bluuebunny/crossref_metadata_2025 huggingface dataset:
INCORPORATION OF C14-LABELED MALEIC HYDRAZIDE INTO THE ROOT-TIP CELLS OF ALLIUM CERNUUM, VICIA FABA, AND TRADESCANTIA PALUDOSA
Authors: Callaghan John J. , Grun Paul
Published: May 2004

Allium cernuum, Vicia faba, and Tradescantia paludosa were treated by root immersion in maleic hydrazide (1 mM/liter) labeled with C14 (C14-MH) for 1 hour to determine the location within the cell to which MH moves during various periods of time after treatment. Root tips were fixed 24 hours, 48 hours, 72 hours, and 3 weeks after treatment. Autoradiographs of root tips squashed 24 to 72 hours after fixation showed that C14-MH was distributed throughout the nuclei and was particularly concentrated in the nucleoli. The nucleolar localization of the chemical was transitory, fixations made 3 weeks after treatment showing well labeled nuclei many of which completely lacked label in the nucleoli. The chromosomes seen in mitotic divisions of all three species had the same amount of label in euchromatic as heterochromatic areas. Since the chemical was not accumulated preferentially in heterochromatic areas, it seems likely that the reported specificity of MH for the breakage of heterochromatin can not be due to preferential heterochromatic incorporation.
THE ORGANIZATION OF THE FLIGHT MUSCLE IN A DRAGONFLY, AESHNA SP. (ODONATA)
Authors: Smith David S.
Published: May 2004

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.
SARCOMERE SIZE IN DEVELOPING MUSCLES OF A TARSONEMID MITE
Authors: Aronson John
Published: May 2004

The embryo of a tarsonemid mite was found to be suitable for in vivo observations of muscle development by polarization microscopy. The four dorsal muscles of the metapodosoma each contain three sarcomeres, the anterior two of which can be seen clearly. These sarcomeres can be identified and followed during much of their development. Sarcomeres are about 2.5 micra long when first detected and increase in length until they are about 10 micra long. The change in length is associated with a slow, approximately constant rate of increase in the length of the A region, and an initially slow then much more rapid increase in the length of the I band. Preceding the period when the I band elongates rapidly there is an increase in the diameter of the muscle fibers and an increase in the retardation of the A band. A, I, Z, and H bands are visible during most of these changes. The change in A band length has been interpreted in terms of the growth of the A filaments which have been observed by electron microscopy in muscles of other animals. It is suggested that the exceptionally long sarcomeres in this mite result from the early fixing of the number of sarcomeres in a given muscle fiber.
THE LOCALIZATION OF ACID PHOSPHATASE IN RAT LIVER CELLS AS REVEALED BY COMBINED CYTOCHEMICAL STAINING AND ELECTRON MICROSCOPY
Authors: Holt S. J. , Hicks R. Marian
Published: May 2004

Discrete localization of stain in pericanalicular granules was found in 10 µ frozen sections of formol-phosphate-sucrose-fixed liver stained by the Gomori acid phosphatase technique and examined in the light microscope. The staining patterns, before and after treatment with Triton X-100 and lecithinase, were identical with those previously reported for formol-calcium-fixed material treated in the same way, and it can be assumed that the stained granules are identical with "lysosomes." Examination in the light microscope of the staining patterns and lead penetration in fixed blocks and slices of various dimensions showed nuclear staining and other artefacts to be present, produced by the different rates of penetration of the various components of the staining medium into the tissue. A uniform pericanalicular staining pattern could be obtained, however, with slices not more than 50 µ thick, into which the staining medium could penetrate rapidly from both faces. The staining pattern produced in 50 µ slices was the same both at pH 5.0 and pH 6.2, and was not altered by subsequent embedding of the stained material in butyl methacrylate. Electron microscopy showed the fine structure of fixed 50 µ frozen slices to be well preserved, but it deteriorated badly when they were incubated in the normal Gomori medium at pH 5.0 before postfixing in osmium tetroxide. After incubation in the Gomori medium at pH 6.2, the detailed morphology was substantially maintained. In both cases lead phosphate, the reaction product, was found in the pericanalicular regions of the cell, but only in the vacuolated dense bodies and never in the microbodies. Not every vacuolated dense body contained lead, and stained and unstained bodies were sometimes seen adjacent to each other. This heterogeneous distribution of stain within a morphologically homogeneous group of particles is consistent with de Duve's suggestion (9) that there is a heterogeneous distribution of enzymes within the lysosome population. It is concluded from these investigations that the vacuolated dense bodies seen in the electron microscope are the morphological counterparts of the "lysosomes" defined biochemically by de Duve.
THE DISTRIBUTION OF MUSCLE ANTIGENS IN CONTRACTED MYOFIBRILS DETERMINED BY FLUORESCEIN-LABELED ANTIBODIES
Authors: Tunik Bernard , Holtzer Howard
Published: May 2004

Chick myofibrils in different states of contraction were treated with fluorescein-labeled antibodies. The rabbit antibodies were prepared against chick myosin, light and heavy meromyosins, and actin. For any one state of contraction, a single myofibril was photographed through the phase contrast microscope, stained with one of the antisera, and photographed through the fluorescence microscope. The cytological changes in the sarcomeres accompanying contraction as observed under phase were correlated with changes in the distribution of the precipitated antibodies as observed under the fluorescence microscope. The changing patterns observed through the fluorescence microscope were compared with those predicted by the sliding filament model of contraction.
s-cyclophilin is retained intracellularly via a unique COOH-terminal sequence and colocalizes with the calcium storage protein calreticulin.
Authors: Arber S , Krause K H , Caroni P
Published: May 2004

Cyclophilins (cyclosporin A-binding proteins) are conserved, ubiquitous, and abundant proteins that accelerate the isomerization of XaaPro peptide bonds and the refolding of proteins in vitro. s-Cyclophilin is a member of the cyclophilin family with unique NH2- and COOH-terminal extensions, and with a signal sequence. We now report that s-cyclophilin is retained in the cell, and that the conserved s-cyclophilin-specific COOH-terminal extension VEKPFAIAKE is sufficient to direct a secretory protein to s-cyclophilin containing structures. Antibodies to s-cyclophilin-specific peptides were produced and the location of the protein was determined by an immunocytochemical study at the light microscopic level. s-Cyclophilin colocalized with the Ca(2+)-binding protein calreticulin and, to a lesser extent, with the microsomal Ca(2+)-ATPase in the myogenic cell line L6, and with the Ca(2+)-binding protein calsequestrin in skeletal muscle. In activated platelets, s-cyclophilin immunoreactivity was detected in a ring-like structure that might correspond to the Ca(2+)-storing and -releasing dense tubular network. In spreading cells, s-cyclophilin containing vesicular structures accumulated at actin-rich protrusion sites. While s-cyclophilin consistently codistributed with Ca2+ storage site markers, the distribution of s-cyclophilin immunoreactivity was not identical to that of ER markers. To determine whether the COOH-terminal extension of s-cyclophilin was involved in its intracellular transport we added this sequence to the COOH-terminus of the secretory protein glia-derived nexin. Appropriate constructs were expressed transiently in cultured cells and proteins were detected with specific antibodies. We found that glia-derived nexin with the COOH-terminal sequence VEKPFAIAKE (but not with the control sequence GLVVMNIT) colocalized with endogenous s-cyclophilin, indicating that the sequence contained retention information. These results indicate that s-cyclophilin is a retained component of an intracellular organelle and that it may accumulate in specialized portions of the ER, and possibly in calciosomes. Because of its conserved structure, widespread distribution, and abundance s-cyclophilin may be a useful marker to study the biogenesis and distribution of ER subcompartments.
cAMP and Ca(2+)-mediated secretion in parotid acinar cells is associated with reversible changes in the organization of the cytoskeleton.
Authors: Perrin D , Möller K , Hanke K , Söling H D
Published: May 2004

The potential involvement of actin and fodrin (brain spectrin) in secretory events has been assessed in primary cultured guinea pig parotid acinar cells, using as a tool affinity purified anti-alpha-fodrin antibody, phalloidin, and immunofluorescence techniques. In resting parotid acinar cells fodrin and actin appeared as a continuous ring under the plasma membrane of most of the cells. Upon stimulation with secretagogues fodrin and actin labeling at the level of the plasma membrane disappeared almost completely. To establish a correlation between secretion and cytoskeletal changes at the individual cell level, anti-alpha-amylase-antibodies were used to label secreted amylase exposed at the surface of secreting cells. The number of cells expressing alpha-amylase on their surface followed bulk secretion of alpha-amylase. A strict correlation between secretion and alteration of the actin-fodrin labeling was observed at the individual cell level. The cytoskeletal changes occurred in parallel with secretion independently of the secretagogue used (carbamoylcholine in the presence of Ca2+, isoproterenol in presence or absence of Ca2+, forskolin, or dibutyryl-cyclic-AMP). The changes were reversible upon removal of the secretagogue. Since Ca2+, as well as cAMP-mediated secretion, was associated with the same kind of cytoskeletal changes, a reorganization of the cytoskeleton may play an essential part in regulated secretion.
Reducing inositol lipid hydrolysis, Ins(1,4,5)P3 receptor availability, or Ca2+ gradients lengthens the duration of the cell cycle in Xenopus laevis blastomeres.
Authors: Han J K , Fukami K , Nuccitelli R
Published: May 2004

We have microinjected a mAb specifically directed to phosphatidylinositol 4,5-bisphosphate (PIP2) into one blastomere of two-cell stage Xenopus laevis embryos. This antibody binds to endogenous PIP2 and reduces its rate of hydrolysis by phospholipase C. Antibody-injected blastomeres undergo partial or complete arrest of the cell cycle whereas the uninjected sister blastomeres divided normally. Since PIP2 hydrolysis normally produces diacylglycerol (DG) and inositol 1,4,5-triphosphate (Ins[1,4,5]P3), we attempted to measure changes in the levels of DG following stimulation of PIP2 hydrolysis in antibody-injected oocytes. The total amount of DG in antibody-injected oocytes was significantly reduced compared to that of water-injected ones following stimulation by either acetylcholine or progesterone indicating that the antibody does indeed suppress PIP2 hydrolysis. We also found that the PIP2 antibodies greatly reduced the amount of intracellular Ca2+ released in the egg cortex during egg activation. As an indirect test for Ins(1,4,5)P3 involvement in the cell cycle we injected heparin which competes with Ins(1,4,5)P3 for binding to its receptor, and thus inhibits Ins(1,4,5)P3-induced Ca2+ release. Microinjection of heparin into one blastomere of the two-cell stage embryo caused partial or complete arrest of the cell cycle depending upon the concentration of heparin injected. We further investigated the effect of reducing any [Ca2+]i gradients by microinjecting dibromo-BAPTA into the blastomere. Dibromo-BAPTA injection completely blocked mitotic cell division when a final concentration of 1.5 mM was used. These results suggest that PIP2 turnover as well as second messenger activity influence cell cycle duration during embryonic cell division in frogs.
Nuclear protein import is inhibited by an antibody to a lumenal epitope of a nuclear pore complex glycoprotein.
Authors: Greber U F , Gerace L
Published: May 2004

Gp210 is a major transmembrane glycoprotein associated with the nuclear pore complex that is suggested to be important for organizing pore complex architecture and assembly. A mouse monoclonal IgG directed against an epitope in the lumenal domain of rat gp210 was expressed in cultured rat cells by microinjection of mRNA prepared from a hybridoma cell line. The expressed IgG, which becomes assembled into a functional antibody in the lumen of the endoplasmic reticulum, bound to the nuclear envelope in vivo. Expression of anti-gp210 antibody in interphase cells specifically reduced approximately fourfold the mediated nuclear import of a microinjected nuclear protein (nucleoplasmin) coupled to gold particles. The antibody also significantly decreased nuclear influx of a 10-kD dextran by passive diffusion. This transport inhibition did not result from removal of pore complexes from nuclear membranes or from gross alterations in pore complex structure, as shown by EM and immunocytochemistry. A physiological consequence of this transport inhibition was inhibition of cell progression from G2 into M phase. Hence, binding of this antibody to the lumenal side of gp210 must have a transmembrane effect on the structure and functions of the pore complex. These data argue that gp210 is directly or indirectly connected to pore complex constituents involved in mediated import and passive diffusion.
Stromelysin generates a fibronectin fragment that inhibits Schwann cell proliferation.
Authors: Muir D , Manthorpe M
Published: May 2004

Our previous report (Muir, D., S. Varon, and M. Manthorpe. 1990. J. Cell Biol. 109:2663-2672) described the isolation and partial characterization of a 55-kD antiproliferative protein found in Schwann cell (SC) and schwannoma cell line-conditioned media and we concluded that SC proliferation is under negative autocrine control. In the present study the 55-kD protein was found to possess metalloprotease activity and stromelysin immunoreactivity. The SC-derived metalloprotease shares many properties with stromelysin isolated from other sources including the ability to cleave fibronectin (FN). Furthermore, limited proteolysis of FN by the SC-derived protease generated a FN fragment which itself expresses a potent antiproliferative activity for SCs. The active FN fragment corresponds to the 29-kD amino-terminal region of the FN molecule which was also identified as an active component in SC CM. Additional evidence that a proteolytic fragment of FN can possess antiproliferative activity for SCs was provided by the finding that plasmin can generate an amino-terminal FN fragment which mimicked the activity of the SC metalloprotease-generated antiproliferative FN fragment. Both the 55-kD SC metalloprotease and the 29-kD FN fragment could completely and reversibly inhibit proliferation of SCs treated with various mitogens and both were largely ineffective at inhibiting proliferation by immortalized or transformed SC lines. Normal and transformed SC types do secrete the proform of stromelysin, however, transformed cultures do not produce activated stromelysin and thus cannot generate the antiproliferative fragment of FN. These results suggest that, once activated, a SC-derived protease similar to stromelysin cleaves FN and generates an antiproliferative activity which can maintain normal SC quiescence in vitro.
Biological activities of peptides and peptide analogues derived from common sequences present in thrombospondin, properdin, and malarial proteins.
Authors: Tuszynski G P , Rothman V L , Deutch A H , Hamilton B K , Eyal J
Published: May 2004

Thrombospondin (TSP), a major platelet-secreted protein, has recently been shown to have activity in tumor cell metastasis, cell adhesion, and platelet aggregation. The type 1 repeats of TSP contain two copies of CSVTCG and one copy of CSTSCG, per each of the three polypeptide chains of TSP and show homology with peptide sequences found in a number of other proteins including properdin, malarial circumsporozoite, and a blood-stage antigen of Plasmodium falciparum. To investigate whether these common sequences functioned as a cell adhesive domain in TSP, we assessed the effect of peptides corresponding to these sequences and an antibody raised against one of these sequences, CSTSCG, in three biological assays which depend, in part, on the cell adhesive activity of TSP. These assays were TSP-dependent cell adhesion, platelet aggregation, and tumor cell metastasis. We found that a number of peptides homologous to CSVTCG promoted the adhesion of a variety of cells including mouse B16-F10 melanoma cells, inhibited platelet aggregation and tumor cell metastasis, whereas control peptides had no effect. Anti-CSTSCG, which specifically recognized TSP, inhibited TSP-dependent cell adhesion, platelet aggregation, and tumor cell metastasis, whereas control IgG had no effect. These results suggest that CSVTCG and CSTSCG present in the type I repeats function in the adhesive interactions of TSP that mediate cell adhesion, platelet aggregation, and tumor cell metastasis. Peptides, based on the structure of these repeats, may find wide application in the treatment of thrombosis and in the prevention of cancer spread.
Carboxy terminally truncated forms of ribophorin I are degraded in pre-Golgi compartments by a calcium-dependent process.
Authors: Tsao Y S , Ivessa N E , Adesnik M , Sabatini D D , Kreibich G
Published: May 2004

Two COOH terminally truncated variants of ribophorin I (RI), a type I transmembrane glycoprotein of 583 amino acids that is segregated to the rough portions of the ER and is associated with the protein-translocating apparatus of this organelle, were expressed in permanent HeLa cell transformants. Both variants, one membrane anchored but lacking part of the cytoplasmic domain (RL467) and the other consisting of the luminal 332 NH2-terminal amino acids (RI332), were retained intracellularly but, in contrast to the endogenous long lived, full length ribophorin I (t 1/2 = 25 h), were rapidly degraded (t 1/2 less than 50 min) by a nonlysosomal mechanism. The absence of a measurable lag phase in the degradation of both truncated ribophorins indicates that their turnover begins in the ER itself. The degradation of RI467 was monophasic (t 1/2 = 50 min) but the rate of degradation of RI332 molecules increased about threefold approximately 50 min after their synthesis. Several pieces of evidence suggest that the increase in degradative rate is the consequence of the transport of RI332 molecules that are not degraded during the first phase to a second degradative compartment. Thus, when added immediately after labeling, ionophores that inhibit vesicular flow out of the ER, such as carbonyl cyanide m-chlorophenylhydrazone (CCCP) and monensin, suppressed the second phase of degradation of RI332. On the other hand, when CCCP was added after the second phase of degradation of RI332 was initiated, the degradation was unaffected. Moreover, in cells treated with brefeldin A the degradation of RI332 became monophasic, and took place with a half-life intermediate between those of the two normal phases. These results point to the existence of two subcellular compartments where abnormal ER proteins can be degraded. One is the ER itself and the second is a non-lysosomal pre-Golgi compartment to which ER proteins are transported by vesicular flow. A survey of the effects of a variety of other ionophores and protease inhibitors on the turnover of RI332 revealed that metalloproteases are involved in both phases of the turnover and that the maintenance of a high Ca2+ concentration is necessary for the degradation of the luminally truncated ribophorin.
CHROMATOPHORE DEVELOPMENT, PITS, AND OTHER FINE STRUCTURE IN THE RED ALGA, LOMENTARIA BAILEYANA (HARV.) FARLOW
Authors: Bouck G. Benjamin
Published: May 2004

Thin sections of the red alga, Lomentaria baileyana, a tubular member of the Rhodymeniales, were examined after permanganate fixation and Araldite embedding. Many of the cellular structures in Lomentaria were found to be similar to analogous structures in animals and higher plants. However, in the walls between cells are modified areas generally known as pits which are unique to the higher orders of red algae (Florideae). In this study the pits were found to consist of a plug-like structure surrounded by an uninterrupted membrane apparently continuous with the plasma membrane. Examination of the chromatophore revealed a characteristic limiting membrane, a relatively sparse distribution of plates, no grana, and a single disc apparently oriented parallel to the limiting membrane. In addition to their origin from non-lamellate proplastids, chromatophores were found capable of division by simple constriction. Floridean starch grains were observed outside the chromatophore and the possibility of an association of the first formed grains with portions of the endoplasmic reticulum is considered. Gland cells seem to have a high proportion of Golgi components (dictyosomes), and are believed to have some kind of secretory function. Many of the Golgi vesicles seem to open on the wall and presumably discharge their contents.
THE ELONGATION OF MYOFIBRILS FROM THE INDIRECT FLIGHT MUSCLE OF DROSOPHILA
Authors: Aronson John
Published: May 2004

Myofibrils which lengthen by several per cent in the presence of ATP and magnesium ions were prepared by teasing indirect flight muscle of Drosophila in solutions containing ethylenediaminetetraacetate. A study was made of the hydrogen ion, magnesium ion, ATP, and potassium chloride concentrations with which this effect could be observed. The lack of elongation with pyrophosphate and several nucleoside triphosphates suggests that the lengthening is ATP specific. A relaxing factor system comparable to that described for rabbit muscle was not demonstrable, as elongated fibrils did not shorten with calcium ions, carnosine, or digitonin.
SWELLING OF FISH MITOCHONDRIA
Authors: Richardson Thomas , Tappel A. L.
Published: May 2004

The physical properties of fish liver and rat liver mitochondria were compared as a function of temperature and osmotic pressure. The data indicate that fish mitochondria are more flexible and swell at a more rapid rate over a 0 to 30°C temperature range, whereas the rates of swelling at 30 to 40°C are comparable. The swelling rates of both fish and rat mitochondria vary with temperature and approximate the Arrhenius relationship. Apparent energies of activation for swelling averaged 26.5 kcal and 12.9 kcal for rat and fish, respectively. Fish mitochondria were less stable than rat mitochondria to osmotic variation, and the disparity in initial swelling rates became increasingly greater with lower osmotic pressure. The hypotonic swelling of both fish and rat mitochondria was readily reversed osmotically; however, there was a very rapid decay of reversal in fish mitochondria and only a very slow decay in the case of rat. All the data indicate that under comparable conditions the fish mitochondrial membranes are more flexible and presumably more permeable and labile than rat mitochondrial membranes. The findings are discussed in relation to the general metabolic implications and the possible contributions of the membrane constituents to membrane behavior.
ELECTRON MICROSCOPE STUDIES OF THE STRUCTURE OF THE MICROVILLI ON PRINCIPAL EPITHELIAL CELLS OF RAT JEJUNUM AFTER TREATMENT IN HYPO- AND HYPERTONIC SALINE
Authors: Millington P. F. , Finean J. B.
Published: May 2004

Immersion of the intestinal tissue, from rat jejunum, in hypertonic saline produced very rapid changes in all regions of the epithelial cells, but the apical region was apparently unaffected by hypotonic solutions for at least ½ hour. In both cases, blistering of the microvilli was taken as the first sign of degenerative changes which finally resulted in a breakdown to large vesicular particles. Consideration of both normal and modified tissue indicates that the core of the microvillus contains either paired strands or tubular structures. Lateral cross-fibres extended from the core to the microvillus membrane and may be an essential part of the supporting structure of the microvillus. Densitometer traces across the microvillus membrane at various stages of modification indicated that this membrane might include a 75 A unit membrane structure with additional components associated at either surface. Interruptions in the membrane were apparently expanded by the hypotonic solutions and these might possibly be distinguished from preparative artefacts.
THE NUCLEAR ANNULI AS PATHWAYS FOR NUCLEOCYTOPLASMIC EXCHANGES
Authors: Feldherr Carl M.
Published: May 2004

Colloidal gold particles, 25 to 55 A in diameter, which had been coated with polyvinylpyrrolidone, were microinjected into the ground cytoplasm of amebas (Chaos chaos). At time intervals of 1 minute, 2 minutes, 10 minutes, and 24 hours after injection the cells were fixed for electron microscopy. After 24 hours, gold particles were found in both the nuclei and the ground cytoplasm, the concentration being higher in the nuclei. Colloidal particles were also present in the nuclei after 10 minutes, but at this time interval the concentration did not appear to be greater than that in the ground cytoplasm. One and 2 minutes after injection, the gold particles were located almost exclusively in the ground cytoplasm; however, individual particles were often found within the annuli of the nuclear envelope, and were located specifically in the centers of these structures. The results suggest that at least some of the gold particles which enter the nuclei pass through the annuli, and that passage through these structures may be restricted to a central channel.
THE BASE COMPOSITION OF NUCLEIC ACIDS IN CHROMOSOMES, PUFFS, NUCLEOLI, AND CYTOPLASM OF CHIRONOMUS SALIVARY GLAND CELLS
Authors: Edström Jan-Erik , Beermann Wolfgang
Published: May 2004

The base composition of RNA from individually isolated giant chromosomes, puffed chromosome segments, nucleoli, and samples of cytoplasm from Chironomus salivary gland cells was determined by microelectrophoresis. Data on the adenine: guanine quotient of the chromosomal DNA were also obtained. The results show that: 1) Chromosomal, nucleolar, and cytoplasmic RNA's differ significantly from each other in base composition. 2) Nucleolar and cytoplasmic RNA's, in spite of the difference, show great similarities with regard to the base composition and are both rich in adenine and uracil. 3) The RNA extracted from chromosome I differs significantly from the RNA's extracted from different segments of chromosome IV, and the latter differ significantly from each other. 4) The values for the RNA: DNA quotients of chromosome segments parallel the development of synthetically active genes, so-called Balbiani rings. 5) The chromosomal RNA does not show a base symmetry in any of the investigated cases, nor is the content of guanine + cytosine the same as that for DNA. The first of these two facts excludes the possibility that the chromosomal RNA is a complete copy of both strands of the chromosomal DNA. In spite of the difference in guanine + cytosine content between the two nucleic acids the RNA may still partly or completely be a single strand copy depending upon how representative the DNA values are for the synthetically active DNA.
PREVENTION OF PHOSPHATE - INDUCED MITOCHONDRIAL SWELLING
Authors: Kroll Arnold J. , Kuwabara Toichiro
Published: May 2004

The prevention of phosphate-induced mitochondrial swelling in the whole retina of the rabbit was studied with the electron microscope. It was found that a mixture of ATP, Mg++, and bovine serum albumin protected the mitochondria in vitro. This finding confirmed the results obtained spectrophotometrically with isolated rat liver mitochondria by Lehninger.
CHANGES IN THE BASE COMPOSITION OF NUCLEAR RIBONUCLEIC ACID OF NEURONS DURING A SHORT PERIOD OF ENHANCED PROTEIN PRODUCTION
Authors: Hydén Holger , Egyházi Endre
Published: May 2004

Nuclei from isolated nerve cells were sampled by microdissection. The content and composition of the nuclear RNA was studied and compared with that of the cytoplasmic RNA of Deiters' nerve cells of rabbits. Analyses were made of control nerve cells and of cells in which an enhanced RNA and protein production had been induced by chemical means, tricyano-amino-propene, for 60 minutes. The nuclear RNA content of the control nerve cells was 56 µµg, i.e. 3 per cent of the total RNA content of the nerve cell. The base ratios were: adenine 21.3, guanine 26.6, cytosine 30.8, uracil 21.3. Purine-pyrimidine analyses showed that the nuclear RNA differed significantly from the cytoplasmic RNA in having higher adenine and uracil values. The guanine and cytosine values were high, however, and the ratio G/C was 0.86 as compared with 1.16 for the cytoplasmic RNA. The composition of the nuclear RNA was interpreted as reflecting the extraordinarily strong development of the nucleolus in these neurons. During the 60 minutes of enhanced neuronal RNA production (+25 per cent) the guanine value increased and the uracil value decreased significantly in the nuclear RNA. In the cytoplasmic RNA the guanine value also increased although not so much as the nuclear guanine. The cytoplasmic cytosine value decreased. The result indicated that the production of the characteristic cytoplasmic RNA had been influenced by the change in the nuclear RNA
FINE STRUCTURE OF CELLS ISOLATED FROM ADULT MOUSE LIVER
Authors: Berry M. N. , Simpson F. O.
Published: May 2004

Suspensions of isolated cells in various media were prepared from mouse liver which had been perfused via the portal vein with a buffered medium containing 0.40 M sucrose, and the cells were fixed with osmium tetroxide. Their fine structure was compared with that of cells from perfused and unperfused intact liver. Perfusion brought about some separation of the cells with little or no damage to cell membranes. When cells were dispersed in 0.40 M sucrose medium the plasma membranes partially broke down, and this disintegration was increased by transfer of the cells to media of lower osmolarity. This is presumed to account for the loss of permeability barriers which occurs in isolated liver cells. The mitochondria in cells of perfused liver and in isolated cells remained elongated, but the layers of many mitochondrial cristae became separated by clear spaces. When cells were transferred to a medium containing 0.20 M sucrose, the mitochondria swelled and became spherical, often with displacement of the swollen cristae to the periphery. In a medium containing 0.06 M sucrose and 0.08 M potassium chloride the outer chamber of many mitochondria became swollen with displacement of the mitochondrial body to one side to give a crescent-shaped appearance. These changes in mitochondrial morphology are discussed in relation to the metabolic activity of isolated liver cells.
ISOLATION AND BIOCHEMICAL STUDY OF SECRETORY GRANULES FROM RAT PITUITARY GLANDS
Authors: Perdue J. F. , McShan W. H.
Published: May 2004

Secretory granules from anterior pituitary glands of young adult male castrate rats were isolated by differential centrifugation, microfiltration, and discontinuous density gradient centrifugation. The granules were obtained as pellets, sectioned, and studied with the electron microscope. A major part of the gonadotropin and a substantial amount of the TSH were associated with the isolated granules. Negligible amounts of growth hormone and prolactin were present as contaminants. Succinic dehydrogenase, glucose-6-phosphatase, acid protease, and acid and alkaline phosphatases were not found in the granules. Alkaline protease was the only enzyme found to be associated with the granules, and it is suggested, in the light of these results, that the alkaline protease may be involved in the release of the hormones.
ENZYME DISTRIBUTION IN THE RAT FETUS AND PLACENTA FOLLOWING THE ADMINSTRATION OF ETHIONINE
Authors: Schultz Richard L. , Schultz Phyllis W.
Published: May 2004

Enzyme changes which accompany ethionine-induced resorption of the rat conceptus have been studied by both histochemical and biochemical techniques. Pregnant rats were injected with ethionine over a 3-day period prior to autopsy on day 12 of pregnancy. Sections of the whole conceptus were studied for acid phosphatase with both the Burstone and Gomori methods and for succinoxidase activity with nitro-BT. Biochemical determinations of cathepsins, acid phosphatase, and succinoxidase were performed on homogenates of the fetuses, placentae, and deciduas basalis from ethionine-treated and saline-treated rats. The histochemical study has shown that resorption is accompanied by an increase in the size and number of acid phosphatase granules in the decidual tissues and a concurrent loss of acid phosphatase granules in the fetal tissues. Biochemical methodology indicated that there was no increase in total cathepsin or acid phosphatase activities in the resorbing tissues. No change in succinoxidase activity was found with either histochemical or biochemical techniques. The significance of these results was discussed with reference to the lysosome hypothesis.
CENTRIOLES AND THE FORMATION OF RUDIMENTARY CILIA BY FIBROBLASTS AND SMOOTH MUSCLE CELLS
Authors: Sorokin Sergei
Published: May 2004

Cells from a variety of sources, principally differentiating fibroblasts and smooth muscle cells from neonatal chicken and mammalian tissues and from organ cultures of chicken duodenum, were used as materials for an electron microscopic study on the formation of rudimentary cilia. Among the differentiating tissues many cells possessed a short, solitary cilium, which projected from one of the cell's pair of centrioles. Many stages evidently intermediate in the fashioning of cilium from centriole were encountered and furnished the evidence from which a reconstruction of ciliogenesis was attempted. The whole process may be divided into three phases. At first a solitary vesicle appears at one end of a centriole. The ciliary bud grows out from the same end of the centriole and invaginates the sac, which then becomes the temporary ciliary sheath. During the second phase the bud lengthens into a shaft, while the sheath enlarges to contain it. Enlargement of the sheath is effected by the repeated appearance of secondary vesicles nearby and their fusion with the sheath. Shaft and sheath reach the surface of the cell, where the sheath fuses with the plasma membrane during the third phase. Up to this point, formation of cilia follows the classical descriptions in outline. Subsequently, internal development of the shaft makes the rudimentary cilia of the investigated material more like certain non-motile centriolar derivatives than motile cilia. The pertinent literature is examined, and the cilia are tentatively assigned a non-motile status and a sensory function.
A METHOD FOR THE QUANTITATIVE ESTIMATION OF CYTOPLASMIC STRUCTURES
Authors: Loud Alden V.
Published: May 2004

A sampling procedure and calculations are described by which electron micrographs of cytoplasmic structures may be quantitatively analyzed. The relative areas occupied by formed bodies and by the "membrane space," the remainder of the cytoplasm, are evaluated. A method for making a measurement of the quantity of endoplasmic reticulum or other membrane profiles is described. The technic and results are illustrated with normal rat liver cells.
N-VINYLPYRROLIDONE AS A WATER COMPATIBLE CONSTITUENT OF EMBEDDING RESINS FOR SECTIONING IN ELECTRON MICROSCOPY
Authors: Fabergé A. C. , Lewis C. Willard
Published: May 2004

The use as an embedding resin for ultrathin sectioning of a cross-linked triple copolymer of N-vinylpyrrolidone, acrylonitrile, and ethylene glycol dimethacrylate is described. The first of these components is miscible with water, in all proportions, and can be used as a dehydrating agent, or, alternatively, ethanol may be used in the standard way. Polymerization is carried out at 37°C or even lower temperatures. This resin is unsuitable for use after osmium fixatives, but after permanganates it gives results similar to epoxy. Photographs of rye root-tip cells fixed in permanganate and sectioned in this resin are presented. Because of the water-permeable nature of the product and low polymerization temperature, this resin appears to have possibilities for histochemistry.
CHEMICAL AND MORPHOLOGICAL STUDIES ON THE IN VITRO CALCIFICATION OF AORTA
Authors: Martin G. R. , Schiffmann E. , Bladen H. A. , Nylen Marie
Published: May 2004

After a lag period, rat aortas incubated in rat serum in vitro accumulated substantial quantities of calcium and phosphate. Examination of the tissue by x-ray diffraction, microradiography, electron diffraction, and electron microscopy indicated that the calcium-phosphate phase which formed was hydroxyapatite and that the crystals were localized almost exclusively in elastin. Selective elimination of various components of the aorta with proteolytic enzymes indicated that the presence of elastin was required for mineralization. Collagen fibers did not appear to be required for the initiation of calcification, nor did they seem to undergo appreciable calcification in the time periods studied. Analysis of the initial lag period suggested that at least two changes occurred in serum prior to the mineralization of this tissue. Inhibitors of the reaction were destroyed, and the level of dialyzable calcium was elevated owing to its release from serum protein.
BETA GLUCURONIDASE-RICH CYTOPLASMIC PARTICLES IN ANDROGEN-STIMULATED MOUSE KIDNEY
Authors: Plaut Andrew G. , Fishman William H.
Published: May 2004

Androgens produced by stimulating mouse testis with gonadotropic hormones cause a rise in renal ß-glucuronidase but not an increase in acid or alkaline phosphatase. All subcellular components increase in ß-glucuronidase activity, with a relatively greater increment in particulate enzyme as compared with that free in the cytoplasm (non-sedimentable). A small percentage of recovered ß-glucuronidase, acid phosphatase, and alkaline phosphatase is found in material which rises to the surface during centrifugation in sucrose media (fraction I). The specific activity of ß-glucuronidase and acid phosphatase in this fraction is normally quite high with respect to the homogenate, while that of alkaline phosphatase is not. On the other hand, the fraction I material from androgen-stimulated mice exhibits a further increase in specific activity with respect to ß-glucuronidase and not acid phosphatase. It thus appears that there is an independence in the behavior of individual enzymes in response to physiologic stimuli in spite of obvious morphologic proximity.
FORMATION OF MITOCHONDRIA IN NEUROSPORA CRASSA
Authors: Luck David J. L.
Published: May 2004

Cells of a choline-requiring mutant of Neurospora crassa, labeled with radioactive choline, were transferred to unlabeled medium. At various times during their subsequent logarithmic growth, a highly purified mitochondrial fraction was prepared by sucrose density gradient centrifugation, and the distribution of label among individual mitochondria was determined by quantitative autoradiography. Preliminary experiments indicated that, under the conditions of this "washout" experiment, choline served as a stable mitochondrial label. Radioautographic analysis showed that, in fully labeled mycelia and for three mass doubling cycles in the unlabeled medium, radioactivity was randomly distributed among all mitochondria; i.e., the distribution of autographic grains among individual mitochondria followed a Poisson distribution. In experiments in which pulse labeling for 10 minutes was used, the label was randomly distributed among all mitochondria. The data suggest that the mitochondrial mass is increased by a continuous process of addition of new lecithin units to the already existing mitochondrial framework.
ELECTRON MICROSCOPIC STUDIES ON THE INDIRECT FLIGHT MUSCLES OF DROSOPHILA MELANOGASTER
Authors: Shafiq S. Ahmad
Published: May 2004

The myofibrils in Drosophila have thick and thin types of myofilaments arranged in the hexagonal pattern described for Calliphora by Huxley and Hanson (15). The thick filaments, along most of their length in the A band, seem to be binary in structure, consisting of a dense cortex and a lighter medulla. In the H zone, however, they show more uniform density; lateral projections (bridges) also appear to be absent in this region. The M band has a varying number of granules (probably of glycogen) distributed between the myofilaments. The myofilaments on reaching the Z region appear to change their hexagonal arrangement and become connected to one another by Z filaments. The regular arrangement of the filaments found in most regions of the fibrils is not seen in the terminal sarcomeres of some flight muscles; the two types of filaments appear to be intermingled in an irregular pattern in these parts of the fibrils. The attachment of myofibrils to the cuticle through the epidermal cells is described.
FINE STRUCTURE IN FROZEN-ETCHED YEAST CELLS
Authors: Moor H. , Mühlethaler K.
Published: May 2004

The freeze-etching technique, which is a special kind of freeze-drying, allows electron microscopic investigation of cells and tissues in the frozen state. In regard to yeast cells (Saccharomyces cerevisiae) a freeze-fixation technique has been developed which does not kill the object. The electron micrographs therefore are considered to impart an image of high fidelity. The cutting of the frozen object, which actually consists of a fine splintering, produces not only cross-sectional views (cross-fractures) of the structures but also surface views of the membranes and organelles. Many surface structures are described which have not been shown by the usual sectioning techniques. The cytoplasmic membrane contains hexagonal arrangements of particles which are apparently involved in the production of the glucan fibrils of the cell wall. Alterations of the distribution of nuclear pores are shown in cells of different ages. Freeze-etching enables a clear distinction of endoplasmic reticulum and vacuoles in yeast cells. The membranes of the vesicular systems are covered by ribosomes arranged in circular patterns. The mitochondrial envelope shows small perforations which could allow the exchange of macromolecules. The storage granules consist of concentric layers of lipid, presumably phosphatide. A Golgi apparatus has been detected which may be involved in the storage of lipid. The structure of the unit membrane and the membrane structures of all organelles as revealed by chemical fixation are confirmed in principle. Glycogen agglomerations are identified in the ground plasm of older cells. Insight into artifacts introduced by common chemical fixation and embedding techniques is obtained and discussed.
THE SARCOPLASMIC RETICULUM OF STRIATED MUSCLE OF A CYCLOPOID COPEPOD
Authors: Fahrenbach Wolf H.
Published: May 2004

The fine structure of the abdominal musculature of the copepod Macrocyclops albidus was investigated by electron microscopy. Tubules penetrate into the muscle fibers from the sarcolemma, continuity between the wall of the tubules and the sarcolemma being clear. A dense network of tubules envelops the myofibrils, its interstices being occupied by cisternal elements. At the Z lines the tubules traverse the interior of myofibrils, giving off branches which course longitudinally within the substance of the myofibrils. These branches are also accompanied by elongate, non-intercommunicating cisternae. Comparison of this fast acting copepod muscle with other vertebrate and invertebrate muscles indicates that the complexity of the tubular system is a function of the myofibrillar geometry, whereas the degree of development of the cisternal system is related to the contraction speed of the muscle.
AN ANALYSIS OF DESMOSOME SHAPE, SIZE, AND ORIENTATION BY THE USE OF HISTOMETRIC AND DENSITOMETRIC METHODS WITH ELECTRON MICROSCOPY
Authors: Tamarin Arnold , Sreebny Leo M.
Published: May 2004

The probable shape, size, and orientation of desmosomes of the cells comprising the secretory tubules in rat submaxillary gland was determined by statistical and algebraic methods applied to electron micrographs. It was concluded that these desmosomes are discrete ellipsoidal discs whose principal axes are in the order of 4100 and 2500 angstrom units, and that they are preferentially oriented with their long axis more or less parallel to the base-apex axis of the cell. Densitometric interpretation agrees with the statistically based reconstruction of desmosomal shape. By densitometric analysis it was also determined that the peak to peak distances between layers within these desmosomes are in essential agreement with other reported findings. The approach described may have general applications to problems in the analysis of submicroscopic morphology.
STUDIES ON ISOLATED NUCLEI
Authors: Maggio Rachele , Siekevitz Philip , Palade George E.
Published: May 2004

This article describes a method for the isolation of nuclei from guinea pig liver. It involves the homogenization of the tissue in 0.88 M sucrose-1.5 mM CaCl2 followed by centrifugation in a discontinuous density gradient in which the upper phase is the homogenate and the lower phase is 2.2 M sucrose-0.5 mM CaCl2. Based on DNA recovery, the isolated fraction contains 25 to 30 per cent of the nuclei of the original homogenate. Electron microscopical observations showed that ∼88 per cent of the isolated nuclei come from liver cells (the rest from von Kupffer cells and leucocytes) and that ∼90 per cent of the nuclei appear intact, with well preserved nucleoli, nucleoplasm, nuclear envelope, and pores. Cytoplasmic contamination is minimal and consists primarily of the nuclear envelope and its attached ribosomes. The nuclear fraction consists of ∼22.3 per cent DNA, ∼4.7 per cent RNA, and ∼73 per cent protein, the DNA/RNA ratio being 4.7. Data on RNA extractibility by phosphate and salt and on the base composition of total nuclear RNA are included.