The demonstration of close nerve-Purkinje fibre contacts in false tendons of sheep heart

Summary An observation of intimate nerve-Purkinje fibre associations in false tendons of sheep heart is reported. Nerve bundles were observed in deep clefts of Purkinje fibres, in channels running between coupled Purkinje cells and embedded within Purkinje cells, as well as in the outer connective tissue sheath. Most nerve terminals in these areas were filled with small clear vesicles and a few large dense-cored vesicles. Only a few axons with many small dense-cored vesicles were observed.Intimate associations (separation, 60 to 90 nm) between the Purkinje cell and nerve varicosity were observed in the deep clefts. Similar close appositions were also present where nerves were embedded in Purkinje cells. In these cases the Purkinje cell enclosing the nerve bundle formed intercellular junctions with its own sarcolemma.Elaborate sarcolemmal folds with multi-vesicular bodies were also frequently observed near nerve bundles and varicosities. The identity of the transmitter is unknown although the nerves forming intimate associations with Purkinje cells have a morphology typical of cholinergic nerves.     

The autonomous innervation of the buffalo (Bubalus bubalis) testis. An immunohistochemical study

The innervation pattern in the buffalo testis was determined by using histochemical and immunohistochemical methods. Nerves were concentrated in the tunica albuginea and septula testis, and did not show an uniform distribution. The tunica albuginea at the lateral and medial sides and at the free border of the testis is most densely innervated than at the epididymal border. At the cranial pole thick nerve bundles were observed between albugineal vessels and muscle bundles. Rare parenchymal nerves were found in perivascular position between seminiferous tubules and their occurrence is confined to lobules at the cranial and caudal testicular poles. An intense NPY immunoreactivity occurred in nerve bundles and in solitary varicose fibres. Nerves were concentrated in the tunica albuginea at the lateral and medial side and at the free border of the testis, and in the lobules at the cranial and caudal testicular poles. Sub P immunoreactivity was occasionally detected in some thicker nerve bundles and solitary fibers, in the tunica albuginea and in the wall of blood vessels, showing a similar distribution but less intensity and density than NPY immunoreactivity. TH immunoreactivity stained nerve fibers in the buffalo testis with a distribution pattern similar to that obtained with general neuronal markers. The histochemical reaction for AchE was negative, so cholinergic fibers cannot be detected in the buffalo testis. The histochemical NADPHd reaction stained rare nitrergic nerve bundles and solitary fibers. The majority of NADPHd activity was confined to the vascular endothelium, and rarely to the interstitial Leydig cells, whereas the Sertoli and germ cells did not show any reaction.     

Distribution of actin-filament bundles in myoid cells,Sertoli cells,and tunica albuginea of rat and mouse testes

Summary Frozen sections of the rat and mouse testes were stained with either FITC-phalloidin or NBD-phallacidin and viewed with conventional fluorescence and confocal laser microscopes in order to demonstrate the arrangment of actin-filament bundles in myoid cells, Sertoli cells and tunica albuginea. Myoid cells are rich in actin-filament bundles crossing at right angles. These bundles running in different directions can also be visualized by means of electron microscopy. Nerve fibers occur in the vicinity of myoid cells, suggesting a neural control of the cell. At Sertoli cell junctions actin filaments occur at the circumference of the cell, where they show a honeycomb pattern. The ratio of the number of Sertoli cells per myoid cell can be calculated by means of confocal microscopy; this technique may provide a new parameter for determining spermatogenic activity. In the tunica albuginea of the juvenile mouse testis, actin filaments are arranged in an alternate fashion.     

Scanning electron microscopy of the muscle coat of the guinea-pig small intestine

Summary We have studied the layers of the muscular coat of the guinea-pig small intestine after enzymatic and chemical removal of extracellular connective tissue. The cells of the longitudinal muscle layer are wider, have rougher surfaces, more finger-like processes and more complex terminations, but fewer intercellular junctions than cells in the circular muscle layer. A special layer of wide, flat cells with a dense innervation exists at the inner margin of the circular muscle layer, facing the submucosa. The ganglia of the myenteric and submucosal plexuses are covered by a smooth basal lamina, a delicate feltwork of collagen fibrils, and innumerable connective tissue cells. The neuronal and glial cell processes at the surface of ganglia form an interlocking mosaic, which is loosely packed in newborn and young animals, but becomes tightly packed in adults. The arrangement of glial cells becomes progressively looser along finer nerve bundles. Single varicose nerve fibres are rarely exposed, but multiaxonal bundles are common. Fibroblast-like cells of characteristic shape and orientation are found in the serosa; around nerve ganglia; in the intermuscular connective tissue layer and in the circular muscle, where they bridge nerve bundles and muscle cells; at the submucosal face of the special, flattened inner circular muscle layer; and in the submucosa. Some of these fibroblast like cells correspond to interstitial cells of Cajal. Other structures readily visualized by scanning electron microscopy are blood and lymphatic vessels and their periendothelial cells. The relationship of cellular elements to connective tissue was studied with three different preparative procedures: (1) freeze-cracked specimens of intact, undigested intestine; (2) stretch preparations of longitudinal muscle with adhering myenteric plexus; (3) sheets of submucosal collagen bundles from which all cellular elements had been removed by prolonged detergent extraction.     

Morphological and histochemical peculiarities of the gut in the white sturgeon, Acipenser transmontanus.

The gut of adult sturgeon was examined. The oesophageal mucosa contained numerous caliciform cells, synthesizing both neutral and acidic glycoconjugates, the latter of the sialylated type. The deep tunica propria-submucosa contained lobules of multilocular adipose tissue, specially abundant during the cold season. The oesophageal tunica muscularis was made up of a large sheath of striated muscle fibres, arranged orthogonally to a thin, subserous smooth muscle layer. The siphon-shaped stomach showed a ciliated epithelium in cardiac and gastric proper gland zones, where tubular glands were present in the tunica propria. The columnar cells which composed the superficial epithelium and gastric pits were demonstrated to synthesize almost exclusively neutral glycoconjugates. Appendices pyloricae constituted a glandular body equipped with intestinal mucosa. The intestinal mucosa was organized in folds, containing numerous caliciform cells which synthesized neutral or acidic glycoconjugates, the latter either of the sialylated and sulphated type. The sulphoglycoconjugates were more abundant in the caliciform cells of the distal intestinal tracts. The tunica propria-submucosa of the spiral valve (medium intestine) contained lymphatic tissue and large lymphatic follicles. A muscularis mucosae was present only in the rectum, where in addition a peculiar granular cell type was present in the superficial tunica propria-submucosa, possibly related to defensive properties. The subserous connective tissue contained pancreatic lobules all along the stomach and intestine. The enteric nervous system showed some special aspects, the most intriguing of which was the presence of large, longitudinally oriented nerve bundles in the t. propria-submucosa of oesophagus and cardiac stomach. The nerve bundles contained, near unmyelinated nerves, some myelinated nerves, as well as neuronal bodies. Both these aspects are exceptional in vertebrates and obscure in their significance. The structural and histochemical aspects we here describe are in part different from those described for other fish. Some of these special features are possibly related with special functional roles, others require a deeper insight and different approaches to clarify them functionally.     

Organization of rat mesenteric artery after removal of cells or extracellular matrix components

Summary Rat mesenteric arteries, perfusion fixed in relaxed or contracted conditions, were digested with acid and elastase, bleach (sodium hypochlorite), or alkali to selectively remove collagen, elastin, or cells. Scanning electron microscopy was used to study the three-dimensional organization of the remaining cells or extracellular components. Smooth muscle cells of the tunica media were elongated and circumferentially oriented. Superior mesenteric artery cells had an irregular surface with numerous projections and some ends were forked. Small mesenteric artery cells were spindle shaped with longitudinal surface ridges, and showed extensive corrugations upon contraction. Elastin was present both as laminae and as an interconnected fibrous meshwork. Collagen was arranged in an irregular network of individual fibrils and small bundles of fibrils that formed nests around the cells in both arteries. This irregular arrangement persisted, with no apparent reordering or loss of order, upon contraction. The lack of an ordered arrangement or specialized organization at the cell ends suggests mechanical coupling of the cells to elastin or collagen throughout the length of the cell, allowing for force transmission in a number of directions. The tunica media is thus a composite material consisting of cells, elastin, and collagen. The isotropic network of fibers is well suited for transmitting the shearing forces placed on it by contraction of smooth muscle cells and by pressure-induced loading.     

Three-dimensional organization of the collagen fibrils in the rat sciatic nerve as revealed by transmission- and scanning electron microscopy

Summary The organization of collagen fibrils in the rat sciatic nerve was studied by scanning electron microscopy after digestion of cellular elements by sodium hydroxide treatment, and by conventional transmission electron microscopy. The epineurium consisted mainly of thick bundles of collagen fibrils measuring about 10–20 m in width; they were wavy and ran slightly obliquely to the nerve axis. Between these collagen bundles, a very coarse meshwork of randomly oriented collagen fibrils was present. In the perineurium, collagen fibrils occupied the interspaces between the concentrically arranged perineurial cells; in each interspace, they formed a sheet of characteristic lacework elaborately interwoven by thin (about 3 m or less in width) bundles of collagen fibrils. In the subperineurial region, there was a distinct sheet of densely woven collagen fibrils between the perineurium and underlying endoneurial fibroblasts. In the endoneurium, collagen fibrils surrounded individual nerve fibers in two layers as scaffolds: the inner layer was made up of a delicate meshwork of very fine collagen fibrils, and the outer one consisted of longitudinally oriented bundles of about 1–3 m in width. The collagen fibril arrangement described above may protect the nerve fibers against external forces.     

Morphology of the proboscis of Hubrechtella Juliae (nemertea,pilidiophora): Implications for pilidiophoran monophyly

The proboscis of Hubrechtella juliae was examined using transmission electron microscopy, scanning electron microscopy, and confocal laser scanning microscopy to reveal more features of basal pilidiophoran nemerteans for morphological and phylogenetic analysis. The proboscis glandular epithelium consists of sensory cells and four types of gland cells (granular, bacillary, mucoid, and pseudocnidae‐containing cells) that are not associated with any glandular systems; rod‐shaped pseudocnidae are 15–25 μm in length; the central cilium of the sensory cells is enclosed by two rings of microvilli. The nervous plexus lies in the basal part of glandular epithelium and includes 26–33 (11–12 in juvenile) irregularly anastomosing nerve trunks. The proboscis musculature includes four layers: endothelial circular, inner diagonal, longitudinal, and outer diagonal; inner and outer diagonal muscles consist of noncrossing fibers; in juvenile specimen, the proboscis longitudinal musculature is divided into 7–8 bands. The endothelium consists of apically situated support cells with rudimentary cilia and subapical myocytes. Unique features of Hubrechtella's proboscis include: acentric filaments of the pseudocnidae; absence of tonofilament‐containing support cells; two rings of microvilli around the central cilium of sensory cells; the occurrence of subendothelial diagonal muscles and the lack of an outer diagonal musculature (both states were known only in Baseodiscus species). The significance of these characters for nemertean taxonomy and phylogeny is discussed. The proboscis musculature in H. juliae and most heteronemerteans is bilaterally arranged, which can be considered a possible synapomorphy of Hubrechtellidae + Heteronemertea (= Pilidiophora). J. Morphol. 274:1397–1414, 2013. © 2013 Wiley Periodicals, Inc.     

Electron microscopic observations on the formation of elastic fibres in cultures of aortic medial cells and adventitial cells

The formation of elastic fibres was observed in the cultured cells derived from the tunica media and the tunica adventitia of mouse aorta. Bundles of myofilaments with dense bodies were abundantly observed in the cytoplasm of the cultured medial cells, and numerous bundles of microfibrillar components were present in the intercellular spaces. Fine granules of approximately 50 nm in diameter were observed in the bundles of microfibrillar components. It was supposed that these fine granules of elastin fused with each other and formed elastic aggregates and then formed large elastic clumps. Numerous bundles of microfibrillar components were also present in the intercellular spaces of the cultured adventitial cells. Elastic aggregates were scarcely observed in the bundles of microfibrillar components. However, large elastic clumps as observed in the medial cell culture could not be found in the adventitial cell culture. It is suggested that the formation of large elastic clumps might be related to the sheet structures or lamellae of elastic fibres in the tunica media.     

Cytoskeletal arrangement and its intercellular connec-tion in wheat young leaf cells

By using the techniques of partial digestion of cell wall and selective extraction,we examined the cytoskeleton of wheat yong leaf cells under scanning electron microscope(SEM).A 3-dimensional cytoskeletal system,showing some new features,was observed.The cortical network located beneath the cross wall was an anastomosing organization.The association of nucleus with the cell wall by some skeletal filaments was also found.It is notice able that there were cytoskeletal filaments,which passed through cell wall and connected together with cytoskeletal arrays of adjacent cells,Thus,it is possible that an integral skeletal network existed within the yong leaf tissue of wheat.     

Ultrastructural features of the gut in the white sturgeon, Acipenser transmontanus

Electron-microscopic examinations of the sturgeon gut were performed. Oesophageal goblet cells were abundant in the stratified epithelium. The ultrastructural features of the secretory granules of the oesophageal and intestinal goblet cells were quite similar to those of other vertebrates. Lobules of multilocular adipose tissue were observed in the deep tunica propriasubmucosa of the oesophagus, in close association with vasculature and large fibre bundles of myelinated and unmyelinated axons. Similarly composed nerve fibre bundles were observed in the cardiac stomach, too. The presence of myelinated axons is an unusual feature in the vertebrate enteric nervous system. Cardiac and fundic zones of the stomach showed an epithelium with columnar ciliated and non-ciliated cells, the latter equipped with fuzzy microvilli. Cells lining the tubular gastric proper glands were markedly granulated. Intestinal superficial epithelium was columnar and contained ciliated, as well as non-ciliated and goblet cells. In the tunica propria all over the intestine, the presence and ultrastructure of granulated cells was in addition described. Intraepithelial granulated leukocytes were seen throughout the alimentary canal. Various types of endocrine cells were seen both in the stomach and in the intestine, the size of their granules was measured and their ultrastructure described and compared to that of mammalian cell types.     

Microscopic anatomy of the thin-walled vessels leaving the heart of the lobster Homarus americanus: anterior lateral arteries

Abstract. The anterior lateral arteries are paired vessels leaving the anterior end of the lobster ( Homarus americanus ) heart and proceeding to the antennae and eyestalks, the stomach and hepatopancreas, the gonads, and the thoracic and branchial muscles. These vessels have a trilaminar organization, consisting of a tunica interna with elastic fibrils, a tunica intermedia represented by a bilayered cell mass, and a tunica externa with collagen fibrils. In the tunica intermedia, cells flanking the tunica interna (light cells) show less affinity for basic dyes and electron stains than those flanking the tunica externa (dark cells). Each light cell exhibits an irregularly shaped stress fiber (a bundle of closely packed microfilaments) in the region adjoining the tunica interna. Collectively, these bundles have a circumferential or slightly oblique orientation relative to the lumen of the vessel. The role of the stress fibers is unresolved. If they are static structures, they might contribute to the non-linear elasticity shown by lobster arteries. If they generate force, and small bundles of microfilaments do diverge from the stress fibers to enter filamentous mats applied to the plasmalemmata, a coordinated contraction of the cells might reduce the luminal diameter and, thus, retard the flow of hemolymph. Coordination of contraction would have to occur in the absence of nerves and without the benefit of communicating (gap) junctions between the light and dark cells.     

Scanning electron microscopy of echinoid podia

Summary Tube feet of the sea urchin Strongylocentrotus franciscanus were studied with the scanning electron microscope (SEM). By use of fractured preparations it was possible to obtain views of all components of the layered tube-foot wall.The outer epithelium was found to bear tufts of cilia possibly belonging to sensory cells. The nerve plexus was clearly revealed as being composed of bundles of varicose axons. The basal lamina, which covers the outer and inner surfaces of the connective tissue layer, was found to be a mechanically resistant and elastic membrane. The connective tissue appears as dense bundles of (collagen) fibers. The luminal epithelium (coelothelium) is a single layer of flagellated collar cells.There is no indication that the muscle fibers, which insert on the inner basal lamina of the connective tissue layer are innervated by axons from the basiepithelial nerve plexus.The results agree with previous conclusions concerning tube-foot structure based on transmission electron microscopy, and provide additional information, particularly with regard to the outer and inner epithelia.This investigation was supported by the Sonderforschungsbereich 138 of the Deutsche Forschungsgemeinschaft. The work was carried out at the Friday Harbor Laboratories of the University of Washington. The authors are indebted to the Director, Professor A.O.D. Willows for use of the facilities, and to Drs. Christopher Reed and Tom Schroeder for invaluable instruction and assistance     

Observations on the fine structure of the baroreceptors and adrenergic innervation of the guinea-pig carotid sinus

We examined the fine structure of the baroreceptors and the adrenergic innervation of the guinea-pig carotid sinus. The tunica adventitia contained many nerve bundles whose perineuria enclosed unmyelinated nerve fibers, alone or together with myelinated nerve fibers. Baroreceptors, which lay close to elastic and collagen fibers in the adventitia and media, were surrounded by “terminal” cells with ultrastructural features characteristic of Schwann cells and contained inclusions of various types. Morphologic features of the baroreceptors included densely packed mitochondria, osmiophilic lamellated and homogeneous bodies, clear and granular vesicles, lamellar systems, glycogen granules, neurofilaments, neurotubuli, and vacuolated mitochondria. In animals that had been treated with 6-hydroxydopamine, occasional electrondense endings (or fibers) were observed in the adventitial layer. The baroreceptors in the guinea-pig carotid sinus appear to have most of the morphologic features reported for other species.     

Microscopic anatomy of the thin-walled vessels leaving the heart of the lobster Homarus americanus: anterior median artery

Abstract. The anterior median artery is an unpaired vessel that leaves the anterior end of the lobster ( Homarus americanus ) heart and supplies hemolymph to the brain, the antennae, and the eyestalks. This vessel has a trilaminar organization, consisting of a tunica interna with elastic fibrils, a tunica intermedia represented by a bilayered cell mass, and a tunica externa with collagen fibrils. The exposed ends of the medial cells in the tunica intermedia exhibit small, diffuse bundles of microfilaments that are penetrated by microtubules. These bundles have a circumferential or a slightly oblique orientation relative to the lumen of the vessel. The precise role of the microfilaments is unresolved. If the irregularly shaped bundles are static structures, they might contribute to the non-linear elasticity of the artery. Alternatively, if they generate force, a coordinated contraction of the medial cells might reduce the luminal diameter of the artery and, thus, retard hemolymph flow. Microfilaments of the medial cells anchor to subplasmalemmal filamentous mats, some of which are integral to intermediate junctions and some of which are associated with unbounded cell membranes (hemi-intermediate junctions). Contraction of the microfilament-bearing cells would have to occur without the benefit of nervous innervation or the participation of communicating (gap) junctions. If cell contractility is confirmed, a reclassification of the anterior median artery, from capacitance vessel to resistance vessel, is in order, and the bilayered cell mass in the tunica intermedia would likely qualify as the first unstriated muscle found in crustaceans.     

日本沼虾消化道形态和组织学特点

应用石蜡切片和扫描电镜技术对日本沼虾消化道进行了研究。结果表明,食道壁向腔内形成四个纵突,食道上皮由单层柱状细胞构成,上皮下的结缔组织中具有放射肌和皮肤腺,环肌层近于连续。食道和胃连结处的管腔背方具食道瓣,胃内具胃磨、滤器和滤沟等结构,胃的组织学结构中除无皮肤腺分布外与食道相似。中肠较长,约占整个消化道的７１７％,具一对中肠前盲囊。中肠上皮细胞大致有两种类型,基膜着色深,环肌层连续,纵肌成束分散排列。后肠为一短管,管腔呈迷路状,其中部为一球形膨大的直肠。后肠的组织学结构与前肠相似。     

A scanning electron microscopic study of germ cell maturation in the reproductive tract of the male soupfin shark (Galeorhinus galeus)

Germ cell maturation in the reproductive tract of the soupfin shark (Galeorhinus galeus) was studied using scanning electron microscopy (SEM). The SEM showed changes in Sertoli cytoplasm volume during spermatogenic development. In immature spermatocysts in the germinal zone, spermatogonia were embedded in Sertoli cytoplasm. In spermatogonial spermatocysts, Sertoli cells were adluminally located in the spermatocyst, with spermatogonia enveloped in the basal portions of the cytoplasm. During the round spermatid stage, Sertoli cytoplasm was very scanty. Spermatid elongation was accompanied by a progressive increase in the volume of Sertoli cytoplasm, notably around the spermatid heads. In the mature spermatocyst, bundles of spermatozoa are totally enveloped by Sertoli cytoplasm. Spermatozoa occurred randomly in the epididymis. However, in the ampulla ductus deferentis, spermatozoa reaggregated and were embedded in a mucoid substance to form highly ordered spherical bundles. In the sperm bundle, the spermatozoa heads were arranged such that the helical turns of adjacent spermatozoa were precisely aligned, and all the heads in the bundle formed a distinct apex. This study demonstrates the utility of exploring the relationship between germ cells and Sertoli cells in an evolutionarily ancient vertebrate, such as the shark.     

Ultrastructure of the nuchal organs in the polychaete Platynereis dumerilii (Annelida,Nereididae)

Abstract. We examined the nuchal organs of adults of the nereidid polychaete Platynereis dumerilii by means of scanning and transmission electron microscopy. The most prominent features of the nuchal organs are paired ciliary bands located dorsolaterally at the posterior margin of the prostomium. They are composed of primary sensory cells and multiciliated supporting cells, both covered by a thin cuticle. The supporting cells have motile cilia that penetrate the cuticle and are responsible for the movement of water. Subapically, they have a narrowed neck region; the spaces between the neck regions of these supporting cells comprise the olfactory chamber. The dendrites of the sensory cells give rise to a single modified cilium that crosses the olfactory chamber; numerous thin microvillus-like processes, presumably extending from the sensory cells, also traverse the olfactory chamber. At the periphery of the ciliated epithelium runs a large nervous process between the ciliated supporting cells. It consists of smaller bundles of sensory dendrites that unite to form the nuchal nerve, which leaves the ciliated epithelium basally and runs toward the posterior part of the brain, where the perikarya of the sensory cells are located in clusters. The ciliated epithelium of the nuchal organs is surrounded by non-ciliated, peripheral epidermal cells. Those immediately adjacent to the ciliated supporting cells have a granular cuticle; those further away have a smooth cuticle. The nuchal organs of epitokous individuals of P. dumerilii are similar to those described previously in other species of polychaetes and are a useful model for understanding the development of nuchal organs in polychaetes.     

Surface morphology of rat peritoneal mast cells during in vitro regeneration after histamine secretion

Summary Cell-surface morphology of regenerating mast cells was followed over a period of 48 h after histamine release. Control cells (not stimulated to secrete) were characterized by anastomosing folds of membrane of equal depth and width. During exocytosis these folds disappeared and were replaced by deep cup-shaped flaps of membrane evident in cells incubated for 10 min. During the first hours of regeneration these flaps fused mutually or with the plasma membrane. This activity suggests membrane retrieval, maybe specifically recycling the granule-type patches of membrane. Membrane-fusion activity was observed to some degree also after extended incubation. After 48 h of incubation the regeneration process was still not completed, as indicated by the fact that holes leading to intracellular cavities could still be found.     

THE DEVELOPMENT AND POTENTIAL ROLES OF CELL‐WALL TRABECULAE IN CAULERPA MEXICANA (CHLOROPHYTA)1

Siphonous plants represent an alternate scheme to the way most macroscopic plants are constructed. They are single, often large (1–2 m), sometimes morphologically complex, multinucleate (coenocytic) cells where the whole of the cytoplasm is a continuum. Caulerpa mexicana Sond. ex Kütz. is a siphonous tropical marine green alga characterized by four morphologically distinct regions and, as with other members of the genus, by the presence of a dense network of anastomosing cylindrical cell wall in growths called trabeculae. Based on the results of this study, we propose several roles for trabeculae: (i) They are structural components, which likely add some small amount of support in compression but add considerable strength in tension. (ii) As extensions of the cell wall and plasma membrane, they act as diffusion channels from the cell exterior to the interior cytoplasm. It is possible that trabeculae also play a role in determining cell shape through developmental positioning and placement patterns, thus facilitating the diverse shapes found in the morphologically distinct regions of Caulerpa sp.