The tendons of Ophiocomina nigra and their role in autotomy (Echinodermata,Ophiuroida)

Summary In the arm of the ophiuroid Ophiocomina nigra the intervertebral muscles are linked to the vertebral ossicles by tendinous connective tissue fibres. When an arm autotomizes, rupture of the tendons at one end (the autotomy insertion) permits each muscle in the autotomizing segment to separate cleanly from an ossicle while its other attachment (the non-autotomy insertion) remains intact. The anatomical relations, composition and function of the tendons were investigated by histochemical, electron microscopical and experimental methods. The tendons consist of a carbohydrate-rich secreted collagen derived from the basal lamina of the muscles. At autotomy their rupture is preceded and facilitated by an increase in extensibility, which represents the first evidence for variable tensility in an echinoderm connective tissue not composed of interstitial collagen. Granule-containing juxtaligamental cell processes are associated with the tendons of the autotomy insertions but are absent from the non-autotomy insertions. There appears to be widespread release of granules from these processes at autotomy. The results of a simple experiment implicate the juxtaligamental cells in the control of tendon extensibility and a possible mechanism for this control is presented.     

The ultrastructure of the sinus gland of Carcinus maenas (Crustacea: Decapoda)

Summary The sinus gland of Carcinus maenas consists of the swollen axonal endings of the neurosecretory cells of the major ganglia and acts as a storage release centre for the membrane bound neurosecretory material. These neurosecretory granules fall into five different types based on size and electron density. Their contents are released by exocytosis of the primary granules or smaller units budded from the primary granules.I thank Professor E. Naylor for his constant advice and Professor E. W. Knight-Jones, Department of Zoology, University College, Swansea, for the provision of laboratory facilities. I am grateful to the Science Research Council for the financial support. Finally, I thank the Electron Microscope Unit, Southampton General Hospital, where the work was completed.     

Variable tensility of the oral arm plate ligaments of the brittlestar Ophiura ophiura (Echinodermata: Ophiuroidea)

The oral arm plates of the brittlestar Ophiura ophiura L. are connected to lateral arm plates at distal and proximal ligamentous junctions. The distal junction is mobile and is disrupted during arm autotomy; the proximal junction is more rigid and does not participate in autotomy. Aspects of the morphology and mechanical properties of the distal and proximal oral arm plate ligaments have been investigated in order to determine if their tensility is under physiological control. By means of creep tests it was found that elevation of the external potassium (K⁺) ion concentration causes a decrease in the viscosity of the distal ligament which is either transient or continues until rupture intervenes. In forced vibration tests the distal ligament often shows a biphasic stiffening then softening response to excess K ^- ions. Anaesthetics block the softening phase but enhance the stiffening component of this response. This ligament is also softened by repetitive electrical stimuli but stiffened by excess calcium ions and by acetylcholine. The proximal ligament appears to have the capacity for only transient changes in mechanical properties. Both ligaments are penetrated by the processes of juxtaligamental cells whose perikarya are arranged in clusters innervated by hyponeural nerves. These cells are thought to modulate the interfibrillar cohesion of the ligaments. It is concluded that the distal and proximal ligaments are mutable collagenous structures which in their stiffened condition help to maintain arm posture without the need for continuous muscular activity, and that at autotomy the distal ligament undergoes a profound loss of tensile strength which facilitates arm detachment.     

Juxtaligamental cells in the arm of the brittlestar <Emphasis Type="Italic">Amphipholis kochii</Emphasis> Lütken, 1872 (Echinodermata: Ophiuroidea)

We found that ligaments connecting the skeletal elements in the arm of the brittlestar Amphipholis kochii, consisted of mutable collagenous tissue (MCT), whose basichistological character is that of numerous processes of juxtaligamental cells penetrating the extracellular matrix. These cells are located in the hyponeural neuroepithelium associated with hyponeural nerves, and were also recorded in the spine ganglia and basiepithelial nervous plexus in the coelomic epithelium. The distinctive feature of juxtaligamental cells is the presence of electron-dense granules in their cytoplasm and a well developed synthetic apparatus. Based on personal and literature data it is concluded that juxtaligamental cells are a part of the nervous system, and their basic function is related to providing MCT with innervation.     

Fine structure of the eyes of Onithochiton neglectus (Mollusca: Polyplacophora)

Summary Onithochiton neglectus a common littoral chiton possesses large numbers of small eyes embedded in the outer layer of the shell, the tegmentum. These are arranged in a definite pattern on each shell valve. Each eye lies in a pocket, and is surrounded by pigment laid down in the shell. There is a lens, cup of retina cells and an optic nerve running in an optic canal through the shell. Glial elements are present. The retina cells give rise centrally to a packed array of microvilli, a rhabdom. Cilia are present at the edge of the rhabdom; they have a 9 + 2 arrangement of ciliary filaments and do not appear to be involved in the formation of microvilli. Cells at the periphery of the eye cup give rise to large whorls of membranes, lamellate bodies. These bodies are derived from the membranes of cilia having a 9 + 2 pattern, and form into an extra-cellular space. Nerve processes from the retina cells pass into the optic canal. On the basis of previous work it is thought that the lamellate bodies are also sensory. These structures are discussed in relation to other microvillar and lamellate structures described from photoreceptors.I thank Professor J. E. Morton for his advice in the early stages of this work, and Dr. S. J. Bullivant for the fixation and embedding of material for electron microscopy. To Professor G. A. Horridge I am grateful for advice and the facilities of his laboratory, and to Professor M. S. Laverack, Patricia Holborow and Charles Coleman for much help and encouragement. I am supported by the Science Research Council, and in New Zealand held a Commonwealth Scholarship.     

Juxtaligamental system of the disc and oral frame of the ophiuroid Amphipholis kochii (Echinodermata: Ophiuroidea) and its role in autotomy

Abstract. The ophiuroid Amphipholis kochii is able to detach its central disc from the underlying oral frame in response to external stimuli. In this article we supply new observations on the microanatomy and ultrastructure of the autotomy plane, and of the juxtaligamental system which is believed to bring about connective tissue changes that underpin the detachment process. We correct previous confusion over the innervation of juxtaligamental nodes involved in disc autotomy, provide evidence that juxtaligamental cells are a population of specialized nerve cells, and present observations on changes in the ultrastructure of juxtaligamental cells during autotomy, which support the view that they are responsible for connective tissue disruption.     

Responses of single receptor cells in the pine engraver beetle,Ips pini (SAY) (Coleoptera: Scolytidae) to its aggregation pheromone,ipsdienol, and the aggregation inhibitor,ipsenol

Summary Responses of single olfactory cells on the antennal club ofIps pini have been recorded electrophysiologically. The majority of cells were strongly activated by either/or of the two behavior-modifying chemicals, the aggregation pheromone ipsdienol and the aggregation inhibitor ipsenol. Simultaneous stimulation with these terpence alcohols showed that ipsenol had no inhibitory effect on the receptor responses to ipsdienol. It appears that the behavioral inhibition by ipsenol on the attraction of ipsdienol results from central integration of the information from separate receptor cells rather than blockage at the peripheral receptors.This work was carried out at the College of Environmental Sciences and Forestry, SUNY, Syracuse. We are grateful to Professor R.M. Silverstein and his staff for provision of purified compounds and laboratory facilities. We also acknowledge with thanks the provision of equipment and laboratory facilities by Professor D. Tapper, College of Veterinary Medicine, Cornell University.     

The giant neurone system in ophiuroids

Summary The radial nerve cords of members of the class Ophiuroidea consist of two parts, the ectoneural and the hyponeural tissues, which are separated by an acellular basal lamina. The hyponeural tissue is composed entirely of motor fibres. The cell bodies of the hyponeural neurones are arranged in ganglia, one to each segment of the arm, and each containing approximately one hundred cell bodies. Synaptic contact between the two tissues occurs across the basal lamina. Ultrastructural evidence shows that the majority of these synapses operate in the ectoneural to hyponeural direction. Three pairs of nerve bundles, each containing approximately thirty five large motor fibres arise from each ganglion and innervate the intervertebral muscles. The large motor fibres divide into a number of pre-terminal axons in the region in which the motor fibre enters the muscle block. The terminal axons run at right-angles across the muscle fibres and neuromuscular junctions are found at the points of contact between the two; each terminal axon makes contact with a large number of muscle fibres. The hyponeural axons also pass through the juxtaligamental tissue before they reach the muscle blocks and there is some evidence of synaptic contact with the juxtaligamental cells. The juxtaligamental tissue is thought to be associated with changes in the structural properties of the collagenous ligaments of the arm during arm autotomy (Wilkie 1979). Degeneration studies confirmed the layout of the hyponeural motor axons.     

Identification of gastrin-secreting cells and cholecystokinin-secreting cells in the gastrointestinal tract of the human fetus and adult man

Summary Gastrin-and cholecystokinin (C.C.K.)-containing cells were detected by using anti-gastrin and anti-C.C.K. sera in the gastrointestinal tract of human fetuses and premature infants and in the stomach and duodenum of adult man obtained by biopsy from eight patients with normal gastro-duodenal endoscopy. The specificity of immunocytological reactions was ascertained by studying the inhibition of the reaction by gastrin, C.C.K., secretin, somatostatin, glucagon, insulin, serotonin, histamin, caerulein and octapeptide of C.C.K. In adult man, the gastrin cells are located only in the antrum and juxtapyloric region; C.C.K. was detected in the duodenum. In the human fetus, the first gastrin cells are seen in the antrum at 14 weeks of age and in the duodenum as early as 10 weeks; the C.C.K. cells are seen in the small intestine at 10 weeks of age. Acknowledgements. The authors should like to thank Professors Magnin and Liaras, Hôpital Edouard Herriot, M. Dumont, Hôpital de la Croix-Rousse, Notter et Garmier, Hôtel-Dieu, Bethenod, Hôpital Debrousse, Lyon, for their cooperation. We also thank Professor R. Guillemin, the Salk Institute, La Jolla, California, Doctor M.P. Dubois, I.N.R.A., Station de Physiologie de la Reproduction, Nouzilly, Mme Vagne, U. 45, I.N.S.E.R.M. and Professor Y. Minaire, Hôpital Edouard Herriot, Lyon, for their donations. This work was supported by a grant from the Institut National de la Santé et de la Recherche Médicale     

The structures of the tentacles of Rhabdopleura compacta (Hemichordata) with special reference to neurociliary control

Summary The tentacle of Rhabdopleura compacta (Hemichordata) consists of two layers of cells surrounding a central coelomic cavity. The two layers of cells are separated by a cell free basement lamella.The tentacles on the arms of Rhabdopleura bear three longitudinal rows of cilia. The ciliated cells are closely associated with bundles of nerve fibres, and between some of the cells and nerve fibres there are synapses. The peripheral regions of the ciliated cells are joined to one another by desmosomes. Tonofibrils join some of these desmosomes to the kinetosomes of the cilia.The nerve fibres are confined to the ectodermal layer and the muscle cells to the layer of cells within the basement lamella. In the ectodermal layer besides ciliated cells there are mucus cells, densely pigmented cells, and green bodies. The function of these last two types of cells is secretory. Most of the epithelial cells have microvilli upon their free borders.I wish to thank Professor J. Z. Young F. R. S. for enthusiastic advice and encouragement. Dr. R. Bellairs generously provided the facilities for electron microscopy. Mr. R. Moss gave excellent technical and photographic assistance. Dr. A. Stebbing of the Plymouth Marine Biological Laboratory helped me to obtain and to identify the specimens. Professor D. W. James kindly allowed me to use his facilities for interference microscopy.     

Chloroplast isolation from Laminaria digitata (Phaeophyceae): A reproducible methodology yielding photosynthetically active chloroplasts

A technique for chloroplast isolation from the brown algae macrophyte Laminaria digitata (Phaeophyceae) is described which results in stable and photo-synthetically active chloroplasts. Chloroplasts in L. digitata are mainly located in the surface layer meristodermic cells. The expanded intercellular regions of the cortex tissue contain abundant mucilage which is exuded during mechanical tissue disruption. The method described for chloroplast isolation is based on appropriate isolation buffers and the removal of mucilage which is secreted during tissue disruption, and it may be applicable to other macroalgae with large amounts of mucilage. The isolated chloroplasts showed stable oxygen-evolving activities. The reproducibility of this technique was established by analysis of electron transport and evaluation of the activities of photosystems I and II.Abbreviations BSA bovine serum albumin - DCPIP 2,6-dichlorophenol-indophenol - DMBQ 2,6-dimethyl-benzoquinone - MV methyl viologen We thank Mr. D. Cameron and Mr. R. Wilkie (University Marine Biological Station Millport, Isle of Cumbrae, Scotland) for supplying Laminaria digitata and Mr. J. Pacy (King's College London, UK) for assistance with electron microscopy. This work was supported by EC (Brussels) grant CT/0770 and a fellowship from CNPq (Brasilia) to M. A. Rodrigues.     

Histological and electron microscopical observations on the effects of different salinities and heavy metal Ions,on the gills of Jaera nordmanni (Rathke) (Crustacea,Isopoda)

Summary The effects of different salinities and concentrations of copper, mercury and cadmium ions on the gills of Jaera nordmanni are investigated by means of light and electron microscopy. After exposure to 10% and 50% sea water the gill epithelium cells show a marked uniformity in appearance, possessing characteristically large, sub-cuticular spaces which are prominent between microvilli. With exposure to the heavy metal ions a similar sequence of histological and ultrastructural changes occur in all the gill epithelial cells, culminating in cell breakdown. The ultrastructural changes include distended microvilli, dilated endoplasmic reticulum, dissociated ribosomes, diffuse (swollen) cytoplasm, swollen mitochondria and a basal membrane withdrawn from the basal lamina. An increase in the number of haemocytes is also commonly observed in the haemolymph spaces during heavy metal ion exposure. The significance of the morphological changes undergone by the gill epithelial cells after exposure to different salinities and heavy metal ion concentration, are discussed in relation to the physiological functioning of the gill.The author wishes to thank Dr. M.B. Jones and Mr. R.H. Moore for setting up the experiments and to Professor E. Naylor for providing laboratory facilities at the Department of Marine Biology, University of Liverpool, Port Erin, Isle of Man. This work was supported by a Ministry of Defence (Navy) Contract No. AT/2198/010/CDL.     

The motor innervation of the oral plate ligament in the brittlestar Ophiura ophiura (L.)

Summary The innervation of the connective tissue ligaments between the oral plates on the arm of a brittlestar by small branches of the hyponeural motor system is described. These branches arise from the hyponeural part of each segmental ganglion and pass laterally round the nerve cord and then orally across the epineural sinus to penetrate a small intersegmental node of juxtaligamental tissue located centrally. The endings of the nerve branches contain numerous small agranular vesicles and make chemical synapses onto the juxtaligamental cells. Processes from the juxtaligamental cells containing large granular vesicles ramify amongst the connective tissue of the oral ligaments. This innervation is associated with rapid changes in the stiffness of the connective tissue.     

Pituitary response to environmental osmotic changes in the larval guppy,Lebistes reticulatus (Peters)

Summary The pituitary of the larval guppy, Lebistes reticulatus (Peters), was investigated cytologically in relation to the osmotic properties of the environment. In larvae of Stage I, removed from the mother about one week before birth, the sizes of the nuclei of the prolactin, STH, ACTH, and pars intermedia cells were small. The nuclei of these cell types became larger when Stage I larvae were transferred into fresh water (F. W.); however, these animals did not survice. In Stage-II larvae, removed from the mother just before birth, the nuclei of the abovementioned adenohypophysial cells were larger than those of the Stage-I larvae. Stage-II larvae were able to survive in F. W. Thus, it seem that osmoregulatory mechanisms in the larval guppy develop between Stages I and II.The nuclei of prolactin, STH, ACTH and pars intermedia cells decreased significantly in size after birth (0-hr). When larvae of the 0-hr group were immersed in 1/3 sea water (S. W.), only the nuclei of the ACTH cells increased in size. The neurohypophysis of Stage-I larvae contained a very small amount of aldehyde-fuchsin (AF)-positive material, while the neurohypophysis of Stage-II and 0-hr fish was rich in this material. When Stage-I larvae were immersed in F. W., the neurohypophysis showed as great an accumulation of AF-positive material as Stage-II and 0-hr fish.Supported by grants from Japan Society for Promotion of Science and from Ministry of Education of Japan to Professor Kobayashi, and by grants from the Deutsche Forschungsgemeinschaft to Professor Oksche. A part of this investigation was effected while Professor Kobayashi held a Visiting Professorship at the University of Giessen. We are indebted for histological assistance of Miss I. Woltmann, Mrs. H. Schneider and Mrs. T. Bremer.     

Uptake of ferritin by the cephalopod optic gland

Summary Horse spleen ferritin, injected into the blood of the octopus, leaves the capillaries via pericyte junctions and windows. One hour after the administration, ferritin has entered the optic gland main cells; three hours later, it is accumulated in dense-bodies. The evidence for resorption supports earlier papers reporting that the ultrastructure of the organ is unusual for an endocrine gland.We thank Professor R. Martin for his help and for permission to work in his laboratory at the Ulm University (Germany) and the Swiss Government for financial support     

Ultrastructure of the compound eye and first optic neuropile of the photoreceptor mutant ora JK84 of Drosophila

Summary The developmental mutant of Drosophila (ora ^JK84) is characterized by nonfunctional photoreceptor cells (R1–6), while the R7/R8 cells are normal. A fundamental question is: Does the near absence of photosensitive membranes inhibit development of the Rl-6 axons and their synapses at the other end of the cell? The retina and first optic neuropile (lamina ganglionaris) were examined with freeze-fracture technique and high voltage electron microscopy. R1–6 have reduced rhabdomere caps; rhabdomeric microvilli have about 50% of the normal diameter and 20% of the normal length. Affected cells exhibit prominent vacuoles which appear to communicate with some highly convoluted microvillar membranes. Almost no P-face particles (putative rhodopsin molecules) are present in the R1–6 rhabdomeres, and particle densities are lower in R7 than previously reported. Near the rhabdomere caps, microvilli of R1–6 are fairly normal, but at more proximal levels they are greatly diminished in length and changed in orientation, while at still more proximal levels they are lost. R1–6, R7, and R8 axons from each ommatidium are bundled into normal pseudocartridges beneath the basement membrane. No abnormalities are found in the lamina ganglionaris, and all synaptic associations as well as the presumed virgin synapses (of R1–6) appear normal. No glial anomalies are present, and R7/R8 axons project through the lamina in the usual fashion. These fine structural findings are correlated with known electrophysiological, biochemical, and behavioral correlates of both sets of photoreceptors (R1–6, and R7/R8).This study was supported substantially by the UW-HVEM Laboratory, in addition to a Faculty Development Award, a UMC Biomedical Research Support Grant N.I.H. RR07053 to W.S.S., and a Hatch Grant, Project 2100 to S.D.C. Freeze fracture was done at the Wisconsin Regional Primate Research Center, N.I.H. Grant RR00167. We thank Professor Hans Ris, Dr. J. Pawley, Dr. D. Neuberger, and Ms. M. Bushlow, HVEM Laboratory, Dept. of Zoology, UW. We also thank Mrs. K. Srivastava, Mr. M.B. Garment, Mr. G. Gaard, and Mr. D. Liu for technical assistance.     

Fine structure of the ciliated epidermis on the tentacles ofOwenia fusiformis (Polychaeta,Oweniidae)

Summary The ultrastructure of the epithelium on the oral surface of the tentacles ofOwenia fusiformis has been studied by light and electron microscopy. Unspecialized monociliated cells are the dominant cell type of the epithelium; however, biciliated epidermal cells, monociliated mucous cells and non-ciliated cells are also present. In all of the ciliated cells each cilium is provided with a diplosomal basal body and two striated rootlets. The cytological features of this unspecialized epithelium strongly resemble those of similar epithelia in the Phoronida, Brachiopoda and Hemichordata. So far,O. fusiformis is the only polychaete known to possess an unspecialized monociliated epidermis. Other characters ofOwenia are also mentioned. The relationship between the nervous system and the epidermal cells on the tentacles is examined. It is proposed thatOwenia should assume an important role in discussions of the phylogeny of the Polychaeta.I wish to thank Dr. R.M. Rieger for his inspiration and guidance during the course of this study. Ms. Wilma Hanton provided technical assistance for the electron microscopy. This work was supported by NSF Grant # GB-42211 to R.M. Rieger     

Nuclear membranous whorls

Summary Membranous whorls have been seen in the nuclei of peritoneal and testicular cells which had been subjected to various experimental manoeuvres. It seems likely that this is an early manifestation of cell degeneration which is demonstrated readily only by glutaraldehyde fixation, and to that extent can be regarded as a glutaraldehyde artifact. Acknowledgements. This work was supported by grants from the Medical Research Council, and the University of Sheffield Tuberculosis Research Fund, and by a grant to the Department from Unilever Ltd.I am grateful to Professor R. Barer for his advice and criticism, to Dr. G. A. Meek for guidance on electron microscopy, to Dr. E. J. Clegg for permission to use material from joint experiments. Technical and photographic assistance was provided by Messrs. P. GarLick and L. Murgatroyd and by Miss M. Tune.     

Olfactory sensilla on the antennae of the pine weevil,Hylobius abietis

Summary Olfactory sensilla on the antennal club of the pine weevil, Hylobius abietis, have been studied by scanning and transmission electron microscopy. Two main types, sensilla basiconica and sensilla trichodea, have been classified. The former are found in two variants, one containing one sensory cell and the other two. Sensilla trichodea have one sensory cell. The two sensillum types are localized in different fields of the club. The numerous sensilla basiconica are confined exclusively to the two constriction bands. Sensilla trichodea, fewer in number, are mainly restricted to the distal part where they also form a third ring. In addition, they are found immediately distal to the two construction bands. The fine structure of the two types suggests an olfactory function which could be proven by electrophysiological studies. The other hairs on the club are shown to belong to mechano- and taste sensilla and uninnervated setae.I am grateful to the Norwegian Research Council for Science and Humanities NAVF and the Norwegian Forest Research Institute for financial support; to Professor D. Schneider for providing laboratory facilities at Seewiesen; to him and his colleagues, particularly Dr. R.A. Steinbrecht, for helpful discussions and to Miss Barbara Müller for electron microscopy assistance. I also wish to thank Dr. T. Braathen and his staff for help with scanning electron microscopy facilities at the University of Oslo.     

Ultrastructure of the neurosecretory system of the Colorado potato beetle,Leptinotarsa decemlineata (Say)

Summary The ultrastructure of seven types of neurosecretory cells (NSC) in the medial and lateral groups of the protocerebrum is described. The differences among cell types established earlier by light microscopy parallel differences in size and appearance of the neurosecretory particles observed in electron micrographs. No relationship was found between the affinity for Gomori's paraldehyde fuchsin stain and the nature of the particles.The secretions of the A-, A₁-, and C-types of NSC of the medial group are characterized by electron-dense neurosecretory granules of 1250 Å dia., medium-dense granules of 2100 Å, and electron-lucent vesicles of 1700 Å, respectively. The L-type NSCof the lateral group contain smaller (1300 Å) or larger (1700 Å) neurosecretory granules. The medial B- and E-types of NSC and the lateral L_B-type contain granulated vesicles (1200 Å) of the same appearance. These cell types differ in other respects and most likely have separate functions.The author wishes to thank the Laboratory of Virology of the Agricultural University for the use of the electron microscope, Mr. J. Groenewegen and Miss J. van Rinsum for technical assistance, and Professor J. Lattin for correcting the English text. Part of the work has been done while the author was in the service of the Netherlands Organization for the Advancement of Pure Research (ZWO, grant 942-48), and the National Council for Agricultural Research (TNO).