Architecture of the media of the arterial vessels in the dog brain: A scanning electron-microscopic study

Summary The architecture of the media of arterial vessels in dog brain was investigated using scanning electron microscopy. The arrangement and shape of the circularly-oriented smooth muscle cells varied with vessel diameter: The arteries (>100 m in diameter) had 4–10 layers of spindle-shaped smooth muscle cells; the muscular arterioles (30–100 m), 2–3 layers of spindle-shaped smooth muscle cells; the terminal arterioles (10–30 m), a compact layer of spindle-shaped smooth muscle cells with more dominant nodular or rod-like processes and thin lateral processes; and the precapillary arterioles (5–15 m), a less compact layer of branched smooth muscle cells.Longitudinally-oriented muscles were observed in the medio-adventitial border. The distribution and arrangement of these muscles varied with vessel size: in the large arteries (> 300 m in diameter), at the branching sites only; in the small arteries (100–300 m), at both the branching and non-branching sites; in the muscular arterioles, at both the branching and non-branching sites in a reticular arrangement with some muscle cells having an asteroid appearance; in the terminal aterioles, only asteroid-like muscle cells were found at the branching and non-branching sites.     

Pulmonary vascular remodelling in a high-altitude Aymara Indian

A histological study of the pulmonary vasculature in a young male high-altitude Aymara Indian revealed four aspects of interest. There was muscularization of the terminal portion of the pulmonary arterial tree to involve pulmonary arterioles as small as 15 m in diameter, thus forming a basis for the slightly increased pulmonary vascular resistance of native highlanders. Intimal longitudinal muscle was found in pulmonary arteries and arterioles and thought to be due to chronic alveolar hypoxia. Inner muscular tubes similar to those found in chronic obstructive lung disease were present. Pulmonary veins and venules also showed intimal muscularization suggesting that alveolar hypoxia affects vascular smooth muscle cells per se irrespective of their situation. The nature of the remodelling in a pulmonary blood vessel depends on a combination of hypoxia and haemodynamics.     

In vivo restitution of airway epithelium

Epithelial shedding occurs in health and, extensively, in inflammatory airway diseases. This study describes deepithelialisation, reepithelialisation and associated events in guinea-pig trachea after shedding-like epithelial denudation in vivo. Mechanical deepithelialisation of an 800-m wide tracheal zone was carried out using an orotracheal steel probe without bleeding or damage to the basement membrane. Reepithelialisation was studied by scanning- and transmission electron microscopy and light microscopy. Nerve fibres were examined by immunostaining. Cell proliferation was analysed by [³H]-thymidine autoradiography. Immediately after epithelial removal secretory and ciliated (and presumably basal) epithelial cells at the wound margin dedifferentiated, flattened and migrated rapidly (2–3 m/min) over the denuded basement membrane. Within 8–15 h a new, flattened epithelium covered the entire deepithelialised zone. At 30 h a tight epithelial barrier was established and after 5 days the epithelium was fully redifferentiated. After completed migration an increased mitotic activity occurred in the epithelium and in fibroblasts/smooth muscle beneath the restitution zone. Reinnervating intraepithelial calcitonin gene-related peptide-containing nerve fibres appeared within 30 h. We conclude that (1) reproducible shedding-like denudation, without bleeding or damage to the basement membrane, can be produced in vivo; (2) secretory and ciliated cells participate in reepithelialisation by dedifferentiation and migration; (3) the initial migration is very fast in vivo; (4) shedding-like denudation may cause strong secretory and exudative responses as well as proliferation of epithelium, and fibroblasts/smooth muscle. Rapid restitution of airway epithelium may depend on contributions from the microcirculation and innervation.     

Urinary bladder of rat: fine structure of normal and hypertrophic musculature

Summary The fine structure of the muscle of the urinary bladder in female rats is similar to that of other visceral muscles, although it is arranged in bundles of variable length, cross-section and orientation, forming a meshwork. When distended, the musculature is 100–120 m thick, with some variation and occasional discontinuity. Extended areas of cell-to-cell apposition with uniform intercellular space occur between muscle cells, whereas attachment plaques for mechanical coupling are less common than in other visceral muscles. There are no gap junctions between muscle cells. Many bundles of microfilaments and small elastic fibres run between the muscle cells. After chronic partial obstruction of the urethra, the bladder enlarges and is about 15 times heavier, but has the same shape as in controls; the growth is mainly accounted for by muscle hypertrophy. The outer surface of the hypertrophic bladder is increased 6-fold over the controls; the muscle is increased 3-fold in thickness, and is more compact. Mitoses are not found, but there is a massive increase in muscle cell size. There is a modest decrease in percentage volume of mitochondria, an increase in sarcoplasmic reticulum, and no appreciable change in the pattern of myofilaments. Gap junctions between hypertrophic muscle cells are virtually absent. Intramuscular nerve fibres and vesicle-containing varicosities appear as common in the hypertrophic muscle as in controls. There is no infiltration of the muscle by connective tissue and no significant occurrence of muscle cell death.     

The innervation and fine structure of paraneuronic cells in an amphibian pulmonary artery

Summary The pulmonary artery of Bufo marinus contains large numbers of bipolar cells situated in the tunica adventitia and in the outer layers of the media. These cells show a bright green-yellow fluorescence (emission spectra 485 nm) after formaldehyde pre-treatment suggesting that they contain a primary monoamine. The most characteristic fine-structural feature of these cells is the presence of numerous dense-cored vesicles (80—300 nm diameter) in their cytoplasm. The cells are in close contact (20 nm gap) with both agranular and granular nerve fibres. Both EM-cytochemical and formaldehyde-induced fluorescence tests indicate that the granule-containing nerve fibres are adrenergic. The agranular nerve fibres form discrete synaptic contacts with pre-and post-synaptic membrane thickenings on the cells. This was never observed with respect to the adrenergic fibres. Each process of the cells is about 45 m long. The processes do not bear any special relationship to either vessels of the arterial vasa vasorum or medial smooth muscle cells. Their location in the wall of the artery suggests that they are functionally significant with respect to activity of the arterial media.     

Cellular organization of the embryonic lobster heart

Summary The cellular organization of the embryonic heart of the lobster Homarus americanus was examined in 6-week and 6-month-old animals. The heart wall consists of an outer adventitial layer of fibroblast cells and an inner layer of transversely striated myocardial cells. Present in close association with the myocardium are cardiac neurons, hemocytes and so-called storage cells.Adjacent fibroblasts form fasciae adhaerentes and gap junctions. Adherent junctions also occur between fibroblasts and myocardial cells. Intercalated discs and differentiated membrane regions of close apposition (4 nm) occur between adjacent myocardial cells.The cardiac neurons form a ganglion that contains four small and five large somata. Regions of neuropil are present. Motor axons arising from the cardiac ganglion form neuromuscular synapses with the myocardial cells.The storage cells contain large inclusions and form gap junctions with the myocardial cells. They may supply nutritive material to the developing myocardium.The heart at 6 weeks is about 200 m long and 160 m wide. At 6 months, it is about 300 m long and 250 m wide. The myocardium at 6 weeks is one cell layer thick, and the cells are from 2–6 m in maximum width. At 6 months the myocardium is 2–4 cells thick, and the cells are from 6–12 m in width. Therefore, the myocardium grows by an increase in the number and size of the myocardial cells.     

Metabolic variability within individual fibres of the cat tibialis posterior and diaphragm muscles

Summary Variance in succinate dehydrogenase activity along the transverse and longitudinal axes of fibres from the cat tibialis posterior and diaphragm muscles was determined in order to estimate the three-dimensional distribution of mitochondria within single fibres. The variance (coefficient of variation) in succinate dehydrogenase activity along the transverse fibre axis was greatest in type IIB fibres from both muscles. Intracellular compartmentalization (i.e. subsarcolemmal vs central core differences in succinate dehydrogenase activity) was observed only in type II fibres from the tibialis posterior; the succinate dehydrogenase activity of the subsarcolemmal region was significantly greater than that of the central core. The extent of succinate dehydrogenase variance along the longitudinal fibre axis was dependent on the total length of the fibre segment analyzed, the muscle, and fibre type. The coefficient variation for short fibre segments, i.e. 40 m, was significantly lower than that for much longer fibre segments (840 m). Significant differences in the coefficient variation for 840 m fibre segments were observed between the diaphragm and tibialis posterior muscles. The longitudinal variance in succinate dehydrogenase activity was higher in diaphragm muscle fibres. The succinate dehydrogenase variance along the longitudinal axis of type II fibres was significantly greater than that of the type I fibre population. These results indicate that mitochondria are heterogeneously distributed within muscle fibres. Possible functional implications of such intrafibre metabolic variance are discussed.     

Three-dimensional organization of smooth muscle cells in blood vessels of laboratory rodents

Summary Three-dimensional aspects of smooth muscle cells of the microvas-culature were studied ultrastructurally in laboratory rodents by means of serial thin sections and reconstruction of muscle cell models. It was demonstrated that a muscle cell of an arteriole (luminal diameter (LD) 17 m) in hamster striated muscle was spindle-shaped, 70 m long, and wound twice round the vessel axis. The volume of the cell was calculated as 750 m³ and its surface area as 1330 m². A muscle cell in an arteriole (LD 6 m) in the rat retina was irregular in shape, about 22 m long, and had branched processes. The cell volume was calculated as 139 m³ and its surface area as 298 m².     

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.     

The histochemical demonstration of myoglobin and succinic dehydrogenase activity in the tibialis anterior muscle of the rabbit

Summary The staining reactions for myoglobin and succinic dehydrogenase activity in the tibialis anterior of the rabbit demonstrate four types of muscle fibre. These may be distinguished by their intensity of staining for myoglobin and the distribution of the mitochondria shown by the dehydrogenase reaction.The large fibres (70–80 m diameter) which contain many mitochondria evenly scattered throughout the fibre contain much myoglobin. Smaller fibres (45–60 m diameter) which show an identical staining reaction for the dehydrogenase reaction contain less myoglobin. This suggests that myoglobin may be present to aid the diffusion of oxygen into muscle fibres.     

The cytoarchitecture of the medial layer in rat thoracic aorta: a scanning electron-microscopic study

Summary The cytoarchitecture of the medial layer of rat thoracic aorta was examined by scanning electron microscopy after removal of the connective tissue. The outermost lamella showed a lattice-like structure of muscle bundles of closely apposed smooth muscle cells (SMCs), whereas the inner lamellae consisted of more-or-less continuous muscle sheets of vaguely defined subgroups of parallel SMCs. Longitudinal rows of ridges ran along the adventitial surface of these muscle bundles and sheets. The SMCs of the outermost lamella, were 5.1 m wide, and varied in shape, whereas those of the inner lamellae, were 52.7 m long, 2.6 m wide and 4.1 m thick, and were elongated, spindle-shaped cells with serrated outlines. These latter SMCs extended obliquely, and partially overlapped each other. The surface of the SMCs in the outermost lamella exhibited a rugged texture, with nodular protrusions and oblique and longitudinal laminar folds, while the inner lamellar cells showed longitudinal laminar folds and finger-like processes on both sides of the ridges, pointing in opposite directions to the ridges. The angle of deviation from the transverse axis of the vessel, of the muscle bundles and subgroups in the outermost lamella, was 33.6°, in the second and third lamellae, 22.5°, and in the innermost lamellae, 12.8°. The mean angle of the muscle bundle and subgroup arrangement, with respect to the long axis of the vessel, however, was basically 90° in all lamellae.     

The pattern of innervation of a polyneural muscle: Axolotl iliotibialis

Summary The pattern of innervation on individual iliotibialis muscle fibres from axolotl (Ambystoma mexicanum) has been investigated histologically and elctrophysiologically. These polyneural fibres were found to be innervated on average at five end plate sites. The sites were distributed irregularly along each fibre. Average end plate length was found to be approxiamtely 70 m. Most end plates were separated by less than 1000 m; 26% by less than 150 m; the average separation was 516 m. Advantage was taken of the dual innervation of the muscle to investigate the separation between synaptic terminals from different axons. Some individual fibres were found to be innervated by axons from two different spinal nerves. End plate sites on dually innervated fibres were located by ACh iontophoresis. 30% of such sites were found to be innervated by more than one axon terminal. The average separation of such sites was found to be 950 m. Four different axons were found to innervate some individual muscle fibres. It is suggested that the unusual ability of axolotl muscle fibres to accept synaptic terminals from different axons at closely adjacent sites may be a major factor underlying selective reinnervation in this animal.Vacation student assisted by The Dale Fund of the Physiological Society     

The ultrastructure of the sinuatrial ring bundle and of the caudal extension of the sinus node in the right atrium of the rabbit heart

Summary The cells of the sinuatrial ring bundle are smaller than the ordinary myocardial cells; they have a regular outline and a large content of myofibrils exhibiting distinct H-bands and M-lines. Rudimentary T-tubules are found. The cells are connected by frequent nexus junctions, desmosomes and regions of interfibrillar contact, both on the well developed intercalated discs and at the periphery of the cells.The cells in the cauda of the sinus node, which extends alongside the crista terminalis together with the right branch of the sinuatrial ring bundle, are irregularly outlined and have a highly variable diameter (1–10 m). They occur in clusters of closely packed cells. The content of myofibrils is sparse and the fibrils exhibit no M-lines and only weak H-bands. No T-tubules are found. The cells are not connected by intercalated discs and no nexus junctions are found.Both tissues contain unmyelinated nerves and nerve fibres. Varicosities with mitochondria and vesicles are found in close apposition to the muscle cells.     

Cell and endophyte structure of the nitrogen-fixing root nodules ofCeanothus integerrimus H.and A.

Summary The nitrogen fixing root nodules ofCeanothus integerrimus were very similar in appearance to other non-legume nodules. Each nodule was a cluster of small lobes. Each lobe in cross section had a central vascular cylinder and a hypertrophied cortex. The cortex contained very large infected cells, with large nuclei; among these infected cells were scattered small, normal-appearing cortex cells. The actinomycete endophyte consisted of wavy hyphae 0.4 m in diameter which terminated in pear-shaped vesicles 1.6 m×2 m. The vesicles were not septate. The function of the vesicles was unknown. The infected cells had apparently normal nuclei, chloroplasts and mitochondria and were probably alive, except at the base of the nodule where both infected cells and the endophyte they contained were dead.     

Gap junctions of the muscles of the small and large intestine

Summary The distribution of gap junctions (nexuses) in various parts of the small and large intestines of the guinea-pig was studied using the freeze-fracture technique and in thin sections. The percentage area of smooth muscle cell surface occupied by gap junctions varies from 0.50% in the circular muscle of the duodenum to zero in the longitudinal muscle of the ileum. In the circular muscle of the jejunum and ileum the area occupied by nexuses is 0.22% (or about 11 m² per cell). The sizes of junctions range from less than 0.01 m² to 0.20 m², with two-thirds of them being smaller than 0.05 m². In the colon, gap junctions are rare, very small and confined to the circular muscle layer. Even the smallest aggregates of intramembrane particles correspond to areas of close apposition between the membranes of adjacent cells; it is therefore justified to interpret them as being gap junctions. Some gap junctions are formed between a smooth muscle cell and an interstitial cell. Gap junctions are not found in the longitudinal muscle of the small intestine; this is in sharp contrast to the abundance of gap junctions in the adjacent circular layer.In the small intestine of cats and rabbits, gap junctions are abundant in the circular muscle layer, whereas they are very small in size and very few in number in the longitudinal muscle layer.The authors wish to thank Mr Peter Trigg and Miss Eva Franke for help and support. This work was supported by grants from the Medical Research Council and the Central Research Fund of the University of London     

The fine structure of the olfactory tract in the teleost Carassius carassius L.

Summary Electronmicrographic montages of the olfactory tract at two levels in each of two fish (Carassius carassius L.) were constructed and fibre diameters measured using a Zeiss TGZ 3 particle size analyzer. Medial and lateral tract divisions, rhinocele and dorsal tela were identified. Ciliated ependymal cells line the rhinocele. Meninges form the outer covering of both tract divisions and the tela roofing the central canal.The lateral tract consists of 10–14 fasciculi in which myelinated nerve fibres are prominent. These fibres range in diameter between 0.2 and 1.8 (mean 0.7 ) consistent with conduction velocities averaging 0.6 m/sec recorded in the carp lateral olfactory tract.The medial division of the olfactory tract contains two larger fasciculi within which are numerous fine unmyelinated nerve fibres (mean diameter 0.17 ) arranged in bundles partly enveloped by glial cell processes. Myelinated nerve fibres are unevenly distributed within both fasciculi and have mean diameters of 0.6 .An interesting observation is the consistent presence of synapses within the largest bundle of the medial tract at all levels.Supported by Grant 5 Ro5 TW00154-03 from the National Institutes of Health, United States Public Health Service.The authors are indebted to the Fisheries and Wildlife Department who generously provided the fish from Snob's Creek Fish Hatchery, and gratefully acknowledge the technical assistance of Mr. T. Armitage, Mr. J. Simmons and Miss D. Harrison.     

Localization of 3H-estradiol in the uterus of pregnant and non-pregnant armadillos

Summary The uptake and retention of radiolabeled estradiol by the uterus was examined in the armadillo. One pregnant and two non-pregnant armadillos were treated with 1.4 g/kg body weight of ³H-estradiol (E₂) by injection into the left ventricle, and one non-pregnant animal was injected with both the labeled hormone and 140 g/kg body weight of unlabeled E₂. One and a half hour after injection, the animals were sacrificed and the uteri were removed and processed for autoradiography. In the non-pregnant animals, nuclear localization was observed in the interstitial cells and glandular epithelium of the endometrium and the connective tissue cells and smooth muscle of the myometrium. Additionally, there was a gradation of uptake in the epithelial cells of the endometrium in that the glandular cells of the basal region were heavily labeled, while those cells in the sinusoidal, and luminal regions contained successively less label. The luminal cells were poorly labeled. In the pregnant female, the smooth muscle and glandular cells hypertrophied and their nuclei contained less label than was observed in the non-pregnant animals. The arteries of the myometrium were more easily distinguished in the pregnant animals and the nuclei of the endothelial cells and smooth muscle were more consistently labeled than those of the non-pregnant armadillos.     

„Rote“ Muskelfasern

Zusammenfassung Die Fasern des roten und langsamen M. soleus von Ratte, Kaninchen und Katze und des roten, jedoch schnellen, M. vocalis des Kaninchens wurden licht- und elektronenmikroskopisch untersucht und mit den verschiedenen Fasertypen aus dem M. tibialis anterior der Ratte und dem M. gastrocnemius des Kaninchens und der Katze verglichen. M. soleus und M. vocalis (einschließlich M. thyreoarytenoideus) enthalten nur einen mitochondrienreichen Fasertyp. Im schnellen M. vocalis ist der Z-Streifen schmal (50–60 nm), das sarcoplasmatische Reticulum ist gut entwickelt. Die Anordnung von Reticulum und Mitochondrion ist ähnlich wie in Herzmuskelzellen. Wie auch in anderen langsamen Muskeln verschiedener Tiere ist im M. soleus der Z-Streifen breit (100–120 nm), Triaden und Reticulum sind selten, und die Filamente bilden unregelmäßige Areale anstelle von Fibrillen. Hierin gleichen die Fasern des M. soleus den (mitochondrienreichen) C-Fasern eines entsprechenden gemischten Muskels; dagegen zeigen die Zwischentyp-(B-)Fasern schmale Z-Linien (50–70 nm), isodiametrische Fibrillen und mehr Triaden als die C-Fasern. Entgegen der bisherigen Vermutung, die auf der histochemischen Zuordnung der SoleusFasern zum Typ B und der Vocalis-Fasern zum Typ C beruht, ist daher anzunehmen, daß die langsamen motorischen Einheiten eines gemischten Muskels aus C- und nicht aus B-Fasern bestehen.In einigen Muskeln sind die Sarcomere der C-Fasern länger als die der B-(und A-) Fasern. Im M. tibialis anterior der Ratte verschwindet der Unterschied von 8,5% bei 2,6 m Sarcomerlänge bei der Dehnung auf 2,8 m mittlere Sarcomerlänge; vermutlich weil die Ruhedehnungskurve zunehmend steiler wird. Die isometrische Extraspannung im Tetanus ist bei 120% der Ruhelänge, d.h. bei 2,7 m Sarcomerlänge. am größten. Daher muß bei 2,6 m mittlerer Sarcomerlänge die Kraft der C-Fasern die der B-Fasern übertreffen.Rote Muskeln sind besser vaskularisiert als weiße Muskeln. Für die Mm. soleus und gastrocnemius der Katze verhalten sich die Kapillardichten (Kapillaren/mm² Muskelfaserquerschnitt) wie 2,7:1. Dieser Wert entspricht dem Verhältnis zwischen den Größen für die Durchblutung (ml/min × 100 g) in Ruhe und bei maximaler Gefäßerweiterung.

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Redmuscle fibres

Summary Muscle fibres of the red and slow contracting soleus of rat, rabbit and cat and of the red however fast contracting thyreoarytenoid of rabbit are compared with different fibre types in the anterior tibial muscle of rat and in the gastrocnemius of rabbit and cat. With respect to fibre types soleus and thyreoarytenoid (including m. vocalis) are homogeneous and both being rich in mitochondria. The fast thyreoarytenoid shows a narrow Z-line (50–60 nm) and a well developed sarcoplasmic reticulum. The pattern of reticulum and mitochondria resembles more that of heart muscle cells than of skeletal muscle fibres. Like many slow contracting muscles of different animals the soleus fibres display a wide Z-line (100–120 nm), few triads, little reticulum and irregularly shaped areas of myofilaments instead of fibrils. In that soleus fibres equal fibres of type C (rich in mitochondria) in a corresponding heterogeneous muscle, whereas intermediate (type B) fibres reveal narrow Z-lines (50–70 nm), isodiametrically shaped myofibrils and more triads than C-fibres. Therefore it is far more likely that the slow motor units of a mixed muscle consist of C-fibres than of B-fibres. This is at variance with the histochemical designation of soleus fibres as type B and thyreoarytenoid fibres as type C.In some muscles in C-fibres the sarcomeres are longer than in B-(and A-)fibres. In the anterior tibial muscle of rat this difference is 8.5% at a mean sarcomere length of 2.6 m, and disappears at a mean length of 2.8 m, probably due to the steeper slope of the length tension diagram at rest. Since the isometric extratension in a tetanus is highest at 120% resting length (corresponding to about 2.7 m sarcomere length), the force of C-fibres exceeds that of B-fibres at 2.6 m but not at 2.8 m sarcomere length.Red and white muscle differ with respect to vascularisation. The relation between the densities of capillaries in soleus and gastrocnemius of cat is 2.7:1 and equals the relation between the blood flows through these muscles during rest and maximum vasodilatation.

Herrn Prof. Dr. Bargmann nachträglich zum 65. Geburtstag.     

Regeneration of soleus muscles of rat autografted in toto as studied by electron microscopy

Summary Rat soleus muscles were autografted from right to left legs, and regeneration following necrosis of all original myofibres was studied after 7 to 250 days. The best regenerates were from grafts replacing all calf muscles and sutured to the tendon stumps. After 30 days the size of such regenerates was equal to those from minced gastrocnemius muscles: the cross sectional area of muscle tissue was 30% (1.7 mm²) and the number of fibres was 180% (4500) of normal soleus muscles; the fibre diameters were 10 to 40 m. To increase the number of myoblasts before grafting some muscles were injured by Ringer solution of 70° C and transplanted after 2 days. Nevertheless, this did not influence regeneration.After 7 days clusters of myotubes occurred in the periphery of the muscle. These myotubes originated from myoblasts growing like endothelial cells on the inner face of the persisting basal lamina tubes of necrotic fibres. After 30 days the muscles were vascularized. Fibres formed in a common basal lamina detached and so looked split. Satellite cells of new fibres came from undifferentiated cells associated with myotubes, i.e. from myoblasts. After 30 days and more regenerates contained three sorts of fibres. 1. Thin (5 to 20 m) fibres resembling fetal muscle fibres. They were most prominent after 30 days, and probably not yet innervated. 2. Thin (10 m) degenerating fibres as in long-time denervated muscles. 3. Thick (more than 30 m) mature looking fibres which were innervated and revealed end-plates.Half of the grafts studied after 30 and 60 days contained unmyelinated and myelinated axons which had grown along strands of surviving Schwann cells. After 250 days, only two muscles were studied which both lacked innervation. Almost all regenerates contained muscle spindles, which, however, were not innervated. Within the persisting spindle capsules new muscle fibres had been formed from satellite cells of the former intrafusal fibres.This study was supported by grants from the Danish Medical Research Council, and the National Danish Association against Rheumatic Diseases. I wish to thank Miss U. Hellhammer for valuable technical help and Dr. T. Tobias for correcting my English     

Morphology and structural organization of spiracles in femaleArgas (Persicargas) walkerae (Acari: Argasidae)

Scanning electron microscopical investigations of fractures and corrosion casts of the spiracles from femaleA. walkerae ticks revealed a four-part structure, consisting of spiracular plate, ostium and macula forming the external closure, followed by the subostial space and the vestibulum of regulable volume, as well as the atrial chamber as the innermost part from which the main tracheal trunks originate. On the average, the spiracular plate was 158 m long and 188 m at the broadest width. It consisted of a thin, highly perforated external and a thick internal layer, which enclosed the interpedicellar space with numerous stout pedicels. In its posterior region, the spiracular plate was covered by the macula, which was up to 80 m in length and 110 m in width. The interpedicellar cavity opened into the subostial space measuring 95.5 m in length and 159.6m in width, which proceeded into the 112-m long vestibulum. The roof of the vestibulum was flexible and could be everted and inverted. Inverted, the roof formed a quadratic bulge with numerous deep cuticular folds, which confined the lumen of the vestibulum either partially or completely. In corrosion casts, the roof was everted to a length of up to 89.3 m. In the posterior part of the vestibulum, as well as in the initial fourth of the artrial chamber, numerous anvil-, cone-or drop-like cuticular projections were arranged in wedge-like fashion. The atrial chamber was almost spherical with a diameter of 138.4 m. Five main tracheal trunks of different luminal diameter as well as numerous channels opened into the atrial chamber.