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.     

Nature of vesicles associated with the nervous system of cephalopods

Summary The epistellar body on the stellate ganglion of octopods and the parolfactory vesicles on the optic tracts of decapods have been established as potential photoreceptors on the basis of morphology (rhabdomeric processes) and biochemistry (presence of rhodopsin and vitamin A₁). These organs are not to be viewed as neuroglandular, as originally suggested, but rather as possibly contributing to physiological regulation by recording photic information. Layers of nerve fibers are evident around the vesicles in silver-stained preparations, contributing to nerves with central connections. The stellate ganglion of the young Octopus has lens-like optical properties.Dedicated to Professor Berta Scharrer in honor of her 60th birthday.Aided by U.S. National Science Foundation grant GB-2484. We wish to express our thanks to Doctors Pietro Dohrn and Luisa Tosi for the provision of facilities, to Professor A. de Girolamo and Dr. Aldo Cecio of the Istituto di Anatomia, Istologia e Embriologia Veterinaria of the University of Naples for their generous cooperation in allowing us the use of their electronmicroscope laboratory, to Mr. Jeremy Worthington for able technical assistance, and to Miss Margaret Rufener for the drawings.National Science Foundation Senior Postdoctoral Fellow, Stazione Zoologica di Napoli, 1965–1966     

A histochemical study of chondroid tissue in Limulus and Octopus

Summary In a comparative histochemical study of Octopus and Limulus chondroid tissues and mouse tracheal cartilage, it was demonstrated that both invertebrate chondroids behave as less acid mucopolysaccharides than those in mouse cartilage. Octopus chondroid was less reactive than Limulus chondroid. Cetyl pyridinium chloride blockage of toluidin blue metachramasia could be unblocked in mouse cartilage by an hour's treatment in 0.5 M KCl, but even extended periods in 2.0 M KCl failed to accomplish this in the invertebrate chondroids. With a number of methods for demonstrating proteins, Octopus chondroid was less reactive than Limulus chondroid. Limulus chondroid matrix was intensely stained by methods for demonstrating protein thiol groups, but essentially no staining was observed in Octopus chondroid matrix. These studies indicated that the composition and/or structure of matrix material in the invertebrate chondroids differ from one another, and in turn differ from conditions in vertebrate hyaline cartilage.This work was supported by a grant (HD-1499-04) and a Career Development Award (5-K3-6176-04) from the National Institute of Child Health and Human Development of the U.S. Public Health Service.A contribution of the Sea Horse Key Marine Laboratory of the University of Florida.     

Tissue channel morphology in Octopus

Summary The morphology of tissue channels in muscle and neural tissues of Octopus was investigated, at the ultrastructural level, with a technique involving the precipitation of ferrocyanide ions. The numbers, sizes and conductivities of the channels were estimated from quantitative data. No evidence was gained to indicate that the low microvascular density in Octopus is coupled to an especially extensive network of extravascular channels. The tissue channel system in Octopus appears to be broadly comparable with the mammalian system; a lack of information prevents more appropriate comparisons with marine fishes. Probable functions of tissue channels in Octopus and mammals, and reasons for apparent similarities and differences in the channel organization of these divergent groups, are discussed.     

The Hemocyanin from a Living Fossil, the Cephalopod Nautilus pompilius: Protein Structure, Gene Organization, and Evolution

By electron microscopic and immunobiochemical analyses we have confirmed earlier evidence that Nautilus pompilius hemocyanin (NpH) is a ring-like decamer (M_r = ∼3.5 million), assembled from 10 identical copies of an ∼350-kDa polypeptide. This subunit in turn is substructured into seven sequential covalently linked functional units of ∼50 kDa each (FUs a–g). We have cloned and sequenced the cDNA encoding the complete polypeptide; it comprises 9198 bp and is subdivided into a 5′ UTR of 58 bp, a 3′ UTR of 365 bp, and an open reading frame for a signal peptide of 21 amino acids plus a polypeptide of 2903 amino acids (M_r = 335,881). According to sequence alignments, the seven FUs of Nautilus hemocyanin directly correspond to the seven FU types of the previously sequenced hemocyanin “OdH” from the cephalopod Octopus dofleini. Thirteen potential N-glycosylation sites are distributed among the seven Nautilus hemocyanin FUs; the structural consequences of putatively attached glycans are discussed on the basis of the published X-ray structure for an Octopus dofleini and a Rapana thomasiana FU. Moreover, the complete gene structure of Nautilus hemocyanin was analyzed; it resembles that of Octopus hemocyanin with respect to linker introns but shows two internal introns that differ in position from the three internal introns of the Octopus hemocyanin gene. Multiple sequence alignments allowed calculation of a rather robust phylogenetic tree and a statistically firm molecular clock. This reveals that the last common ancestor of Nautilus and Octopus lived 415 ± 24 million years ago, in close agreement with fossil records from the early Devonian. [Reviewing Editor: Dr. Axel Meyer] The sequence reported in this paper has been deposited in the EMBL/GenBank database under accession number AJ619741.     

Identification of adenohypophysiotropic neurohormone producing neurosecretory cells in Rana temporaria

Summary Electron microscopic investigation of neural isolated and normal pars ventralis of the tuber cinereum showed the presence, in Rana temporaria, of a tubero-hypophysial neurosecretory system. A striking structural resemblance between this parvicellular, aldehydefuchsin negative, tubero-hypophysial neurosecretory system and the magnocellular, aldehydefuchsin positive, hypothalamo-hypophysial neurosecretory system was observed. Six different neurosecretory cell types are described, characterized by different shape and size of their respective secretory granules. The nature of the secretory product of these cells is briefly discussed.Of the results of this investigation, a preliminary note has been published (Dierickx, 1971).The authors wish to thank Prof. Dr. H. Steyaert and Dr. W. Moerdijk of the Laboratory for Mathematical Statistics, Ghent University, for invaluable advice and assistance with their computers. They thank Dr. G. De Waele for his help with the scanning electron microscopy.     

The rostral pars distalis of the pituitary gland of the freshwater and marine alewife (Alosa pseudoharengus)

Summary In the alewife the orohypophyseal duct, a remnant of Rathke's pouch, persists in adults as a tube passing from the rostral pars distalis to the pharyngeal region. Its lumen is not open to the buccal cavity. The prolactin cells are situated around the bifurcations of this duct in the rostral pars distalis. Contents from prolactin cells, such as granules, nuclei, mitochondria and Golgi structures were found in these bifurcations. These contents were indistinguishable from those of intact prolactin cells. Evidence of actual release into the duct was often noted. At the presumptive point of release, the cells lining the lumen separate and the contents, probably of an entire prolactin cell, are extruded. The cilia usually found at the point of extrusion arise from prolactin cells. The prolactin cells of freshwater fish were more heavily granulated than those from a marine environment. Prolactin cells of fish entering freshwater streams were not heavily granulated but showed evidence of increased activity. Granule size was not affected by salinity. The ACTH cells are arranged in bands along branches of the neurohypophysis in the rostral pars distalis. No differences in ACTH cells from fish of different salinities were noted.We would like to thank Mr. D. D. Zumwalt of the John G. Shedd Aquarium in Chicago, Dr. E. D. Warner, Mr. R. L. Flayter, Dr. J. G. Stanley of the University of Wisconsin, Milwaukee, Mr. L. Wells of the U. S. Fish and Wildlife Service, Great Lakes Fisheries Laboratory, Ann Arbor, and Mr. L. N. Flagg of the Department of Sea and Shore Fisheries, Augusta, Maine, for their assistance in obtaining the fish used in this study. Dr. T. N. Tahmisian and Mr. G. T. Chubb of Argonne National Laboratories, and Dr. L. M. Srivastava and Dr. V. Bourne of Simon Fraser University, Canada, kindly made electron microscope facilities available. Finally, we wish to thank Mr. W. Goossens and Mr. D. J. DeJong for valuable assistance. This project was supported by a grant from the National Marine Fisheries Service.     

Mapping of neuropeptide Y expression in Octopus brains

Neuropeptide Y (NPY) is an evolutionarily conserved neurosecretory molecule implicated in a diverse complement of functions across taxa and in regulating feeding behavior and reproductive maturation in Octopus. However, little is known about the precise molecular circuitry of NPY-mediated behaviors and physiological processes, which likely involve a complex interaction of multiple signal molecules in specific brain regions. Here, we examined the expression of NPY throughout the Octopus central nervous system. The sequence analysis of Octopus NPY precursor confirmed the presence of both, signal peptide and putative active peptides, which are highly conserved across bilaterians. In situ hybridization revealed distinct expression of NPY in specialized compartments, including potential “integration centers,” where visual, tactile, and other behavioral circuitries converge. These centers integrating separate circuits may maintain and modulate learning and memory or other behaviors not yet attributed to NPY-dependent modulation in Octopus. Extrasomatic localization of NPY mRNA in the neurites of specific neuron populations in the brain suggests a potential demand for immediate translation at synapses and a crucial temporal role for NPY in these cell populations. We also documented the presence of NPY mRNA in a small cell population in the olfactory lobe, which is a component of the Octopus feeding and reproductive control centers. However, the molecular mapping of NPY expression only partially overlapped with that produced by immunohistochemistry in previous studies. Our study provides a precise molecular map of NPY mRNA expression that can be used to design and test future hypotheses about molecular signaling in various Octopus behaviors.     

LD ratios and the entrainment of circadian activity in a nocturnal and a diurnal rodent

Summary The activity rhythms of 5 flying squirrels,Glaucomys volans, and 7 chipmunks,Tamias striatus, were examined under controlled conditions in the laboratory. Free-running, circadian rhythms were demonstrated using a total of 25 LL or DD experiments. With 46 LD schedules the limits of entrainment in a 24-hour day were determined, and the phase angle difference for each schedule measured.Glaucomys was able to synchronize to schedules ranging from 1 second of light per 24-hour day to at least 18 hours light per day with little or no change in the phase angle.Tamias showed an oscillatory type of entrainment when the photoperiod was less than 3 hours per 24-hour day or greater than 23 hours, but in the intervening region was capable of stable entrainment. A tendency was evident for the phase angle difference to become less positive as the LD ratio increased. InGlaucomys single, isolated light pulses of either one second or 24 hours duration were able to bring about relatively large shifts in the phase of the activity rhythm.Dedicated to Professor Jürgen Aschoff on the occasion of his 60th birthday.I wish to acknowledge the hospitality and assistance of the Zoology Department, University of Wisconsin, Madison, the Max-Planck Institute, Erling-Andechs, Germany, and the Belle W. Baruch Coastal Research Institute, University of South Carolina, Columbia. Special thanks are due Dr. John Emlen and Dr. William Reeder of Madison, Prof. J. Aschoff of Erling-Andechs, Dr. John Vernberg and Dr. Winona Vernberg of Columbia, and my husband, Dr. George DeCoursey, for their untiring encouragement and help with these experiments.     

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.     

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.     

Association of particles that contain double-stranded RNAs with algal chloroplasts and mitochondria

Linear dsRNAs (double-stranded RNAs) belonging to several distinct size classes were found to be localized in chloroplasts and mitochondria of Bryopsis spp., raising the possibility that these dsRNAs are prokaryotic in nature. The algal cytosol and nuclei did not contain dsRNAs. The amount of the dsRNAs in the organelles appeared constant, and there were about 500 copies per chloroplast. The four major dsRNAs from Bryopsis chloroplasts were about 2 kbp (kilobase pairs) in length and originated from discrete isometric particles of about 25 nm diameter. These virus-like particles were purified by CsCl density gradient centrifugation after extraction from isolated chloroplasts with chloroformbutanol and subsequent precipitation with polyethylene glycol. They had a buoyant density of about 1.40 g · cm^–3 and contained four major and three minor proteins. Mitochondrial dsRNAs were about 4.5 kbp in length and formed less-stable particles of about 40 nm in diameter with a buoyant density of 1.47 g · cm^–3. Some observations support the hypothesis that vertical transmission of the protein-coated, non-infectious dsRNAs occurs within cell organelles. Double-stranded RNAs of various sizes were found in most green, red, and brown algae. The characteristics of the algal dsRNAs are compared with those of dsRNAs from higher plants and the biological significance of the dsRNAs in cell organelles is discussed.Abbreviations dsRNA double-stranded RNA - kbp kilobase pairs - SDS sodium dodecyl sulfate - SSC 0.15 M NaCl 0.015 M sodium citrate - PAGE polyacrylamide gel electrophoresis The authors would like to express their gratitude to Dr. T. Natsuaki, Utsunomiya University, and Dr. D. Hosokawa, Tokyo University of Agriculture and Technology, for their helpful suggestions throughout this research. They are also much indebted to Dr. B. Wang, Institute of Genetics, Academia Sinica, Beijing, PRC, for his suggestions on rice dsRNA, and to Dr. T. Kohbara, Senshu University, on Bryopsis cells. Sincere thanks are also due to Dr. T. Misonou, Yamanashi University, and Dr. K. Masuda, Akita Prefectural College of Agriculture, for supplying plant materials; to Dr. N. Sonoki, Azabu University, for nucleotide analysis of dsRNAs; and to Y. Koshino for technical assistance. This research was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.     

Secondary sensory cells in the gravity receptor system of the statocyst of Octopus vulgaris

Summary The presence of secondary sensory cells in the Octopus gravity receptor system has been demonstrated. In serial thin sections of the receptor cells (hair cells) no axons were found leaving the cells. Instead, synapses were observed with synaptic vesicles lying inside the receptor cells. Both data clearly indicate that the receptor hair cells represent secondary sensory cells. In addition, efferent contacts to the receptor cells could be confirmed.This work was supported in part by grant Wo 160/5 of the Deutsche Forschungsgemeinschaft to Prof. Dr. H.G. WolffThe experimental work was done in part at the Zoological Station in Naples and at the Sechenov Institute of Evolutionary Physiology and Biochemistry of the USSR Academy of Sciences (Laboratory of Prof. Dr. Ya.A. Vinnikov), Leningrad, USSR. The authors thank Prof. Vinnikov and Dr. Tsirulis for stimulating discussions     

The Vascularization of the Anuran Brain Rhombencephalon and Medulla spinalis: A scanning electron microscopical study of vascular corrosion casts

Albrecht, U., Lametschwandtner, A., Adam, H. 1980. The vascularization of the anuran brain. Rhombencephalon and medulla spinalis. A scanning electron microscopical study of vascular corrosion casts. (Department of Zoology, University of Salzburg, Austria.) — Acta zool. (Stockh.) 61 (4): 239–246. The vascularization of the rhombencephalon and the medulla spinalis of Bufo bufo (L.) is demonstrated by scanning electron microscopy of vascular corrosion casts. The arterial supply of the rhombencephalon is performed by central arteries. The same is shown in the medulla spinalis. The venous pathways are represented by venae craniales occipitales and by a posterior and bulbar group of the encephaloposthypophysial portal vein, by veins draining into the venae craniales occipitales, by venae spinales ventrales (for the rostral regions of the medulla) and by venae spinales laterales (in the caudal medulla). In the regions examined so far a centrifugal course of the arterial vessels is reported.     

Aerobic dissimilation of d-ribulose by yeasts

Aerobic dissimilation ofd-ribulose by various genera and species of yeasts was examined. Most strains tested utilizedd-ribulose fairly well, andd-arabitol was accumulated in the fermented broth but ribitol was not found. Torulopsis famata ATCC 20214,T.mannitofaciens CBS 5981 andT.versatilis CBS 1752 producedd-arabitol with a yield of 26 to 67% of the sugar used. Extracellular and intracellular formation of pentitol fromd-ribulose were compared with those fromd-xylulose.We wish to express our sincerest thanks to Prof. K. Arima and Prof. Y. Ikeda of the University of Tokyo for their kind guidance. We also wish to thank Dr. M. Mogi and Dr. N. Iguchi of this Institute for their encouragement and Mr. K. Kouchi for his technical assistance.     

Spermiogenesis in the brush-tailed possum,Trichosurus vulpecula (Marsupialia)

Summary Acrosome development in the Australian Brush-tailed possum, Trichosurus vulpecula, displays a number of extraordinary features. This is particularly evident in the later stages of spermiogenesis, when the area of the nuclear surface bounded by the nuclear ring, and covered by the acrosome, is reduced considerable. As a result, the acrosomal material becomes located over its definitive position on the anterior third of the dorsal nuclear surface; in this process it is thrown into a series of folds, and a wide subacrosomal space is formed.Further changes around the time of spermiation result in the release of a spermatozoon in which a thin layer of acrosomal material is closely applied to the nucleus over the area of the definitive location of the acrosome, whilst its margins are greatly extended and project freely away from the nucleus. The latter feature does not appear to have been reported for the sperm of other mammals.The authors would like to thank Dr. D.J.H. Cockayne, Director of the Electron Microscope Unit, University of Sydney, for the generous provision of transmission electron microscope facilities, and Dr. M.R. Dickson, Electron Microscopist in charge, Biomedical Electron Microscope Unit, University of New South Wales, for the use of other facilities. Also, thanks are due to Miss Robin Arnold and Mrs. Eva Vassak of the Department of Histology and Embryology, University of Sydney, for their expert assistance. The assistance of the N.S.W. National Parks and Wildlife Service in the provision of permits to work on these native mammals is acknowledged     

Keratin-like fibres in the hemichordate Rhabdopleura compacta

Summary The coenecium of Rhabdopleura is a tube that surrounds the zooid. It is secreted by the cephalic shield of the zooid and contains three sorts of fibres in an electron lucent matrix. One of the fibre types contains a double helix of fine fibrils. Preliminary histochemical investigations suggest that the fibres may be keratinous.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. Dr. A. J. Southward and Dr. A. Stebbing of the Plymouth Marine Biological Laboratory generously gave of their time and expertise, and helped me to obtain and identify the specimens. Dr. R. Willis and Miss Marion Dennison assisted with the preliminary stereoscopic electron microscopy. Mr. R. Moss gave excellent technical and photographic assistance.     

Behavioral analyses of auditory sensitivity inCycnia tenera Hübner (Lepidoptera: Arctiidae)

Summary The auditory characteristics of the arctiid moth,Cycnia tenera were examined using two behavioral criteria, sound production and flight cessation. The majority of the individuals tested indicated a maximum sensitivity to frequencies between 30 and 50 kHz although there is a substantial degree of interindividual variation.Spectral analyses of the echolocation/hunting cries of two species of sympatric, insectivorous bats,Eptesicus fuscus andMyotis lucifugus reveal maximally intense spectral peaks at 29.3 and 41.9 kHz, respectively.The audiograms offer a behavioral correlation to neural studies done with other species of moths and illustrate the existence of behaviorally expressed intensity discrimination in these insects. The variation evident between individuals may represent the differential response characteristics reported for free-flying tympanate moths in field conditions.I am indebted to Ms K. Van Every for her invaluable and patient assistance during the field portion of this study and to Dr. R. Robertson for his permission to use the facilities at the Queen's University Biology Station. Dr. M.B. Fenton provided me with financial support and much appreciated advice for the project. I also thank Dr. Fenton for the tapes ofE. fuscus that he made. The helpful criticisms of Drs. G.K. Morris, G.E. Kerr, A. Michelsen and an anonymous reviewer were gratefully appreciated. I thank Dr. J.A. Simmons for Fig. 1.     

A comparative ultrastructural study on ultimobranchial glands of some Israeli anurans (Bufo viridis,Rana ridibunda and Hyla arborea)

Summary The comparative fine structure of ultimobranchial (UB) glands of adult Israeli anurans (Bufo viridis, Rana ridibunda, Hyla arborea) taken in the wild during the breeding season is presented. Common aspects of the UB secretory cells are considered with especial reference to secretory granules, lipid droplets and tonofilaments. In B. viridis a second cell type with large electron-dense cytoplasmic granules is found in UB follicles. R. ridibunda and H. arborea UB follicles have a second cell type similar to goblet cells in appearance and these appear to be discharging their mucoid contents into the lumina of the follicles. The possible significance of these various cell types is considered.I am indebted to the Central Research Fund of London University for an award for apparatus and travel enabling a short research visit to the Dept. of Zoology, The Hebrew University, Jerusalem and am particularly indebted to Dr. Dvorah Boschwitz and her colleagues for their enormous help during my visit. I would also like to thank Alan D. Phillips for his assistance in the analysis of the material made possible by an award from the Science Research Council. My thanks are also due to Raynor L. Jones for his excellent technical assistance.     

Vascular organization of the catfish gill filament

Summary Light and scanning electron microscopic observations were made on methyl-methacrylate corrosion casts of the blood vessels in the gills of channel catfish (Ictalurus punctatus). The vasculature of the gill filament can be divided into three distinct pathways: 1. the well-known respiratory circulation which includes the afferent filamental artery (AF), afferent lamellar arteriole (AL), lamella (L), efferent lamellar arteriole (EL) and efferent filamental artery (EF), 2. a nutritive pathway from the EF through small nutritive capillaries (NC) and into one of several filamental veins (FV), and 3. an interlamellar circulation in which small prelamellar arterio-venous anastomoses (PAVA) connect the AL into a series of organized vascular spaces (interlamellar vessels, ILV's) that underlie the interlamellar filamental epithelium. Several sinuslike spaces associated with AF, EF and the filamental cartilagenous support were also observed. The physiological significance of these vascular pathways is discussed.Supported in part by NSF Grant No. PCM 76-16840The authors wish to acknowledge the assistance of Mr. P. Holbert, Miss K. Drajus and Mrs. J. Smith. Gratitude is expressed by Kenneth R. Olson to Dr. Janice Nowell for her helpful suggestions with corrosion casting techniques