for the development of involution which involves the exudation of antigens from the capsular capillaries. A comparable mechanism could also account for the development of a particular experimental immune tolerance.     

小鼠卵母细胞成熟和受精过程中Ca^2＋分布变化的研究

用焦锑酸钾原位定位法、膜结合Ｃａ＾２＋荧光探针金霉素标记法,分别在电镜和光镜水平对小鼠卵成熟和卵受精过程中结合态Ｃａ＾２＋的分布及其变化进行了研究,发现：１）Ｃａ＾２＋分布于线粒体、胞质、内质网囊泡、微绒毛和透明带等部位,其中以线粒体基质中分布密度为最大;２）减数分裂Ｉ中、后期于纺锤体极区结合有较多的Ｃａ＾２＋;３）生发泡、纺锤体和原核内膜结合态Ｃａ＾２＋含量很少,但纺锤体和原核周围分布较多;４）     

Myofibrillar ATPase activity of intrafusal fibers in chronically de-afferented rat muscle spindles

Dorsal root ganglia L4, L5 were removed to accomplish long-term (1 year) de-afferentation of the rat soleus muscle. Muscle spindles in the muscles deprived of sensory innervation were morphologically and histochemically abnormal. The spindle periaxial fluid space was greatly diminished with a thicker capsular investment. De-afferented intrafusal muscle fibers lacked either "nuclear bags" "nuclear chains" at their midlengths. The intracapsular myofibrillar ATPase staining pattern of de-afferented nuclear bag fibers resembled that which the bag fibers normally display in their extracapsular regions. These abnormalities are discussed with respect to the regulatory functions of spindle sensory and motor nerves.     

Histochemistry and ultrastructure of the metacercarial cyst of Bolbogonotylus corkumi (Trematoda: Cryptogonimidae).

Histochemical and ultrastructural studies were conducted on the metacercarial cyst of the cryptogonimid trematode Bolbogonotylus corkumi from the muscle tissue of fantail darters Etheostoma flabellare. The metacercarial cyst consisted of an outer host capsule and an inner parasite cyst. The host capsule was composed of an outer region of fibroblasts, collagen, macrophages, and unidentified cells, and an inner region containing degenerating cells. The parasite cyst was thin, homogenous, and noncellular in nature. The host capsule stained strongly for connective tissue and proteins and moderately for lipids, nucleic acids, nonspecific esterase activity, and acid and alkaline phosphatase activities. The parasite cyst stained intensely for acid mucopolysaccharides and moderately for acid phosphatase activity. A thick glycocalyx occurred between the parasite cyst and metacercarial tegument.     

Appearance of sensory nerve terminals in cat muscle spindles stained for NADH-tetrazolium reductase

Cat muscle spindles were studied histochemically in serial transverse sections of the tenuissimus muscle stained for myofibrillar ATPase, cholinesterase or NADH-tetrazolium reductase. The terminal sites of the primary and secondary axons on intrafusal muscle fibers could be demonstrated due to their high NADH-TR activity. This sensory NADH-TR reactivity at the equator and in the juxtaequatorial regions disappeared following spindle chronic de-afferentation, but not after de-efferentation. Spindle poles that carried both primary and secondary sensory endings had a longer periaxial fluid space than poles with primary endings only, and their motor innervation, as determined by staining for ChE, was positioned at the greater distance from the equator. Some of the secondary endings occurred in intrafusal regions that displayed surface fiber ChE activity. The histochemical reaction for NADH-TR represents a simple, rapid and reliable method for studies of the distribution of sensory nerve terminals in the spindle.     

Motor innervation of intrafusal fibers in rat muscle spindles: incomplete separation of dynamic and static systems

Distributions of 53 motor axons to different types of intrafusal fibers were reconstructed from serial 1-micron-thick transverse sections of 13 poles of spindles in the rat soleus muscle. The mean number of motor axons that innervated a spindle pole was 4.1. Approximately 60% of motor axons lost their myelination prior to or shortly after entry into the periaxial fluid space of spindles. Motor innervation to the juxtaequatorial portion of nuclear bag fibers (particularly the bag1) consisted of groups of short, synaptic contacts that were terminations of thin, unmyelinated axons. In contrast, motor endings on both the bag1 and bag2 fibers were platelike in the polar intracapsular region. Chain fibers had a single midpolar platelike ending. The ratio of motor axons that innervated the bag1 fiber exclusively to axons that innervated bag2 and/or chain fibers was 1:1. However, one-fourth of motor axons coinnervated the dynamic bag1 fiber in conjunction with static bag2 and/or chain fibers. Thus the complete separation of motor control of the dynamic bag1 and static bag2 intrafusal systems observed in cat tenuissimus spindles is neither representative of the pattern of motor innervation in all other species of mammals nor essential to normal spindle function.     

Appearance of sensory nerve terminals in cat muscle spindles stained for NADH-tetrazolium reductase

Summary Cat muscle spindles were studied histochemically in serial transverse sections of the tenuissimus muscle stained for myofibrillar ATPase, cholinesterase or NADH-tetrazolium reductase. The terminal sites of the primary and secondary sensory axons on intrafusal muscle fibers could be demonstrated due to their high NADH-TR activity. This sensory NADH-TR reactivity at the equator and in the juxtaequatorial regions disappeared following spindle chronic de-afferentation, but not after de-efferentation. Spindle poles that carried both primary and secondary sensory endings had a longer periaxial fluid space than poles with primary endings only, and their motor innervation, as determined by staining for ChE, was positioned at a greater distance from the equator. Some of the secondary endings occurred in intrafusal regions that displayed surface fiber ChE activity. The histochemical reaction for NADH-TR represents a simple, rapid and reliable method for studies of the distribution of sensory nerve terminals in the spindle.     

Ultrastructure of the capsule of the rabbit adrenal gland

The capsule of the normal rabbit adrenal gland was examined with the electron microscope, and three layers are described. The outermost layer consists of collagen and elastic fibrillae, with cytoplasmic processes of fibroblasts in between. Myofibroblasts with parallel cytoplasmic filaments, dense bodies at the inner surface of the plasma membrane and an irregular nuclear outline were seen in the middle layer. The presence of unmyelinated nerves and an axon terminal near to the myofibroblasts of the middle layer raised the possibility of innervation of these cells which could have a contractile function. The basal lamina of the fenestrated capilaries in the inner vascular layer is occasionally fused with that of the outer zona glomerulosa, suggesting a probable route for blood supply and secretion.     

Histochemical study of an unusual cat intrafusal muscle fiber

Summary A cat tenuissimus muscle spindle that contained two long chain intrafusal fibers in its distal pole is described. One of the fibers (1 c₁) had a histochemical profile (ATPase, NADH-TR, ChE reactions) of the kind which is characteristic for long chain fibers. The other fiber (1 c₂) consisted of two separate segments. The inner 1 c₂ segment included the sensory equatorial region and was histochemically normal. The outer 1 c₂ segment carried a motor plate, and did not stain for NADH-TR in the same way as the inner 1 c₂ segment and the 1 c₁ fiber. It is suggested that the unusual enzyme staining properties of the outer 1 c₂ segment stemmed from its lack of sensory innervation, a situation which may have permitted the full expression of influences mediated by its motor nerve supply.     

Microvasculature of the epididymis in the boar

Summary Microvasculature of the epididymis was investigated by scanning electron microscopy of vascular corrosion casts. The basic structure of blood supply to the boar epididymis consists of two superimposed vascular networks. Capillaries surrounding the epididymal duct constitute the inner level. They form polygonal meshes around the efferent ductules whereas circular capillaries strongly predominate in the subsequent region of the caput epididymidis. This annulate feature is progressively lost from corpus to cauda, where the capillary network once again has a polygonal appearance. The outer network is composed of feeding and draining vessels. Intertubular arteries pass between the loops of the epididymal duct and give rise to longitudinally oriented vessels attributable to only one adjacent duct segment. They feed the capillary network via circular ramifications debouching in different sectors of its circumference. The sparse veins draining the capillaries encircling the efferent ductules give way to a gradually increasing number of confluent veins up to the cauda.