Ultrastructure of crystalloid inclusions in the dog and rat epididymis

Fine structural studies of the epididymis of mature mongrel dogs and of Sprague-Dawley rats were undertaken in conjunction with research dealing with the effects of vasectomy upon this organ. This paper reports the observation of crystalloid and lamellar inclusions present in these species following fixation of the epididymis in 5 % glutaraldehyde, post-fixation in osmium, and routine processing for electron microscopy. In the dog, crystalloid inclusions were observed within the cauda epididymidis of unoperated and vasectomized animals. They were found within the apical cytoplasm of principal cells in association with the Golgi apparatus and endoplasmic reticulum, and in some instances, in close proximity to the nucleus. These crystalloids exhibited a 12 nm periodicity and often measured over 3 μm in length. In the rat, two types of inclusions were found, one within mitochondria of clear cells from unoperated animals and another within membrane-bound bodies of principal cells from the caput epididymidis of unoperated and vasectomized animals. The mitochondria which contained inclusions were basally located and were observed in stacks of up to eight elongate mitochondria each. The mitochondrial inclusions exhibited a complex lamellar structure with an approximate periodicity of 36 nm. In contrast, the crystalloid inclusions found within principal cells were sequestered within supranuclear cytoplasmic bodies which increased in number with age. Such crystalloids exhibited a linear periodicity of 11–13.5 nm, but the precise lattice structure remains to be determined. Although certain aspects of the morphology of these bodies suggests a relationship to microbodies, we have been unable to demonstrate catalase activity within them. At present, neither the origin of crystalloid structures described, nor their relationship to epididymal physiology is clear.     

Morphology of globule leucocyte from bovine bronchopulmonary lavage

It has hitherto been assumed that the globule leucocytes (GL) occur as free cells in the airways of animals. The present study provides definite evidence for the occurrence of these cells in the bronchopulmonary lavage of cattle. At the light-microscopic level, the GL was a round to elongate cell containing the characteristic large, round, metachromatic granules and an eccentric nucleus with a prominent nucleolus. Ultrastructurally, the cell was round, approximately 15 micron in diameter, and contained round, electron-dense granules which measured approximately 1.25 micron in diameter. The cell nucleus was endowed with abundant heterochromatin. The cytoplasm had only inconspicuous organelles. We conclude that the bovine GL is a specific cell which reaches the airway lumen following migration from the lining of the bronchopulmonary mucous membrane.     

Ultrastructure of the milk fat globule membrane with and without triglyceride

Summary The primary milk fat globule membrane (MFGM) around freshly secreted milk fat globules consists of a unit membrane separated from the triglyceride core by a dense material. This dense material may widen to include cytoplasmic organelles or may form small blebs. Preincubation and fixation of the globules at temperatures between 4° C and 60° C has no effect on the width or appearance of the dense material. Isolated MFGM profiles show structures identical to those found on intact globules. The dense material on the isolated MFGM profiles is unaffected by extractions which remove essentially all the triglyceride present in the pellets of MFGM.The structure of the primary MFGM is therefore independent of any triglyceride content and the earlier suggestions that the dark material represented a triglyceride layer of high melting point adsorped during cooling of the globules after milking are not supported by the work described in this paper.     

Ultrastructure of vacuolar inclusions in root tips

Summary Vacuoles in plant cells often contain inclusions which at early stages of development are bounded by a single membrane. The inclusion bodies (IBs) comprise a diversity of forms and various stages of differentiation are recognizable. IBs are divided into two categories: those which have a matrix without internal membranes, and those which contain cytoplasmic organelles and other membranous material. The internal membranes may be tightly coiled or in the form of vesicles. IBs develop from invaginations of the tonoplast which become detached into the vacuole. They are initiated mainly during active cell growth but may remain within the vacuole in differentiated cells. Various components contribute to the contents of IBs: endoplasmic reticulum, nuclear envelope, Golgi vesicles, extruded portions of mitochondria and plastids, ribosomes and groundplasm. In most IBs the limiting membrane and contents eventually disappear within the vacuole. Some IBs prior to their breakdown within the vacuole also function as sites for the formation of material not found elsewhere in the cell. The disappearance of IBs from vacuoles suggests that such vacuoles behave as lysosomes.     

Eosinophilic globule cells in mouse MFH-like sarcomas

Light and electron microscopic observations were made on eosinophilic globule (EG) cells found in 27 subcutaneous malignant fibrous histiocytoma (MFH)-like sarcomas in aged ICR mice. These tumors, which were composed of fibroblast-like cells as the major component, with small numbers of histiocyte-like cells and undifferentiated cells, showed one or more of five growth patterns: storiform, pleomorphic, fascicular, myxoid, and hemangiopericytoma-like. EG cells were interspersed among the tumor cells and were also present in metastatic lesions. They were pleomorphic in shape and contained various numbers of cytoplasmic globules, which were positive with the periodic acid-Schiff reaction and resistant to diastase digestion. Electron microscopic observation revealed that these cells had small finger-like and pseudopodia-like projections, and contained varied numbers of characteristic osmiophilic globules and glycogen particles in the cytoplasm which seemed to become more abundant as the cells differentiated. The osmiophilic globules consisted of a dense homogeneous core and a marginal area rich in small vesicular membranous structures. A small population of EG cells exhibited features suggesting differentiation to fibroblasts; these were characterized by an increased amount of rough endoplasmic reticulum. It is suggested that the EG cell is a neoplastic cell, perhaps derived from a primitive mesenchymal cell, although an inflammatory cell origin is also possible.     

Ultrastructure of blastocyst attachment in the mouse

Summary Mouse embryos have been examined with light and electron microscopy after fixation by perfusion with glutar aldehyde, and embedding in plastic.The Zona pellucida is dissolved gradually around the blastocyst just prior to attachment, and Zona free blastocysts exist only for a very short time.Blastocyst attachment is established when the trophoblast and uterine cell surface membranes lie within 150 Å apart over wide areas. The uterine epithelium does not show any signs of degeneration.Trophoblast attachment probably precedes decidual cell reaction.This work was supported by the Swedish Government Funds for Supporting Medical Research and the Swedish Medical Research Council (Project No. 12 X-70-02).     

Glycosyl transferases in mouse and human milk fat globule membranes

Summary Membranes isolated from mouse and human milk fat globules were found to contain the enzymes responsible for the synthesis of dolichol monophosphate mannose and dolichol monophosphate glucose as well as those involved in the transference of the glycosyl residues from the two dolichol derivatives to dolichol diphosphate oligosaccharides. The levels of most of the enzymes were comparable to those found in mouse mammary gland microsomes. The presence of enzymes involved in protein glycosylation via dolichol derivatives in the milk fat globule membrane provides evidence in favor of an outward flow of membrane components from the rough endoplasmic reticulum, where these enzymes are active in vivo, towards the cell surface.     

Ultrastructure of bacilli and the bacillary origin of the macrophagic inclusions in Whipple's disease

An electron microscopic and cytochemical study of the Whipple bacillus in jejunal biopsies from three untreated patients was made using fixation procedures developed for the satisfactory preservation of bacterial ultrastructure. The envelopes of the normal-looking bacilli present free in the lamina propria consisted of the following layers. (i) A cytoplasmic membrane with a triple-layered profile and a mean thickness (peak-to-peak distance) of 6.08 nm. (ii) A thick (20 nm) cell wall containing peptidoglycan; the wall had a hitherto undescribed inner layer that contained polysaccharides, possibly teichoic acids. (iii) Surrounding the cell wall, a surface membrane with a symmetric profile and a mean peak-to-peak distance of 4.74 nm. The ultrastructural pattern of the Whipple bacillus wall corresponds to that of Gram-positive bacteria, but with an additional surface membrane. This membrane is different from the outer membrane of Gram-negative bacteria because it has a symmetric profile, is thinner and has no periodic acid-Schiff (PAS)-positive components. Normal-looking bacilli were seen very rarely inside jejunal macrophages, but degenerating bacteria were abundant in these phagocytes. Electron microscopy and ultrastructural cytochemistry of Whipple bacilli inside jejunal macrophages of the three untreated patients showed that the degenerative process is a sequence that leads to the loss of bacillary forms and to the accumulation of bacterial remnants resistant to degradation by the macrophage. These remnants correspond to the innermost, polysaccharide-containing portion of the bacillus wall. The progressive accumulation of these PAS-positive wall remnants is the origin of the intramacrophagic inclusions that are important in the histological diagnosis of Whipple's disease. The reported results indicate that in the three patients studied, the Whipple bacillus multiplies extracellularly, the bacteria that are phagocytosed by macrophages being degraded.     

Ultrastructure of the mouse tracheal epithelium

The ultrastructure of mouse tracheal epithelium was examined. The three cell types, basal cells, ciliated cells and goblet cells, described for other mammalian trachea were found to be present although goblet cells occurred only rarely. A cell type, termed the nonciliated cell, not described in other mammalian trachea was frequently found in mouse tracheal epithelium. These cells contained abundant smooth and rough endoplasmic reticulum, free ribosomes, a large Golgi complex, and many mitochondria. There were many vesciles containing an electron dense material near the luminal surface of these cells; these cells were positive for PAS. These features suggested a secretory function for the cells. This, along with the scarcity of goblet cells, suggested that the nonciliated cells of mouse tracheal epithelium fulfill the function of the goblet cells found in other mammalian trachea.     

Intramitochondrial inclusions in the kidney of the pygmy mouse

Intramitochondrial inclusions averaging 1000 A in diameter were observed in the cells of the proximal convoluted tubules in samples of kidneys from pygmy mice, Baiomys taylori. Electron microscopic study of unstained sections and mitochondrial fractions showed that these inclusions are lipid and found at a S.G. 1.37 in a linear sucrose gradient. Renal glycogen, inorganic phosphate and plasma sodium were significantly higher in the pygmy mouse and plasma calcium was lower, as compared with the laboratory mouse. We believe these intramitochondrial inclusions to be lipid which accumulates divalent cations, particularly calcium, which acts as a sodium pump allowing the pygmy mouse to conserve water and adapt to its environment.     

Ultrastructure of pericytes in mouse heart

The pericytes of mouse myocardium are extensively branched cells that form an incomplete layer around the endothelium of capillaries and postcapillary venules. The membranes of pericytes and endothelial cells are connected by specialized junctions. Microtubules, intermediate (10-nm) filaments and microfilaments are oriented within circumferentially-arranged cytoplasmic processes of pericytes so as partially to encircle the endothelial cylinder. The intracellular organization of these myocardial pericytes suggests that they are smooth muscle-like cells which may be capable of influencing microvascular dynamics in the heart.     

Ultrastructure of the endolymphatic sac in the mouse.

The ultrastructure of the endolymphatic sac (ES) in the mouse was examined by light and electron microscopy. This organ was divided into three parts: proximal, intermediate and distal. In the proximal portion of the ES, the epithelium consisted of thin squamous cells. The epithelial cells had acquired basolateral processes, numerous small vesicles and well-developed Golgi apparatus. In the intermediate portion, the epithelium consisted of columnar or cuboidal cells. The epithelial cells could be classified into two types: type I and type II. The type I cells had abundant microvilli, pinocytotic vesicles, vacuoles, multivesicular bodies, lysosomes and mitochondria. The type II cells had fewer numbers of these organelles. A few free-floating cells could be observed in the lumen of this intermediate portion, most of which were macrophages. In the distal portion, the epithelium consisted of squamous or cuboidal cells. The epithelial cells had a few cytoplasmic organelles. In the ultrastructural study, each portion of the mouse ES was found to have a very distinct morphological feature. It was suggested that each of these three portions has a different function.     

Ultrastructure and permeability of lymph node microvasculature in the mouse

Summary The microvasculature of lymph nodes and Peyer's patches consists of arterioles, capillaries and venules. The postcapillary segment comprises high-endothelial venules (HE venules) as well as ordinary venules. In order to study the ultrastructure of the microvasculature, particularly with respect to the nature of intercellular junctions, lanthanum and ruthenium red were used as tracers. Furthermore, to evaluate the permeability properties of the different segments of the microvasculature, intravenously injected horseradish peroxidase (HRP; MW: 40,000) was used.All segments of the microvasculature are permeable to HRP. However, the mechanism of transport across the vascular wall varies in the different segments, apparently correlated with a gradual decrease in number of transport vesicles and a gradual attenuation in the sealing of the endothelial cells. Tight junctions are present in arterioles, and it is assumed that HRP reach the basal lamina exclusively by vesicular transport. Incomplete or focal tight junctions are present in the capillaries, and both intercellular and vesicular pathways are observed. In the venules the intercellular pathway seems to be the dominant one, while vesicular transfer is negligible. However, some micropinocytic vesicles in the HE venule endothelial cells probably represent the initial stage of an intracellular digestion.