The fine structure of the retinal plexus in Octopus vulgaris

Summary The plexiform layer of the octopus retina was investigated by means of electron microscopy. The axonic processes of visual cells contain closely packed microtubules, 300 Å in diameter, running parallel to one another along the long axis of the processes. Visual cells also send out a large number of thin axon collaterals. Each of them forms presynaptic knobs with numerous clear vesicles along their course. These are assumed to be concerned with the reciprocal communication between visual cells. Nerve endings with dense-cored vesicles form synaptic contacts with visual cells. The visual cells show some spherical protrusions into the perivascular spaces.Octopuses, captured off the coast of Onagawa (Miyagi-ken, Japan) in autumn of 1963 and 1964, were offered for this research through the kindness of Dr. Kyoji Tasaki, Assistant Professor of Physiology in Tohoku University School of Medicine.I wish to thank Professor Toshiyuki Yamamoto for his encouragement and suggestions throughout all stages of this work, and also Mr. Masae Kato for his technical assistance in drawing.     

The reorganization of the hypophysial stalk following hypophysectomy in the teleost Porichthys notatus Girard

Summary Porichthys notatus has a long infundibular stalk, measuring 3 to 5 mm. The third ventricle extends into the stalk as a long infundibular funnel.In several hypophysectomised specimens the proximal cut end of the stalk enlarged to form a round or oblong body, having axonal endings with AF-positive material, glial elements, and blood sinuses. This suggests that in the absence of the pituitary the cut end of the stalk is reorganised into a neurohypophysis-like organ.The regenerated stalk seems to have the essential components for neurohypophysial function; axonal endings for the storage of secretory products, and related blood vessels for the release of the stored principles.This work was supported by the Population Council of the Rockefeller Institute, New York. I am thankful to Dr. R. L. Fernald, Director of the Friday Harbor Marine Biological Laboratories, University of Washington, for providing collection and laboratory facilities. I am indebted to Dr. Aubrey Gorbman, Department of Zoology, University of Washington for the generous hospitality in his laboratory and encouragement, which made this work possible. Material support in part for this research was received from U. S. P. H. S. grant NB 04887 awarded to Dr. A. Gorbman.     

The ultrastructure of monkey eccrine sweat glands

Summary The ultrastructure of monkey eccrine sweat glands is described. The secretory portion of the sweat gland is discussed in detail. The morphological differences in the secretory coil using three different fixatives and fixative combinations are emphasized. The secretory product of dark cells is seen to have three distinct appearances depending upon the fixative used. The biochemical significance of the latter finding is discussed. The appearance of clear cell cytoplasmic processes is described using the different fixatives. The similarity of adjacent clear cell processes to those of avian salt glands is pointed out and discussed. Evidence is presented to indicate that dark cells arise from clear cells via an intermediate cell type. The appearance of the clear cell plasma membrane is described and the necessity for the use of the general term multilaminar plasma membrane is discussed.Supported by U.S.P.H.S. grant 5 T 1-GM-29 F-04 AS. The author would like to express his gratitude to the Lederle Laboratories and in particular to Dr.James Vickers for providing the tissue. Sincere thanks is given to Mrs.Dagmar Graham and Mrs.Ditza Springer for technical assistance and also to MissMary Lorenc for preparation of the diagram. In addition, I would like to thank Dr.J. A. G. Rhodin for his criticism and advice.     

Cells with microvillous borders in the cerebral ganglion of Leptodora kindtii focke (Crustacea)

Summary Groups of large cells in the cerebral ganglion of Leptodora kindtii join in intricate patterns to enclose lacunar spaces. The cell surfaces bordering on these lacunae are covered by long, densely packed microvilli that all but fill the spaces. Near their brush borders the cells are joined by adhesion plates; for the rest they are separated from each other by glial septa. The possible significance of these structures is discussed.Dedicated to an inspiring teacher and loyal friend, Prof. F. Wassermann on the occasion of his 80th birthday, August 13, 1964.Supported by Grant No. NB-02145 from the United States Public Health Service. The expert assistance of Mrs. Cynthia Jones, Mrs. Sarah Wurzelmann, and Mr. Stanley Brown is gratefully acknowledged.     

Ultrastructural study on the morphogenesis of the neuromuscular junction in the skeletal muscle of the chick

Summary The morphogenesis of the neuromuscular junction was examined at the ultrastructural level in the skeletal muscle of the lower limb of the chick. The fine structure of the neuromuscular junction of the adult fowl was essentially the same as that in other vertebrates; the junction consists of the axon terminal, the Schwann cell, and the muscle fiber. The first visible sign of neuromuscular junction formation, in embryos of 13 days in ovo, was the membrane thickening of the sarcolemma which develops into the postsynaptic membrane. The axons approaching the muscle fibers were incompletely ensheathed by a Schwann cell and contained vesicles. The subsequent differentiation of the junctional sarcoplasm, the axoplasm, and the Schwann cell cytoplasm takes place from 13 to 18 days in ovo and the junction nearly reaches maturity at around 20 days in ovo. The formation of complicated anastomoses and branching of the junctional infoldings seems to occur after hatching. These ultrastructural observations are in good agreement with histochemical findings (cholinesterase method) in terms of the chronology of the morphogenesis of the junction.This investigation was supported in part by U. S. Public Health Service Grant MH 12269-01, administered by Dr. Kazuo Ogawa. It was initiated on the suggestion of Prof. J. Nakai, Department of Anatomy, Faculty of Medicine, Tokyo University, and a part of it was performed in his laboratory. The author is greatly indebted to Prof. K. Ogawa, Department of Anatomy, Kansai Medical School, for his guidance and encouragement, and to Dr. S. Igarashi, Department of Anatomy, Tokyo University, for some technical advice.     

The fine structure of the megakaryocyte of the bone marrow of the guinea pig

Summary The cytoplasm of megakaryocytes frequently exhibits bullate processes which protrude into the lumen of the sinusoids through small apertures in the reticular cells. These bullae differ morphologically from the platelet demarcation zones and are held to have a different function. It is concluded that the bullae detach from the megakaryocyte in a manner comparable to apocrine secretion and enter the blood stream.This investigation was supported by a PHS research grant (CA 05493) from the U.S. Public Health Service.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his 80^th birthday.     

Formation of giant nuclei in continuously irradiated tumors

Summary Transplantable malignant tumors of mice and rats have been irradiated continuously for several days by means of embedded glass beads containing Sr⁹⁰-Y⁹⁰. Concomitant enlargement of tumor cell nuclei occurs in areas where mitosis is inhibited, i.e. associated with doses of 1200–2000 rads per day and above. Some enlarged cells survive at least a week in areas where the dose rate is much higher than this. Intense incorporation of thymidine into enlarged cells can be seen at least a week after beginning irradiation.Work supported by the U.S. Atomic Energy Commission.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his eightieth birthday.     

On orderly domains of particles associated with cytomembranes during spermatogenesis in Helix aspersa

Summary In an electron microscopic study of the maturation of the spermatid of the snail Helix aspersa, it was found that: 1. there is persistence and utilization of the mitochondrial architecture in the formation of the middle piece, and 2. particles originate in orderly domains within the pseudomatrix of the mitochondria. Other orderly domains of cytomembranes (Golgi system and peripheral tubules of Grassé) are discussed.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his eightieth birthday.Work supported by U.S. Atomic Energy Commission.     

On the glycocalyx,the external coat of the plasma membrane,of some secretory cells

Summary The free surface of epithelial cells of secretory organs (human placenta, lactating mammary gland of the rat, choroid plexus of man and rat) and of the accessory organs of the genital tract of the male rat is characterized by a plasmalemmal differentiation named glycocalyx or surface mucous coat. This structure is built up by filamentous or globular substructures.Two main ultrastructural types of the glyeocalyx were observed: 1) The filamentous type such as in the rat epididymis, which resembles the cat intestinal glyeocalyx (Ito, 1965) and that one of human transitional epithelium (Monis and Zambrano, 1968), and 2) The globular type, as observed in the lumen of the lactating mammary gland of the rat.Sialic acid was demonstrated histochemically in the luminal glyeocalyx of all organs studied. In addition, the glyeocalyx of acinar cells of the lactating mammary gland contains sulfate and phosphate groups which were identified by histochemical technics, using enzymatic digestion procedures, suggesting the chemical heterogeneity of this glyeocalyx.Present investigations follow the working hypothesis that the complex carbohydrates of glycocalyces become part of the product of activity of secreting cells.We thank Mr. Luis Iwakawa, Miss Silvia Falcón, Miss Elsa M. Orgnero for technical help, Miss Graciela Aliaga for secretarial assistance. Photography by Mr. H. Magnani. Dr. Hugo F. Carrer cooperated in the initial stages of this investigation.The authors acknowledge the use of the electron microscope of the Department of Pathology, Córdoba University Medical School, for which they thank Prof. E. Mosquera and Dr. E. Hliba. Dr. Hliba photographed picture number 4.     

Analysis of glial cell differentiation in peripheral nervous tissue: II. Neurons promote S100 synthesis by purified glial precursor cell populations

Isolated sensory neurons in vitro do not contain or synthesize S100, whereas glial cell precursor populations do. These precursor cells, when isolated from other cell types, produce low levels of S100 but never undergo the developmental transition to produce high levels of S100. When glial cell precursors are combined with isolated, live or paraformaldehyde-fixed sensory neurons, the precursor cells do undergo the second transition, and accumulate high levels of S100. Peroxidase-anti-peroxidase immunohistochemical staining for S100 confirms previous conclusions (B. Holton and J. A. Weston, 1982, Develop. Biol.89, 64–71) that only those glial cells which are closely apposed to neurons contain augmented levels of S100. This stimulation appears to be specific to neuronal/glial interactions since live or fixed fibroblasts, when cocultured with glial precursor cells, do not promote accumulation of S100 by the glial cells.     

The fine structure of the mouse adrenal X zone

Summary Electron microscopically the adrenal X zone was examined in the fourteen SMA female mice aged 40 and 70 days. At these ages, the X zone showed no signs of degeneration. The X zone cell was somewhat smaller than the permanent cortical cell.The mitochondria in the X zone cell were quite bizarre in shape, provided with tubules or cristae. Many intramitochondrial bodies very similar to the cytoplasmic lipid droplets were found in the X zone. A few lipid droplets and globules were also noticed in this zone. The lipid droplets may possibly be formed within the mitochondria.The light and dark cells were differentiated. For the light cells, scant mitochondria and tubular granular endoplasmic reticulum were characteristic in contrast to the abundant mitochondria and multi-lamellated agranular endoplasmic reticulum in the dark cells. The cellular variety in density was discussed with regard to steroid synthesis.The author wishes to express his sincere appreciation to Prof. H. Tauchi, The 2nd Department of Pathology, Nagoya University School of Medicine, for kind advice, to Dr. M. Hoshino for helpful suggestion, and to Mr. J. Aoki for excellent technical assistance.     

Theoretical analysis of two models of cell differentiation

Summary Two models of differentiation in renewal systems have been analyzed, and their capacities to regulate production of differentiated cells in the normal steady state and under conditions of excessive and prolonged growth stimulation have been compared. Decompensation of one model leads to depletion of stem cells, the other to accumulation of stem cells. The histological demonstration of accumulation of stem cells in tissues subjected to chronic hyperstimulation by hormones is well explained by the present extension of one of the models. Such hyperplasia of stem cells accompanying the disequilibrium state induced by hyperstimulation is characteristic of precancerous tissues and may be an important causal mechanism of cancer.Work supported by the U.S. Atomic Energy Commission.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his 80th birthday.     

Fine structure of the retina in Haliotis discus

Summary The fine structure of the retina of Haliotis discus has been studied by means of electron microscopy. The visual cell has an apical projection which protrudes beyond the retinal surface into the ocellar cavity. The surface of this projection shows elaborate in- and outfoldings. Besides, two apposed laminae of the folded plasma membrane contact each other to form a quintuple-layered compound membrane. The analogy of this compound membrane system to the rhabdomere of the visual cell of Cephalopoda has been discussed. Subsurface endoplasmic reticulum and giant multivesicular bodies are also notable structures in Haliotis visual cells. The observations include the fine structure of supporting cells, plexiform layer, optic nerve and transitional zone of retina to epidermis.The author wishes to express his appreciation for the encouragement and suggestions given by Dr. Toshi Yuki Yamamoto, Professor of Anatomy, Tohoku University School of Medicine, through all stages of this work.     

Histochemical localization of adenosinetriphosphatase in the testicular and pre-testicular nephridia of the Indian leech Hirudinaria granulosa (Savigny)

Summary Adenosinetriphosphatase has been histochemically demonstrated in the initiallobe cells and the central canal in its first round in the pre-testicular and testicular nephridia of the Indian leech Hirudinaria granulosa. The apical-lobe cells are positive in the pre-testicular and negative in the testicular nephridia. The functional significance of the enzyme at these sites has been discussed.I wish to express my gratitude to Dr. H. B. Tewari, under whose guidance the present work has been carried out. I am also indebted to Dr. M. L. Dhar, Director, Central Drug Research Institute, Lucknow; Dr. P. S. Krishnan, Professor of Biochemistry, University of Lucknow; and Dr. (Miss) Usha Gupta, Department of Pathology, University of Lucknow, for the hospitality which they have accorded me in their laboratories.     

The fine structure of photoreceptors in a marine flatworm

Summary The ocelli or eyes of the marine polyclad turbellarian Notoplana acticola are clustered on the paired dorsal nuchal tentacles and in two longitudinal bands lateral to the cerebral ganglion. The ocelli, studied by electron microscopy, were characterized as rhabdomeric and non-ciliary in origin. There are 60 to 80 ocelli per animal each enclosed in a fibrous capsule to which muscle fibers may attach. An ocellus consists of a pigmented eyecup into which 30 to 50 photoreceptor cells send dendritic processes through interruptions in or among pigment cell projections across the eyecup opening. The dendritic processes terminate in numerous long intertwined microvilli which fill the eyecup. The nucleated cell body of each photoreceptor cell lies outside the eyecup and projects an axonal process to the cerebral mass. Within the dendritic processes are observed mitochondria, ribosomes, neurotubules, multivesicular bodies, vesicles and vacuoles. The cell body contains smaller mitochondria, endoplasmic reticulum, ribosomes, vesicles and prominent Golgi complexes.After dark adaptation, there are some structural alterations in terms of swelling of microvilli, increased numbers of vacuoles associated with the microvilli and dendritic processes, and changes in the pigment cell projections.This work was supported by Grant No. GM 10292 from the U.S. Public Health Service to Professor Richard M. Eakin, Department of Zoology at the University of California, Berkeley, U.S.A., where this investigation was conducted during the author's sabbatical leave of absence from the University of Illinois, and by Grant No. 1 SO 1 FR 5369 from the U.S. Public Health Service to the University of Illinois at the Medical Center.I express appreciation to Professor Eakin for interesting discussions and generous hospitality to me as a guest in his laboratory, and to the John Simon Guggenheim Memorial Foundation for the Fellowship which I held during 1964–65. I thank Dr. John P. Marbarger, Director of the Aeromedical Laboratory for the electron microscope facilities used at the University of Illinois.     

Patterns of segregation of structural elements during cell division

Summary Adaptations of currently available autoradiographic electron microscopic methods to the study of organelle development in single Paramecium cells are described. Cells of known age were pulse-labeled with tritiated leucine and observed either as whole mounts, or as sections of specified age and cell-lineage. Labeled trichocysts appear as early as one hour after uptake and retain the label without dilution for at least 3 fission generations thereafter. Cells grown in unlabeled medium, but derived from labeled cells, show the bulk of the residual label to be conservatively associated with the stable structural organelles of the cell surface. These organelles include those trichocysts and ciliary corpuscles that had been synthesized in the ancestral progenitor cell during or shortly after administration of the isotopically labeled amino acid.Work supported by U. S. Atomic Energy Commission.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occassion of his eightieth birthday.     

The organization of the substantia gelatinosa rolandi in the cat lumbosacral spinal cord

Summary The organization of the substantia gelatinosa and adjacent lamina III in cat lumbo-sacral spinal cord has been studied by light and electron microscopical techniques in normal cord and following dorsal root section.The substantia gelatinosa (lamina II of Rexed) is characterized by bundles of small, non-myelinated axons, principally oriented longitudinally. The substantia gelatinosa cells are small, spindle shaped, with a cytoplasm generally devoid of Nissl substance. There are extensive axo-dendritic and axo-axonal contacts within the substantia gelatinosa and less frequent axo-somatic contacts.Larger marginal cells oriented horizontally on the surface of the substantia gelatinosa and containing Nissl substance are also seen.Lamina III is somewhat similar to the substantia gelatinosa, but lacks the complex bundles of non-myelinated axons.Following dorsal root section, heavy degeneration is seen by light and electron microscopy in lamina III, but is rarely seen in the substantia gelatinosa. It is concluded that the substantia gelatinosa and lamina III are distinct anatomically and therefore may differ functionally.The possible physiological role of the substantia gelatinosa is discussed.This work was supported by a Special Fellowship 2F11 NB 1140 02 NSRB from the National Institute of Neurological Diseases and Blindness, United States Public Health Service.The author is indebted to Dr. E. G.Gray for his excellent advice. I thank Dr. R. W. Guillery, Dr. L. E. Westeum and Dr. B. G. Cragg for their assistance, and Prof. J. Z. Young, F. R. S. for his kind suggestions. I also wish to thank Mr. S. Waterman for the photography.     

Uptake of radioactive thymidine and cytidine by Ehrlich ascites tumor cells in different stages of growth

Summary In Strong A female mice, the Ehrlich ascites tumor inoculated into the peritoneal cavity grows exponentially for the first 7 days with a doubling time of about 36 hours. The tumor enters then into a late stage during which the number of tumor cells in the peritoneal cavity does not increase. The uptake of intraperitoneally injected thymidine decreases from the exponential to the late stage, mostly because of a decrease in the fraction of cells in DNA synthesis. During the exponential phase, the uptake of thymidine is a function of the amount of radioactive thymidine injected per tumor cell, the utilization decreasing with increasing cell dose. The uptake of intraperitoneally injected cytidine decreases slightly with time after inoculation although the fraction of tumor cells in RNA synthesis remains constant.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his 80th birthday.This investigation was supported by U.S.P.H.S. Grant CA-05667. The author is a U.S.P.H.S. Research Career Development Awardee.     

The effect of metopirone and aldactone on the interrenal tissue of intact and pars distalis-ectomized common frogs Rana temporaria

Summary The effects of metopirone (SU-4885) and aldactone (SC-9420) on the histology and histochemistry of the interrenal tissue of male common frogs, Rana temporaria, have been studied. From the changes in nuclear and cellular size, mitotic rate, and lipid and cholesterol content it has been concluded that in intact animals the administration of metopirone as well as of aldactone resulted in an increase of the interrenal secretory activity. The effects were very similar to that of ACTH. Ablation of the pars distalis prevented the interrenal activation by metopirone. The effect of aldactone was markedly reduced in operated frogs, but some stimulation remained. It is suggested that both substances act by way of enhancing pituitary ACTH ouput. The dependence of the secretion of corticosteroids on pituitary ACTH is discussed.I wish to thank Dr. W. J. van Dongen for his cooperation and criticism and for enabling me to do this work in his laboratory. I am endebted to Prof. Dr. P. G. W. J. van Oordt for his valuable and constructive suggestions. Thanks are also due to Miss Mieke Roelofsen and Miss Ineke Wienen for their conscientious technical assistance.     

Thyroidal radioiodine uptake and thyrotropic potency of the pituitary in a freshwater catfish,Mystus vittatus (Bloch), in response to L-thyroxin,antithyroid drugs,and heavy doses of 131I

Summary Experiments were conducted to ascertain the thyroidal ¹³¹I uptake and thyrotropic potency of the pituitary gland in a freshwater catfish, in response to L-thyroxine, antithyroid drugs and heavy doses of radioiodine. L-thyroxine treatments slightly lowered thyroidal radioiodine uptake, and there was at least a trend of lowered TSH content in the pituitaries of these animals. Administration of antithyroid drugs (propylthiouracil, thiourea, KSCN) caused a significant decrease in radioiodine uptake and a highly significant increase in TSH content of the pituitary. Heavy doses of I¹³¹ almost completely blocked thyroidal iodine uptake but they were as effective as antithyroid drugs in elevating TSH content of the pituitary.I am greatly indebted to Dr. G. E. Pickford, Yale University, U.S.A. for her helpful suggestions; to Dr. A. G. Sathyanesan, Banaras Hindu University, India, for encouragements; to Professor S. P. Ray-Chaudhuri, Banaras Hindu University, India, for providing laboratory facilities. I am also grateful to Baxtor Laboratories Inc., Morton Grove, Illinois, U.S.A. for the gift of Crystalline L-thyroxine which was made available through the courtesy of Professor Paul Starr and Dr. Thomas Garrett.