Some aspects of the fine structure of the statocysts of the molluscsPecten andPterotrachea

Summary The statocyst ofPecten is composed of hair cells and supporting cells. The hair cells bear kinocilia and microvilli at their distal ends and the supporting cells bear microvilli. The cilia have a 9+2 internal filament content, and arise from basal bodies that have roots, basal feet and microtubular connections. Two different ciliary arrangements are described, one with a small number of cilia arranged in a ring, and another with many more cilia arranged in rows. Below the hair cells are probable synapses. A ciliated duct connects to the lumen of the static sac and passes through the centre of the static nerve. The hair cells in the statocyst ofPterotrachea bear kinocilia and microvilli. The possible importance of cilia and microvilli in the transduction process is discussed.We would like to thank ProfessorJ. Z. Young for bringing specimens ofPterotrachea from Naples and also the staff of the Stazione Zoologica for the provision of specimens, Dr.M. Land for providing specimens ofPecten, the Science Research Council (U.K.) for providing the electron microscope used in much of the study and also for a grant to one of us (V.C.B.), and Mrs.J. Parkers and Mr.R. Moss and Mrs.J. Hamilton for much photographic and technical assistance.     

Cilia and other surface structures of the trochophore of Harmothoë imbricata (Polychaeta)

Summary Ciliary aggregations occur as the prototroch, neurotroch, apical system and as tufts associated with the eyes and superficial glands. The major collection of cilia is the locomotory organ or prototroch that runs around the equatorial plane of the larva. This band is composed of four contiguous rows of cells, the two medial rows bearing the long locomotory cilia. The cilia occur in clumps, with several clumps arising from each prototroch cell while both the main cells contribute to each clump. The central filaments of these cilia are orientated at right angles to the long axis of the clump, the direction of ciliary beat being at right angles to the progression of the metachronal wave along the prototroch. The neurotroch, extending from the mouth to the posterior pole of the larva, beats away from the mouth. The rate of beating is rapid, and the cilia are short. The apical area of the larva is bordered by five single lines of compound cilia that surround a few stiff cilia. All the cilia beat occasionally. A further line of cilia, the akrotroch, exists at a position halfway between the apical area and the prototroch on the same side as the mouth. These cilia beat towards the prototroch. Some of these cilia are associated with sets of glandular openings. The fine structure of the glands and cuticle is described. The glands are small mucous glands that open via a projecting pore which is encircled by rings of microvilli. They often occur in groups of four or in pairs. The cuticle is similar to that described previously for adult polychaetes.This work was started under a Science Research Council (U.K.) grant (B/SR/1871) for a Research Assistantship to Dr. M. S. Laverack and grateful acknowledgement is made for this. We would like to thank Dr. A. Boyde for all his advice and use of apparatus. The scanning electron microscope used in this study was provided by a Science Research Council (U. K.) grant to Dr. Boyde. We would like to thank Mrs. J. Parkes for photographic assistance.     

The fine structure of sensory receptor processes in the auricular epithelium of the planarian,Dugesia tigrina

Summary The marginal epithelium of the lateral auricles of the planarian, Dugesia tigrina, includes a cell type with surface cilia and microvilli, a basal nucleus, and dense cytoplasm containing secretory vacuoles, Golgi elements, mitochondria and ribosomes. Through channels within the epithelial cytoplasm, cellular processes, interpreted as extensions of neurosensory receptor cells located in the subepidermis, project to the surface. The receptor processes, containing microtubules, mitochondria, vesicles and an agranular tubular reticulum, project beyond the epithelial cell surface; one or two cilia each emerge from a basal body in the apex of the projection. Close to the point of emergence to the epithelial surface, each cylindrical receptor process is surrounded by a collar-like septate junction between adjacent plasma membranes. The cilia of the projections differ from those of the epithelial cells in diameter, density of matrix and in the banding patterns of the rootlets. A few projections appear with the apex and basal body retracted below the epithelial surface. The possible function of these ciliated processes in sensory reception is discussed.This work was supported by Grant No. SO 1 FR 5369 from the U.S. Public Health Service to the University of Illinois at the Medical Center.I thank Dr. J. P. Marbarger, Director of the Research Resources Laboratory, for use of the electron microscope facilities, Miss Irena Kairys for technical help, Miss Marie Jaeger for assistance with photography, and Mr. Robert Parshall for the drawing.To Professor Arthur Wagg Pollister, I respectfully dedicate this article on the occasion of his retirement from Columbia University.     

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.     

Fine structure of the Paraventricular organ of Xenopus laevis tadpoles

Summary The ultrastructure of the Paraventricular organ in the hypothalamus of Xenopus laevis tadpoles is described. It appeares that the Paraventricular organ of this anuran species is homologous with the Organon vasculosum hypothalami or the Paraventricular organ of other vertebrates.The Paraventricular organ of Xenopus laevis is characterized by an ependymal lining with only few cilia and by two types of nerve cells. Both types of nerve cells have ventricular processes, protruding into the lumen of the third ventricle and forming a network. The protrusions bear cilia of the 8+1 pattern. It has been possible to distinguish both types of nerve cells on account of their dense-core vesicles. A secretory function of both cell types is suggested.In a region close to the Paraventricular organ, another granulated type of nerve cell has been observed. A relationship between these cells and the preoptic nucleus is discussed.The author thanks Prof. Dr. P. G. W. J. van Oordt for his helpful comments and criticism, Mr. H. van Kooten for photographic assistance and Mr. F. Dijk 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 cyclic nucleotide-activated conductance in olfactory cilia: Effects of cytoplasmic Mg2+ and Ca2+

Summary Olfactory receptor neurons depolarize in response to odorants. This depolarization is mediated by an increase in intracellular cyclic AMP, which directly gates channels in the membranes of the neuronal cilia. Previous evidence suggests that a Ca²⁺ influx during the odorant response may ultimately play a role in terminating the response. One way Ca²⁺ inside the cell could terminate the odorant response would be to directly inhibit the cAMP-gated channels. In this report the effects of cytoplasmic Ca²⁺ and Mg²⁺ on the cAMP-activated current were measured in single olfactory cilia. Near the neuronal resting potential, cytoplasmic Ca²⁺ and Mg²⁺ only slightly reduced the cAMP-activated current. Even at high levels (1.0mm Ca²⁺ or 5.0mm Mg²⁺), the average inhibition was only around 20%. It is therefore unlikely that an influx of divalent cations terminates the odorant response by a direct effect on the cAMP-gated channels.     

The fine structure of the olfactory mucosa and nerve in the teleost Carassius carassius L.

Summary Morphological studies on teleost olfactory mucosa confirm the findings of previous authors regarding the general arrangement of conventional cell types, viz. receptor, sustentacular, mucous and basal, but teleosts show certain distinct differences. The receptor cells have the general mammalian bipolar shape but their peripheral dendrite does not project beyond the epithelial surface. In addition to numerous typical cilia, an exceptional ciliary formation was observed in which the filaments, instead of forming individual cilia, are grouped together in clusters and are enveloped in a single limiting membrane.At the junction between the finger-like process and the mucosal fold myelinated nerve fibres are observed within the subepithelial stroma.Within the postero-medial zone of the mucosa is a conspicuous well-differentiated new cell type. A thick rim of electron-dense cytoplasm, bounded by an outer trilaminar membrane, encloses prominent foliate (leaf-like) organelles, a basal nucleus, numerous mitochondria and vacuolar spaces. These foliaceous cells communicate with the external environment through a small stoma, their close association with epithelial components suggesting a possible secretory or absorptive function. Their intricate morphology, however, suggests that they may be receptors, but their role and neural connections still require definition.Supported by Grant 5 RO 5 TWOO 154-02 from the National Institutes of Health, United States Public Health Service.The authors are indebted to Dr. A. S. Wilson for his helpful criticism and gratefully acknowledge the photographic technical assistance of Mr. J. Simmons and Mr. S. Frank.     

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.     

Stimulation of olfactory receptors alters regulation of [Cai] in olfactory neurons of the catfish (Ictalurus punctatus)

Summary Intracellular calcium was measured in single olfactory neurons from the channel catfish (Icatalurus punctatus) using the fluorescent Ca²⁺ indicator fura 2. In 5% of the cells, olfactory stimuli (amino acids) elicited an influx of calcium through the plasma membrane which led to a rapid transient increase in intracellular calcium concentration. Amino acids did not induce release of calcium from internal stores in these cells. Some cells responded specifically to one stimulus (l-alanine,l-arginine,l-norleucine andl-glutamate) while one cell responded to all stimuli. An increase in intracellular calcium could also be elicited in 50% of the cells by direct G-protein stimulation using aluminum fluoride. Because the fraction of cells which respond to direct G-protein stimulation is substantially larger than the fraction of cells responding to amino acids, we tested for possible damage of receptor proteins due to exposure of the olfactory neurons to papain during cell isolation. We find that pretreatment with papain does not alter specific binding ofl-alanine andl-arginine to olfactory receptor sites in isolated olfactory cilia. The results are discussed in terms of their relevance to olfactory transduction.     

Ependymal cells in tissue culture

Summary Ciliated ependymal cells from various locations of the ventricular system of mammals showed outgrowth either in the form of epithelial sheets or in the form of pools and elongated double rows, dependant on the site from which the ependyma was taken and on the original orientation of the cells in respect to the cover slip.The ependymal cilia in such cultures were shown to be moving at a rate of 51/2 to 6 times per second. The movement of the cilia of adjacent cells was apparently well coordinated causing the surrounding fluid to flow in a particular direction.The effect of physiological saline, morphine sulfate, ethyl and methyl alcohol on the ciliary motility has been studied in living cells with phase contrast cinematography.Grateful acknowledgement is made to Messers G. Lefeber, E. Pitsinger and J. Carnes for indispensible aid with photographic work. Mrs. J. Finerty and Mrs. W. Hild contributed generously in the management of the tissue cultures and in executing staining procedures.This investigation was supported by a research grant [PHS B-364 (C3)] from the National Institute of Neurological Diseases and Blindness, of the National Institutes of Health, Public Health Service, administered by C. M. Pomerat.     

Differential effects of antimitotic agents on the stability and behavior of cytoplasmic and ciliary microtubules

Summary When sea urchin gastrulae are treated with colchicine or hydrostatic pressure the cytoplasmic microtubules disappear, but the ciliary microtubules which make up the ciliary axoneme (9+2) remain. With calcium-free sea water the cytoplasmic microtubules are reduced in number yet the 9+2 complex in the cilia is unaffected. Furthermore during the administration of any of these agents the cilia continue to beat so that functionally as well as morphologically the ciliary microtubules are normal even though the cytoplasmic microtubules are broken down and their presumed function in development is interrupted.Available evidence indicates that these two types of microtubules appear to be made up of similar subunits. Since there are morphological connections between the microtubules of the ciliary axoneme, and since the ciliary microtubules appear to stain more intensely than the cytoplasmic microtubules, we conclude that the ciliary microtubules are stabilized either by the addition of material or through interactions between adjacent tubules or both.Supported by Grant #5T 1-GM-707 from the National Institutes of Health to ProfessorKeith R.Porter.     

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.     

Über das optische Auflösungsvermögen der Facettenaugen von Limulus

Summary The resolving power of the human eye and the apposition eye in insects is discussed on the basis of Fraunhofer's diffraction theory. It is then shown that diffraction does not play an important role in the Limulus facet eye. In spite of this the visual fields of neighboured ommatidia overlap strongly as Waterman has shown. A mathematical relation which describes the process of imaging the optical surroundings onto the generator potentials of the excentric cells of the receptors is presented. This relation takes into account the overlap of the visual fields and the logarithmic relation between light intensity and generator potential (MacNichol, Fuortes). On the basis of Hartline and Ratliff's reports on lateral inhibition in the Limulus eye it is shown that this process corrects the overlap and therefore increases the resolving power of the eye. The functional mechanism of lateral inhibition is in principle able to create an image of the optical surroundings in the optic nerve. It therefore can compensate for the dioptric apparatus in front of the receptor mosaic. The correction process in the Limulus eye is studied quantitatively and other cases of principle interest are investigated by means of an analog computer. The results are discussed and other inhibitory processes in the visual and auditory system etc. are mentioned.     

Changes in the fine structure of the human testicular interstitial cells after treatment with human gonadotrophins

Summary Study of the fine structure of the human interstitial cells after prolonged stimulation with human gonadotrophin reveals a striking increase in the quantity of the agranular endoplasmic reticulum. This is accompanied by an increase in the number of mitochondria which exhibit more extensive cristae, collections of intramitochondrial lipid and aggregations of electron-dense granular deposits. A rise is also evident in the number of lipofuscin pigment deposits and granular membrane-bounded bodies, both of which exhibit acid phosphatase activity. These changes after gonadotrophic stimulation are discussed in relation to steroid biosynthesis.In the pretreatment biopsies of these patients aged between 25–35 years, some interstitial cells contain intranuclear crystals which exhibit a hexagonal structure. The relationship of these intranuclear crystals to the cytoplasmic crystals of Reinke is discussed.The author is indebted to Dr. J. W. Johnstone and Dr. A. Long for the human material used in this study. Thanks are also due to Dr. H. P. Taft for helpful suggestions in the management of these patients, to Professor B. Hudson for the estimations of plasma testosterone and to Dr. J. B. Brown for the supply of human pituitary gonadotrophin and the estimations of urinary oestrogens. The technical help of Mr. T. Mezciems and the photographic assistance of Mr. J. S. Simmons F. R. P. S. and Miss S. Flett is gratefully acknowledged.     

Observations on neurons and neuroglia from the area of the mesencephalic fifth nucleus of the cat in vitro

Summary Neurons from the mesencephalic root of the fifth nucleus from the kitten have been maintained in vitro for periods up to 70 days. During this time they retained their characteristic form and structure as seen in sectioned material. Axonic processes as long as 8 mm have been measured. The occurrence of recurrent axons intertwining with dendritic arborizations of the same neuron is discussed. Modulations are described for astrocytes in relation to their possible dependence on the scarcity of connective tissue elements and a lack of an outer mesenchymal tissue zone in cultures of mesencephalic explants.Grateful acknowledgment is made to Mrs. Walther Hild and Mrs. C. C. Morris for their generous contribution in the management of the tissue cultures and in executing staining procedures. Messrs G. Lefeber and E. Pitsinger offered indispensable aid with photographic work.This investigation was supported by a grant [PHS B-364 (C3)] from the National Institute of Neurological Diseases and Blindness, of the National Institutes of Health, Public Health Service, administered by C. M. Pomerat.     

Effect of mercury vapour on the non-protein thiol content and respiration rate of eggs of the cattle tickBoophilus microplus (Acari: Ixodiae)

With continuous and ultimately lethal exposure of eggs ofboophilus microplus to mercury vapour, respiration and non-protein thiol decreased to low levels. With sub-lethal exposure, respiration decreased initially and then increased, whereas non-protein thiol (largely glutathione) increased initially and then decreased. This initial increase in glutathione (GSH) which probably resulted from an attempt by the organism to control or reverse the biochemical lesion, did not arise from the oxidized form (gssg) through a shift in the equilibrium from right to left thus: 2gshgssg+2H⁺+2e^–.The significance of the results are discussed in relation to theories ofgsh function and of Hg toxicity. The possible application of mercurials in tick control is discussed.     

The distribution of monoamine oxidase and acetylcholinesterase in the hypothalamus and its relation to the hypothalamo-hypophysial neurosecretory system in the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The distribution of monoamine-oxidase and acetylcholinesterase activities in the hypothalamus of the White-crowned Sparrow has been studied in relation to the hypothalamohypophysial neurosecretory system. The enzyme activities, as revealed by the methods employed, are unaffected during photoperiodically induced testicular growth. Monoamine oxidase has a distribution distinctly different from that of the aldehyde-fuchsin positive neurosecretory material in that there is high activity in the peripheral palisade layers of both the anterior and posterior divisions of the median eminence. Intimate contact is made between these areas with the primary vessels of the hypophysial portal system. A second concentration of activity lies in a layer between the ependymal cells and the neurosecretory material of the fiber tract. In general, monoamine oxidase appears to be associated with glial elements and non-neurosecretory axons. The pars nervosa has little or no monoamine-oxidase activity. The distribution of acetylcholinesterase activity in the anterior division of the median eminence is very similar to that of the aldehyde-fuchsin positive neurosecretory neurons; however, acetylcholinesterase also occurs in the posterior division without associated neurosecretory fibers. These distribution of enzyme activities are considered in relation to possible adrenergic and cholinergic mechanisms in the median eminence.Dedicated to Professor Berta Scharrer in honor of her 60th birthday.Supported by grant NB-01353 from the National Institutes of Health to Professor Farner. This investigation was conducted while Doctor Kobayashi was a National Science Foundation Senior Foreign Scientist at Washington State University. We are indebted to Doctor Christian Da Lage, Laboratoire de Histologie, Falculté de Médecine de Paris, for the preliminary development of some of the techniques used in these investigations.     

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.     

Location of amino acid and carbohydrate transport sites in the surface membrane of normal and transformed mammalian cells

Summary Amino acid and carbohydrate transport in normal and malignant transformed hamster cells was studied after binding of the protein Concanavalin A (Con. A) to the surface membrane. Experimental conditions were used so that a similar number of Con. A molecules were bound to both types of cells. The transport of amino acids was inhibited after Con. A binding in the transformed cells but not in normal cells. This was found with the metabolizable amino acidsl-leucine,l-arginine,l-glutamic acid, andl-glutamine, and with the non-metabolizable amino acids cycloleucine and -aminoisobutyric acid. Transport ofd-glucose andd-galactose was more inhibited by Con. A in transformed than in normal cells, and in both types of cellsd-glucose was inhibited more thand-galactose. The inhibition by Con. A on transport was specific, since there was no effect on the transport ofl-fucose in either normal or transformed cells. Con. A also did not effect the entry of 3-0-methyl-d-glucose.These observations can be used to locate amino acid and carbohydrate transport sites in the surface membrane in relation to the binding sites for Con. A. The results indicate that Con. A sites are associated in normal cells with transport sites ford-glucose and to a lesser extentd-galactose, and in transformed cells with transport sites for amino acids and to a greater extent than in normal cells withd-glucose andd-galactose. Malignant transformation of normal cells therefore results in a change in the location of amino acid and carbohydrate transport sites in the surface membrane in relation to the binding sites for Con. A.