Fine structure of cilia in rat cerebral cortex

Summary Cilia have been demonstrated on granular neurons and astroglial cells in the fascia dentata, a part of the hippocampal region, in the rat. Every granular cell seems to possess one cilium, which shows an 8+1 pattern in the greater part of its length. This 8+1 pattern is shown to result from the displacement of one peripheral doublet of a 9+0 cilium into the middle of the cilium. The neuronal cilia have a two-centriole basal organization, and fine rootlets radiate from the basal body proper into the cytoplasm. The possible function and significance of these cilia are discussed on the basis of earlier literature.This study was supported in part by Grant NB 02215 of The National Institute of Neurological Diseases and Blindness, U.S. Public Health Service, in part by The Norwegian Research Council for Science and the Humanities. This aid is gratefully acknowledged. I am greatly indebted to Dr. Th. Blackstad for encouragement and advice during this study, to Mrs. J. L. Vaaland, Mr. B. V. Johansen and Mr. E. Risnes for technical assistance, and to Dr. B. Afzelius for valuable discussions.     

On the alleged presence of non-argyrophile argentaffin cells in the human gastro-intestinal tract

Summary The relationship between the argentaffin and argyrophile cells of the human gastrointestinal tract has been studied, in foetal and adult material, by a technique involving the staining of sections first by an argentaffin method (Gomori-hexamine silver, Schmorl, Diazonium) and subsequently by an argyrophile method (Bodian). A comparison of the cells staining by the two methods shows that all argentaffin cells of the human gastrointestinal tract are also argyrophile and that there is no evidence to support the claim of Hellweg (1952) and of HamPerl (1952) regarding the presence of non-argyrophile argentaffin cells.W. H. O. fellow from the Department of Anatomy, Medical College, Rohtak, India. — I am very grateful to Professor J. D. Boyd and to the World Health Organisation for having made it possible for me to carry out this research at the Anatomy School, Cambridge. I am indebted to Dr. G. A. Gresham and his staff for their very willing cooperation in providing material from surgical resections. My thanks are also due to Mr. J. F. Crane for the photographs and to Mr. J. W. Cash and Mr. R. Smith for helpful discussions on staining techniques.     

Fluorescence studies on neural structures and endocrine cells in the pancreas of the cat

Summary With the use of the Falck-Hillarp histochemical technique for the detection of monoamines, nerve fibre fluorescence is observed throughout the tail of the pancreas of the cat and the arrangement and distribution of the nerve fibres can be studied in both the exocrine and endocrine tissue. In the exocrine pancreas, adrenergic nerve fibres innervate arterioles, larger veins and major pancreatic ducts. Adrenergic nerve fibres also appear to terminate on the non-adrenergic nerve cell bodies of the intrapancreatic ganglia. In the islets of Langerhans, adrenergic nerve fibres innervate both the endocrine cells and blood vessels. Some of the islet cells exhibit fluorescence with the Falck-Hillarp technique and these cells have been identified as alpha cells. In animals treated with reserpine, the fluorescence in nerve fibres and in alpha cells is absent.The author wishes to thank ProfessorG. C. Schofield and Dr.G. C. Smith for their encouragement and valuable criticism during the course of this study. The assistance of MissJ. Bennett and MissW. Kemp and the photographic help of Mr.J. S. Simmons, F.R.P.S., are gratefully acknowledged. The diagram was drawn by MissS. Flett.     

Fine structure of synapses in the dorsal nucleus of the lateral geniculate body of normal and blinded rats

Summary Degenerating boutons, observed from 2 to 60 days after eye enucleation, displayed decreased plasma membrane density, increased axoplasmic density, and enlarged mitochondria with deformed cristae when compared with boutons from normal animals. There was also a loss of synaptic plasma membrane specialization and the boutons abnormally indented contiguous dendrites. The number and appearance of synaptic vesicles in some degenerating boutons were notably altered. Phagocytosis of boutons in most instances appeared to be accomplished by astrocytes. When degeneration was first apparent in some boutons, the subsynaptic organelle in the adjacent dendritic cytoplasm was enlarged, somewhat less dense and was associated with small granular and circular profiles. Subsynaptic organelles in experimental animals were absent from contiguities between dendrites and other cell processes, except in a few instances when only small portions of boutons remained at their synaptic sites, suggesting that the organelles disappeared when boutons had been completely phagocytized.Degenerating myelinated axons, observed from 2 to 300 days after enucleation, exhibited the same triad of features as degenerating boutons. They appeared to be phagocytized in most instances by dense glial processes, presumably oligodendrocytic, which were normally situated between the axon and its myelin sheath and were related to the inner mesaxon.This investigation was supported by U.S.P.H.S. Training Grants Nos. 2 T1 GM 202 T1 CA 505506, and 2RO 1 AM 368806.The author expresses his appreciation to Dr. A. J. Ladman for acquainting him with the techniques used in the study and to Dr. R. J. Barrnett for valuable criticism of this report. Gratitude is also extended to Mr. E. Z. Rutkowski for making the drawing.     

Histochemical demonstration of monoamine oxidase in the hypothalamo-hypophysial system of the tree sparrow and the rat

Summary Localization of monoamine oxidase (MAO) was investigated essentially according to the method of Glenner et al. (1957) in the hypothalamo-hypophysial system of the tree sparrow and the rat. The hypothalamic neurosecretory cells of both species showed relatively weak MAO activity. A similar localization of MAO activity was observed in the median eminence of both species: (1) slight or no MAO activity was observed in the ependymal layer, (2) relatively strong activity was revealed in the tissue just beneath the ependymal layer, (3) strong activity was revealed in the outer layer, particularly in the tissues surrounding capillary loops of the primary plexus. It is suggested that an adrenergic mechanism functions in the median eminence. In the pars nervosa, strong reaction was observed in the rat, while a weak reaction occurred in the tree sparrow. However, the color and the size of formazan crystals deposited in the rat pars nervosa differed from those in the hypothalamus. As a whole, the distribution of the neurosecretory material differed from the localization of MAO activity in the hypothalamo-hypophysial system. It is discussed that the neurosecretory neuron is not adrenergic but cholinergic.Aided by Grant A-3678 from the United States Public Health Service. The authors are indebted to Dr. S. Kambara, Zoological Institute, and Dr. H. Hirano, Department of Anatomy, University of Tokyo, for their valuable advice, and also to Assoc. Prof. S. Yamamoto, Department of Hygiene, University of Tokyo, for making available some facilities. They also wish to thank Dr. L. M. Barbato, University of Illinois, and Mr. K. Asami, National Institute of Radiological Sciences, Chiba, and Mr. Suzuki, Research Laboratory, Chugai Pharmaceutical Company Ltd., Tokyo, for the kind supply of MAO inhibitors.     

Scanning electron microscopic studies of cilia

Summary A simple method for the preparation of ciliated epithelia for study with the scanning electron microscope is described. Ciliary groups are well preserved and it is possible to discern individual cilia and work out their numbers and orientation. Following scanning electron microscopical study some of the material was prepared for transmission electron microscopy and the ultrastructure of the tissue was found to be surprisingly well preserved. The tracheal epithelium of the rabbit, the olfactory epithelia of the goldfish and the rabbit, and the sensory epithelia in the statocyst of a cephalopod mollusc were examined with the scanning electron microscope to demonstrate the possibilities of the method. Acknowledgements. We would like to thank Professor J. Z. Young for his continued interest and support. The scanning electron microscope was purchased with a grant provided by the Science Research Council to Dr. Boyde, Mr. R. Willis helped in the initial stages of the study, Mr. G. Savage provided help with the goldfish material, Mr. S. Waterman provided much photographic assistance, and Mrs. N. Finney the secretarial assistance.     

Comparative immunochemical studies with antisera to sheep prolactin and bovine growth hormone on anterior pituitaries of ox,sheep, rats and mice

Summary Rabbit antisera to sheep prolactin and bovine growth hormone were used in the indirect fluorescent antibody technique on cryostat sections of anterior pituitaries of sheep, ox, rats and mice. It is demonstrated that in sheep and ox prolactin and growth hormone are manufactured by different acidophilic pituitary cells. Though the antisera do not precipitate the analogous hormones of rats and mice in gel diffusion tests, evidence is given for the specificity of the cross-reaction of the antisera with the analogous murine hormones in situ as found with the fluorescent antibody technique.We are grateful to Dr. J. D. H. Homan of Organon, Oss (The Netherlands) for the supply of bovine growth hormone and for kindly giving us the information concerning this preparation.We should like to thank Dr. F. J. A. Prop for the prolactin assays, Dr. H. G. Kwa for providing the mouse pituitary tumours and Dr. L. M. Boot for the mouse pituitary isograft in the kidney, Mrs. J. J. Geiger-Koedijk for the conventional staining of pituitary sections, and Mr. J. van der Kamp for the photography.     

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.     

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.     

Stem cell renewal and duration of spermatogonial cycle in the goldhamster

Summary The duration of the cycle of the seminiferous epithelium of the hamster is estimated to be 9.0 days. The duration of the stages was also determined.The stem cells were found to be daughter cells of the A₃ spermatogonia. This confirms the hypothesis that in general stem cells for the next spermatogonial cycle arise on the moment of morphological differentiation of the A spermatogonia.The author wishes to thank Prof. Dr. M. T. Jansen and Dr. M. F. Kramer for helpfull discussions and Mr. J. G. van Essen for technical assistance.     

The yield of synaptosomes from the cerebral cortex of guinea pigs estimated by a polystyrene bead “tagging” procedure

Summary The number of synaptosomes (pinched-off nerve endings) produced/g guinea pig cortex on homogenizing this tissue under defined conditions is estimated to be in the region of 4×10¹¹ using two different polystyrene bead tagging procedures. This is the same order of magnitude as the number of nerve endings/g cortex calculated from histological estimates given in the literature of the number of neurones in the cortex and the extent of their cortical connexions.This investigation was supported by U.S. Public Health Service Grant no. NB 03928 from the National Institute of Neurological Diseases and Blindness. Dr. Sheridan was a Postdoctoral fellow of the U.S. Public Health Service during the period of his participation in this investigation (1963/64). The electron microscope facilities were provided by the Wellcome Trust. Mr. T.F.J. Hobson, of the ARC Statistics group, Cambridge, kindly advised on statistical aspects of the work. We are most grateful to him, to Miss L. Swales and Mr. G.H.C. Dowe for their skilled technical assistance and to Dr. E.G. Gray for helpful discussions.     

Further evidence for the proposed way of spermatogonial stem cell renewal in the rat and the mouse

Summary By means of ³H-thymidine and autoradiography it could be established that in rats and mice the A₁, A₂ and A₃ spermatogonia do not give rise to a significant number of stem cells.The number of A₀ spermatogonia was found to be circa 20% of the number of A₀+A₁ spermatogonia in the rat and circa 10% in the mouse.The author wishes to thank Prof. Dr. M. T. Jansen and Dr. M. F. Kramer for helpful discussions and Mr. J. G. van Essen for technical assistance.     

Localization of melanin synthesis within the pigment cell: Determination by a combination of electron microscopic autoradiography and topographic planimetry

Summary Localization of melanin synthesis within the pigment cells of the Cloudman S-91 mouse melanoma was determined by means of a combination of high resolution autoradiography and topographic planimetry. Initial melanin biosynthesis occurred predominantly in the endoplasmic reticulum and associated ribonucleoprotein particles of the melanocytes. By measuring a number of cell organelles and employing the index of relative specific localization it could be shown that the nucleus and mitochondria are of little or no importance in the process of melanogenesis.This investigation has been supported by the following research grants: CA 06548 CB, NIH, PHS and an Institutional Grant from the American Cancer Society (to H. M. H.); CA-05887, NIH, PHS (to A. S. Z.); M-00388 and NB-00782, NIH, PHS (to J. F. H.). One of us (H. M. H.) holds a Research Cancer Development Award (5-K 3-GM-2634) of the National Institute of General Medical Sciences, Public Health Service.We are grateful to Mr. Ronald Abler for his help with the topographic measurements; to Dr. Erhard Haus for help and advice; to Mr. J. Thornby and Mr. A. P. Basu for assistance with the statistical aspects of this study; and to Mrs. Lenore Mottaz, Miss Bernice Uittenbogaard, and Mrs. Judith Strong for careful technical assistance.     

Farbenpolymorphismus bei einigen Feldheuschrecken

Ohne ZusammenfassungMit 5 TextabbildungenWir danken Mr.M. J. R. Healy vom Research Statistical Service, Rothamsted Experimental Station, für die elektronische Berechnung der Inversion der Streuungsmatritzen sowie Dr.W. Loher für die Übersetzung dieser Arbeit.     

Hypothalamic control of the pars intermedia in Xenopus laevis tadpoles

Summary In Xenopus laevis tadpoles the relation between a paired nucleus of bio-amine producing neurons in the caudal hypothalamus and the pars intermedia of the hypophysis was studied.Treatment of the animals (stage 49 to 50 of Nieuwkoop and Faber's normal table) with reserpine caused aggregation of the skin melanophores within one hour, followed by redispersion five to six hours after the beginning of the experiment. This was at exactly the same time as the bio-amines in the caudal hypothalamus disappeared. However, the drug was ineffective if the nuclei had been removed. This indicates that reserpine acts via these nuclei and does not influence the skin melanophores directly.It was concluded that the initial aggregation of the melanophores may be the result of a reduced extrusion of MSH from the pars intermedia, caused by an increased output of a MIF by the bio-amine producing nuclei. The redispersion was explained by assuming that the bio-amines were depleted and the nuclei stopped with the extrusion of the MIF. This does not mean that the production of a MIF is the only function of the paired bio-amine producing nucleus in the caudal hypothalamus.The author thanks Prof. Dr. P. G. W. J. van Oordt for his helpful comments and criticism. Mr. J. H. I. J. M. ten Berge and Mr. E. W. A. Kamperdijk provided great assistance during the course of the experiments. Mr. H. van Kooten made the diagram and the photograph.     

Electron microscope observations of some peripheral synapses in the sensory pathway of the sucker of Octopus vulgaris

Summary A synaptic axo-dendritic linkage is described between primary receptors lying in the epithelia of the sucker of Octopus and encapsulated nerve cells found near the rim of the sucker in the subepithelial connective tissue. These synapses are postulated to perform a drastic reduction of inputs between the primary receptors of the order of more than ten thousand and the subjacent encapsulated nerve cells of the order of some hundreds. The morphology of these cells as well as that of the synaptic structures are described from electron microscope studies. Aknowledgement. This work was done at University College London, while I was in receipt of a Medical Research Council grant.I am deeply indebted to Prof. J. Z. Young F. R. S. for support and criticism, and to Dr. E. G. Gray for advice and discussion. My thanks are due to Mr. A. Aldrich for the photographs.     

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.     

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.     

The ultrastructure of the nuclei and the behaviour of the sex chromosomes of human spermatogonia

Summary The nuclear structure of human spermatogonia has been studied with electron microscopical and histochemical methods. Type B spermatogonia have chromatin clumps without any special ultrastructure and several nucleoli. Five different types of nuclear bodies, and besides, a nuclear vacuole, have been observed in type A spermatogonia. Type I bodies are typical nucleoli consisting of three regions: amorphous, fibrillar and granular. Type II, III and V are considered to be atypical nucleoli. Type IV bodies are small chromatin condensations. Type I bodies are the only ones in which RNA was demonstrated by light histochemical techniques and no PAS positive material was found inside the nuclei. The absence of any special ultrastructure in the chromatin from spermatogonia, and the small mass of the chromatin condensations, show that the human X chromosome and perhaps the Y chromosome are not heteropycnotic in the interphasic nuclei of human spermatogonia.Abbreviations Used RNA ribonucleic acid - gonia spermatogonia This work has been supported by a grant (No. 2623) of the Consejo Nacional de Investigaciones Cientificas y Tecnicas, and partially by a grant (C.M. 6522) from the Population Council.We wish to thank Professor R. E. Mancini for his suggestions during this investigation and his support for its achievement, and to Dr. J. C. Lavieri for providing the biopsies.     

The rhabdomeric microvilli of several arthropod compound eyes kept in darkness

Summary Electron microscopical studies were made on the fine structure of the rhabdomeric microvilli of the compound eyes of seven species of arthropods (Procambarus, Neocaridina, Caridina, Potamon, Artemia, Diestrammena, Drosophila) raised in complete darkness for 1–8 months or for successive generations, using various fixation techniques.The rhabdomeric microvilli of the individuals kept in darkness for a long period were regularly arranged as in normal eyes in the material prepared by double fixation with glutaraldehyde and OsO₄, whereas in those fixed solely by OsO₄ various forms of vesiculations were seen. The structural changes of the rhabdomere in darkness, which have been reported by several workers, were conceived to be an artefact caused by OsO₄ fixation.This work is supported by a grant from the U.S. Army Research and Development Group (Far East), Department of the Army (DA-CRD-AG-S92-544-67-G61).The authors wish to express their gratitude to Dr. S. Mori and Mr. T. Kuramoto for their help in supplying some of the materials. We are also indebted to Dr. C. J. C. Rees for correcting English of this paper.