Ultrastructural study of the pericardial organ-anterior ramifications complex neurosecretory terminals

Summary The pericardial organ-anterior ramifications complexes of Uca pugilator and Callinectus sapidus were studied by transmission electron microscopy. Five morphologically distinguishable groups of granules and two groups of vesicles were identified. These granules and vesicles are present in approximately the same proportions in the pericardial organs and anterior ramifications of both species. Two of the granule groups are never mixed in the same axon terminals and are believed to represent different hormone-protein complexes. The remaining granule and vesicle groups are believed to be products of neurosecretory hormone release. Evidence that at least some of these granules and vesicles arise from intraaxonal release of neurosecretory material is presented.This work was supported by USPHS-NIH Training Grant GM-00669 and by the University of Texas Institutional Grant No. 5 SO 1 RR 05426-11.     

Central projections of the frontal organ of Rana pipiens,as demonstrated by the anterograde transport of horseradish peroxidase

Summary The central projections of the frontal organ of Rana pipiens are more widespread, and more similar to those of the epiphysis, than previously realized. Fibers labeled with horseradish peroxidase were traced to the amygdala, the epiphysis, the pretectal region, and several nuclear areas of the mesencephalic and diencephalic central gray. When peroxidase reaction product was carefully distinguished from neuromelanin by means of polarization microscopy, no unequivocally labeled cell bodies of centrifugal fibers could be found.Supported by NIH grants GM-09181, EY-02083, and by BRSG RR-05357 awarded by the Biomedical Research Grant Program, Division of Research Resources, N.I.H.     

Ultrastructural development of the neurohemal organ joined to the ecdysial gland after imaginal moulting in the male isopod Sphaeroma serratum fab. (Crustacea Flabellifera)

Summary The present ultrastructural study deals with the lateral cephalic nerve plexus of Sphaeroma serratum, a neurohemal organ joined to the Y organ (ecdysial gland). This plexus acts as a storage centre for neurosecretory products from two sources: the two autochtonous cells (plexus cells) within the plexus itself, and the neurosecretory cells in various parts of the central nervous system, particulary the mandibular ganglion (A-cells).In prepuberal animals, plexus cells and subesophageal A-cells produce neurosecretory granules of two types measuring 1550±50Å and 1570±40Å respectively. Five categories of axon terminals were distinguished in the plexus. The granules found in two of these terminal types are believed to come from the plexus cells and from the mandibular ganglion A-cells.Cessation of production of neurosecretory granules in these A cells and plexus cells was observed in puberal animals, in the plexus with concomitant depletion and disappearance of different granule categories. The first axon terminals affected by this process are the two categories containing granules originating in the plexus and mandibular ganglion A-cells. Degeneration of the ecdysial gland in male Sphaeroma serratum might be connected with the cessation of granule formation in these two types of cell.

---

Résumé Chez Sphaeroma serratum, la mue de puberté est suivie d'une dégénérescence de l'organe Y (glands de mue). Le plexus nerveux céphalique latéral, organe neurohémal accolé à cette glande a été l'objet de la présente étude ultrastructurale. Cet organe représente un centre de stockage de neurosécrétions qui proviennent d'une part, de deux cellules autochtones (cellules plexales) situées au sein même de ce plexus, d'autre part, de cellules neurosécrétrices situées dans le ganglion mandibulaire (cellules de type A).Chez les individus pubères, les cellules plexales et les cellules A du ganglion sous-oesophagien synthétisent des granules de neurosécrétion dont la taille est respectivement 1550±50Å et 1570±40Å. Il a été reconnu au sein du plexus 5 catégories de terminaisons dont les granules proviendraient pour deux d'entre elles des cellules plexales et des cellules A du ganglion mandibulaire. Chez les animaux pubères on observe un arrêt de la synthèse des granules de neurosécrétion au sein des cellules plexales et des cellules A du ganglion mandibulaire. Simultanément on enregistre dans le plexus la raréfaction puis la disparition des divers types de granules. Ce processus atteint en premier les terminaisons correspondant aux cellules plexales et aux cellules A du ganglion mandibulaire. La dégénérescence de la glande de mue chez les mâles pourrait être en relation avec l'arrêt de synthèse de ces cellules.

     

Vomeronasal receptors in Turtles

Summary The functional similarities observed with electrophysiological techniques between olfactory and vomeronasal receptors allow speculation that morphological details essential to the common function should be observed in both cases. Both mucosae have primary receptors within the epithelium which are surrounded, but not completely isolated, by so-called supporting cells. These last secrete a granular product. In both epithelia receptor cells contact each other at the axonal, perikaryal, dendritic and junctional complex levels. The axons of the two types of receptors are unmyelinated and their diameter ranges from 0.1 to 0.4 micron. The most interesting difference between the two types of receptors lies at the level of their exposed endings. The olfactory vesicle, as it is classically represented in olfactory receptors and is common in those of turtles in the form of a ball-like protrusion above the epithelial surface, is usually missing in the vomeronasal receptors. These have a tapering cone-shaped irregular projection always complicated by a set of branched microvilli. They do, furthermore, consistently lack cilia. This observation is in agreement with recent TEM observations. The assumption that cilia are essential in the mechanism of olfactory transduction is discussed on the basis of these anatomical findings.     

Ciliary-structure precursor bodies as stable constituents in the sensory cells of the vomero-nasal organ of reptiles and mammals

Summary The sensory cells of the vomero-nasal organ in reptiles and mammals do not develop cilia. In several species they contain centrioles together with cilium-structure precursor bodies measuring 400–700 Å in diameter. These structures resemble axonemal precursor bodies which are known to occur in developing ciliated cells. They are enclosed in a fibrogranular matrix. The precursor bodies are resistant to pepsin digestion in Araldite sections. In Tupaia precursor bodies may join periodically in a row. In the vomero-nasal receptor cells the precursor bodies can be considered stabilized with a corresponding reduction of cilia. The periodically arranged precursor bodies could represent a special storage form.     

Efferent connections from the brain to the frontal organ in Rana temporaria demonstrated by labeling with horseradish peroxidase

After injection of horseradish peroxidase into the frontal organ of Rana temporaria, labeled perikarya were found in the medial part of the amygdala, the preoptic area, the nucleus rotundus, the pretectal area, and the lateral parts of the midbrain central griseum.     

Hormonal control of seasonal morph determination in the swallowtail butterfly, Papilio xuthus L. (Lepidoptera: Papilionidae)

Seasonal morphs (spring and sumer forms) of Papilio xuthus L. are determined coincidentally with diapause and non-diapause in pupae by larval exposure to short days and long days respectively. The neuroendocrine principle underlying seasonal-morph determination was studied using surgical operations in P. xuthus.When recipient 0-day old or chilled diapause pupae were joined to donor 0-day old non-diapause pupae, the recipients developed into summer or intermediate morphs. When the same kinds of recipients used above were joined to 0-day old or chilled diapause pupae, there were no significant effects on the adult morph. In contrast, recipient non-diapause pupae all developed into summer morphs, regardless of groups of the type of donors.Furthermore, when the brain of 5th-instar larvae, pharate pupae or pupae, predetermined to be diapause, was transplanted into the abdomen of 0-day old, 30-day old or chilled diapause pupae, the recipients developed into summer or intermediate morphs.The results indicate that the brain of non-diapause pupae secretes a humoral factor producing the summer morph. In non-diapause pupae, the factor may be secreted at about the stage of larval-pupal ecdysis coincidentally with that of prothoracicotropic hormone.     

20-Hydroxy-ecdysone as a modulator of electrical activity in neurosecretory cells of Rhodnius prolixus

Ovariectomy of the adult female Rhodnius results in a reduction of the electrical activity of the corpus cardiacum. Injection of 20-hydroxy-ecdysone into ovariectomized animals increases the activity to control values. Incubation of heads isolated from ovariectomized females in 1.0 × 10^?7 M 20-hydroxy-ecdysone elicits a recruitment of large-amplitude spikes and the appearance of bursting activity in the corpus cardiacum of mated females which is characteristic of neurohormone release. Simultaneous recordings from the median neurosecretory cells and the corpus cardiacum demonstrate that the action neurosecretory cells.     

Evidence for a frontal-organ homologue in the pineal complex of the salamander,Hynobius dunni

The pineal complex of larval and adult salamanders, Hynobius dunni, was examined by light and scanning electron microscopy. This pineal complex displays an anterior and a posterior portion, both of which possess a lumen. The anterior lumen is small and closed, whereas the posterior lumen is in open communication with the third ventricle. Cell processes of the photoreceptor cells and microvilli of the supportive cells are visible in both lumina. The anterior part of the complex is formed by an independent, second evagination from the common pineal anlage; this process takes place immediately after hatching. The anterior body of the pineal complex of H. dunni appears to be homologous to the frontal organ of anurans.     

中国多目涡虫属四新种记述（三肠目：淡水亚目：扁涡虫科）

本文记述多目涡虫属４新种：五台山多目涡虫,标本采自山西繁峙县和五台县;静乐多目涡虫,山西静乐县,泾源多目涡虫,宁夏泾源县;哈密多目涡虫,新疆哈密县。     

中国多目涡虫属两新种记述

本文记述多目涡虫属两新种;中华多目涡虫,新种Polycelis sinensis sp.nov.和林周多目涡虫,新种Polycelis lhunzhkubica sp.nov前者标本采自西藏拉萨市管辖的达孜县、当雄县、林周县和墨竹工卡县;后者标本采自西藏林周县旁多乡.     

The hypothalamo-hypophysial system in acipenseridae

Summary The neurohypophysis of sexually mature male and female Acipenser güldenstädti Brandt and Acipenser stellatus Pallas was studied light and electron microscopically. The recessus hypophysei lined with ependymal cells of two main types, narrow and wide, are in the center of the neurohypophysial roots. Processes from both cell types run radially to the basement membrane of the connective tissue layers abutting on the hypophysial intermediate lobe. Protrusions penetrating deep into the recessus hypophyseus are found in the apical parts of the wide cells. Pituicytes are rare in the neurohypophysis. The ultrastructure of both ependymal cell types and of the pituicytes is described. Nonmyelinated Gomori-positive (peptidergic) neurosecretory A₁ and A₂ type fibres and their terminals containing elementary neurosecretory granules (1400–1800 Å and 1000–1400 Å respectively) are the main structural elements of the neurohypophysis. Some dark and single myelinated neurosecretory fibres have been found. The adrenergic fibres (type B) were described earlier (Polenov et al., 1972a). The structural peculiarities of the neurohypophysis are discussed in functional and comparative-morphological terms.     

Luminescent metalloreceptors with pendant macrocyclic ionophore: Synthesis, characterization, electrochemistry and ion-binding study

Metalloreceptors containing ruthenium(II) bipyridine unit as fluorophore and pendant macrocyclic units as ionophore have been synthesized and their luminescence and electrochemical properties have been investigated. Ion-binding study of these fluoroionophore with the anions F⁻, Cl⁻, Br⁻, I⁻, , , , , CH₃COO⁻, and and cations Na⁺, K⁺, Mg²⁺, Ca²⁺, Zn²⁺, Ba²⁺, Sr²⁺ Cd²⁺, Hg²⁺, Pb²⁺ and Cu²⁺, monitored by luminescence and ¹H NMR spectral changes, reveal strong interactions of and F⁻ for 2 and 3 and of Cu²⁺ only for 3. Luminescence titrations for 2 and 3 have been carried out to determine binding constants (K_s), and the calculated values are in the range 2.85 × 10² to 4.48 × 10⁴ M⁻¹. The ¹H NMR spectral changes for 2 and 3 with the addition of increasing concentration of F⁻ and exhibit substantial low-field shift of the CONH proton indicating its involvement in complex formation with the anions. The adduct of 2 and 3 have been isolated and characterized by ¹H and ³¹P NMR, mass and IR spectroscopy. The results are discussed in light of selectivity, structures of the anion bound complexes and their luminescence property.     

Fluoreszenzmikroskopische Untersuchungen mit Pseudoisocyaninchlorid am Schlundring einiger stylommatophorer Pulmonaten

Zusammenfassung Durch Anwendung von Pseudoisocyaninchlorid lassen sich im zentralen Nervensystem von stylommatophoren Pulmonaten in allen Schlundringganglien Neurosekretzellen fluoreszenzmikroskopisch nachweisen. In den Cerebralganglien bilden sie mit ihren ausleitenden Axonen das bereits bei einigen Gastropoden bekannte geschlossene neurosekretorische System. In den übrigen Ganglien liegen die neurosekretorischen Zellen einzeln oder in Gruppen zusammen und bilden kein System. Es konnte nachgewiesen werden, daß in Visceral-Parietaganglien der untersuchten Arten die meisten sekretorischen Neurone vorkommen. Die Zahl der neurosekretorischen Zellen in den einzelnen Ganglien ist zumindest für die hier untersuchten Arten wahrscheinlich artspezifisch, ihre topographische Lage in den Ganglien aber bei allen Arten identifizierbar ähnlich. Bei allen untersuchten Pulmonaten fanden sich nur im Procerebrum keine neurosekretorischen Zellen. Die vielfältigen, über die Darstellung der Neurosekretzellen hinaus feststellbaren fluoreszenzmikroskopischen Erscheinungen in den Nerven- und Gliazellen, in der Fasermasse der Ganglien, den Kommissuren und Nerven und im Bindegewebe um die Ganglien, wurden mit Hilfe anderer lichtmikroskopischer Färbungen vergleichend untersucht; die Ergebnisse werden diskutiert.

---

Fluorescence-microscopic investigations with pseudoisocyanin on the central nervous system of some stylommatophoran pulmonates

Summary Using fluorescence microscopy with pseudoisocyaninchloride one can demonstrate the well known cells and axons of the neurosecretory system in the cerebral ganglia of some gastropod pulmonates. In the other ganglia of the central nervous system-except procerebrum-single secretory neurones or groups of neurosecretory cells occur. These secretory neurones do not build up a neurosecretory system. The majority of these cells is concentrated in the visceral-parietal ganglia. The specific elements are topographically identifiable relative to each other in the ganglia of each species of pulmonates investigated. Other fluorescence-microscopic investigations of the neurones, glial cells and the neuropil of the ganglia, commissures and nerves and in the connective tissue surrounding the ganglia are described in comparison with other light microscopical findings obtained by staining methods; the results are discussed.

     

Fluorescent granules in the guinea-pig hypothalamus and their possible relation to neurosecretory substance

Summary Sections of guinea-pig hypothalamus stained by various methods have been studied under bright-field and fluorescence microscopy to determine the characteristics of refractile granules originally observed in fixed tissue stained for cholinesterases but also seen in unfixed, unstained material.When sections of fresh tissue or tissue fixed in formaldehyde: Na₂SO₄ were examined by fluorescence microscopy, the granules emitted with a wave-length between 600 and 620 m. The density and distribution of the granules closely paralleled that of one of the components that stained with the aldehyde-fuchsin and chrome-alum techniques for neurosecretory substance; granules were absent, however, from the area characterized by Herring substance, and from the stained perikarya and beaded axons of the supraoptic and paraventricular nuclei.The granules were most numerous in the median eminence and infundibular nuclei. Their concentration was about average in mature males and in lactating and hypophysectomized females; they were more abundant in late pregnancy and after ovariectomy and were particularly plentiful some weeks after hysterectomy. They were virtually absent from immature males and females.The possible relation of the granules to releasing factors or to vasotocin-like neurosecretory material is discussed.The authors are most grateful to Dr. J. S. Perry who carried out the surgery involved in these experiments. They also wish to express their thanks to Dr. R. B. Heap who, with Dr. Perry, gave invaluable help in planning the experiments and assessing the data; to Mr. S.P. Mann and Dr. D. F. Sharman for advice on the fluorescence technique and to Miss M. Hamon for skilled technical assistance.Publication of the colour pictures was made possible by a generous grant from the Wellcome Foundation; this support is gratefully acknowledged.Thanks are also due to the University of Alberta for a contribution towards the cost of reprints.     

Studies on the chemoreception of the nemertean, Procephalothrix simulus Iwata, 1952

The feeding responses of Procephalothrix simulus Iwata to the whole-body extracts of different animals (Tubifex sp., Saccocirrus gabrillae) and single compounds (including 17 amino acids, 2 compounds related to amino acids and 2 sugars) were tested, and the response of the nemerteans with and without a frontal organ to stimulus was compared experimentally. Results showed that P. simulus is able to detect food from a distance by chemoreception. It preferred extracts to live prey, and boiled extracts were more attractive than extracts that were not boiled. Of the 21 compounds tested, 6 amino acids (glycine, alanine, isoleucine, phenylalanine, glutamine, asparagine) and betaine induced a feeding response, suggesting that they were possibly the chemical signals in the food location of P. simulus. When the frontal organ was removed, P. simulus apparently lost its selectivity, which indicated that in this nemertean chemoreception for the detection of prey was performed by the frontal organ.     

G-Proteins mediate inhibition and activation of Ca2+-induced exocytosis from SLO-permeabilized peptidergic nerve endings

In SLO-permeabilized isolated nerve endings from the rat neurohypophysis, GTP, guanosine 5[y-thio]triphosphate (GTPyS) and guanosine 5(ßy-imido]triphosphate (GMPPNP) inhibit the Ca²⁺-evoked vasopressin release. Pretreatment with pertussis toxin enhances the inhibitory effects of both GTP-analogues. Omission of Mg²⁺ overcomes the effect of GMPPNP and reverses the inhibitory effect of GTP and GTPyS. In the absence of Mg²⁺, GTP and GTPyS now potentiate Ca²⁺-evoked secretion.