Ultrastructure of the Circulatory System of the Phoronid Phoronopsis harmeri Pixell, 1912. 2. Main vessels

The ultrastructure of the wall of the main blood vessels of the phoronid Phoronopsis harmeri is described. The walls of the lophophoral and left lateral vessels consist of myoepithelial cells of the coelomic lining (peritoneal cells), a thin basal lamina, and an incomplete endothelial lining. In the head region of the body, the wall of the medial vessel consists of myoepithelial cells of the coelomic lining (peritoneal cells), a basal lamina, and true muscular endothelial cells. The anterior part of the medial vessel functions as the heart. In the anterior part of the body, the medial vessel wall consists of five layers: the external nonmuscular coelothelium, a layer of the extracellular matrix, the internal muscular coelothelium, an internal layer of the extracellular matrix, and an incomplete endothelial lining. The complicated structure of the medial vessel wall may be explained by the superimposition of the lateral mesentery on the ordinary vessel wall.     

Structure of the Digestive Tube in the Holothurian Eupentacta fraudatrix (Holothuroidea: Dendrochirota)

In the holothurian Eupentacta fraudatrix,the gut wall exhibits trilaminar organization. It consists of an inner digestive epithelium, a middle layer of connective tissue, and an outer mesothelium (coelomic epithelium). The pharynx, esophagus, and stomach are lined with a cuticular epithelium composed of T-shaped cells. The lining epithelium of the intestine and cloaca lacks a cuticle and consists of columnar vesicular enterocytes. Mucocytes are also encountered in the digestive epithelium. The connective tissue layer is composed of a ground substance, which houses collagen fibers, amoebocytes, morula cells, and fibroblasts. The gut mesothelium is a pseudostratified epithelium, which is dominated by peritoneal and myoepithelial cells and also includes the perikarya and processes of the neurons of the hyponeural plexus and vacuolated cells.     

Heart and the peritoneal cover of the gut sinus in the polychaete Arenicola marina: an ultrastructural and autoradiographic study

To elucidate the cellular mechanism underlying the growth of the peritoneal cover of the gut sinus and the heart in the polychaete Arenicola marina, cellular organization of these structures and proliferative potential of their cells were investigated using electron microscopy and electron microscopic autoradiography. Arenicola has a pair of dorsolaterally situated hearts connected to the gut sinus via a short duct and composed of two muscular layers separated by a layer of the extracellular matrix (ECM). The peritoneal cover of the gut sinus and the outer muscular layer of the heart present a myoepithelial layer resting on the ECM. The inner muscular layer of the heart is composed of myofibril-containing cells lacking well-defined polarity in arrangement of organelles. However, their persistent connection to branches of the ECM and the adherens-like intercellular junctions allow for considering the inner layer a modified myoepithelium. In the peritoneal cover of the gut sinus and in both myoepithelial layers of the heart, noncontractile epithelial cells have been observed. As determined by thymidine labeling, these epithelial cells are capable of DNA synthesis, while myoepithelial cells are not. Some suggestions are made about the myogenic nature of the epithelial cells in the investigated structures of A. marina.     

Ultrastructure of the body cavities in juveniles and adults of the appendicularian Oikopleura gracilis (Tunicata,Chordata) suggests how the heart may have evolved in tunicates

The organization of the body cavities is an important morphological trait that can be used for establishing the phylogenetic relationships between different groups of animals. In the present study, the hemocoel and coelomic systems of 10‐hr‐old juveniles and adults of the hermaphroditic oikopleurid Oikopleura gracilis were examined using light and transmission electron microscopy. The trunk hemocoel in 10‐hr‐old juveniles was represented by small clefts containing layers of extracellular matrix of adjacent tissues or interstices with migrating primordial germ syncytium. The wide hemocoel in the tail contained extracellular strands, subdividing the hemocoel into hemal sinuses. In adults, a large hemocoel appeared in the trunk and tail, and also contained extracellular strands. The hermaphroditic gonad was surrounded by its own lining, separating it from the hemocoel. The gamete‐filled cavity in the ovary and testis appeared only at late‐stage gonadogenesis, when the pre‐spawning reduction of syncytium occurred in the gonads. The true coelom in 10‐hr‐old juveniles and adults was represented by the pericardium. The lining of the pericardium consisted of myoepithelial and peritoneal cells. In the myoepithelial cells of 10‐hr‐old juveniles, myofibrils had been formed. The myoepithelial cells of adults had several parallel rows of completely differentiated myofibrils. The substantial reduction of the coelomic and circulatory systems in O. gracilis evidently results from the extreme shortening of ontogeny in appendicularians. Development in O. gracilis from early juvenile to adult involves the following steps, which also suggest how the tunicate heart may have evolved: a single‐layered coelomic sac gives rise to a grooved pericardium with an open hemal sinus (simple heart). In ascidians, this simple heart in turn gives rise to a closed tubular, double‐layered heart–pericardial complex, with a separate pericardial cavity and a closed heart, whose wall is formed by specialized myocardium.     

The periesophageal celom of the articulate brachiopod Hemithyris psittacea (Rhynchonelliformea,Brachiopoda)

The celomic system of the articulate brachiopod Hemithyris psittacea is composed of the perivisceral cavity, the canal system of the lophophore, and the periesophageal celom. We study the microscopic anatomy and ultrastructure of the periesophageal celom using scanning and transmission electron microscopy. The periesophageal celom surrounds the esophagus, is isolated from the perivisceral cavity, and is divided by septa. The lining of the periesophageal celom includes two types of cells, epithelial cells and myoepithelial cells, both are monociliary. Some epithelial cells have long processes extending along the basal lamina, suggesting that these cells might function as podocytes. The myoepithelial cells have basal myofilaments and may be overlapped by the apical processes of the adjacent epithelial cells. The periesophageal celom forms protrusions that penetrate the extracellular matrix (ECM) of the body wall above the mouth and the ECM that surrounds the esophagus. The canals of the esophageal ECM form a complicated system. The celomic lining of the external circumferential canals consists of the epithelial cells and the podocyte‐like cells. The deepest canals lack a lumen; they are filled with the muscle cells surrounded by basal lamina. These branched canals might perform dual functions. First, they increase the surface area and might therefore facilitate ultrafiltration through the podocyte‐like cells. Second, the deepest canals form the thickened muscle wall of the esophagus and could be necessary for antiperistalsis of the gut. J. Morphol., 2011. © 2010 Wiley‐Liss, Inc.     

Reinvestigation of epithelial lining of the genital coelomic sinus in asteroids. An ultrastructural study

Ultrastructural study of gonadal muscles in sea star, Asterina pectinifera, showed that myoepithelial cells were located only in the epithelial lining of the genital coelomic sinus. No myoepithelial cells were found in the visceral peritoneal epithelium or within connective tissue layer of the outer sac. Morphology of the myoepithelial cells in gonads of A. pectinifera varies during the reproductive cycle. During the gametogenic phase of the reproductive cycle, the myoepithelial cells get an elongated, spindle-like shape having a length of 20–30 μm. In prespawning gonads, many of the myoepithelial cells form cytoplasmic extensions of 3–5 μm in length, filled with myofilaments and penetrating into the underlying connective tissue of the outer sac or haemal sinus. Besides, myoepithelial cells, simultaneously anchored in the inner and outer sacs, were also observed. These changes result in development of more elaborated musculature and increase in contractility of the gonadal wall in prespawning gonads as compared to that during other stages of the reproductive cycle.     

Breeding and post-breeding structure of the vas deferens of the long-toed salamander Ambystoma macrodactylum

The vas deferens of Ambystoma macrodactylum is composed of a peritoneal epithelium, connective tissue layer with fibroblasts, circular smooth muscle, capillaries, cells containing lipid, and a luminal epithelium composed of a single layer of cuboidal cells covered by a net of interconnected ciliated squamous cells. The cuboidal cells have abundant rough endoplasmic reticulum, mitochondria, and PAS + secretory vesicles. Squamous cells of breeding males consistently have tufts of ～100 cilia located at one end of the long axis of each cell. These cilia may help distribute secretory products. The squamous cells, absent in post-breeding males, are apparently sloughed into the lumen. Lipid vesicles are present throughout the cytoplasm of the cuboidal and squamous epithelial cells and are also in some cells of the connective tissue layer. These vesicles increase dramatically in number during the first 4 weeks after breeding and may serve as an energy pool for the next breeding season. Enzyme-histochemical tests for testosterone synthesis were negative. In addition to the accumulation of lipid and the loss of squamous cells in the vas deferens, after breeding PAS + vesicle production is terminated. These alterations appear to represent energy conservation strategies employed by the sperm-depleted vas deferens.     

Ultrastructural study of the proboscis endothelium of Riseriellus occultus (Nemertea, Heteronemertea)

We have examined with transmission electron microscopythe epithelial layer exposed to the rhynchocoel fluidof the proboscis in the heteronemertine Riseriellus occultus. This epithelium is organized asa monociliated, pseudostratified myoepitheliumconsisting of two cell types: apically situatedmonociliated supportive cells and subapical myocyteslacking cilia. The low supportive cells form acontinuous adluminal sheet and reach with numerouscytoplasmic processes into the extracellular matrix;these cells are characterized by numerous, irregularlyshaped, apical folds projecting into the rhynchocoelfluid, delimiting broad extracellular spaces. Theauthors suppose that both apical and basal folds couldaccommodate stretching of the endothelium when theproboscis is everted. The apical folds of thesupportive cells increase the interface of these withthe rhynchocoel fluid; this feature, together with thepresence of pinocytotic vesicles in such cells,suggest that they could be involved in the exchange ofsubstances between the rhynchocoel fluid and theproboscis. The myocytes are scattered singly withinthe monociliated pseudostratified myoepithelium. Theyare situated between the supportive cells and thesubjacent extracellular matrix. Basement membraneseparating both cells types is lacking. Myofibrillarparts protrude basally from the myocyte somata. Themyofibrillar parts lie in direct apposition to theextracellular matrix, and are oriented circular to thelongitudinal axis of the proboscis. We consider themyocytes to be intra-epithelial, myoepithelial cells.     

Brachiopod tentacles: Ultrastructure and functional significance of the connective tissue and myoepithelial cells in Terebratalia

Summary The fine structure of the tentacles of the articulate brachiopod Terebratalia transversa has been studied by light and electron microscopy. The epidermis consists of a simple epithelium that is ciliated in frontal and paired latero-frontal or latero-abfrontal longitudinal tracts. Bundles of unsheathed nerve fibers extend longitudinally between the bases of the frontal epidermal cells and appear to end on the connective tissue cylinder; no myoneural junctions were found. The acellular connective tissue cylinder in each tentacle is composed of orthogonal arrays of collagen fibrils embedded in an amorphous matrix. Baffles of parallel crimped collagen fibrils traverse the connective tissue cylinder in regions where it buckles during flexion of the tentacle.The tentacular peritoneum consists of four cell types: 1) common peritoneal cells that line the lateral walls of the coelomic canal, 2) striated and 3) smooth myoepithelial cells that extend along the frontal and abfrontal sides of the coelomic canal, and 4) squamous smooth myoepithelial cells that comprise the tentacular blood channel.Experimental manipulations of a tentacle indicate that its movements are effected by the interaction of the tentacular contractile apparatus and the resilience of the supportive connective tissue cylinder. The frontal contractile bundle is composed of a central group of striated fibers and two lateral groups of smooth fibers which function to flex the tentacle and to hold it down, respectively. The small abfrontal group of smooth myoepithelial cells effects the re-extension of the tentacle, in conjunction with the passive resiliency of the connective tissue cylinder and the concomitant relaxation of the frontal contractile bundle.The authors wish to express their appreciation to Professor Robert L. Fernald for his advice and encouragement throughout the course of this study. Some of the work was conducted 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. Part of this study was supported by NIH Developmental Biology Training Grant No. 5-T01-HD00266 and NSF grant BMS 7507689     

Ultrastructural changes in the connective tissue of the gastric region during the metamorphosis of Xenopus laevis

Development of the gastric connective tissue of Xenopus laevis during metamorphosis was investigated by electron microscopy. Throughout the larval period to stage 60, the layer of connective tissue underlying the gastric epithelium consists of immature fibroblasts surrounded by a sparse extracellular matrix. At the beginning of the transition from the larval to the adult epithelial form, at about stage 60, extensive changes occur in the connective tissue. The number of cells suddenly increses and different cell types appear. Numerous contacts between epithelial and connective tissue cells are established through random gaps in the thickened basal lamina. During stages 62–63, just after the beginning of the morphogenesis of adult-type glands, the basal lamina lining the glandular epithelium becomes thinner, and the number of contacts decreases rapidly except near the tips of the glands. After the glandular cells begin to produce zymogen granules at stage 64, contacts become rare. From stage 63, when the muscularis mucosae develops, until the completion of metamorphosis, the connective tissue consists mainly of typical fibroblasts. Outside the muscularis mucosae, the fibroblasts of the lamina propria are aligned in parallel with the curvature of the glands. These observations indicate that developmental changes in the connective tissue are closely related spatiotemporally to those of the epithelial transition from larval to adult form during metamorphic climax. Although some changes are similar to those in the intestine (Ishizuya-Oka and Shimozawa, '87b), others are specific to the gastric region, which suggests that connective tissue may have a role in organ-specific differentiation of the gastric epithelium.     

Light,electron microscopical,and immunocytochemical investigation of the stomach wall of the tigerfish Hydrocynus forskahlii

The wall of the stomach of the tigerfish is described and compared with that of other vertebrates. Light microscopic and ultrastructural characteristics of the stomach wall correspond to a large extent to those of other vertebrates, although some differences are found. The mucosa contains (1) surface epithelium characterized by narrow columnar cells with abundant mucous granules; (2) gastric glands consisting of pepsinogenic cells of variable height, containing tubulovesicles and bearing microvilli; (3) five granulated cell types located basally in the epithelium (types 1–5); and (4) lamina propria and muscularis mucosae. Connective tissue separating smooth muscle fibers of the muscularis mucosae constitutes a stratum compactum. The submucosa contains a loose connective tissue, a tunica muscularis of inner circular and outer longitudinal layers, and a serosa of mesothelium and subjacent connective tissue. Immunocytochemical tests with antisera to five polypeptides show gastrin/cholecystokinin (CCK), vasoactive intestinal polypeptide (VIP) immunoreactivities in some cells of the gastric glands, and somatostatin in cells lying among epithelial cells lining the gastric luminal surface or gastric pits.     

Ultrastructure of the Alimentary Canal in Five-Month-Old Pentactulae of the Holothurian Eupentacta fraudatrix

Five-month-old pentactulae (juveniles) of the holothurian Eupentacta fraudatrixpossess a well-developed alimentary canal comprising an esophagus, a stomach, an intestine, and a rectum. The intestine in turn consists of five parts. The esophagus, stomach, and rectum are lined with a cuticular epithelium. The intestinal lining lacks a cuticle and is composed of mainly polyfunctional vesicular enterocytes. Granular enterocytes are less abundant; their cytoplasm contains electron-dense granules, which are probably zymogenic. The gut connective tissue consists of electron-lucent ground substance with collagen fibers and embedded coelomocytes. The gut mesothelium is composed of myoepithelial and peritoneal cells and contains the neurons of the hyponeural nerve plexus.     

Ultrastructural and chemical changes in the cell wall of Haematococcus pluvialis (Volvocales,Chlorophyta) during aplanospore formation

Changes in the ultrastructure and chemistry of the cell wall of the unicellular volvocalean green alga Haematococcus pluvialis were investigated during the transformation of flagellates into aplanospores. The motile biflagellated state exhibited a distinct gelatinous extracellular matrix. Its ultrastructure resembled the typical volvocalean multilayered architecture with a median tripartite crystalline layer. The transformation into the non-motile cell state was characterized by formation of a new layer, a primary wall, within the extracellular matrix. During this process, the initial extracellular matrix remained intact except for the outer layers of the tripartite crystalline layer, which decomposed. Further morphogenesis of the aplanospore resulted in the formation of a voluminous multilayered cell wall. A trilaminar sheath was formed inside the primary wall and the innermost and thickest part was an amorphous secondary wall, consisting mostly of a mannan. Results obtained by staining with the fluorescent dye primuline as well as by acetolysis suggest the occurrence of sporopollenin-like material (algaenan) within the trilaminar sheath of the aplanospore cell wall. The primary wall and the outer remnants of the extracellular matrix disintegrated as the aplanospores aged, and were completely absent in the resting cell state.     

The Role of the Microenvironment in Mammary Gland Development and Cancer

The mammary gland is composed of a diverse array of cell types that form intricate interaction networks essential for its normal development and physiologic function. Abnormalities in these interactions play an important role throughout different stages of tumorigenesis. Branching ducts and alveoli are lined by an inner layer of secretory luminal epithelial cells that produce milk during lactation and are surrounded by contractile myoepithelial cells and basement membrane. The surrounding stroma comprised of extracellular matrix and various cell types including fibroblasts, endothelial cells, and infiltrating leukocytes not only provides a scaffold for the organ, but also regulates mammary epithelial cell function via paracrine, physical, and hormonal interactions. With rare exceptions breast tumors initiate in the epithelial compartment and in their initial phases are confined to the ducts but this barrier brakes down with invasive progression because of a combination of signals emitted by tumor epithelial and various stromal cells. In this article, we overview the importance of cellular interactions and microenvironmental signals in mammary gland development and cancer.The mammary gland is composed of a combination of multiple cell types that together form complex interaction networks required for the proper development and functioning of the organ. The branching milk ducts are formed by an outer myoepithelial cell layer producing the basement membrane (BM) and an inner luminal epithelial cell layer producing milk during lactation. The ducts are surrounded by the microenvironment composed of extracellular matrix (ECM) and various stromal cell types (e.g., endothelial cells, fibroblasts, myofibroblasts, and leukocytes). Large amount of data suggest that cell-cell and cell-microenvironment interactions modify the proliferation, survival, polarity, differentiation, and invasive capacity of mammary epithelial cells. However, the molecular mechanisms underlying these effects are poorly understood. The purification and comprehensive characterization of each cell type comprising normal and neoplastic human breast tissue combined with hypothesis testing in cell culture and animal models are likely to improve our understanding of the role these cells play in the normal functioning of the mammary gland and in breast tumorigenesis. In this article, we overview cellular and microenvironmental interactions that play important roles in the normal functioning of the mammary gland and their abnormalities in breast cancer.     

Ultrastructure of somatic tissues in the ovaries of a chaetognath (Ferosagitta hispida)

Transmission electron microscopy reveals that the ovaries of Ferosagitta hispida contain four somatic tissues. A myoepithelial ovary wall, continuous with a thin layer of peritoneocytes lining the coelomic cavity, encloses a fluid-filled ovarian space in which oocytes develop. Lamellar extensions of a “follicular reticulum” branch throughout the ovarian space and ensheath developing oocytes. This tissue has been overlooked in most previous studies of chaetognath ovaries. A bipartite oviductal complex extends the length of each ovary just within the lateral ovary wall. It consists of a flattened, blindly ending cellular tube, herein referred to as the cellular sheath, and an enclosed syncytium. Sheath cells secrete an electron-dense product into the ovarian space. Those sheath cells directly bordering the syncytium are contractile and are joined to the to the syncytium by gap junctions and microvillar interdigitations. The syncytium contains a complex of membrane-bounded lumina. The latter sometimes enclose sperm received during mating or ovulated eggs. Thus the syncytium serves both as a seminal receptacle and as a duct for passage of eggs to the outside. Contrary to several classical reports, the cellular sheath and syncytium of the oviductal complex do not separate at ovulation to form a temporary oviductal lumen.     

Myoepithelial molecular markers in human breast carcinoma PMC42-LA cells are induced by extracellular matrix and stromal cells

Summary The microenvironment plays a key role in the cellular differentiation of the two main cell lineages of the human breast, luminal epithelial, and myoepithelial. It is not clear, however, how the components of the microenvironment control the development of these cell lineages. To investigate how lineage development is regulated by 3-D culture and microenvironment components, we used the PMC42-LA human breast carcinoma cell line, which possesses stem cell characteristics. When cultured on a two-dimensional glass substrate, PMC42-LA cells formed a monolayer and expressed predominantly luminal epithelial markers, including cytokeratins 8, 18, and 19; E-cadherin; and sialomucin. The key myoepithelial-specific proteins α-smooth muscle actin and cytokeratin 14 were not expressed. When cultured within Engelbreth-Holm-Swarm sarcoma-derived basement membrane matrix (EHS matrix), PMC42-LA cells formed organoids in which the expression of luminal markers was reduced and the expression of other myoepithelial-specific markers (cytokeratin 17 and P-cadherin) was promoted. The presence of primary human mammary gland fibroblasts within the EHS matrix induced expression of the key myoepithelial-specific markers, α-smooth muscle actin and cytokeratin 14. Immortalized human skin fibroblasts were less effective in inducing expression of these key myoepithelial-specific markers. Confocal dual-labeling showed that individual cells expressed luminal or myoepithelial proteins, but not both. Conditioned medium from the mammary fibroblasts was equally effective in inducing myoepithelial marker expression. The results indicate that the myoepithelial lineage is promoted by the extracellular matrix, in conjunction with products secreted by breast-specific fibroblasts. Our results demonstrate a key role for the breast microenvironment in the regulation of breast lineage development.     

Effect of components of the extracellular matrix on the accumulation of lipids in human cells

The effect of extracellular matrix components on cholesterol accumulation in different human cells was studied. Insoluble LDL-Heparin-Fibronectin-Gelatin complexes were incubated with human cells: fibroblasts, monocytes, peritoneal macrophages, cells of the aortic wall--endothelial, subendothelial intimal, medial; and total cholesterol content in these cells was determined. It has been demonstrated that components of extracellular matrix being complexed with LDL enhance total cholesterol accumulation in all cell types studied: the highest amount of cholesterol was accumulated by subendothelial intimal cells and peritoneal macrophages. It is suggested that components of extracellular matrix can play an important role in the development of lipid-laden foam cells that are accumulated in the arterial wall in atherosclerotic lesions.     

To create the correct microenvironment: three-dimensional heterotypic collagen assays for human breast epithelial morphogenesis and neoplasia

The normal human breast comprises an inner layer of luminal epithelial cells and an outer layer of myoepithelial cells separated from the connective tissue stroma by an intact basement membrane. In breast cancer, tumor cells are in direct contact with the surrounding highly activated collagenous stroma, with little or no discernible myoepithelial fence from the original double-layered structure. To understand the evolution of these two scenarios, we took advantage of a three-dimensional hydrated collagen gel approach. The contribution of myoepithelial cells to normal morphogenesis was studied by ablation and rescue experiments, and genes regulated on tumor cell-fibroblast interaction were identified in a tumor environment assay. In normal breast morphogenesis, the ability to correctly polarize sialomucin to the luminal membrane of emerging acini was used as a criterion for apical polarity and functional differentiation. In the assay of breast neoplasia, the consequence of reciprocal tumor cell-fibroblast interaction was addressed morphologically as well as by a differential display approach. Normal breast epithelial cells were purified immunomagnetically and an established cell line, MCF-7, was used as a surrogate tumor cell. With regard to the importance of myoepithelial cells in normal breast epithelial morphogenesis, the collagen gel assay elucidated the following subtleties: In contrast to culturing in basement membrane gels, luminal epithelial cells when cultured alone made structures that were all inversely polarized. This aberrant polarity could be rescued by co-culture with myoepithelial cells. The molecular activity of myoepithelial cells responsible for correct morphogenesis was narrowed down to the laminin-1 component of the basement membrane. As for the consequence of interaction of tumor cells with connective tissue fibroblasts, the assay allowed us to identify a hitherto undescribed gene referred to as EPSTI1. The relevance of the assay-based identification of regulated genes was confirmed in a series of breast carcinomas in which EPSTI1 was highly upregulated compared with normal breast. Few if any of these observations would have been possible on two-dimensional tissue culture plastic.     

The ultrastructure of the atrium in the adult newt Notophthalmus viridescens (Amphibia,Salamandridae)

The atrial wall of Notophthalmus viridescens is 25–75 μm thick and is trabeculated sparsely. Coronary vessels are absent. The endocardial endothelium is continuous and has 50–60 nm-wide fenestrae with diaphragms, rests on a discontinuous basal lamina and lacks occluding junctions. Cells found in the subendothelial connective tissue are xanthophores, melanophores, mast cells, fibroblasts, macrophages, and unmyelinated nerve fibers with Schwann cell investments. Epicardial mesothelial cells contain numerous 6–7 nm filaments and lamellar bodies which resemble myelin figures. Mesothelial cell junctions include maculae adhaerentes diminutae, desmosomes, and interdigitations. The epicardial connective tissue layer is more extensive than that of the endocardium, with xanthophores and melanophores rarely present and nerve fibers never observed. The myocardium consists of a mesh-work of myocytes 3–5 cell layers thick with little intervening connective tissue. Myocytes are 6–10 μm in diameter and have two or three peripheral myofibrillae. Typical A, I, H, Z, and M bands are present with a sarcomere length of 2.5 μm. T tubules are not observed. The sarcoplasmic reticulum has subsarcolemmal dilations. The nuclear pole region contains abundant mitochondria and atrial granules, extensive Golgi, and elements of smooth and rough-surfaced endoplasmic reticulum. Lateral intercellular junctions consisting of dense plaques, frequently continuous with Z-line material, are common. Oblique and transversely oriented junctions consisting of primarily of fascia adhaerentes, are present. It appears that amphibian atrial myocytes more closely resemble those of the amphibian ventricle than those of the mammalian atrium. Structural differences between amphibian atrial and ventricular myocytes seem to be quantitative rather than qualitative in nature.     

Connective tissue is involved in adult epithelial development of the small intestine during anuran metamorphosis in vitro

Summary The role of connective tissue in metamorphic changes of the small intestinal epithelium inXenopus laevis tadpoles was investigated by using organ culture techniques and electron microscopy. Tissue fragments isolated from various parts of the small intestine at stage 57 were cultivated. Larval cell death of the epithelium was induced by thyroid hormone in all fragments, whereas adult epithelial development was observed only in fragments isolated from the anterior intestinal region containing the typhlosole where most of the larval connective tissue was localized. The epithelium was then cultivated in recombination with homologous or heterologous non-epithelial components. The adult epithelium developed only in recombinants containing a thick connective tissue layer from the typhlosole. There was no regional difference in the developmental potency of the epithelium itself. In all explants where adult epithelium developed, the connective tissue increased in cell density just beneath the epithelium, which was rapidly proliferating and forming typical islets. At the same time, fibroblasts possessing well-developed rough endoplasmic reticulum differentiated close to epithelial cells and often made contact with them. These results indicate that the connective tissue originating from the typhlosole plays an important role in adult epithelial development of the anuran small intestine, probably via direct cell-to-cell contacts or some factor(s) synthesized by the fibroblasts.