American Journal of Anatomy最新文献

Ultrastructural alterations induced by colchicine were investigated to determine the secretory activities of odontogenic cells during formation of tooth enameloid matrix in skates. Treated skate inner dental epithelial (IDE) cells did not display dilated cisternae of the granular endoplasmic reticulum (GER) nor accumulate Golgi-associated secretory granules at any dose level or time interval examined. This response was markedly different from that observed in teleost IDE cells synthesizing the enameloid collagen matrix. Treated skate IDE cells did show increased accumulations of glycogen-containing vesicles and intercellular glycogen associated with amorphous material, compared to controls. Additionally, the aberrant occurrence of large intracellular glycogen pools and amorphous material suggested that carbohydrate processing was a major function of skate IDE cells. Treated odontoblasts associated with enameloid matrix formation sometimes showed dilated GER cisternae, but procollagen secretory granules were not observed. Instead, electron dense material was present within the Golgi cisternae, tubular granules, and large granules. Some electron-dense material appeared to be shunted to a resorptive pathway via multivesicular bodies in treated odontoblasts. The continuity of tubular granules with the enameloid matrix suggested that they contained precursors of the enameloid matrix, and possibly the periodic, 17.5-nm cross-striated, "giant" fibers. Treated odontoblasts associated with predentin collagen matrix deposition showed dilated GER cisternae and accumulations of procollagen secretory granules, features consistent with the function of active collagen synthesis and secretion. The findings indicate that (1) skate IDE cells do not synthesize enameloid collagen as found in bony fish tooth development; (2) skate IDE cells do process glycogen for secretion into the enameloid matrix; (3) collagen, although present, is not a major constituent of skate enameloid matrix; (4) enameloid "giant" fibers are unique to elasmobranchs; and (5) odontoblasts synthesize and secrete proteins other than collagen into the enameloid matrix.

In male Fischer rats, a class of follicles with flat epithelium is present as a minor component of thyroid glands in which most of the follicles have cuboidal epithelium. These follicles occur in thyroids that have been made hyperplastic by feeding the rats thiouracil for 21 days and then allowing involution for 21 days or more. They also occur in older control rats. The follicles resemble in morphology, at the light-microscope level, the so-called "cold" follicles that occur in aged mice. We have examined the ultrastructure of the flat cells in these follicles and compared it with that of the flat cells occurring in the thyroid follicles of hypophysectomized rats. The cells in involution have abundant rough endoplasmic reticulum (RER) and few lysosomes and, in these respects, differ markedly from cells in hypophysectomized rats. The follicles with flat cells are surrounded by a normal incidence of blood capillaries, so that the cells do not appear to be deprived of access to an adequate supply of materials necessary to satisfy their metabolic requirements. Their abundant RER suggests that they have thyroid-stimulating hormone (TSH) receptors, so that the flat cell may be the result of some process occurring at a step distal to receptor coupling with TSH. Their occurrence in young rats after the induction of hyperplasia may be a consequence of cell multiplication producing a clone of neighboring abnormal cells that have an abnormally small cell height.

Mitochondrial structure has been examined in three dimensions using high-resolution scanning electron microscopy in cells from rat liver, retina (photoreceptors and retinal pigment epithelium), and kidney (proximal convoluted tubular cells and podocytes). Tissues were prepared by aldehyde-osmium fixation and freeze cleavage using a cryoprotectant, followed by removal of the cytosol by immersion in a dilute osmium tetroxide solution. The microscope used (Hitachi S-570) was equipped with a secondary electron detector located in the column above the specimen, situated within the objective lens. Mitochondria in all tissues examined were found to have only tubular cristae, which in some instances could be seen to span the entire diameter of the organelle. The walls of the tubular cristae, when unfractured, were in contact with the inner mitochondrial membrane; and their lumens were open to the intermembranous space. We hypothesize that in cells of many, perhaps most tissues, mitochondrial cristae are not shelf-like but are, in fact, tubes which span the mitochondrial matrix and are continuous with the inner mitochondrial membrane at both ends.

The endodermal layer of the human yolk sac was examined three-dimensionally with light microscopy on serial sections using scanning electron microscopy and transmission electron microscopy to find the origin of hemopoiesis in the yolk sac. Cell-labelling techniques were also employed using the monoclonal anti-transferrin receptor antibody. Orifices of the endodermal and intracellular tubules facing the yolk-sac cavity were demonstrated on the endodermal surface. Various-sized blood cells in various stages of differentiation and maturation were distributed in the yolk-sac cavity and tubules and were observed also at the orifices of the tubules. The morphological and the immunological findings suggest that blood cells with large nuclei in the endodermal layer are the most immature. The present results suggest that blood cells originate from the endodermal layer and are carried to the embryo through the yolk sac cavity and the vitelline duct. It is probable that the endodermal and intracellular systems of tubules have an important role in the transport of blood cells, including stem cells.

An immunohistochemical and ultrastructural study of human melanoma colonies grown in soft agar for up to 50 days was performed. Three morphological variants of developing tumor colonies are reported: 1) large light colonies, 2) small dark colonies, and 3) smooth-edged colonies. The large light colony variant is the most frequently observed in the soft agar assay (approximately 70%), followed by the dark colony variant (approximately 27%), and the smooth-edged colony variant (approximately 3%). Major morphological characteristics are associated with each variant, as shown with light microscopy (LM) and transmission electron microscopy (TEM). Both LM and TEM analyses demonstrated that the large light colony variant was hypomelanotic and contained a microfibrillar extracellular matrix (ECM). The small dark colony variant was found to be hypermelanotic and contained a less demonstrable ECM. The smooth-edged variant has an encapsulated periphery, no demonstrable ECM, and tightly packed cells with desmosome-like junctions. In order to characterize further the ECM in the most commonly observed variant, the large light colony, specific antibodies to fibronectin (FN) and collagen types IV and V (COLs IV and V) were applied and observed with immunofluorescence microscopy and immunoperoxidase. In paraffin sections of melanoma colonies, FN was observed associated with both the cell surface and the ECM. However, no specific staining was seen for COLs IV and V. In addition, ruthenium red was used to preserve and selectively bind to glycosaminoglycans (GAGs) and proteoglycans (PGs). TEM studies reveal GAG-like granules stained with ruthenium red in the fibrillar ECM and a dotted, punctate staining of the cell surface. Understanding the biological and architectural composition of developing melanoma tumor colonies in soft agar could contribute to the development of more efficient chemotherapeutic strategies.

Cryostat- and vibratome-cut sections of rat kidneys were singly or doubly labeled to visualize immunoreactive tyrosine hydroxylase (THI), dopamine beta-hydroxylase (DBHI), vasoactive intestinal peptide (VIPI), and neuropeptide Y (NPYI). Rats were perfusion fixed with 2-4% paraformaldehyde with or without 0.15% picric acid and rinsed in buffer for 18-48 hr. Single antigens were labeled with horseradish peroxidase in vibratome sections, whereas cryostat sections were used to label one antigen with peroxidase and another with a fluorophore in the same tissue section. A dense plexus of DBHI noradrenergic nerves innervates the renal arterial tree, and such nerves innervate the interlobar veins and renal calyx as well. Immunoreactive NPY is colocalized in most of these nerves, but some intrarenal noradrenergic nerves do not contain NPY but do contain VIP immunoreactivity. The distribution of NPYI nerves resembles that of DBHI nerves, whereas most perivascular noradrenergic nerves immunoreactive for VIP innervate selected arcuate and interlobular arteries. A small population of nonadrenergic, VIPI nerves innervates the renal calyx.

The postnatal development of the Sertoli cell barrier, tubular lumen, fluid flow, and cytoskeletal elements in Sertoli and myoid cells was investigated in the Sprague-Dawley rat. With the aid of hypertonic fixatives, a barrier to the rapid entry of fluid was noted in the majority of tubules on the 15th and 16th postnatal (p.n.) days and was completely formed in all tubules prior to p.n. day 18. The actin forming the ectoplasmic specialization (ES), a cytoskeletal complex related to the occluding junctions composing the barrier, began its development during the period of initial barrier formation (16 p.n. day) and progressively attained its adult prominence. The ES developed its characteristic adult pattern and adult fluorescent intensity at about p.n. day 22. Some seminiferous tubules showed very small lumina as early as p.n. day 10. All tubules were not open until p.n. day 30. The size (diameter) of the lumen increased slowly from p.n. day 10 until p.n. day 30 when it started to increase rapidly until about p.n. day 50. Fluid flow in seminiferous tubules was detected as early as p.n. day 20 and increased in amount thereafter. Myoid cell actin filament bundles, running in parallel, were present at p.n. day 10. Actin formed a meshwork pattern characteristic of the adult on, or slightly prior to, p.n. day 22. These data indicate that there is a temporal relationship between the development of the actin cytoskeleton within the Sertoli cell and initial formation of the Sertoli cell barrier. Similarly, there is a temporal relationship between the development of the actin cytoskeleton of myoid cells and tubular fluid flow. The rapid increase in tubular lumen diameter, however, does not correlate with the initial development of Sertoli and myoid cytoskeletal elements.

Electron micrograph composites of tangenital sections of the fovea centralis of three cynomolgus monkeys (Macaca irus) and one baboon (Papio anubis) were used to determine the spatial density of the principal retinal cells. In the center of the foveola, the density of cones ranged from 113,000 to 230,000/mm2, and pigment epithelial cells from 4,900 to 7,000/mm2. At a distance of 500 microns from the foveolar center the density of the cone cell pedicles ranged from 29,000 to 36,300/mm2, and the density of horizontal cells ranged from 19,000 to 25,100/mm2. Densities of bipolar, Müller, and amacrine cells were determined in only two monkeys and in the baboon. The fact that the cone cell pedicles have a larger diameter than the foveolar cones explains the geometry of the fovea. The morphology of the junction between foveolar cone outer segments and the pigment epithelium reflects the complex metabolism of this functional unit. The comparison with the peripheral primate retina suggests that the densities of horizontal and bipolar cells, but not of amacrine and Müller cells, are correlated with the density of cone cell pedicles.

This study describes the hypophyseal angioarchitecture found in 79 adult New Zealand white rabbits. The pituitary glands and attached hypothalami were removed and carefully processed following routine histological methods, and the vascular organization was studied by light microscopy. Whole mounts of the pituitary median eminence complex were prepared and studied with a binocular dissecting microscope employing transmitted and epi-illumination. Arterial blood was found to be directed primarily to the neurohypophysis by the superior hypophyseal artery (SHA) and the inferior hypophyseal artery (IHA). A direct arterial blood supply was found to the adenohypophysis, but was limited solely to the pars intermedia by branches of the anterior hypophyseal artery (AHA) and the IHA. Capillaries of the pars intermedia were subdivided into an intermediate and a superficial plexus. The superficial plexus was situated between the intermediate plexus and the capillaries of the infundibular process. Capillaries of the superficial plexus did not form anastomoses between themselves, but ramified into the intermediate plexus to form a dense network of anastomosing capillaries that were continuous with capillaries of the pars distalis. A direct arterial blood supply was found only to the superficial plexus.

Upon release from the seminiferous epithelium, spermatoza show a small droplet of cytoplasm attached to the neck region. During transit of spermatozoa in the caput epididymidis, this cytoplasmic droplet migrates along the middle piece of the flagellum. In the corpus epididymidis, the droplet shows a lateral displacement, while in the cauda epididymidis it detaches from the spermatozoon. In the electron microscope, cytoplasmic droplets attached to spermatozoa were seen to contain numerous, short, straight or C-shaped, flattened membranous elements referred to as lamellae, small vesicles, and small particles (35-nm diameter) with a diffuse wall showing no apparent unit membrane. The lamellae were stacked closely on one another or arranged in a loose array. Structurally as well as cytochemically, with different cytochemical markers, the lamellae and vesicular elements failed to show any evidence of being components of the Golgi apparatus or elements of the endoplasmic reticulum. The lamellae, vesicular elements, and 35-nm particles were also seen free in the lumen of the corpus epididymidis but were especially prominent in the cauda epididymidis at a time when droplets were being released from spermatozoa. The lumen of the epididymis, as spermatozoa passed from the caput to the cauda epididymidis, was also noted to acquire progressively a flocculent background material. The epididymal epithelium is composed predominantly of principal and clear cells. The endocytic activity of clear cells was examined in rats at different time intervals after a single injection of cationic ferritin into the lumen of the cauda epididymidis. At 2 min the tracer was bound to the microvilli of these cells and was also observed within large coated and uncoated pits, subsurface coated vesicles, and numerous subsurface small uncoated vesicular membranous elements (150-200-nm diameter). At 5 min, in addition to the above structures, the tracer was present in endosomes, while at 15 and 30 min, pale and dense multivesicular bodies appeared labeled, respectively. At 1 and 2 hr, but more so at 6 hr large dense membrane-bound bodies identified cytochemically as secondary lysosomes became labeled. All of the above endocytic structures were also seen to contain the 35-nm particles, flattened or vesicular membranous profiles, and a fine flocculent background material reminiscent of those seen free in the lumen or found in cytoplasmic droplets attached to spermatozoa. (ABSTRACT TRUNCATED AT 400 WORDS)