The Journal of Biophysical and Biochemical Cytology最新文献

In mitotic cells of Walker 256 carcinoma some four-layered lamellar structures were observed which had the appearance of paired cysternae of the ER. Two inner membranes were regular, smooth surfaced, and closely applied to each other. The two outer membranes were somewhat irregularly placed in relation to the inner pair; they showed attached RNP particles and connections with cysternae of the ER. The membranes often appeared to radiate from the region of the centrosphere towards the compact mass of chromosomes. Thus, they lay amid the spindle fibres and are referred to as "spindle lamellae." They approached the centrioles closely but were not observed to be continuous with them. They appeared to terminate in the pole of the spindle by joining smooth surfaced membranes in the centrosphere. Their equatorial termination was in relation to the chromosomes. At the surface of the chromosome mass they frequently split into two double membranes, which were closely applied to the chromosome substance. The most prominent and complicated membranes were seen in anaphase cells. An hypothesis is advanced which ascribes the development of the nuclear membrane to the spindle lamellae.

HeLa and conjunctiva tissue culture cells were incubated for various intervals with tritiated nucleosides and the incorporation into RNA was localized in different parts of the cell by means of autoradiography. In order to obtain quantitative measurements of incorporation from grain count data the influence of cell geometry on the absorption of the tritium beta ray was considered. Relative correction factors, E = g/g*, relating an idealized grain count in the absence of absorption, g, to the actual grain count, g*, were derived for the different cell compartments. For the average HeLa cell the factors for the nucleolus, n, non-nucleolar parts of the nucleus, N, and the cytoplasm, C, are in the ratio E(n)/E(N)/E(C)= 2.3/1.6/1.0. The kinetics of incorporation for cytidine and adenosine are similar. The n and N curves are characterized by a rapid rise and early saturation, whereas the C curves show an appreciable lag and no evidence of saturation for intervals as long as one generation time. Estimates of the relative amounts of RNA in each compartment were obtained from ultraviolet micro absorption measurements and used together with the kinetic data to calculate specific activities. For incubation periods of short duration the ratio of specific activities n/N for cytidine is approximately twice that for adenosine. Three hypotheses for the mechanism of ribonucleoside incorporation and RNA synthesis are discussed, and arguments favoring a transport of RNA or an RNA by-product from the nucleus and nucleolus to the cytoplasm are presented.

The structure of the flight muscle of a dragonfly (Aeshna sp.) has been studied with the light and electron microscopes, and the organization of this specialized tubular muscle is described. This tissue is characterized by the great development of the sarcosomes, which are slab-like and are arranged within the fiber opposite each sarcomere of the radially oriented lamellar myofibrils. A well developed and highly ordered sarcoplasmic reticulum is present, consisting of perforated curtain-like cisternae extending across the face of each fibril, together with tubular invaginations of the fiber plasma membrane situated within indentations in the sarcosomes and traversing the fibril surface midway between the Z and M levels. The structure of these fibers, and notably the organization of the reticulum, is compared with that of other types of muscle, and the possible role of the two components of the sarcoplasmic reticulum in the contraction physiology of the dragonfly muscle fiber is discussed.

Physiological investigations have shown that the synaptic input to the sensory neuron of the stretch receptor in the abdominal muscles of the crayfish is purely inhibitory. This neuron was chosen, therefore, as a site in which to study the fine structure of inhibitory synaptic endings. It was hoped that this fine structure might (a) provide a morphological prototype for the study of more complex synaptic systems and (b) reflect the inhibitory mechanisms. Stretch receptors were fixed in situ in buffered OsO(4), dehydrated, and embedded in Araldite. Both cross and longitudinal sections were examined after staining with phosphotungstic acid. The inhibitory endings were easily identified by their great similarity to previously described excitatory endings. Small circular profiles (synaptic vesicles) about 460 A in diameter and an accumulation of mitochondria were consistently observed within the presynaptic endings. An increased osmiophilia of pre- and postsynaptic membranes, where they were in apposition, was also seen. The only observed difference between these inhibitory endings and excitatory endings, described by other authors, was the variable presence of a latticework of 230 A tubules in the connective tissue immediately adjacent to the inhibitory endings. Inhibitory endings were observed on all parts of the sensory neuron except the axon.

This paper describes the fine structure and its relationship to the direction of beat in four types of cilia on the gill of the fresh-water mussel Anodonta cataracta. The cilia contain nine outer, nine secondary, and two central fibers, such as have been described previously in other material. Each outer fiber is a doublet with one subfiber bearing arms. One particular pair of outer fibers (numbers 5 and 6) are joined together by a bridge. The two central fibers are enclosed by a central sheath; also present in this region is a single, small mid-fiber. The different groups of fibers are connected together by radial links that extend from the outer to the secondary fibers, and from the secondary fibers to the central sheath. The basal body consists of a cylinder of nine triplet fibers. Projecting from it on one side is a dense conical structure called the basal foot. The cylinder of outer fibers continues from the basal body into the cilium, passing through a complex transitional region in which five distinct changes of structure occur at different levels. There are two sets of fibers associated with the basal bodies: a pair of striated rootlets that extends from each basal body down into the cell, and a system of fine tubular fibers that runs parallel to the cell surface. The relationship between fine structure and direction of beat is the same in all four types of cilia examined. The plane of beat is perpendicular to the plane of the central fibers, with the effective stroke toward the bridge between outer fibers 5 and 6, and toward the foot on the basal body.

The fine structure of fibrillar flight muscle of the mature adult beetle Tenebrio molitor is described. Although the very high frequency of contraction of fibrillar muscle has previously been in part accounted for as the result of mechanical specialization of the wing-bearing segment rather than of a correspondingly high rate of motor impulse supply, the problem of the nature of the pathway by which excitation is conducted into these large fibers remained. Therefore, particular attention has been given to the disposition and relationships of the plasma membrane and sarcoplasmic reticulum in this tissue. The invading tracheoles draw with them a sheath of plasma membrane from the surface to all depths in the fiber, and it is suggested that these sheaths, together with the extensive tubular arborisations arising from them, reduce the maximum plasma membrane-to-fibril distance from the radius of the fiber to a value of less than 2 micro. The evidence presented here confirms Veratti's contention that in fibrillar muscle the "reticulum" is associated with, though entirely distinct from the fibrils. Unlike other muscles so far examined, these flight muscle fibers contain a plasma membrane reticulum only, but it is possible that elsewhere the general "sarcoplasmic reticulum" includes a component derived from the plasma membrane, likewise acting as the pathway for inward conduction of excitation. Profiles of the internalised plasma membrane in Tenebrio showing the usual triple-layered 25-25-25 A organization are frequently seen, in sections, in close association with isolated vesicles (defined by "simple" 50 A membranes) which are here considered to represent, in vestigial form, the portion of the sarcoplasmic reticulum which in other types of muscle is complex and highly developed. Such associations, in Tenebrio, between these two dissimilar elements are here termed "dyads" and the possible morphological and functional homology between these and the "triads" of other types of fiber is considered.

A technic is described for high resolution intracellular autoradiography in the electron microscope. Cultures of LLC-MK(2) monkey kidney cells were incubated for 72 hours in a medium containing 0.4 microcurie per ml of thymidine-H(3). After labeling, the cells were fixed with osmium tetroxide and embedded in methacrylate. Ultrathin sections of the labeled tissue were taken up on Formvar-coated and carbon-stabilized electron microscope grids. A 150 to 450 A layer of silver metal was then evaporated onto the tissue. The coated grids were exposed to bromine vapor for 1.5 to 2 minutes under red light, allowed to dry for 1 minute, and then covered with a thin film of 1 per cent aqueous gelatin applied by means of a fine wire loop lowered over the grid supported on a glass peg. For autoradiographic exposure, the grids were stored 50 days in a light-proof container at 4 degrees C with calcium chloride desiccant. Development was carried out for 5 minutes at 20 degrees C in Promicrol (May and Baker, England) diluted 1:1 with water, followed by a 1 minute water wash and fixation for 2.5 minutes in 15 per cent aqueous sodium thiosulphate. After removal of the gelatin by immersion for 16 hours in water at 37 degrees C, the autoradiograms were dried and examined in the electron microscope. Ultrastructural detail was fairly well defined and the cytoplasm of each labeled cell was covered with an electron opaque deposit of silver, suggesting that a polynucleotide containing thymidine may be synthesized in the cytoplasm. The matter is discussed.