Elektronenmikroskopische Untersuchungen am Pinealorgan von Pterophyllum scalare Cuv. et Val. (Cichlidae,Teleostei)

Zusammenfassung Das Pinealorgan (Epiphysis cerebri) des Knochenfisches Pterophyllum scalare besteht aus nervösen und gliösen Zellelementen. Sehr stark ausgebildet sind die ependymalen Stützzellen. Sie umhüllen mit ihren Ausläufern, die sich überlappen können, andere Zellelemente, z.B. Rezeptorzellen und marklose Nervenfasern. Neben dieser Neuroglia-Art finden sich auch noch oligodendrocytenähnliche Gliazellen. In ihrer Grundstruktur entsprechen die Rezeptoren den Epiphysensinneszellen anderer Knochenfische. Vom cilientragenden Teil des Außenglieds geht ein schürzenartiger Lamellenstapel aus. Dieser besteht aus 50–70 Lamellenplatten von etwa 6 m Länge. Im basalen Teil der Rezeptorzelle sind neben schlanken Mitochondrien mit unregelmäßigen Cristae und Tubuli auch noch große, runde Mitochondrien mit einer regelmäßigen Cristastruktur zu beobachten. Der basale Fortsatz der Rezeptorzelle ist auf die axial verlaufenden Axonbündel ausgerichtet. Synapsenartige Kontakte sind selten. Die Zahl der marklosen Axone nimmt hirnwärts zu; dieser Befund wurde in partiellen Rekonstruktionen gesichert. Am Übergang in den Epiphysenstiel treten einige markhaltige Axone auf. Zur Verteilung der Zelltypen und zum Verlauf der Axonbündel im Epiphysenstiel des Skalars liegen detailliertere Angaben vor als bei anderen bisher untersuchten Knochenfischepiphysen. In der Diskussion werden nach Vergleich der pinealen Rezeptoren verschiedener Fische drei Außengliedformen unterschieden: Bürsten-, Schürzen- und Kappentyp. Diese Varianten werden in Verbindung mit den bekannten physiologischen Reaktionsformen der Pinealorgane diskutiert. Die elektrophysio logischen Unterschiede lassen sich nicht mit verschiedenen Strukturtypen des Außenglieds erklären.

---

Electron microscopic studies of the pineal organ in Pterophyllum scalare cuv. et val. (Cichlidae, Teleostei)

Summary The pineal organ (epiphysis cerebri) of Pterophyllum scalare is formed by neuronal and glial elements. Ependymal supportive cells are very abundant, and their cytoplasmic processes envelop adjacent receptor cells and unmyelinated nerve fibers by an intertwining network. In addition to this type of neuroglia, oligodendrocytic cells have also been identified. The receptor cells show the general structural pattern (outer segment, inner segment, basal process) of teleostean pineal receptors. The ciliary part of the outer segment bears a dome-like stack of 50–70 curved saccules each of average length of 6 m. In the basal part of the receptor cell, slender mitochondria containing irregular cristae and tubules, and also some more spherical mitochondria with a highly regular arrangement of cristae, can be observed. The basal cytoplasmic process radiates into neuropile-like areas that contain axial bundles of axons. Synaptoid contacts rarely occur. The number of unmyelinated axons of the pineal stalk, increases in a proximad direction (towards the brain). This finding has been verified in partial reconstructions. In the transitional zone leading from the pineal body into the pineal stalk, a few myelinated fibers become visible. With respect to cell types and the axonic bundles of the pineal stalk in Pterophyllum scalare, more detailed data are presented than for most other teleostean pineal organs examined thus far. The comparison of pineal sensory cells in several fishes allows a distinction among three different types of outer segments, i.e., a slender type, a dome-like type, and a cap-like type. These structural types are discussed with respect to the relevant physiological results. The existence of particular structural types of the outer segment does not explain the different electrophysiological reactions observed in different pineal organs.

Ein Druckkostenzuschuß wurde von beiden Instituten zur Verfügung gestellt.     

Elektronenmikroskopische Untersuchungen am Pinealorgan von Passer domesticus

Zusammenfassung Das Pinealorgan von Passer domesticus enthält Zellen mit innengliedartigem mitochondrienreichen Stift, der eine bulböse Zilie mit 9+0 Zilienfibrillen entsendet. Randzipfel solcher Zilien, die vielfach in Gruppen anzutreffen sind, können sich in etwa 200 Å starke Lamellen fortsetzen. Konzentrische und wirbeiförmige Lamellenzüge, die von mehreren solchen Zilien ausgehen, bilden im Lumen markscheidenartige oder auch ungeordnete Membranenkomplexe. Ein direkter Zusammenhang mit Zilien konnte nur bei einem Teil dieser Lamellenkörper ermittelt werden. Der Bauplan der bulbösen Zilien entspricht den frühen Entwicklungsstadien des Photorezeptoren-Außengliedes. Es fehlen aber die für die letzteren so charakteristischen Membraninvaginationen; die Lamellenkomplexe der Vogelepiphyse haben eine ektopische Lage zur bulbösen Zilie. Oft finden sich an den Lamellenkörpern Degenerationszeichen. Diese degenerativ veränderten Strukturen erinnern an die Gebilde, die in den pinealen Lichtsinnesorganen der niederen Vertebraten aus zerfallenden Außengliedplättchen hervorgehen. Im Vergleich zu voll differenzierten pinealen Sinneszellen erscheinen die rezeptorenähnlichen Pinealocyten von P. domesticus rudimentär. Definitive funktionelle Schlüsse sind aus solchen morphologischen Vergleichen aber nicht möglich. Im Epiphysenstiel von P. domesticus verlaufen kräftige Nervenbahnen, die im Material dieser Studie ausschließlich aus marklosen Nervenfasern (Durchmesser 0,12–1,5 m) bestehen. Diese Faserzüge sind von autonomen Nervenstämmchen zu unterscheiden, die perivasculär die bindegewebige Hülle des Pinealorgans durchsetzen und stellenweise an das Parenchym vordringen. Im Pinealorgan von P. domesticus findet sich auch ein Pinealocytentyp mit 800–1200 Å großen granulierten Vesikeln, die im Golgi-Apparat dieser Zellen entstehen. Die elektronenmikroskopischen und neurohistologischen Befunde werden mit Hinweis auf verhaltensphysiologische (Gaston und Menaker) und elektrophysiologische (Ralph und Dawson) Ergebnisse diskutiert. Da das Pinealorgan von P. domesticus nach Menaker eine zentrale Komponente der biologischen Uhr beherbergt, sind Fragen nach einem sensorischen Eigenapparat und nach der sekretorischen Aktivität dieses Organs besonders aktuell.

---

Electron microscopic studies of the pineal organ in Passer domesticus

Summary The pineal organ of Passer domesticus contains cells with an inner segment, rich in mitochondria, from which a bulbous 9+0 type cilium originates. These cilia are often grouped together and may form 200 Å thick lamellae. Concentric or irregular whorl-like lamellar complexes arise from a number of such cilia. The structure of the bulbous cilia corresponds to that of early developmental stages of the photoreceptor outer segment. The characteristic membrane invaginations of the retinal cones are absent in the bulbous cilia of the avian pineal organ; the lamellar complexes have a position ectopic to the cilia. Signs of degeneration are present at the lamellar bodies. The degenerated forms resemble structures that arise from disintegrating outer segment plates in the pineal photoreceptor cells of lower vertebrates. The receptor-like pinealocytes of P. domesticus appear rudimentary when compared with the fully differentiated pineal sensory cells. Definitive functional interpretations are not possible from such morphological comparisons. In the pineal stalk of P. domesticus, nerve tracts are present consisting of unmyelinated fibers of 0.12–1.50 m diameter. These nerve tracts differ from autonomic nerves that traverse perivascularly the connective tissue of the pineal capsule and in places enter the pineal organ. In the pineal organ of P. domesticus, a cell type is also found containing 800–1,200 Å diameter granular vesicles which originate in the Golgi complex. The electron microscopic and neurohistological findings are discussed with reference to experiments by Gaston and Menaker (i.e. the effect of pinealectomy on the circadian locomotor rhythm of P. domesticus) and to electrophysiological results of Ralph and Dawson. Since, according to Menaker, the pineal organ of P. domesticus is a crucial component of the endogenous time-measuring system, questions concerning the presence of a sensory apparatus and secretory activity in this organ assume a special significance.

Eine kurze Zusammenfassung der Ergebnisse wurde im Seminar on Hypothalamic and Endocrine Functions in Birds, Tokio (19.–24. Mai 1969), vorgetragen.     

Opsin-immunoreactive outer segments and acetylcholinesterase-positive neurons in the pineal complex of Phoxinus phoxinus (Teleostei,Cyprinidae)

Summary The pineal complex of the teleost, Phoxinus phoxinus L., was studied light-microscopically by the use of the indirect immunocytochemical antiopsin reaction and the histochemical acetylcholinersterase (AChE) method.Opsin-immunoreactive outer segments of photoreceptor cells were demonstrated in large numbers in all divisions of the pineal end-vesicle and in the pineal stalk. Moreover, they were found in the roof of the third ventricle, adjacent to the orifice of the pineal recess as well as scattered in the parapineal organ. These immunocytochemical observations provide direct evidence of the presence of an opsin associated with a photopigment in the photosensory cells of the pineal and parapineal organs of Phoxinus. By means of the AChE reaction (Karnovsky and Roots 1964) inner segments of pineal photoreceptors, intrinsic nerve cells, several intrapineal bundles of nerve fibers, and a prominent pineal tract were specifically marked. The pineal neurons can be divided into two types: one is located near the pineal lumen, the other near the basal lamina. The latter perikarya bear stained processes directed toward the photoreceptor layer. A rostral aggregation of two types of AChE-positive nerve cells occurs in the ventral wall of the pineal end-vesicle. The main portion of the AChE-positive pineal tract, which lies within the dorsal wall of the pineal stalk, can be followed to the posterior commissure where some of the nerve fibers course laterally. A few AChE-positive pineal nerve fibers run toward the lateral habenular nucleus via the habenular commissure. In the region of the subcommissural organ single AChE-positive neurons accompany the pineal tract. The nerve cells of the parapineal organ exhibit a moderate AChE activity.These findings extend the structural basis for the remarkable light-dependent activity of the pineal organ of Phoxinus phoxinus. To the memory of Professor Karl von Frisch, pioneer and master in the field of photoneuroendocrine systemsThis investigation was supported by grants from the Deutsche Forschungsgemeinschaft to A.O. (Ok 1/24; 1/25: Mechanismen biologischer Uhren) and to H.-W. K. (Ko 758/1; 758–2)On leave from the 2nd Department of Anatomy, SOTE, Budapest, Hungary     

Rasterelektronenmikroskopische Beobachtungen an pinealen Sinneszellen der Forelle,Salmo gairdneri (Teleostei)

Zusammenfassung Im rasterelektronenmikroskopischen Bild des Pinealorgans vonSalmo gairdneri kann man drei verschiedene Außengliedtypen der Photorezeptoren unterscheiden. Diese Ergebnisse werden im Hinblick auf die Ultrastrukturkonzepte von Rüdeberg (1969) und Bergmann (1971) diskutiert. Rasterelektronenmikroskopische Studien erleichtern die anatomische Klassifizierung von pinealen Sinneszellen aufgrund ihrer Außengliedform.

---

Scanning electron microscopic observations of pineal photoreceptor cells in the trout,Salmo gairdneri (teleostei)

Summary The outer segments of pineal photoreceptor cells ofSalmo gairdneri were investigated with the scanning electron microscope. The scanning electron micrographs showed three different types of outer segments. These results are discussed with respect to the ultrastructural concepts of Rüdeberg (1969) and Bergmann (1971). Scanning electron microscopy permits better anatomical classification of pineal photoreceptor cells according to the form of their outer segments.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft (Arbeitsgruppe Prof. Dr. A. Oksche). Herrn Prof. Dr. G. Pfefferkorn, Direktor des Instituts für Medizinische Physik an der Westfälischen Wilhelms-Universität Münster, danke ich für einen Arbeitsplatz am Rasterelektronenmikroskop, Herrn Prof. Dr. H. G. Fromme, Münster, für die Unterstützung bei der präparativen Aufbereitung des Materials.     

Physiologische Untersuchungen und weitere Bemerkungen zur Struktur des lichtempfindlichen Pinealorgans von Pterophyllum scalare Cuv. et Val. (Cichlidae,Teleostei)

Zusammenfassung Im Einklang mit neuen Befunden von Hafeez (1971) bei Symphysodon aequifasciatus (Cichlidae) gehört das Pinealorgan von Pterophyllum scalare1 zum tubulärsensorischen Typ der Knochenfischepiphyse. — In Versuchen mit elektrophysiologischer Ableitung vom freigelegten Epiphysenstiel wurde eine direkte Lichtempfindlichkeit des Pinealorgans von Pterophyllum scalare beobachtet. Die Wirkung bestand in der Regel in einer Hemmung der Spontanaktivität, für die ein Photopigment mit einem Absorptionsmaximum bei 530 nm verantwortlich sein dürfte. Die Bestimmung der Lichtschwelle ergab Werte, die bei Berücksichtigung der Lichtverluste dafür sprechen, daß das Pinealorgan von Pterophyllum scalare bei Dunkelheit und in der Dämmerung aktiv ist, während die Impulsaussendung im vollen Tageslicht gehemmt wird. — Die Ableitungsstellen wurden an Hand von elektrolytisch gesetzten Eisenmarken histologisch lokalisiert.

---

Physiological studies and some further remarks on the structure of the photosensitive pineal organ of Pterophyllum scalare Cuv. et Val. (Cichlidae, Teleostei)

Summary The pineal organ of Pterophyllum scalare was classified as tubular and sensory. Our anatomical findings are in agreement with recent observations of Hafeez (1971) in Symphysodon aequifasciatus (Cichlidae). Electrical recordings from the exposed pineal stalk of Pterophyllum scalare, after removal of the lateral eyes and upon direct illumination of the diencephalic roof, indicated that: (1) the pineal organ of P. scalare is photosensitive, and (2) light mainly inhibits the spontaneous discharge of its nervous elements. Maximum sensitivity was observed at 525 nm. Prom estimates of the light absorbed by the tissue in front of the pineal organ and measurements of the light threshold of the exposed organ, it was concluded that the pineal organ of Pterophyllum scalare functions optimally in dim light, and that its activity is inhibited in daylight. The site of the recording electrodes was identified histologically by electrolytically-deposited iron marks.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft (Arbeitskreise E. Dodt und A. Oksche).     

Putative cholinergic elements in the photosensory pineal organ and retina of a teleost,Phoxinus phoxinus L. (Cyprinidae)

Summary Putative cholinergic neurons in the photosensory pineal organ of a cyprinid teleost, the European minnow, were studied by use of choline acetyltransferase (ChAT) immunocytochemistry and acetylcholinesterase (AChE) histochemistry. Pinealofugally projecting neurons were visualized using retrograde HRP-filling through their cut axons. For comparison, the distribution of choline acetyltransferase immunoreactivity (ChAT-IR) and AChE-positive elements in the retina was investigated.While the distributional patterns of ChAT-IR and strongly AChE-positive perikarya in the retina are similar and may represent the same neuronal population, ChAT-IR and AChE-positive elements in the pineal organ appear to belong to separate populations. In the retina, small- to medium-sized perikarya in the inner nuclear layer, and small perikarya in the ganglion cell layer are ChAT-IR and AChE positive. The entire inner plexiform layer is AChE positive, while only sublaminae 1, 2 and 4 are ChAT-IR. No indication of cholinergic activity was observed in the optic axon layer.In the pineal organ, ChAT-IR is restricted to small perikarya situated rostrally and dorsally in the pineal end-vesicle. AChE-positive neurons are present throughout the pineal end-vesicle and the pineal stalk. The pineal tract (the pinealofugally projecting axons of intrapineal neurons) is strongly AChE positive, but displays no ChAT-IR. The distribution of pinealofugally projecting neurons, labeled with retrogradely transported HRP, is markedly dissimilar to that of the ChAT-IR elements. It is proposed that the photosensory pineal organ transmits photic information to the brain via a non-cholinergic pathway. The possibility that the ChAT-IR neurons represent small local interneurons is discussed in the light of comparative physiological and anatomical findings.     

Pattern of synaptic connections in the pineal organ of the ayu,Plecoglossus altivelis (Teleostei)

Summary Synaptic connections were studied by means of electron microscopy in the sensory pineal organ of the ayu, Plecoglossus altivelis, a highly photosensitive teleost species. Three types of specific contacts were observed in the pineal end-vesicle: 1) symmetrically organized gap junctions between the basal processes of adjacent photoreceptor cells; 2) sensory synapses endowed with synaptic ribbons, formed by basal processes of photoreceptor cells and dendrites of pineal neurons; 3) conventional synapses between pineal neurons, containing both clear and dense-core vesicles at the presynaptic site. Based on these findings, the following interpretations are given: (i) The gap junctions may be involved in an enhancement of electric communication and signal encoding between pineal photoreceptor cells. (ii) The sensory synapses transmit photic signals from the photoreceptor cells to pineal nerve cells. (iii) The conventional synapses are assumed to be involved in a lateral interaction and/or summation of information in the sensory pineal organ. A concept of synaptic relationships among the sensory and neuronal elements in the pineal organ of the ayu is presented.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany     

Substance P-like-immunoreactive neurons in the photosensory pineal organ of the rainbow trout,Salmo gairdneri Richardson (Teleostei)

Summary Restricted numbers of substance P-like-immuno-reactive (SPL-IR) neurons were demonstrated in the photosensory pineal organ of the rainbow trout. The small parapineal organ of this teleost species receives a distinct SPL-IR innervation via the habenular nuclei, but displays no intrinsic SPL-IR neurons. Intrapineal SPL-IR neurons were located in the rostral portion of the pineal end-vesicle. Neuronal somata were found in a lateral position with smooth axonal processes extending mediad. Immunoreactive somata and axonal processes were observed intraparenchymally as well as in the pineal lumen. The pattern of immunoreactivity was not changed in excised pineal organs that had been incubated in tissue culture medium in the dark for 18 h. The possibility that the intrapineal SPL-IR neurons are not part of the neural circuitry involved in the transduction of photic information, but may have other functions, is discussed.Fellow of the Alexander von Humboldt-Stiftung, Bonn, Federal Republic of GermanySupported by research funds from the Deutsche Forschungsge-meinschaft (Ko 758/2-4)     

Synapse-like contacts between axons of the pineal tract and the subcommissural organ in Rana perezi (Anra) and their absence in Carassius auratus (Teleostei): ultrastructural tracer studies

The present study was designed to investigate the controversial subject of the existence of a neural input from the pineal organ via the pineal tract to the subcommissural organ (SCO) in teleosts and anurans. Horseradish peroxidase was injected into the pineal organ and pineal tract of Carassius auratus and Rana perezi. Within the pinealofugal fibers the tracer was visualized at the light-and electron-microscopic levels either by immunocytochemistry using an anti-peroxidase serum, or by revealing the enzymatic activity of peroxidase. In both species, labeled myelinated and unmyelinated fibers of the pineal tract were readily traced by means of electron microscopy. In R. perezi, numerous terminals contacting the SCO cells in a synapse-like (synaptoid, hemisynaptic) manner bore the label, whereas a different population of endings was devoid of the tracer, indicating that in this species the SCO receives a dual neural input, one of pineal origin, the other of unknown source and nature. In the SCO of C. auratus, neither labeled nor unlabeled synapse-like contacts were found. Thus, in this latter species, a direct neural input to the SCO is missing. It is concluded that the secretory activity of the SCO can be controlled by different mechanisms in different species, and that more than one neural input mechanism may operate in the same species.     

Weitere Untersuchungen zur Feinstruktur und Innervation des Pinealorgans von Passer domesticus L.

Zusammenfassung Im Parenchym der Epiphysis cerebri von Passer domesticus kommen Nervenzellen vor. Ihre Neuriten ziehen im langgestreckten Epiphysenstiel zur Commissura habenularum. Im proximalen Endabschnitt des Epiphysenstiels wird ein Teil dieser Fasern myelinisiert. Zwischen die Nervenfasern schieben sich zahlreiche Pinealocytenausläufer; synaptische Bänder helfen die letzteren eindeutig zu identifizieren. Im Bereich der synaptischen Bänder liegen: 1. 300 Å Vesikel, 2. 300 Å Vesikel und 800–1200 Å Granula, 3. nur 800–1200 Å große granulierte Vesikel. Die Tatsache, daß in Pinealocytenausläufern nebeneinander synaptische Bänder und Granula vorkommen, und daß apikal in zilientragenden Zellen ebenfalls Granula nachweisbar sind, spricht dafür, daß bei Passer domesticus ein Pinealzelltyp sensorische und sekretorische Strukturmerkmale besitzen kann. Außerdem werden Kontaktsynapsen beobachtet; ihre praesynaptischen Fasern enthalten die gleichen Strukturelemente wie die Fasern mit synaptischen Bändern. Die Zahl der Mikrofibrillen und Mikrotubuli variiert in den Pinealocytenausläufern, in den postsynaptischen Dendriten und in den Neuriten so stark, daß es mitunter schwierig ist, diese Fortsatztypen einwandfrei zu unterscheiden und die Zahl der zum Gehirn ziehenden Neuriten exakt zu ermitteln.Efferente sympathische Nervenfasern dringen in die Bindegewebssepten der Epiphyse ein. Sie enthalten Granula mit einem Durchmesser von 300–500 Å und 800–1200 Å. Nach Injektion von Nialamid zeigen beide Granulatypen einen elektronendichten Kern. Mikrospektrographisch ist Serotonin und Noradrenalin in diesen Nervenfasern nachweisbar. Das Material dieser Studie enthält keinen fluoreszenzmikroskopischen oder elektronenmikroskopischen Hinweis darauf, daß die sympathischen Nervenfasern durch die Basalmembran in den Zellverband des Epiphysenparenchyms eintreten. Im elektronenmikroskopischen Bild haben manche Pinealocytenausläufer eine Ähnlichkeit mit autonomen Nervenfasern.Die funktionelle Bedeutung der Vogelepiphyse als photo-neuro-endokrines Organ wird diskutiert.

---

Further investigations on the structure and innervation of the pineal organ of Passer domesticus L.

Summary The pineal organ of Passer domesticus contains nerve cells within its parenchyma. Axons of the nerve cells run within the elongated stalk of the pineal organ to the habenular commissure. At the proximal end of the stalk, some axons become myelinated. In the stalk, the axons intermingle with pinealocyte processes containing synaptic ribbons. The synaptic ribbons are in contact with (1) vesicles with a diameter of 300 Å; (2) 300 Å diameter vesicles and 800–1,200 Å diameter dense-core granules; or (3) the dense-core granules only. Dense-core granules are also present in pinealocytes with 9+0 type cilia. These results suggest that sensory and secretory structures are present in the same pineal cell type. Furthermore, conventional synapses are present between receptor and nerve cells: The presynaptic fibers have the same structure as the fibers containing synaptic ribbons. The numbers of microfibrils and microtubules vary among postsynaptic fibers (dendrites), the pinealocyte processes, and the neurites. Thus it is difficult to obtain an exact count of the number of axons running to the brain.Efferent sympathetic nerve fibers enter the pineal organ associated with the connective tissue surrounding blood vessels. The fibers show granules of 300–500 Å diameter or 800–1,200 Å diameter. After nialamide injection, both types of granules contain a dense core. Microspectrographically serotonin and noradrenaline are demonstrated in the sympathetic nerve fibers. There is no evidence found in the material of this study to suggest that sympathetic nerve fibers perforate the basement membrane and enter the parenchymal cell complexes of the pineal organ. Pinealocyte processes and sympathetic nerve fibers often show a very similar ultrastructural pattern.The role of the avian pineal organ in photo-neuro-endocrine regulation is discussed.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Photoreceptor development in the pineal organ and the eye of Plecoglossus altivelis and Paralichthys olivaceus (Teleostei)

Summary The ontogenetic apperance of pineal photo-receptors was compared with that of retinal photoreceptors in the ayu Plecoglossus altivelis and the lefteye flounder Paralichthys olivaceus, which hatched 10 days and 3 days after fertilization, respectively. Despite the disparity in incubation time, the outer segments (containing membranous lamellae) of the pineal photoreceptors first appeared from 3 to 4 days after fertilization in both species. In contrast, the outer segments of the retinal photoreceptors first became visible 5 to 6 days after fertilization, although a characteristic retinal stratification and the optic tract leaving the ganglion cell layer were already found 4 days after fertilization in both species. The functional significance of these temporal disparities and/or similarities in photoreceptor development are discussed with special reference to the timing of daily rhythmic activities during the early developmental period of the teleosts.The results were presented at the Annual Meeting of the Japanese Society of Scientific Fisheries on April 2, 1990 (Tokyo)     

Epidermale Organe am Kopf von Morulius chrysophakedion (Cyprinidae,Ostariophysi, Pisces)

Zusammenfassung Im Kopfbereich von Morulius chrysophakedion liegen epidermale Organe in Gruppen beisammen. Im Gegensatz zu den oberflächlichen Perlorganen anderer Cyprinidae handelt es sich hierbei um voluminöse Einsenkungen der Epidermis in das darunter gelegene Bindegewebe, von dem aus gefäßführende Papillen in das Organinnere hineinziehen. Elektronenmikroskopische und histochemische Befunde sprechen für eine relativ hohe Stoffwechselaktivität der das Organ aufbauenden Polsterzellen, als deren Ergebnis eine Art Verhornung an der Organspitze vorliegt. Hinweise auf eine Funktion der epidermalen Organe von Morulius als Sinnesorgane wurden nicht gefunden; sie müssen daher von den Fahrenholzschen Organen der Dipnoi und Brachiopterygii unterschieden werden.

---

Epidermal organs on the head of Morulius chrysophakedion (Cyprinidae, Ostariophysi, Pisces)Histochemical and electron microscopical study of the so called flask-shaped organs of Labeo

Summary There are groups of epidermal organs in the head region of Morulius chrysophakedion. In contrast to the superficial perlorgans of some other Cyprinidae these organs are formed by an infolding of epidermis into the connective tissue beneath. From this tissue vessels reach into the interior of the organ. Electron microscopical and histochemical data give evidence of a relatively high metabolic activity in the Polsterzellen from which the organ is built up. These cells give rise to a keratinized area at the top of the organ. There is no evidence that the epidermal organ has the function of a sense organ as in the organs of Fahrenholz in Dipnoi and Brachiopterygii.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Vergleichende histophysiologische Tracer-Untersuchungen an verschiedenen Organsystemen von Branchiostoma lanceolatum (Cephalochordata) und Brachydanio rerio (Teleostei)

Zusammenfassung In vergleichend autoradiographischen Untersuchungen wurde der Einbau von percutan applizierten 3H-Uridin, 3H-Histidin und 3H-Glucose in die wichtigsten Organsysteme (Epidermis, ZNS, Muskeln, Chorda, Leber, Kiemendarm, Darm) von Branchiostoma lanceolatum (Acranier) und Brachydanio rerio (Teleosteer) nach Inkorporationszeiten von 11 min bis zu 7 Tagen verfolgt. Der Stoffwechsel der markierten Substanzen in den einzelnen morphologisch miteinander zu homologisierenden Organen war bei den beiden Spezies sehr ähnlich, bei Branchiostoma allerdings (mit Ausnahme des ZNS) 4–5mal stärker als bei Brachydanio. Bei dem letzteren wurde außerdem eine zeitliche Verzögerung in der Tracer-Aufnahme (lag-Phase) beobachtet. Insbesondere der ZNS-Stoffwechsel von Acraniern zeigte ähnliche Charakteristika wie der von Vertebraten: Verbleib des Hauptanteils der neusynthetisierten RNS im Perikaryon, axonalen Protein-Transport, Vorwiegen der Glykogensynthese in den Nervenfaserendformationen. Allerdings fanden sich im ZNS von Branchiostoma niedrigere Stoffwechselraten als im ZNS von Brachydanio.

---

Comparative histophysiological investigations of different organs in Branchiostoma lanceolatum (Cephalochordata) and Brachydanio rerio (Teleostei)

Summary Incorporation of 3H-uridine, 3H-histidine and 3H-glucose into some organs (epidermis, CNS, muscles, spinal cord, notochord, liver, gills, intestine) of Branchiostoma lanceolatum (Acrania) and Brachydanio rerio (Teleostei) was investigated by means of comparative autoradiograms following incorporation periods of 11 min to 7 days. The metabolism of the labeled substances in the various homologous organs examined was quite similar, although it was 4 to 5 times higher in Branchiostoma than in Brachydanio; in the latter there was also a delay of tracer incorporation of about 3 hrs, a so-called lag-phase. In particular the metabolism of the CNS of Branchiostoma showed the same characteristics as the CNS of vertebrates, e. g. storage of neuronally synthesized RNA in the neuronal perikarya, axonal flow of proteins, glycogen synthesis in nerve endings. However, metabolic activity of the CNS was lower in Branchiostoma.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Pineal neurons projecting to the brain of the rainbow trout,Salmo gairdneri Richardson (Teleostei)

Summary The morphology of intrapineal neurons that give rise to the pineal tract and project to the brain in the rainbow trout was visualized by the use of neuronal backfilling with horseradish peroxidase (HRP). The tracing was performed on excised pineal organs under in-vitro conditions at 4° C, with filling times ranging from 6 to 24 h. Large multipolar, bipolar and unipolar neurons were visualized in the rostral tip of the pineal organ (pineal ganglion). These neurons possessed extended dendritic trees participating in the formation of a circumscribed neuropil-like area. Throughout the pineal organ small bipolar elements were the most ubiquitous type of neuron, however, with markedly smaller numbers in the proximal portion of the pineal end-vesicle. In the pineal stalk, some bipolar neurons were observed to contact the pineal lumen, which is continuous with the third ventricle, via dendritic processes of various types. It could not be established whether any of these CSF-contacting processes were identical with photoreceptor outer segments. The basal processes of the bipolar neurons sometimes possessed distally projecting collaterals. In conclusion, it has been shown that (i) different types of neurons displaying varied patterns of regional distribution contribute to the pineal tract, and (ii) certain CSF-contacting neurons in the pineal organ send axonal processes directly toward the brain.Supported by Research grant Ko 758/2-4 from the Deutsche ForschungsgemeinschaftFellow of the Alexander von Humboldt Foundation, Bonn, Federal Republic of Germany     

The outer segments of photoreceptive pinealocytes in the pineal organ of the funa,Carassius gibelio langsdorfi

Summary The form and size of the outer segments of photoreceptive pinealocytes in the pineal organ of the funa, Carassius gibelio langsdorfi, were observed with the scanning electron microscope. The height of the outer segments measures between 1 and 3 m and the diameter varies widely from 1.5 to 8 m. Various forms of outer segments, i.e. a slender type, a dome-like type, a cap-like type and a helical type, were demonstrated. The parallel-oriented filamentous processes of the inner segments have the same length as the outer segments and a diameter of approximately 100 nm; they are projections from the apical border of the inner segment and surround the cone-like outer segments. The processes make a right angle with the lamellar disks. The distance between two processes averages 100 nm. The lamellar disks of the outer segments are oriented at right angles to the modified cilium in the basal part, but the angle often changes in the peripheral part, where the lamellar disks are raised and become parallel to the cilium.Supported by a fellowship from the Japan Society for the Promotion of Science and a grant from the Deutsche Forschungsgemeinschaft to M. UeckSupported by a grant from the Ministry of Education of Japan to K.Wake     

GABA-immunoreactive neurons in the photosensory pineal organ of the rainbow trout: two distinct neuronal populations

Summary The distribution of putative GABA-ergic neurons in the photosensory pineal organ of the rainbow trout was investigated by use of a specific antiserum against -aminobutyric acid (GABA). GABA-immunoreactive (GABA-IR) neurons were located in the rostral portion of the pineal end-vesicle, presumably constituting a population of interneurons. GABA-IR neurons were also found in the pineal stalk. The axons of these neurons were traced along the pineal stalk toward the brain. The terminal areas of these axons could not be established. GABA-IR glial cells were observed in the pineal end-vesicle, but not in the pineal stalk.     

Phylogenetic relationships of the enigmatic ornate shiner,Cyprinella ornata,a species endemic to Mexico (Teleostei: Cyprinidae)

The systematics and taxonomy of North Americancyprinid fishes has historically been said tobe in a chaotic state of affairs. Much of theconfusion as to relationships of species restsin the lack of explicit phylogenetic hypothesesof species and reliance upon degree ofdistinctiveness of taxa or their overallsimilarity for generic placement. Some specieshave had more turbulent or variable taxonomichistories than others. The ornate shiner, Cyprinella ornata, is one of those species,having been placed in the genera Notropis,Codoma, and Cyprinella within the last20 years and found in current texts in any ofthese three genera. Most of the confusionregarding placement of this species has beenrelated to lack of explicit phylogenetichypotheses to formulate its classification, butto a certain degree some researchers haveignored phylogenetic studies and preferred torely upon its morphological distinctiveness orreproductive behavioral traits of questionablehomology to place it in a monotypic genus andalign it with the genera Pimephales andOpsopoeodus.I present a phylogenetic analysis based oncomplete Cytochrome b sequences fromornate shiners, other species of Cyprinella, purported relatives, and severalother species of the Shiner Clade to determinethe phylogenetic affinities of this enigmaticspecies. Molecular analysis reveals the ornateshiner to be more closely related to species ofCyprinella than to a Pimephalesplus Opsopoeodus clade as previouslydiscussed in one morphological analysis, or asargued by Page and Johnston (1990), Johnstonand Page (1992) and Page and Ceas (1989) basedon observations of spawning behaviors andhypotheses of homology between crevice-spawningand egg-clumping behaviors. This molecularanalysis is more consistent with earliermorphological phylogenetic hypotheses of Mayden(1989) wherein these two clades are notconsidered to be closely related and the ornateshiner is a member of the genus Cyprinella.     

Mitochondrial diversity of Opsariichthys bidens (Teleostei, Cyprinidae) in three Chinese drainages

We describe the phylogeographic structure of 28 Chinese populations of the cyprinid Opsariichthys bidens across three main Chinese river drainages. Our study is based on the phylogenetic analysis of the complete mitochondrial cytochrome b gene (1140 bp). We combined this analysis with population processes inferred from nested clade analysis (NCA) and mismatch distributions. Both analyses showed that Chinese O. bidens consists of five mtDNA lineages (Opsariichthys 1-5) with high genetic divergence among them. Molecular divergences (TrN+G) higher than 20% among the Opsariichthys 1-5 mtDNA lineages suggest a taxonomic underestimation at the species level. About 92% of the genetic variance among samples was explained by differences among Opsariichthys mtDNA lineages. Drainage-restricted haplotypes with high frequencies and moderate nucleotide diversity show that Opsariichthys populations have evolved independently. NCA results were congruent with the phylogeny, and unimodal mismatch distributions with negative Tajima's D values suggest population expansions in some Opsariichthys lineages. The phylogeographic structure of the Opsariichthys 1-5 mtDNA lineages appears to be related to their long-term interruption of gene flow (theta(ST)>0.97). Our results suggested that fragmentation of ancestral ranges might have caused Opsariichthys diversification in Chinese waters. However, current distribution of common haplotypes across the Yangtze and Pearl drainages suggests a recent river connection that could have favoured gene flow across drainages. Overall, the results indicated that the richness of current Asian widespread species might have been underestimated, and that the cyprinid populations of O. bidens in the Yangtze, Pearl and Hai He drainages may correspond to five species.