Manfred Ueck

Uptake of peroxidase from the third ventricle by ependymal cells of the median eminence

SummaryPeroxidase injected into the subarachnoid space in mice is absorbed by ependymal cells of the median eminence. The ependymal cells of the median eminence of the rat and Japanese quail absorb peroxidase injected into the third ventricle. The processes of these ependymal cells terminate at the capillaries of the primary plexus or those surrounding the ventromedial nucleus of the hypothalamus. In all three species, peroxidase is absorbed by the ependymal cells of the paraventricular organ and by those in close proximity to it. Some ependymal cells send processes to the capillaries in the lateral nucleus of the hypothalamus. These phenomena are discussed in relation to adenohypophysial function.

Acetylcholinesterase-containing nerve cells in the pineal complex and subcommissural area of the frogs, Rana ridibunda and Rana esculenta.

SummaryIn Rana esculenta and Rana ridibunda the frontal organ and the pineal organ (epiphysis cerebri) form a pineal complex. Approximately 60 nerve cells of the frontal organ and 220–320 nerve cells of the pineal organ display a positive acetylcholinesterase reaction (Karnovsky and Roots, 1964). The dorsal wall of the pineal organ is considerably richer in acetylcholinesterase-positive neurons than the ventral wall (ratio 3∶1); a group of unusually large-sized nerve cells occurs in the rostral portion of the frog pineal. Two different types of nerve cells were observed in the pineal complex: multipolar and pseudounipolar elements. The former are embedded in the pineal parenchyma and their processes penetrate radially into the plexiform layer, whereas the latter are distributed along the roots of the pineal tract near the basal lamina. The ratio of the multipolar to pseudounipolar neurons is 1∶4 for the frontal organ and 3∶5 for the pineal organ. The multipolar elements may be interneurons; the pseudouni-polar cells send one of their processes into the pineal tract. At the caudal end of the pineal organ 30–50 unipolar nerve cells are clustered in juxtaposition with the pineal tract, and other 30–50 unipolar neurons are scattered along the basis of the subcommissural organ. Some of these nerve cells emit their processes toward the mesencephalon and others toward the pineal organ via the pineal tract. The results are discussed with respect to previous physiological and morphological findings on the pineal complex of Anura.

Vergleichende Untersuchungen über acetylcholinesterase-haltige Neurone im Pinealorgan der Vögel

SummaryThe acetylcholinesterase reaction of the pineal organ (epiphysis cerebri) varies among avian species. Acetylcholinesterase-positive neurons with axons running along the pineal stalk to unidentified brain centers are seen in the pineal organ of passeriform birds such as Passer domesticus, Serinus canaria, and Amadina faciata alexander. A different kind of acetylcholinesterase-positive fibers are observed running along the blood vessels to the pineal organ. The acetylcholinesterase-positive structures are much reduced in Columba livia. The pineal parenchyma in Columba livia shows a slightly brown color in the light microscope; spot-like deposits of the reaction product are seen between the pinealocytes in the electron microscope. In Passer domesticus, the reaction products are seen in the pinealocyte endoplasmic reticulum as well as at the outer membrane of the perikaryon and axon of the nerve cells. The results in three-day old chicken and in Excalfactoria chinensis are similar to those obtained in Columba livia. In Coturnix coturnix japonica, a heavy reaction is evident in the basal part of the follicles. A small number of nerve cells and fibers are seen in the pineal stalk of Melopsittacus undulatus. The results are discussed in respect of sensory and secretory functions of the pineal organs in passeriform birds.ZusammenfassungIm Pinealorgan der Vögel zeigen die acetylcholinesterase-positiven Strukturen speziesabhängige Unterschiede. Die passeriformen Arten Passer domesticus, Serinus canaria und Amadina faciata alexander besitzen acetylcholinesterase-positive Nervenzellen, deren Axone im Epiphysenstiel hirnwärts ziehen; außerdem kommen an den Blutgefäßen spezifisch tingierbare Nervenfasern vor, die offenbar zur Epiphyse verlaufen. Bei Columba livia sind diese Nervenstrukturen sehr stark reduziert. Dagegen zeigt das Parenchym der Taubenepiphyse lichtmikroskopisch eine schwache Anfärbung; diese Stellen manifestieren sich im elektronenmikroskopischen Bild als punktförmige interzelluläre Reaktionsorte zwischen einzelnen Pinealzellen. Ferner finden sich bei Passer domesticus elektronenmikroskopisch nachweisbare Reaktionsprodukte im endoplasmatischen Reticulum und im perinukleären Raum der Pinealocyten (=Pinealzellen) sowie am Plasmalemmüberzug (Außenmembran) der Perikaryen und der Axone der Nervenzellen. Die Befunde bei 3 Tage alten Küken (Gallus domesticus) und bei Excalfactoria chinensis ähneln denen bei Columba livia, während bei Coturnix coturnix japonica das Reaktionsprodukt vermehrt im basalen Abschnitt der Follikel liegt. Melopsittacus undulatus zeigt im Epiphysenstiel eine begrenzte Zahl von hirnwärts ziehenden Nervenfasern und nimmt zwischen den Passeriformes und der Taube etwa eine Mittelstellung ein. Die sensorische und sekretorische Kapazität des Pinealorgans von Passeriformes wird diskutiert.

Ultracytochemical localization of Ca++-ATPase activity in the paraphyseal epithelial cells of the frog, Rana esculenta.

SummaryCa++-ATPase activity was studied ultracytochemically (cf. Ando et al. 1981) in the paraphysis cerebri of the frog. An intense reaction was demonstrated on the plasmalemma of the microvilli at the apical pole of paraphyseal cells; in contrast, the basolateral plasmalemma showed only a slight staining. In addition, mitochondria, gap junctions, cilia, and cytoplasmic elements (e.g., microfilaments) displayed Ca++-ATPase activity. Variation of the Ca++-concentration in the incubation medium from 0.1 mM to 100 mM altered the Ca++-ATPase activity of the cell organelles. The substitution of Ca-by Mg-ions resulted in a conspicuous decrease in the enzyme activity, especially on the apical plasmalemma. Ca++-ATPase activity is claimed to be involved in a number of extra-and intracellular functions. In comparison to the epithelium of the adjacent choroid plexus the paraphyseal epithelial cell is thought to be a principal Ca-ion regulator of the cerebrospinal fluid in frogs.

Fluoreszenzmikroskopischer Nachweis biogener Monoamine in der Epiphysis cerebri von Rana esculenta und Rana pipiens

SummaryThe pineal organ of Rana pipiens and Rana esculenta was studied by fluorescence microscopy for the histochemical demonstration of certain biogenic monoamines. Under normal conditions, no fluorogenic amines were visible in the organ. After pretreatment of the animal with a monoamine-oxidase inhibitor, nialamide, an intense yellow formaldehyde-induced fluorescence appeared both in the sensory cells and in the supporting cells. Microspectrofluorometric analysis indicated that the fluorophore is identical with 5-hydroxytryptamine; the presence of other closely related indoles, such as 5-hydroxytryptophan and melatonin, however, cannot be excluded. Fluorescent adrenergic nerves were found in the connective tissues surrounding the pineal organ; fluorescent fibers were observed also in the pineal parenchyma.The presence of 5-hydroxytryptamine in the anuran pineal organ is discussed with regard to the role that the amine plays in melatonin synthesis and with regard to a possible functional relation to some as yet unidentified protein- or polypeptid-hormone within the pineal parenchyma.ZusammenfassungDie Epiphysis cerebri von Rana pipiens und Rana esculenta wurde fluoreszenzmikroskopisch auf das Vorhandensein bestimmter biogener Amine untersucht. Unter normalen Bedingungen lassen sich im Parenchym der Froschepiphyse keine Amine fluoreszenzmikroskopisch darstellen. Nach Vorbehandlung der Tiere mit dem Monoaminoxydase-Hemmer Nialamid ist aber eine intensive, durch Formaldehyd-Behandlung induzierte Gelbfluoreszenz in den Sinnes- und Stützzellen zu beobachten. Mikrospektrofluorometrische Messungen zeigen, daß das Fluorophor mit 5-Hydroxytryptamin identisch ist, obwohl das Vorkommen von anderen verwandten Indolen — z.B. 5-Hydroxytryptophan und Melatonin — nicht ausgeschlossen werden kann. Grünfluoreszierende adrenerge Nervenfasern sind im meningealen Hüllbindegewebe der Epiphyse zu erkennen; einige dieser Fasern scheinen auch in die Epiphyse einzudringen.Der Nachweis von 5-Hydroxytryptamin wird im Zusammenhang mit der Frage einer Melatoninsynthese diskutiert. Das 5-Hydroxytryptamin könnte außerdem in einer funktionellen Beziehung zu einem bisher noch unbekannten Protein- der Polypeptid-Hormon des Epiphysenparenchyms stehen.

Weitere experimentelle und neuroanatomische Untersuchungen an den Nervenbahnen des Pinealkomplexes der Anuren

SummaryThe pineal systems of 32 frogs of the species Rana temporaria, Rana esculenta, and Rana catesbeiana were studied first electrophysiologically and then with histological and electron microscopical methods. The nervus pinealis (= tractus frontalis) showed two kinds of responses to photic stimulation. Uninterrupted conduction of nerve impulses from the frontal organ to the midbrain was demonstrated by antidromic electrical stimulation of the intracranial pineal tract and recording of action potentials from the extracranial pineal nerve. Beside the uninterrupted fibres from the frontal organ to the tractus pinealis, other fibres of the nervus pinealis either did not reach the pineal organ or ended in the pineal organ. The extracranial part of the nervus pinealis contained afferent as well as efferent fibres.Morphologically it was shown in 32 investigated animals that the nervus pinealis penetrates the skull and runs to the pineal organ. During its course, the diameter of the pineal nerve became smaller, which was related to a decrease of the number of myelinated and unmyelinated fibres as revealed by the electron microscope. In comparison with the extracranial portion of the pineal nerve, the intracranial portion of the nerve near the pineal organ contained only half of the myelinated and one third of the unmyelinated fibres counted in one specimen of Rana catesbeiana. The thickness of the myelin sheats of the residual myelinated fibres was much reduced. The nervus pinealis entered the rostral region of the pineal organ, and single nerve fibres were seen to penetrate the basal lamina of the pineal organ reaching its parenchyma dorsomedially. These fibres, and axons of the nerve cells of the pineal organ, constituted the pineal tract (see also Paul et al., 1971). In one specimen of Rana esculenta, only ∼2% of the nerve fibres of the tractus pinealis originated in the frontal organ. The results have been discussed in relation to the generation and conduction of the chromatic and achromatic responses in the pineal system.ZusammenfassungDer Epiphysenkomplex von 32 Fröschen der Arten Rana temporaria, Rana esculenta und Rana catesbeiana wurde zuerst elektrophysiologisch, anschließend neuro-histologisch bzw. elektronenmikroskopisch untersucht. Auf Belichtung des Stirnorgans lassen sich vom Nervus pinealis (= Tractus frontalis) zwei Arten von Antworten ableiten. Antidrome elektrische Reizung des Tractus pinealis zeigt im Nervus pinealis fortgeleitete Aktionspotentiale und damit Nervenfasern, die vom Stirnorgan bis zum Mittelhirn durchgehen. Elektrophysiologisch wurden außerdem Fasern ermittelt, die die Epiphysis cerebri nicht erreichen. Weitere Fasern enden in der Epiphyse. Neben den vom Stirnorgan zum Gehirn ziehenden afferenten Fasern kann man im extracranialen Teil des Nervus pinealis auch efferente Fasern nachweisen.Histologisch ist in allen Fällen der Durchtritt des Nervus pinealis durch das Schädeldach zu beobachten. Der Durchmesser des Nerven nimmt jedoch wahrend seines Verlaufs kontinuierlich ab. Elektronenmikroskopisch konnte gesichert werden, daß diese Durchmesserabnahme auf einem Faserverlust beruht. Bei einer Rana catesbeiana wurden kurz vor Eintritt des Nervus pinealis in die Epiphyse — im Vergleich zum extracranialen Nervenabschnitt —nur noch die Hälfte der markhaltigen und 1/3 der marklosen Nervenfasern gezählt. Bei den verbleibenden markhaltigen Fasern war außerdem die Dicke der Markscheide stark reduziert. Der Nervus pinealis fächert sich in Epiphysennähe auf; seine Fasern treten dorsomedian einzeln durch die Basalmembran in das Parenchym des rostralen Epiphysenabschnittes ein. Gemeinsam mit den Neuriten der Epiphysennervenzellen bilden sie den Tractus pinealis (s. auch Paul u. Mitarb., 1971). Eine synaptische Umschaltung der Fasern des Nervus pinealis konnte in der Epiphyse nicht nachgewiesen werden. Bei einer Rana esculenta betrug der Faseranteil des Nervus pinealis am Tractus pinealis nur etwa 2%. Die Befunde werden im Zusammenhang mit der Entstehung und Leitung der chromatischen und achromatischen Antwort im Epiphysenkomplex diskutiert.

Histochemical and cytochemical demonstration of Ca++-ATPase activity in the stellate cells of the adenohypophysis of the guinea pig

SummaryThe histo- and cytochemical localization of Ca++-ATPase activity in the adenohypophysis of the guinea pig was studied utilizing a newly developed method (Ando et al. 1981). An intense reaction was observed in the wall of the blood vessels and between non-secretory cells (stellate cells) and endocrine cells of the pars distalis. Under the electron microscope the Ca++-ATPase reaction product was located extracellularly in relation to the plasmalemma of the stellate cells. This reaction was dependent on Ca++ and the substrate, ATP, and reduced by the addition of 0,1 mM quercetin to the standard incubation medium. Preheating of the sections before incubation completely inhibited the enzyme activity. When Mg++ in different concentrations were substituted for Ca++ in the incubation medium the reaction was always reduced. Both Ca++ and Mg++ in the incubation medium also reduced the reaction. The plasmalemma of the endocrine cells contains no demonstrable amount of Ca++-ATPase activity. The function of the Ca++-ATPase activity is discussed in relation to the regulation of the extracellular Ca++ concentration which seems to be important with respect not only to the secretory process of the endocrine cells but also to the metabolism of the adenohypophysis.

Ultracytochemical localization of Ca++-ATPase activity in pituicytes of the neurohypophysis of the guinea pig.

SummaryCa++-ATPase activity (cf. Ando et al. 1981) was examined both light- and electron-microscopically in the neurohypophysis of the guinea pig. Apart from a strong activity within the walls of the blood vessels, in the parenchyma of the neurohypophysis the reaction product of the Ca++-ATPase activity was restricted to the plasmalemma of the pituicytes. This reaction was completely dependent upon Ca++ and the substrate, ATP; the reaction was inhibited by 0.1 mM quercetin, an inhibitor of Ca++-ATPase. A reduction of the enzyme activity occurred by 1) adding Mg++ to the standard incubation medium, and 2) substituting Ca++ with Mg++ at varing concentrations. In all experiments the neurosecretory fibers were devoid of Ca++-ATPase activity. The function of the Ca++-ATPase activity in the plasmalemma of the pituicytes is discussed in connection with the regulation of the extracellular Ca++ concentration, which seems to be important with respect to the discharge of secretory material from the neurosecretory fibers.

Cerebrospinal fluid-contacting neurons and other somatostatin-immunoreactive perikarya in brains of tadpoles of Xenopus laevis

SummaryIn untreated tadpoles of Xenopus laevis, stage 60 (Nieuwkoop and Faber 1956), somatostatin-immunoreactive perikarya and fiber networks are already discernible in the pallium and the septum, as well as in the anterior and posterior hypothalamus. In addition, somatostatin-immunoreactive cerebrospinal fluid (CSF)-contacting neurons were found in the periventricular gray of the most caudal division of the ventral tuber cinereum. The results are discussed with respect to the inhibitory influence of the system of CSF-contacting and other somatostatin-immunoreactive neurons on the anterior pituitary and other target sites, especially during the climax of metamorphosis.

A new histo-and cytochemical method for demonstration of cyclic 3′, 5′-nucleotide phosphodiesterase activity in retinal rod photoreceptor cells of the rat

SummaryCyclic 3′, 5′-mononucleotide phosphodiesterase (cyclic nucleotide PDEase) activity was studied histo-and cytochemically in the retinal rod photoreceptor cells of the rat by means of a newly developed technique utilizing the intrinsic 5′ nucleotidase activity instead of an exogenous 5′ nucleotidase source (snake venom). Cyclic GMP and cyclic AMP were used as substrates. When cyclic GMP was used as a substrate, the intense activity of phosphodiesterase (PDEase) was distributed over the entire rod outer segments; reaction product was observed on the plasmalemma and on the disk membranes of the outer segments. A slight reaction was also observed on the plasmalemma of the inner segments. However, no precipitate was found in the perinuclear and synaptic regions of the rod photoreceptors. In contrast, when cyclic AMP was utilized as a substrate, a moderate reaction was seen in the synaptic region of the plexiform layer. The intensity of the reaction in the outer segments was much reduced in comparison to the results with cyclic GMP. The enzyme activity was almost completely inhibited by 2 mM 3-isobutyl-1-methylxanthine (IBMX) or 2 mM theophylline, which were potent inhibitors of PDEase.To confirm the propriety of our new cytochemical method, the localization of 5′ nucleotidase was also studied utilizing 5′ AMP or 5′ GMP as substrates. In contrast to the activity of cyclic nucleotide PDEase, the activity of 5′ nucleotidase was distributed on all membranes of the photoreceptors from the synaptic outer plexiform layer to the tip of outer segments. After inhibition of the intrinsic 5′ nucleotidase activity with the use of 1 mM Ni-ions or 10 mM NaF no demonstration of cyclic nucleotide PDEase activity was possible; the existence of intrinsic 5′ nucleotidase activity is necessary for the release of free phosphateions from 5′ AMP (5′ GMP), which are a prerequisite for the histochemical reaction. For comparison, some sections were incubated with the conventional cyclic nucleotide PDEase incubation medium containing snake venom from Ophiophagus hannah. With this conventional method, morphological preservation was extremely poor, and moreover, the reaction itself was weaker than that with the presently described method.