Fine Structure of the Tunic of Ciona intestinalis L. II. Tunic morphology,cell distribution and their functional importance

Ciona intestinalis L. tunic architecture and cell distribution were investigated with the electron microscope. The observations showed that the ascidian covering is formed by a thin outer cuticle, a subcuticle of variable width and a large single layer of ground substance. “Large granule”, morula, phagocyte and granulocyte are the cellular types encountered; they appear mainly in highly vacuolated states and are distributed throughout the whole tunic. The “large granule” cells, however, are mainly seen in the cuticle layer and the morula cells appear mostly in the outer zone of the ground substance. The role of these cells in tunic construction, repair and regeneration as well as their scavenging function are discussed.     

The structure of the integument of the sea cucumber,Thyone briareus

The integument and podia of the sea cucumber Thyone briareus were examined by bright field and electron microscopy. The epidermal surface was found to be covered by an acellular, PAS positive cuticle which appeared to be secreted by the underlying epidermal cells. Although the superficial portion of the cuticle contains numerous fine filaments, their ultrastructure bears no resemblance to collagen fibers. The epidermal cells are widely spaced and have long apical processes that extend along the under surface of the cuticle forming a contiguous epithelium. The apical expansions of the epidermal cells are attached to one another by means of septate desmosomes which may run continuously around all epidermal cells. Special attachment structures within these apical expansions appear to bind the cuticle to the dermis. The epidermal cells and their apical expansions are separated from the dermis by an 800 Å thick basement membrane. Granule containing cells in the upper dermis send processes up to the cuticle where they are bound to the typical epidermal cells by septate desmosomes. The abundant membrane bound granules of the cells enter villous-like processes which pass through the cuticle. The function of these cells may be to produce an adhesive material on the podia or they may be pigment cells. The thick dermis consists of a superficial zone, containing largely ground substance; a middle or laminated zone containing laminae of collagen fibers arranged in an orthogonal fashion; and a hypodermis consisting largely of ground substance and reticular fibers. Fibroblasts are abundant in the superficial dermis and between the collagen laminae. Wandering coelomocytes, or morula cells, accumulate between the collagen laminae and in the hypodermis. They may also become an integral part of the epidermis by forming septate desmosomes with epidermal cells. Morula cells contain highly specialized spherules whose tinctorial properties and electron microscopic appearance suggest that they contain protein and mucopolysaccharide.     

Electron microscopic study of the giant cells in the olfactory bulb of labyrinth fish (Belontiidae,Perciformes)

Summary The giant cells in the olfactory bulb of labyrinth fish, most likely belonging to the nervus terminalis, show ultrastructural features of active synthesis of secretory material. Dense core vesicles (70–100 nm in diameter), found in the perikaryon as well as in the axon, are the possible storage sites of the secretory substance. Its chemical nature is unknown. In some of these vesicles acid phosphatase is demonstrable. Large membrane-bound bodies (up to 2000 nm in diameter), either containing an electron dense matrix or debris of cytoplasmic organelles, are also acid phosphatase-positive, suggesting their lysosomal nature. Some other ultrastructural characteristics of these cells are also described.     

Further evidence for the protective role of sub-parietal cell membranous secretory product on the cuticle of a pentastomid arthropod parasite developing in its rodent intermediate host

The pentastomid parasite Porocephalus crotali, develops to an infective stage within a granulomatous lesion in the tissues of rodent intermediate hosts. A conspicuous layer of sub-parietal cell (SPC) secretory product, which coats the intermoult cuticle up to a depth of 12 microns, is described. Around the first five nymphal instars this material consists of an amorphous matrix with distinctive electron-lucid lacunae, but that around later instars (six and seven), while retaining much of the original morphology, possesses a significant membranous component. Host effector cells, most notably eosinophils and macrophage/epithelioid cells, are frequently completely enveloped by SPC secretion but invariably appear unreactive to it. Host cells may penetrate to the outermost layer of the epicuticle but again but again cytotoxic activity is absent. During ecdysis, effector cells are recruited to the intercuticular space where widespread degranulation is evident. Some of this is specifically directed against the underside of the cast cuticle, but not against the newly exposed cuticle. Protracted degranulation eventually reduces the cast cuticle to fragments which are endocytosed by giant cells. 1 cm long infective (seventh-stage) nymphs, which retain the sixth stage cuticle as a protective sheath, are largely devoid of membranous secretion and these were dissected from cysts, washed, and surgically transplanted into the body cavities of naive and infected mice. Pronounced differences in the onset and intensity of the subsequent inflammatory response in the two categories of host indicate some form of specific recognition. In both groups of mice though, the cuticle is an eventual target for attack by effector cells, and parasites are killed. The protective function of SPC secretion is discussed.     

B-cells of the synovial membrane

Summary Comparative ultrastructural study of the B-cells in the intimai layer of the synovial membrane in mouse, rat, rabbit, guinea-pig and man clearly distinguishes these cells from both the histiocytic A-cells (macrophage-like cells) and the fibroblasts. In addition to the marked development of the rough endoplasmic reticulum and Golgi apparatus, typical dense secretory vesicles apparently of Golgi origin are always found in mouse B-cells and frequently in those of the rat. These secretory characteristics clearly relate these cells to glandular cells engaged in polypeptidic secretion. The variations in the other species studied concern only the figuration of secretory material. Thus, the B-cells appear to constitute a category of secretory cells specific to the synovial membrane, but the function of which has yet to be determined.     

Localization of cholinesterases in structures of the alveolar portion of the mammary gland

Cholinesterase (ChE) localization in the alveolar compartment of the mammary gland has been studied by light microscopy and ultrahistochemical methods. The product of reaction to ChE is associated with micropinocytotic vesicles of the capillar endothelium and myoepithelial cells, separate nerve terminals in interalveolar connective tissue and perivascular nerve terminals. In secretory cells, the ChE activity is revealed in the sites of fat droplet and on the membranes of endoplasmatic reticulum.     

Cytological differentiation in the uropygial gland.

Ultrastructural changes were studied in the cells undergoing secretory differentiation in zone I of the tubules of the uropygial gland of White Plymouth Rock chickens. A layer of basal cells and four secretory stages are recognized as the cells migrate from the periphery to the lumen of tubules and progressively elaborate a secretion product. Basal cells, containing rough endoplasmic reticulum and free ribosomes, rest on the basement membrane and are the source from which secretory cells arise. Dilated perinuclear cisternae and the proliferation of smooth endoplasmic reticulum in the form of vesicles, invaginated sacs and cusp-shaped cisternae indicate the onset of lipgenesis in stage I cells. The perinuclear cisternae are more dilated and the endoplasmic reticulum is composed on saccules and cisternae in stage II cells. Stage III cells are characterized by concentric lamellae of endoplasmic reticulum surrounding secretory droplets. Dilated cisternae of endoplasmic reticulum and secretory droplets both contain a reticular substance. The perinuclear cisternae of stage III cells have returned to normal dimensions. Large mature lucent secretory droplets, lined with electron-dense material, fill the cytoplasm ostage IV cells which degenerate and release their secretory product into the tubule lumen. Spherical membrane-bound compartments containing a mottled substance of moderate electron density occur in basal cells and all subsequent secretory stages. These mottled bodies are surrounded by saccules of endoplasmic reticulum in stage II cells and are intimately associated with secretory droplets in stage III cells, but there is no evidence that they give rise to secretory droplets and their role in secretory differentiation is unknown.     

The fine structure of the collar cells in the optic tentacles of Helix aspersa

Summary At the base of the optic tentacular ganglion there is a group of large monopolar cells containing numerous secretory inclusions. These are the collar cells. Secretory material can be seen accumulating in swollen portions of the granular endoplasmic reticulum. It is postulated that this material is transported to the Golgi bodies and thus the limiting membrane of the inclusions is derived from the Golgi membranes. The Golgi bodies appear to be polarized and small vesicles resembling secretory inclusions are associated with one face of these organelles. The secretory inclusions fuse together to form large membrane-bound secretory pools in the perikaryon. The collar-cell processes are packed with secretory inclusions. These processes traverse the digital extensions of the tentacular ganglion and pass into the epithelium covering the tip of the tentacle. The secretory inclusions do not resemble neurosecretory inclusions in other situations. The collar cell processes receive a nerve supply from single axons containing granular and agranular vesicles. The evidence that these cells may be modified neurons is only minimal.This work was supported by the Australian Research Grants Committee.     

Ultrastructure of the sexual segments of the kidney in male and female lizards,Cnemidophorus l. lemniscatus (L.)

Summary Kidneys of adult male and female lizards were studied by electron microscopy, in order to understand the ultrastructure of the collecting duct and a differentiated part thereof, the sexual segment, which is an important accessory sexual organ. First portion of sexual segment in males: The cells are filled with large secretory granules of a wide range of opacities. The granular endoplasmic reticulum is abundant; basal formations of superimposed flat cisternae are frequent. Distended vesicles and microvesicles prevail in the supranuclear, well developed Golgi apparatus. Evidences indicate that secretion of these cells is holocrine. Second portion of sexual segment in males: All of the secretory granules are apical in location and relatively electron-opaque; they show a denser core. This core is formed by a substance which, after lying in contact with ribosomes, enters the secretory vesicles of the highly developed Golgi apparatus. A lighter substance is then condensed around it. The secretion of the granules is merocrine. The granular endoplasmic reticulum is very abundant in these cells, but basal ergastoplasmic formations are lacking. Sexual segment in females: The cells show features similar to those of the male first portion, but they are smaller. Undifferentiated collecting duct: Most of the cells are mucigenic. They have small ovoid, apical secretory granules. The density of the granules varies from cell to cell; when they are electron-lucent, they exhibit laminar or dotted opaque figures. Moderately developed Golgi apparatus and granular endoplasmic reticulum, as well as elongated mitochondria, occur in mucigenic cells. Intercalated among the latter are non-secretory cells. They have very abundant mitochondria, numerous microvilli, many pinocytic and smooth-membrane vesicles, whereas the organelles participating in synthetic processes are poorly developed; their function is most likely related to active solute transport.     

Modulation of the mitotic activity and population of the mast cells in the oral mucosa by substance P

To assess the effect of substances inducing mast cell degranulation (substance P and granuliberin R) on the mitotic indices of the gingiva stratified epithelium, basal cells from rats were studied in vivo. Seventy Lewis male rats were used in the study. The rats received injections of either 0.1 ml 0.9% NaCl (l0 rats), or substance P (10(-4), l0(-6), 10(-8) g/ml) (30 rats), or granuliberin R (10(-4), l0(-6), 10(-8) g/ml) (30 rats) into their mandibular gingiva in the vicinity of the right mental foramen. The mitotic index of keratinocytes was established after the kolchicine arrest (2 hours prior to material collection i.p. injection). The number of cells in metaphase was counted on 1000 consecutive basal layer cells after hematoxilin and eosin section staining. Mast cells were revealed using pinocyanol erythrosinate according to Bensley. Numerical density and morphometric features were analyzed. Substance P and granuliberin R injected into the gingiva affect the mast cells and the basal cell proliferation of the gingival epithelium. The diminished mitotic activity of basal layer cells was accompanied by degranulation and/or migration of mast cells under the basal membrane of the epithelium. After administration of high doses of granuloliberin R, mast cells were found in the deep connective tissue alligned towards the epithelium. A neuromediator from the trigeminal nerve (substance P) and substances from mast cells actively interfere in the proliferation of oral keratinocytes and the activity of connective tissue cells.     

The pineal gland of the gerbil,Meriones unguiculatus

Summary Electron microscopy was employed in a study of the pineal gland of the Mongolian gerbil (Meriones unguiculatus). It was determined that the gerbil pineal gland contains pinealocytes and glial cells with the pinealocytes being the predominant cell type. The pinealocytes contain numerous organelles traditionally considered as being either synthetic or secretory in function such as an extensive Golgi region, smooth (SER) and rough (RER) endoplasmic reticulum, secretory vesicles and microtubules. Other cytoplasmic components are also present in the pinealocytes (synaptic ribbons, subsurface cisternae) for which no function has been assigned. Dense-cored vesicles are rare. Vacuolated pinealocytes are present and appear to be intimately associated with the formation of the pineal concertions. Evidence presented supports the proposal that the concretions form within the vacuoles. Once the concretions reach an enlarged state, the vacuolated pinealocytes break down and the concretions are thus extruded into the extracellular space where they apparently continue to increase in size. The morphology of the glial cells was interpreted as indicative of a high synthetic activity. The glial cells contain predominantly the rough variety of endoplasmic reticulum and form an expansion around the wide perivascular area.Supported by NSF grant PCM 77-05734     

Studies on the tardigrades. 3. Fine structure of the esophagus of Milnesium tardigradum Doyère

A fine structural analysis of the esophagus of the tardigrade Milnesium tardigradum Doyère is presented. This foregut structure exhibits pronounced secretory activity with discharge of accumulated material by fusion of secretory vesicles with luminal and lateral surfaces of the esophageal cells. The possible function of this secretory material as well as the organization of esophageal cells and structure of the cuticular lining are discussed.     

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.     

The ultrastructure of the zymogen cells in Hydra viridis

Summary The zymogenic secretory cells of Hydra viridis are scattered between the digestive muscle cells of the gastric region. The mature zymogenic cells are located along the apical surface of the gastrodermal epithelium and contain numerous spherical secretory droplets. They appear to differentiate from stem cells located near the mesoglea. These stem cells resemble epidermal interstitial cells and are filled with free ribosomes. They differ from the interstitial cell in that they usually possess a small amount of granular endoplasmic reticulum. During the process of differentiation they elaborate a highly organized system of granular endoplasmic reticulum. This system becomes dispersed into vesicles as the secretory product is synthesized. There is no indication that the Golgi apparatus participates directly in the formation of the secretory droplets, and there is no indication of a membrane bounding the mature secretory droplet.The fate of the zymogenic cell following the discharge of its secretory product was not determined. It is possible that these cells revert back to a stage resembling the stem cell before resynthesizing a new supply of secretion. In this case the normal secretory process would be very similar to the events described in the dedifferentiation of the zymogenic cells during regeneration.This work was supported by Grant number GB-3262 from the National science Foundation.     

Tunichrome content in the blood cells of the tunicate, Ascidia callosa Stimpson, as an indicator of vanadium distribution

Morula, compartment, signet ring, orange, lymphocyte and amoebocyte (granular and agranular) cells have been identified in the blood of A. callosa; in addition, nephrocytes have been described. Blood cell lysates contain a yellow chromogen with spectrophotometric and fluorimetric properties similar to tunichrome. The fluorescent characteristics of each of the seven blood cell types were determined using microspectrofluorimetry. Vanadium in A. callosa blood cells is primarily associated with tunichrome extracts, although lesser amounts are measurable in blood plasma and blood cell residues; both vanadium and tunichrome concentrations are in the order morula greater than compartment greater than signet ring cells.     

Light and electron microscopy of the ducts and their subepithelial tissue in the rat ventral prostate

Summary The ducts of the rat ventral prostate have been studied by light and electron microscopy for elucidation of their role in prostatic function. The epithelium of the main duct consists of simple columnar cells and polymorphic basal cells. The columnar cells show no indication of secretory activity. The basal cells contain bundles of filaments of 5–6 nm thickness and numerous pinocytotic vesicles. The ducts are surrounded by layers of circular smooth muscle cells interspersed with nerve axons. On ultrastructural grounds the ducts do not appear to secrete material into the seminal fluid, but apparently the muscular coat actively helps drain the gland during ejaculation.     

北柴胡分泌道的超微结构及其挥发油分泌的研究

利用透射电子显微镜技术研究了北柴胡营养器官中分泌道的发育以及挥发油的积累.并重点探讨了挥发油的分泌机制.结果显示.分泌道细胞的质体、细胞基质以及线粒体参与挥发油或其前体物质的合成,而内质网参与挥发油向分泌道腔隙的转运.在分泌道发育的后期,大量小泡与分泌细胞相邻壁的质膜融合,并将其内的物质释放入此部分细胞壁中,与此同时,此部分壁在靠近腔隙的位置结构变得松散.然后围绕腔隙的壁在相邻两分泌细胞相接的位置面向腔隙外突形成许多小泡,并将小泡释放入腔隙中.由此可见.北柴胡分泌道中挥发油主要以胞吐方式被排入分泌道的腔隙中.     

Immunoelectron microscopic study of somatostatin biosynthesis in dog pancreas

The biosynthesis of somatostatin has been studied at the ultrastructural level in pancreatic islets by using rabbit antiserum against synthetic somatostatin. To document that the antiserum specifically bound preprosomatostatin, we have tested the ability of the antiserum to precipitate the product synthesized in vitro. Poly(A) enriched RNA isolated from catfish islets was translated in both the wheat germ extract and nuclease-treated reticulocyte lysate systems. It was found that the in vitro translation product, preprosomatostatin, could be recognized by the antibody against synthetic somatostatin. The morphological study was then performed by immunoelectron microscopy by using the Fab-peroxidase conjugate technique. In dog pancreatic islets, somatostatin immunoreactive reaction product was seen only in the delta cells. In these cells, they were detected on bound ribosomes, in the cisternae of the rough endoplasmic reticulum (ER) and Golgi apparatus, in the Golgi associated vesicles, and in secretory vesicles. These findings suggest that somatostatin precursor molecules are synthesized on bound ribosomes and discharged into the cisternae of the rough ER. They are then transported to the Golgi apparatus and transferred to the secretory vesicles for secretion. The different staining intensities in the secretory vesicles would suggest that the processing of the precursor molecules of somatostatin probably takes place in the secretory vesicles.     

The ultrastructure of secretion in the spermatheca of the salamander, Manculus quadrigitatus (Holbrook)

Spermathecae of mature salamanders (Manculus quadridigitatus) were studied by light and electron microscopy. Gross morphology exhibits a complex of muscle, connective tissue and pigment cells surrounding a cluster of tubules, which empty into a ciliated central duct leading to the cloacal cavity. The tubules are composed of myoepithetial cells and secretory cells, anchored to an encompassing basal lamina. Secretion products appear to be initiated as crystalline deposits, seen in mitochondrial repositories, which are subsequently sequestered in the perinuclear region. Observations show these vesicles to be precursors of the Golgi synthesis of carboxylated polysaccharides, as determined by histochemical tests using toluidine blue, ruthenium red, and alcian blue. The secretory products are emptied into the tubule lumen. thereby bathing the stored sperm and maintaining them in a viable state for approximately 8 months. The storage tubules appear to be a complete complex of varying epithelial cells specifically designed to support viable sperm and to resorb non-functional forms.     

ULTRASTRUCTURAL DEVELOPMENT OF CAPITATE GLANDULAR HAIRS OF CANNABIS SATIVA L. (CANNABACEAE)

The capitate-sessile and capitate-stalked glands of the glandular secretory system in Cannabis, which are interpreted as lipophilic type glandular hairs, were studied from floral bracts of pistillate plants. These glands develop a flattened multicellular disc of secretory cells, which with the extruded secretory product forms the gland head and the auxiliary cells which support the gland head. The secretory product accumulates beneath a sheath derived from separation of the outer wall surface of the cellular disc. The ultrastructure of secretory cells in pre-secretory stages is characterized by a dense ground plasm, transitory lipid bodies and fibrillar material, and well developed endoplasmic reticulum. Dictyosomes and dictyosome-derived secretory vesicles are present, but never abundant. Secretory stages of gland development are characterized by abundant mitochondria and leucoplasts and by a large vacuolar system. Production of the secretory product is associated with plastids which increase in number and structural complexity. The plastids develop a paracrystalline body which nearly fills the mature plastid. Material interpreted as a secretion appears at the surface of plastids, migrates, and accumulates along the cell surface adjoining the secretory cavity. Extrusion of the material into the secretory cavity occurs directly through the plasma membrane-cell wall barrier.