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.     

Ultrastructure of spermatozoa of Peregrinus maidis (Homoptera,Delphacidae)

Summary The spermatozoa of Peregrinus maidis Ashm. are thread-like, approximately 650 long and 1 wide including the head (approximately 28 ).The main part of the spermatozoa consists of two mitochondria derivatives, a central body between them, the axial filament complex, and a newly found element consisting of two wing-shaped bodies. Each mitochondrion derivative shows a peripheral and an inner part. The peripheral part is formed by cristae arranged perpendicularly to the long axis of the spermatozoon. The cristae are approximately 70 Å wide. The dense layers between them measure approximately 280 Å. The inner part of the mitochondrion derivative shows a crystalline array, formed by sub-units of approximately 100 Å diameter. The wing-shaped bodies consist of tubular elements.The head has an elongated nucleus with an electron transparent space inside. At the anterior end of the nucleus lies a tapered acrosome. This appears fibrous and parts of the acrosome fibers seem to run along the nucleus. Acknowledgements. The authors wish to thank Dr. G. H. Bergold for suggestions and support, Drs. J. André, D. W. Fawcett, P. Maillet and G. F. Meyer for very helpful discussion. They are also grateful to Mr. O. Suárez for assistance in the preparation of the organs of P. maidis and to Mrs. M. de Pingarrón for technical assistance.     

Photoperiodic and osmotic influences on the ultrastructure of the hypothalamic neurosecretory system of the White-crowned sparrow,Zonotrichia leucophrys gambelii

Summary An attempt was made to correlate functional changes in the neurohypophysis of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, with morphologic features on the light- and electron-microscope levels. The aldehyde-fuchsin-staining anterior median eminence possesses essentially the same ultrastructural features as the non-staining posterior median eminence. The axon terminals are characterized by the presence of a large number of small vesicles (approximately 400 Å in diameter) and occasional electron-dense granules. The more-or-less depleted anterior median eminence occasionally evident in the photosensitive bird showing testicular development is indistinguishable ultrastructurally from the more intensely staining median eminence generally characteristic of the photorefractory bird. In the median eminence, stainability and functional state do not seem to be correlated with changes in the type, size or number of vesicles. A slight increase in the number of granules was noted in the photorefractory bird but this was considered insufficient basis to account for differences in stainability.The pars nervosa, on the other hand, responded to osmotic stimuli (saline drinking water) by loss of stainability and decrease in numbers of elementary neurosecretory granules. Small vesicles are also present in the pars nervosa axon terminals, but are intermingled with neurosecretory granules in normal birds. Acute-osmotic birds, however, had axon terminals almost entirely occupied by small vesicles.It is to be emphasized that the pars nervosa and the median eminence are two structurally very different regions of the neurohypophysis. The basis for aldehyde-fuchsin staining in the median eminence appears to differ from that in the pars nervosa. The implications of these findings are considered in regard to hypothalamic control over gonadotropic activity in the White-crowned Sparrow.Dedicated to Professor Dr. W. Bargmann in honor of his 60th birthday.This investigation was supported by grant GB-2484 from the National Science Foundation to Professor Bern, grant GB-2819 from the National Science Foundation to Professor Mewaldt, and grant NB-01353 from the National Institutes of Health to Professor Farner. The authors wish to express their appreciation of the technical assistance of Mrs. Irene Brown, Mr. John Butchart, Sally S. Kibby, Mrs. Carol Nicoll, and Mr. John Striffler. Mrs. Emily Reid kindly prepared the histograms.     

Light and dark metabolism ofl- andd-amino acids in 5 strains ofChlorella vulgaris and 4 strains of the genusAnkistrodesmus

In the light, 2 out of the 4 newly testedChlorella vulgaris strains were found to use about as many amino acids as a source of nitrogen as the previously investigated strain Delft; the other 2C. vulgaris strains and 3 of theAnkistrodesmus strains used only a few. The 4thAnkistrodesmus strain used none.On the average,l-amino acids supported better growth thand-amino acids, butd-serine was preferred tol-serine by 3Ankistrodesmus strains.In the dark, growth was only obtained withC. vulgaris strain Delft, and only on a few of thel-amino acids,l-leucine in particular.The author is indebted to the Direction of the Academic Hospital Dijkzigt, Rotterdam and to Prof. Dr. H. Esseveld, Head of the Central Bacteriological Laboratory, Rotterdam, for providing facilities for the performance of this study.He thanks Mrs. Dr. H. J. Leijnse-Ybema for her help in making the chromatograms, and Mr. J. B. Lenstra, pharmacist, for advice in matters of organic chemistry.     

The ultrastructural basis for the identification of cell types in the pancreatic islets I. Guinea pig

Summary The pancreatic islets of the guinea pig have been studied by light and electron microscopy. The B granules in glutaraldehyde-fixed tissue often are cup-shaped with an indentation visible on one side of the granule. Phosphotungstic acid hematoxylin (PTAH) positive cells have been characterized by electron microscopy as three subtypes based on the size of the secretory granules. A_a cells are the most common and have secretory granules around 200 m in diameter. A_b cells have large secretory granules around 300 m in diameter and are relatively infrequent. A_c cells are the least common and have small (160 m) granules. Characteristic D cells are identifiable by electron microscopy and, on the basis of the subsequent study (Munger, Caramia, and Lacy, 1965), are identified as the silver positive cells observed by light microscopy.This investigation was supported in part by United States Public Health Service research grants GM-10102 and GM-03784 from the Institute of General Medical Sciences, and AM-01226 from the Institute of Arthritis and Metabolic Diseases.-The authors wish to acknowledge the valuable technical assistance of Mrs. Aileen Sevier and Mrs. Lidia Donohue.     

Synapses in the cerebral ganglion of adult Ciona intestinalis

Summary Electron microscopy of the synaptic morphology of synapses in the cerebral ganglion of the adult ascidian (sea squirt) Ciona intestinalis reveals that the synapses are restricted to the central neuropil of the ganglion. Many of the synapses show a polarity of structure such that pre and post synaptic parts can be identified. The vesicles in the presynaptic bag are of two main diameters 80 and 30 nm respectively. The large vesicles have electron dense contents that vary both in their capacity and dimensions.The pre and postsynaptic membranes are more electron dense than the surrounding membranes, but they are only slightly thicker. Both the pre and post synaptic membranes have electron dense dots some 10 nm in diameter associated with their cytoplasmic surfaces. Sometimes the presynaptic membrane has larger peg-like projections between the vesicles. Associated with the post synaptic membrane are tubules some 10 nm in diameter. These tubules may be the dots cut obliquely.The synaptic cleft material is more electron dense than the surrounding intercellular material, and in it there is a dense line made up of granules about 3–5 nm in diameter. This dense line is usually mid way between the pre and post synaptic membranes, but may be nearer the postsynaptic membrane.No tight junctions between adjacent nerve process profiles have been observed.I wish to thank Professors J. Z. Young, F. R. S. and E. G. Gray for much advice and encouragement, also Dr. R. Bellairs for the use of electron microscope facilities and Mr. R. Moss and Mrs. J. Hamilton for skillful technical assistance.     

The fine structure of cephalopod blood vessels II. The vessels of the nervous system

Summary Blood vessels of the perioesophageal nerve ganglia (brain) of Octopus vulgaris and the stellate ganglia of Sepia officinalis are described. The vessels have an incomplete endothelium, a complete basement membrane and a complete investment of pericytes. The pericytes are joined by specialised membrane junctions but these are not tight junctions. The main type of neuron/vessel arrangement is one where there is a collagen-filled space between the pericytes and the surrounding glial cells. Axons or neurons are sometimes applied directly to the vessel pericytes and in the neuropil, pericytes contact glial cells that ensheath bundles of axons. Blood spaces between neurons are also present.We would like thank Professor J. Z. Young and Dr. E. G. Gray for encouragement and advice, Mrs. Jane Astafiev for drawing Fig. 11, Mr. S. Waterman for photographic assistance and Miss Cheryl Martin for secretarial and other assistance.     

Electron microscope studies on collagen

Summary The present paper describes the effect of intensive tryptic digestion of native tropocollagen (TC) macromolecules in solution. Contrary to earlier investigations it has been found that the trypsin treatment results in a fragmentation of the TC molecules. The addition of ATP to solutions exposed to the enzyme yields SLS fragments, 2250 Å in length. Comparison between these and normal SLS type aggregates shows that the scission occurs in a well-defined locus adjacent to the -1/2-line seen after positive staining. The significance of this finding is discussed.This study was supported by grant NB-02215-04 from the National Institute of Neurological Diseases and Blindness, Public Health Service, U.S.A. and a Student research fellowship from the Norwegian Research Council for Science and the Humanities. This aid is gratefully acknowledged.I am indebted to Mrs. J. Line Vaaland and Mr. B. V. Johansen for technical assistance.     

The fine structure of the eye of the mollusc Pecten maximus

Summary The retina of Pecten maximus is divided into two light sensitive layers forming the distal and proximal retinae. The cells from these layers have different electrophysiological responses, the distal cells giving primary off responses, and the proximal cells giving on responses. The receptor surfaces of the distal retinal cells are formed from lamellae produced by the outer membranes of flattened cilia. These cilia have a basal body, basal foot, no root system and a 9 + 0 internal filament content. Each cell gives rise to an axon from its distal side, and this process goes up to the basement membrane, which is present below the cellular lens, passes along beneath it, and joins the distal optic nerve. The receptor part of the proximal retinal cells is formed from a vast array of microvilli. Each of these cells also bears one or two cilia with a probable 9 + 0 internal filament complement and no roots. The proximal cells give rise to axons, forming the proximal optic nerve. Below the proximal retina is a reflecting layer, the argentea, and below this is a pigment cell layer.We would like to acknowledge the advice and encouragement of Professor A. F. Huxley, Professor J. Z. Young and Dr. E. G. Gray. — We would like to thank Mrs. J. I. Astafiev for drawing Fig. 1, Mr. S. Waterman for photographic help and Miss C. Martin for clerical assistance.     

The effects of cyanide plus β-hydroxybutyrate on changes in mitochondrial absorbancy and ultrastructure induced by hypotonicity and inorganic phosphate

Summary Absorbancy measurements and electron microscopy of isolated rat liver mitochondria show that simple osmotic swelling is not prevented by cyanide plus -hydroxybutyrate. Similarly, electron microscopy shows that these two agents do not prevent the swelling induced by inorganic phosphate. However, the absorbancy decrease induced by inorganic phosphate is inhibited by cyanide plus -hydroxybutyrate.These findings cast doubt on the validity of the absorbancy method as an index of mitochondrial morphology. The evidence also indicates that both simple osmotic swelling and that induced by inorganic phosphate are independent of respiration. An osmotic mechanism is proposed as an alternative.Dedicated to Prof. Berta Scharrer on her 60^th birthday.This study was supported by Research Grants GM-08900, NB-02145, NB-05219 from the U.S.P.H.S. and a Lederle Medical Faculty Award. Fine technical assistance was provided by Mrs. Cynthia Jones, Miss Ursula Moeller and Mr. Stanley Brown.     

Particles resembling microbodies in normal and neoplastic perianal glands of dogs

Summary By electron microscopy, the parenchymal cells of the perianal glands of dogs contain granules which have the morphological features of microbodies (peroxisomes) including marginal plates and, occasionally, dense nucleoids. Like microbodies, they are occasionally attached to the endoplasmic reticulum. Histochemical evidence is presented suggesting that they contain at least one of the peroxisomal enzymes, L--hydroxy acid oxidase. The granules of a perianal gland adenoma showed abnormal morphologic variations.Mrs. Murtie Still, Mrs. Bertha McClure and Mr. Bob White gave valuable technical assistance.     

The structure of a photoreceptor organelle in the eye of Pterotrachea mutica

Summary The photoreceptor cell of Pterotrachea consists of an elongated cell some 100 m long with recogniseable inner and outer segments. The photoreceptor membranes point towards the light. There are about 300 discs per photoreceptor, a small number of discs arising from a single ciliary base. There are bout 75–100 such bases on each receptor cell. The receptor cells themselves (the inner segments) have four recognisable regions. The vacuolated region, the region of mitochondria, the nuclear region, and the axonal region.The photoreceptor cells are organised in five roughly parallel rows, and separated from one another by pale supporting cells.My thanks are due to Professor J. Z. Young, F. R. S. for his enthusiastic support and help during this work. Dr. R. Bellairs kindly provided electron microscope facilities. Mr. R. Moss, Mrs. J. Hamilton and Mr. A. Aldridge provided excellent technical and photographic assistance.     

Two sites for adenine-nucleotide regulation of ATP-sensitive potassium channels in mouse pancreatic β-cells and HIT cells

Summary ATP-inhibited potassium channels (K(ATP)) were studied in excised, inside-out patches from cultured adult mouse pancreatic -cells and HIT cells. In the absence of ATP, ADP opened K(ATP) channels at concentrations as low as 10 m and as high as 500 m, with maximal activation between 10 and 100 m ADP in mouse -cell membrane patches. At concentrations greater than 500 m, ADP inhibited K(ATP) channels while 10 mm virtually abolished channel activity. HIT cell channels had a similar biphasic response to ADP except that more than 1 mm ADP was required for inhibition. The channel opening effect of ADP required magnesium while channel inhibition did not. Using creatine/creatine phosphate solutions with creatine phosphokinase to fix ATP and ADP concentrations, we found substantially different K(ATP)-channel activity with solutions having the same ATP/ADP ratio but different absolute total nucleotide levels. To account for ATP-ADP competition, we propose a new model of channel-nucleotide interactions with two kinds of ADP binding sites regulating the channel. One site specifically binds MgADP and increases channel opening. The other, the previously described ATP site, binds either ATP or ADP and decreases channel opening. This model very closely fits the ADP concentration-response curve and, when incorporated into a model of -cell membrane potential, increasing ADP in the 10 and 100 m range is predicted to compete very effectively with millimolar levels of ATP to hyperpolarize -cells.The results suggest that (i) K(ATP)-channel activity is not well predicted by the ATP/ADP ratio, and (ii) ADP is a plausible regulator of K(ATP) channels even if its free cytoplasmic concentration is in the 10–100 m range as suggested by biochemical studies.We would like to thank Mr. Louis Stamps for expert technical assistance and Dr. Wil Fujimoto and Ms. Jeanette Teague for generously providing HIT cells obtained from Dr. Robert Santerre at Eli Lilly. We would also like to thank Dr. Michel Vivaudou for providing the program ALEX. Support was provided by the NIH and the Department of Veterans Affairs.     

Ultrastructural study on the release of neurosecretory material from the sinus gland of the land crab,Gecarcinus lateralis

Summary Sites of release of neurosecretory material were examined in a neurohemal organ of decapod crustaceans, the sinus gland of the land crab, Gecarcinus lateralis. Such discharge into the circulation seems to occur primarily at interfaces between the neurosecretory axons and the acellular stromal sheath which is interposed between parenchyma and hemolymph. The evidence obtained from electron micrographs of adult specimens indicates that more or less intact secretory granules are released into the extraaxonal space primarily by the process of exocytosis. Synaptic-type vesicles are clustered in parts of neurosecretory axons facing the stromal sheath. Such vesicles are thought to result from rearrangement of membranes temporarily fused at the release site and to a minor degree from fragmentation of neurosecretory vesicles within the axon. The presence of nonmarginal vesicles and the occasional appearance of free intraaxonal dense material are interpreted as indications of a second, probably less frequent, mode of release of neurosecretory material.Supported by grants AM-3984, NB-00840, and NB-05219 from the U.S.P.H.S., administered by Dr. Berta Scharrer.I am greatly indebted to Mrs. Sarah Wurzelmann for her excellent technical assistance. I thank Dr. Dorothy E. Bliss for contributing the animals used in this study, and Mr. Murray Altmann for technical advice.     

Studies on the receptors in Ciona intestinalis

Summary A photoreceptor type structure not previously described has been found in the dorsal wall of the cerebral vesicle of the tadpole larva of Ciona intestinalis. The membranes of this receptor are organised as tubules some 60–100 nm in diameter and up to 1.5 m long. The tubules are confined in bundles about 1.5 m in diameter, which extend from the cell surface into the cavity of the cerebral vesicle. These tubules are similar to those in the rhabdomeric type of photoreceptor. However, in the cells from which the tubule processes arise are structures typical of the bases of cilia, and found in ciliary type photoreceptors.I should like to thank Professor J. Z. Young, F. R. S. for his continuing encouragement and help, and Dr. R. Bellairs for the use of electron microscope facilities. Mr. R. Moss and Mrs. J. Hamilton gave excellent technical assistance.     

The fine structure of the olfactory tract in the teleost Carassius carassius L.

Summary Electronmicrographic montages of the olfactory tract at two levels in each of two fish (Carassius carassius L.) were constructed and fibre diameters measured using a Zeiss TGZ 3 particle size analyzer. Medial and lateral tract divisions, rhinocele and dorsal tela were identified. Ciliated ependymal cells line the rhinocele. Meninges form the outer covering of both tract divisions and the tela roofing the central canal.The lateral tract consists of 10–14 fasciculi in which myelinated nerve fibres are prominent. These fibres range in diameter between 0.2 and 1.8 (mean 0.7 ) consistent with conduction velocities averaging 0.6 m/sec recorded in the carp lateral olfactory tract.The medial division of the olfactory tract contains two larger fasciculi within which are numerous fine unmyelinated nerve fibres (mean diameter 0.17 ) arranged in bundles partly enveloped by glial cell processes. Myelinated nerve fibres are unevenly distributed within both fasciculi and have mean diameters of 0.6 .An interesting observation is the consistent presence of synapses within the largest bundle of the medial tract at all levels.Supported by Grant 5 Ro5 TW00154-03 from the National Institutes of Health, United States Public Health Service.The authors are indebted to the Fisheries and Wildlife Department who generously provided the fish from Snob's Creek Fish Hatchery, and gratefully acknowledge the technical assistance of Mr. T. Armitage, Mr. J. Simmons and Miss D. Harrison.     

Scanning electron microscopy of primary membrane bone

Summary The organization and the development of primary bone were studied in a number of newborn or foetal mammals. Specimens were derived from long bones, calvaria and mandibles. The surface of the mineral component was exposed by treatment with an organic solvent (hot 1,2 ethane diamine or cold sodium hypochlorite solution) and in the case of some internal surfaces (endosteal surfaces of long bone shafts, endocranial aspects of calvaria and the lining of the dental groove or crypts of mandibles) some success was achieved in dissecting the soft tissues from the bone matrix surface. These surface were studied by scanning electron microscopy. Ground sections of similar material were studied by microradiography.Primary membrane bone was found to be most different from adult lamellar bone on rapidly growing (periosteal) surfaces and in the following respects. 1) Mineralization of both the ground substance and collagen occured close to the mineralizing front, so that a higher level was reached rapidly and without a maturation phase. 2) The mineral clusters at the mineralizing front did not reflect the collagen fibre distribution. 3) The collagen fibre bundles at the matrix surface, at least at internal surfaces, were irregular in size and random in orientation.Mineralizing fronts intermediate in character between those of rapidly forming, periosteal primary bone and adult lamellar bone were found on the uncommon internal forming surfaces and in the walls of vascular channels.Most internal surfaces were resorbing. There was a wide range in the size of the resorption lacunae — the many which were only as large as the osteocyte lacunae exposed by the resorption process may have been caused by uninucleated cells.This work has been supported by a grant from the Medical Research Council. The Stereoscan scannning electron microscope was provided by the Science Research Council, and the microradiographic equipment by Mr. R. V. Ely and the Ely-Webster trust. We would like to thank Mr. P. S. Reynolds, Miss A. J. Sutton and Mr. P. F. Farmer for technical and photographic assistance and Mrs. Jeanne Mills for secretarial assistance.     

Association of glial cells with the terminal parts of neurite bundles extending from chick spinal cord in vitro

Summary The terminal parts of radially directed neurite bundles growing out from chick embryo spinal cord in vitro have been examined by phase and electron-microscopy,A type of ending is described in which the terminal parts of the neurites are associated with a glial cell. The latter sends a single major process proximally towards the explant. Distally it is attached to the substrate, and the neurite ends are related to its dorsal (nonsubstrate) aspect. Appearances suggesting a mechanism of adhesion of neurites to each other and to the gial cell are described.Growth vesicles were found in both neurites and glia.It is suggested that movements of terminal glial cells may affect the pattern of outgrowth of their attached neurite bundles.We are grateful to the Medical Research Council for financial assistance, to Mr. A. Aldrich and Mr. D. Gunn for photography, to Mr. P. Howell and Miss 0. Chmyliwsky for technical assistance, and to Mrs. B. Fisher for valued secretarial help.     

Cellular and subcellular localization of 3H-diethylstilboestrol in the central nervous system

Summary The cellular and subcellular localization of radioactivity in the brain of immature female rats was determined by dry-mount autoradiography 2 h after iv injection of 1.0 g of (monethyl-³H) diethylstilboestrol per 100 g body weight. A specific topographic pattern of nuclear concentration of the synthetic oestrogen was obtained similar to that for ³H-oestradiol-17 in specific neurons of the basal hypothalamus, preoptic region and amygdala. In competition experiments, the nuclear concentration of radioactivity in all areas studied was inhibited by unlabeled oestradiol, while unlabeled testosterone had no effect. These data suggest that although oestradiol can bind to androgen receptors, the oestrogen receptor itself can account for the localization seen after the injection of ³H-oestradiol.This research was supported in part by US PHS Grant No. NS12933NIH Career Development Awardee No. NS00164The expert technical assistance of Ms. Riki Ison and Ms. Linda Furr is gratefully acknowledged.     

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.