A B S T R A C TDifferentiating white adipose tissue from presumptive and developing fat pads of newborn and young rats was fixed in buffered osmium tetroxide, embedded in Vestopal W, and examined in an electron microscope. Pre-adipose cells were found to be fibroblasts characterized by their spindle shape, long tenuous cytoplasmic extensions, and profuse endoplasmic reticulum. The developmental stages traced from fibroblast to mature adipose cell show a gradual change in cell shape, an accumulation of cytoplasm and non-membrane-bounded lipid, a decrease in the endoplasmic reticulum, and a change in shape of mitochondria. Transitory glycogen appears at mid-differentiation. Numerous smooth-membraned vesicles occur in the cytoplasm throughout differentiation. Pinocytosis is constantly evident. Cells of the mnltilocular stage are shown to differ from brown fat ceils, particularly with respect to cytoplasmic membrane systems and mitochondria. No transport of particulate lipid from the lumen of the capillary to, or within, the adipose cell was detected, nor could any cell organelle be demonstrated to be visibly related to lipid synthesis and/or deposition.

This report presents an electron microscope study of white adipose cells depleted of their lipid inclusion by prolonged low food intake. The tissue was fixed in phosphate-buffered 1% osmium tetroxide, embedded in Vestopal W, and sections were stained with lead hydroxide. Such cells differ in shape and size from signet-ring cells. They are ovoid, diminished in size, and the cell surface has become indented in all planes of the cell. The intercellular space is expanded with concentrations of collagen fibers in close association with the cell surface. These changes are presumed to be mechanical events associated with loss of lipid from the cell. Subcellular changes include the appearance of a n extremely large number of pinocytotic vesicles and cytoplasmic vesicles similar in appearance, membrane-bound dense bodies, five-layered membranous structures, vesiculated bodies, and areas of flocculent material. Mitochondria and nuclei appear to be similar to their counterparts in signet-ring cells. Observations are discussed with relation to ultrastructural changes reported in other studies in which adipose cells were depleted of fat by acute starvation or experimental deprivation of insulin. It is suggested that intracellular structural changes are probably associated with adaptive alterations in the metabolism of the cell.

The electron microscopic appearance of glycogen has been studied in the organs of several animal species. Glycogen almost always appears as roughly circular granules from 150 to 400 A in diameter. The intrinsic electron density of glycogen varies from tissue to tissue; however, treatment with lead hydroxide as described by Watson deeply stains the granules. Glycogen pellets were isolated from some of the tissues studied by centrifugation. Such pellets were shown to be glycogen by chemical and histochemical criteria. When thin sections of the pellet are examined under the electron microscope they can be seen to consist of densely packed granules similar to those found in the intact tissues. Such pellets are also stained for electron microscopy by short exposure to lead hydroxide.

Interscapular fat from newborn rats and mice was fixed in buffered 1 per cent osmium tetroxide and thin sections of the methacrylate-embedded tissue were studied with the electron microscope. The findings have reaffirmed the epithelioid character of brown adipose tissue, and have provided additional information on the relation of its cells to each other and to the rich capillary bed. For the most part, the earlier description of the fine structure of brown adipose cells by Lever, has been confirmed, but our observations on the mitochondria and their relation to fat droplets have led us to different conclusions concerning the role of these organelles in lipogenesis. Mitochondria were often found to be very closely associated with lipide inclusions, but no actual communication between the two was observed and no evidence was found to support the hypothesis that mitochondria are transformed into lipide droplets. Large dense bodies which showed a highly ordered fine structure suggesting a crystalline protein were seen in the matrix of some mitochondria. The cytoplasm of the adipose cells contained fine granules that seemed to be of two kinds: particles of uniform size (~150 A) and appreciable density that are believed to be ribonucleoprotein, and granules of lower density and more variable size that are tentatively interpreted as a form of glycogen. The Golgi complex of the adipose cells was small and the endoplasmic reticulum almost entirely absent. The significance of the poor development of these organelles is discussed in relation to current concepts of their function.The interscapular brown fat of rodents was formerly thought to have a function related to winter dormancy and it was therefore called the hibernating glan~, but it is now generally believed to be a special form of adipose tissue. Although it has been a subject of renewed interest to histologists, biochemists, and virologists in the past few years, its physiological significance remains largely unknown. For a review of the early literature on this tissue, a description of its histology, and a tabulation of its occurrence in various" mammalian species, the reader is referred to the classical paper by Rasmussen (17) in 1923. Later work on the microscopic structure, histochemistry, and metabolism of brown adipose tissue, and its

The anterior leaflet of the mitral valve of the dog contains blood vessels, nerve fibers, and cardiac muscle in addition to elastic fibers and collagen. When studied in a myograph, the electrically stimulated mitral valve actively developed tension and shortened. Active tension was found to be a function of initial length of the valve and was increased by norepinephrine and decreased by acetylcholine. The presence of neuronally releasable norepinephrine stores in the valve was indicated by responsiveness to tyramine. The negative inotropic response of the mitral valve to acetylcholine was consistent with an atrial origin of the tissue. Possible functional roles for mitral valve muscle and the potential significance of its neural control are discussed.

The fine structure of myelin was studied in glutaraldehyde-fixed rat sciatic nerves depleted of lipid by acetone, chloroform:methanol (2:1 v/v), and chloroform:methanol:concentrated HC1 (200:100:1, v/v/v). One portion of each of these nerves, plus the extracts, was saponified and analyzed by gas-liquid chromatography for fatty acids. The remainder of each nerve was stained in osmium tetroxide in CC1 4 (5g/100cc) and was embedded in Epon 812. Thin sections, examined in the electron microscope, revealed the preservation of myelin lamellar structure with a 170 A periodicity in nerves depleted of 98% of their lipids. Preservation of myelin lamellar structure depended on glutaraldehyde fixation and the introduction of osmium tetroxide in a nonpolar vehicle (CC14) after the lipids had been extracted. It is concluded that the periodic lamellar structure in electron micrographs of myelin depleted of lipid results from the complexing of osmium tetroxide, plus uranyl and lead stains, with protein.

No abstract

Among mammals prolonged survival of spermatozoa in the reproductive tract of the female (circa 7-t months) is known in only a few species of hibernating bats, but its physiological basis remains obscure. Presumably survival involves special physiological adaptations of the spermatozoa, the uterus, or both. We have initiated studies of sperm survival mechanisms in the hibernating bat Myotis lucifugus involving analysis of the cytology and fine structure of intra-uterine sperm, the morphology and histochemistry of sperm-uterus interactions, and selected biochemical parameters of the reproductive tract. This paper summarizes our findings concerning the morphology of epididymal and intra-uterine spermatozoa, and emphasizes those aspects not detailed by others. The microscopic organization of epididymal sperm is described in detail with particular attention to peculiarities of the head, middle piece and cytoplasmic droplet. The latter contains amylase-resistant PAS + granules and acid phosphatase, both possibly of lysosomal origin. Most ultrastructural features elucidated by Fawcett and collaborators are confirmed, but additional details concerning the head, post-nuclear sheath, origin of axial filament complex, and the outer coarse fibrils are provided. A functional interrelationship between coarse fibrils 3 and 8 and the longitudinal columns of the fibrous sheath is also postulated.Structural differences between epididymal and uterine sperm are minor, suggesting that uterine environmental factors are most important for sperm survival.