Effect of long-term suppression and long-term stimulation of ACTH secretion on the ultrastructure of tanycytes[]

The ultrastructure of the tanycyte ependyma in males of Wistar albino rats (160--180 g of weight) was studied under experimental long-term suppression of ACTH secretion and its long-term stimulation. The former effect was accomplished by daily (for 8 days) intraperitoneal administration of dexamethasone phsophate; the stimulation of ACTH secretion was achieved by bilateral adrenalectomy. The long-term suppression of the hypophyseal adrenocorticotrophic function was followed by the ultrastructural markers of an increased transport and secretory activity in the tanycytes (mainly in beta-tanycytes, i. e. the ependyma lining floor of the third ventricle): the increased amount of microvesicles in the Golgi area and in the vicinity of the particular membranous cavity system (MCS, according to the authors' definition), the appearance of some figures of exocytosis, the elevated amount of lipid inclusions. On the contrary, the long-term stimulation of ACTH secretion was followed by the ultrastructural changes of the opposite character. The data obtained are in agreement with the previously established fact that there is a negative correlation between the tanycyte activity and the hypophyseal adrenocorticotrophic activity. It is supposed that this correlation is regulated by a feedback mechanism and attest to the involvement of tanycytes in the inhibitory control of ACTH secretion. A possible substrate inhibiting ACTH secretion by the tanycytes is discussed.     

Tanycyte and vascular patterns in the basal hypothalamus of Coturnix quail with reference to their possible neuroendocrine significance

Summary A well developed system of ependymal glial cells with long basilar processes stretching to the surface of the brain (tanycytes, Horstmann, 1954) has been described in the basal hypothalamus of Coturnix quail. The tanycytes both in the median eminence and the ventro-lateral hypothalamus form a link between the third ventricle and the hypophysial circulation. The processes of the ventro-lateral tanycytes terminate in the region of the infundibular sulcus in apposition to a loose network of vessels which are continuous with the primary plexus of the hypophysial portal system.Within the median eminence, the subependymal capillary network connects the vasculature of the contra-lateral sides of hypothalamus. There are no direct connections with the hypophysial portal vessels.With the aid of the light and electron microscope the ventricular ependyma was divided into a dorsal typical region and two glandular regions (ventro-lateral and ventral). Each region contains different forms of tanycyte. One of the two forms of tanycyte (designated type 3) associated with the ventro-lateral glandular ependyma has no contact with the third ventricle.Ultrastructural studies on the glandular ependyma failed to show any obvious differences between castrated, oestrogen or testosterone implanted, and sexually mature or immature quail.The possibility that the tanycyte-vascular system may have a neuroendocrine role is discussed.I am indebted to Professor A. Oksche, Dept. of Anatomy, University of Giessen for providing research facilities and to The Royal Society for additional financial support.     

Modulation of adrenal cell functions by cadmium salts. 1. Cadmium chloride effects on basal and ACTH-stimulated steroidogenesis

Cultured Y-1 mouse adrenal tumor cells, which secrete 20--hydroxy-4-pregnen-3-one (20-DHP), were used to investigate the acute nonlethal effects of incremental cadmium chloride (CdCl₂) concentrations on basal and maximally stimulated steroid secretion. In addition, cumulative CdCl₂ effects during 4-hr incubations, effect reversibility, and viability were determined. Cells were incubated in 1 ml serum-free Eagle's Minimal Essential Medium (FMEM) with or without 0.5 IU (ca. 1.5 M) adrenocorticotropin (ACTH) in the presence or absence of CdCl₂. Following incubation, cell viability was quantitated using trypan blue exclusion. The 20-DHP secreted into the experimental incubation medium was measured by radioimmunoassay. CdCl₂ levels of 10.0 g/ml or greater significantly inhibited basal 30 min steroid secretion in a dose-dependent manner; ACTH-stimulated steroid secretion was significantly inhibited by levels 5.0 g/ml or greater. At least 80% of all control and stimulated cells in the presence or absence of cadmium ions excluded trypan blue. The reduction in ACTH-stimulated steroid secretion was greater than the reduction in basal steroid secretion at any cadmium concentration level. The CdCl₂ concentration that reduced stimulated steroid hormone secretion by 50% (IC₅₀) was 45.0 g/ml. Exposing Y-1 cells to either 5.0, 10.0, 45.0 or 500.0 g CdCl₂/ml FMEM for periods ranging from 0.5 to 4 hr inhibited ACTH-stimulated steroid secretion in a time-dependent manner. After 30 min exposure to 10.0, 45.0 or 500.0 g CdCl₂/ml FMEM with or without ACTH, cadmium inhibition was irreversible. When 5.0 g CdCl₂/ml was used, basal and stimulated inhibition was reversible by reincubating in medium containing ACTH alone. The relatively greater cadmium effects on ACTH stimulated steroidogenesis might suggest that cadmium modulated the rate-limited transducing system between the ACTH plasma membrane receptor complex and cholesterol side-chain cleaving mitochondrial enzymes. However, cadmium influences on basal secretion indicated effects on the non-rate-limited steroidogenic pathway.Abbreviations ACTH adrenocorticotropin - ANOVA analysis of variance - CdCl₂ cadmium chloride - Ci Curie - DNA deoxyribonucleic acid - FMEM serum-free Eagle's Minimum Essential Medium - HEPES N-2-hydroxyethylpiperazine-N-1,2-ethanesulfonic acid - IC₅₀ concentration inhibiting stimulated steroid secretion by 50% - IU international unit - MEM Eagle's Minimum Essential Medium - RIA radioimmunoassay - RNA ribonucleic acid - SEM standard error of the mean - SMEM serum-containing Eagle's Minimum Essential Medium - 20-DHP 20--hydroxy-4-pregnen-3-one     

Quantitative analysis of rod-cored vesicles and dense granules of large granular lymphocytes in the liver,spleen, and peripheral blood of rats

Large granular lymphocytes (LGL) comprise a natural defense system in the liver and exert an inhibitory effect on tumor cell metastasis. In order to demonstrate the maturation of LGL in the liver from the morphological aspect, we evaluated electron-microscopically the frequency of 0.2 m vesicles (rod-cored and empty vesicles) and dense granules in LGL from the liver, spleen, and peripheral blood of the rat. Both of these cell organelles are characteristic to LGL and may relate to natural killer-mediated cytolysis. On the average, there were 12.7 of the 0.2 m vesicles and 4.3 rod-cored vesicles (RCV) per cell section in the liver, 6.6 0.2 m vesicles and 1.6 RCV in the spleen, and 8.6 0.2 m vesicles and 0.9 RCV in the peripheral blood. The number of 0.2 m vesicles per cell section ranged from 0 to 19 with the exception of a few higher instances. Therefore, LGL were divided into vesicle-rich(>9 0.2 m vesicles per cell section) and vesicle-poor (<8 per cell section) populations. Hepatic LGL consisted mainly of a vesicle-rich population while splenic LGL consisted mainly of a vesicle-poor population, and peripheral blood contained equal proportions of both populations. In addition to diversity with regard to the number of 0.2 m vesicles, LGL obtained from various organs also displayed heterogeneity in the number and size of dense granules. Since the number of dense granules per cell section usually ranged from 1 to 13, LGL were diveded into 2 populations, i.e., LGL with many (>7 per cell section) granules and those with a few(<6 per cell section) granules. Specifically, splenic LGL had a few small (average diameter, less than 400 nm) dense granules, while sections of LGL from the liver and peripheral blood displayed many small dense granules and a few large (>400 nm) ones, respectively, in addition to the populations seen in the spleen. Thus, the present study has demonstrateda difference in the distribution of 0.2 m vesicles in LGL based on the tissue of origin. The present study has revealed the difference in the distribution of 0.2 m vesicles of LGL by tissue and indicated that immature LGL are predominant in the spleen, while hepatic LGL are generally more mature as defined by the number of vesicles. These data suggest that the microenvironment of the liver may contribute to the increased expression of these vesicles in LGL.     

An ACTH-Like Peptide Immunocortin: Effect on the Activity of Cells from the Rat Adrenal Cortex

The effect of immunocortin, an ACTH-like decapeptide VKKPGSSVKV corresponding to the 11–20 sequence of the variable part of the human IgG1 heavy chain on the content of 11-hydroxycorticosteroids (CS) in rat adrenal glands and blood serum in vivo was studied. An intramuscular injection of immunocortin at a dose of 10 g/kg was found in an hour to induce a twofold decrease in CS content in the adrenal glands and a 1.8-fold increase in the blood serum CS content. At the same time, an immunocortin dose of 100 g/kg exerted practically no effect on the CS content and its dose of 1000 g/kg increased the CS content both in adrenal glands and in blood serum by 1.6 and 2.2 times, respectively. Four hours after the injection of any of the three doses of immunocortin, the CS content in adrenal glands did not differ from the control value, and after 24 h the content decreased threefold. Immunocortin was shown to be bound by the ACTH receptors in the membranes of the rat adrenal cortex with a high affinity and specificity (inhibiting the specific binding of ¹²⁵I-labeled ACTH-(11–24) peptide with K _i of 1.2 nM).     

Über die Entwicklung von Ependym und Plexus chorioideus der Ratte

Zusammenfassung Formentwicklung der Seiten- und des 3. Ventrikels, Histogenese und Chemodifferenzierung des Ventrikelependyms und der Plexus chorioidei der Ratte (163 Tiere) vom 14. Embryonaltag bis zum 40. Lebenstag und bei erwachsenen Kontrolltieren werden lichtmikroskopisch untersucht. Die wesentlichsten Formmerkmale der Ventrikel sind bis zur Geburt ausgeprägt. Die Anpassung an die weitere Größenzunahme des Gehirns ist jedoch erst etwa am 30. Lebenstag abgeschlossen. — Die Bildung der Plexus chorioidei beginnt im Seitenventrikel am 15., im 3. Ventrikel am 17. Embryonaltag. Der Glykogengehalt des Plexusepithels nimmt bis zur Geburt zu und wird bis zum 15. Lebenstag vollständig reduziert. Die postnatale Aktivitätszunahme der verschiedenen Dehydrogenasen (NADH, NADPH, SDH, LDH, -HB-DH, Glu-6-DH), Cytochromoxidase und sauren Phosphatase im Plexusepithel ist etwa am 30. Lebenstag beendet. — Die Histogenese des zunächst mehrreihigen Ependyms aus der Matrix beginnt am 18. Embryonaltag. Die Aktivität der verschiedenen Fermente steigt pränatal nur mäßig an. Im Seitenventrikel tritt Wimpernependym auf; hier ist die morphologische Differenzierung an der medialen Wand (Hippocampusformation) am 3. Lebenstag, an der lateralen Wand (gleichzeitig mit der Rückbildung der subependymalen Zellschicht) zwischen dem 11. und 21. Lebenstag, die Chemodifferenzierung bis zum 30. Lebenstag abgeschlossen. Im 3. Ventrikel beginnt am 19. Embryonaltag die Differenzierung in Wimpernependym und Tanycytenependym; letzteres besitzt keine Wimpern, entsendet jedoch lange Zellfortsätze in den Hypothalamus und zur basalen Oberfläche des Zwischenhirns. Es kleidet nur den ventralen Ventrikelbezirk (Radix und Rec. infundibuli, Rec. inframammillaris) aus und kommt auch in einer schmalen mittleren Zone, dem Wimpernependym unterlagert, vor. Das Wimpernependym des 3. Ventrikels erreicht etwa gleichzeitig mit der morphologischen Reifung zwischen dem 5. und 10. Lebenstag die Fermentaktivität erwachsener Kontrolltiere. Morphologische und histochemische Differenzierung des Tanycytenependyms sind dagegen erst am 34. Lebenstag abgeschlossen. Wimpernependym und Tanycytenependym unterscheiden sich nicht nur morphologisch, sondern auch in ihrem Fermentmuster: ATPase fehlt im Wimpernependym, saure Phosphatase, Bernsteinsäuredehydrogenase, Cytochromoxidase und -Hydroxibuttersäuredehydrogenase zeigen keine Aktivität im Tanycytenependym. Die anderen nachgewiesenen Fermente besitzen — mit Ausnahme der Glucose-6-phosphat-Dehydrogenase — die relativ höchste Aktivität im Wimpernependym. Die Fermentaktivität tritt im Wimpernependym vorwiegend apikal, in den Tanycyten über die ganze Zelle verteilt und in den Fortsätzen auf. Das Enzymmuster des Wimpernependyms wird als Ausdruck eines oxidativen Energiestoffwechsels im Dienst der Wimpernmotorik gewertet. Die Fermentausstattung des Tanycytenependyms deutet auf möglicherweise hier ablaufende Synthesevorgänge hin.

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Summary The development of the lateral and third ventricle, histogenesis and chemodifferentiation of the ventricle ependyma and plexus chorioidei in rats (163 animals) are investigated by means of light microscopy. The age of the animals investigated ranged from the 14th day of embryonic life to the 40th day of life. Adult control animals were included in the investigation.The most essential structural elements of the ventricles are developed before birth. The adjustment to the increasing size of the brain, however, is completed only on the 30th day of life. The formation of the plexus chorioidei begins in the lateral ventricle on the 15th, in the third ventricle on the 17th day of embryonic life. The glycogen content of the plexus epithelium increases steadily until birth; from there on a reduction takes place until, on the 15th day of life, the glycogen is completely reduced. The post-natal increase in the activity of various dehydrogenases (NADH, NADPH, SDH, LDH, -HB-DH, Glu-6-DH), cytochrome oxidase, and acid phosphatase in the plexus epithelium is terminated on the 30th day of life approximately. The histogenesis of the originally stratified ependyma deriving from the matrix starts on the 18th embryonic day. The increase of enzymatic activity before birth is negligible. Ciliated ependyma is observed in the lateral ventricle; there the morphological differentiation of the medial wall (hippocampus formation) is terminated on the 3rd day of life, of the lateral wall (together with the reversion of the subependymal cell layers) between the 11th and 21st day of life. The chemodifferentiation is terminated on the 30th day of life. On the 19th embryonic day the differentiation of the ciliated and tanycyte ependyma starts in the third ventricle. The tanycyte ependyma is not ciliated; it sends long cell processes into the hypothalamus and to the basal surface of the diencephalon. It only lines the ventral region of the ventricle (Radix and Rec. infundibuli, Rec. inframammillaris) and is present in a narrow central zone, where it is demonstrable underneath the ciliated ependyma. Together with the morphological differentiation the ciliated ependyma of the third ventricle obtains an enzymatic activity comparable to adult control animals between the 5th and 10th day of life. The morphological and histochemical differentiation of the tanycyte ependyma is only completed on the 34th day of life. Ciliated ependyma and tanycyte ependyma differ not only morphologically but also in their enzymatic pattern. ATP-ase is not present in ciliated ependyma. There is no activity of acid phosphatase, succinate dehydrogenase, cytochrome oxidase, and -hydroxi-butyric acid dehydrogenase in the tanycyte ependyma. With the exception of glucose-6-phosphate dehydrogenase, all the enzymes investigated show the relatively highest activity in the ciliated ependyma. The enzymatic activity of the ciliated ependyma is predominantly found in the apical part of the cell, whereas in tanycytes it is evenly distributed over the cell and the processes. The enzymatic pattern of the ciliated ependyma is to be regarded as the expression of an oxidative energy metabolism which serves the motoricity of the cilia. The enzymatic pattern of the tanycyte ependyma seems to indicate a certain synthesizing activity.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft und den Universitätsbund Würzburg.

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Die Arbeit hat der Medizinischen Fakultät Würzburg als Inauguraldissertation vorgelegen.     

Fine structure and peroxidatic activity of rat blood monocytes

Summary Transmission and scanning electron microscopy of the rat epithalamus shows a regional variation in the distribution of Supraependymal nerves (SN) which correlates well with Supraependymal yellow fluorescence reported by Richards et al. (1974). The medial habenular nucleus, the intercommissural and suprahabenular recesses, the habenular commissure and the fibrae periventriculares thalami have the greatest density of SN/100 m of ependymal surface. The floor of the suprahabenular and intercommissural recesses is covered by non-ciliated ependyma.The significance of these findings is discussed with respect to (1) a direct functional relationship of SN with ependyma, and (2) a possible participation of the non-ciliated ependyma of the suprahabenular and intercommissural recesses in secretory activity whereby the CSF serves as a vehicle for neuroendocrine communication.The author thanks Drs. Sanford Palay, Milton Brightman, and Constantino Sotelo for their advice in the preparation of nervous tissue for electron microscopy; Dr. Gunter Bahr for the use of scanning electron microscopy facilities; and Messrs. Cyril Wingfield and Walter Engler and Mrs. Michie Vane for their technical support     

α-adrenergic receptor regulation of Ca2+/Mg2+-ATPase in brain synaptic membranes

The effects of ₁ and ₂ receptor ligands on Ca²⁺/Mg²⁺-ATPase have been studied using synaptosomal plasma membranes isolated from rat brain cortex. Both phenylephrine and clonidine inhibited Ca²⁺/Mg²⁺-ATPase, in a concentration-dependent fashion. IC₅₀ values for half-maximal inhibition for phenylephrine and clonidine were 29 M and 18 M, respectively. The inhibitory effect of phenylephrine was reversed by the alpha antagonist prazosin while yohimbine and rauwolscine reversed the inhibition of enzyme activity by clonidine. The two antagonist subtypes were effective only against the respective agonist subtypes, demonstrating distinct subtype preferences. Analysis of the kinetics of enzyme inhibition indicate both agonists to be noncompetitive. Some evidence suggests that yohimbine may exhibit mixed agonist/antagonist properties which depend on [Ca²⁺]. The present study provides biochemical evidence to support auto receptor adrenergic receptor regulation of neurotransmitter release.     

Cellular organization of the lateral and postinfundibular regions of the median eminence in the rat

Summary The structural organization of the rostral, lateral and postinfundibular regions of the median eminence (ME) of 5-day cyclic diestrous rats was studied with light and electron microscopic methods. The ependymal cells lining (i) the floor of the infundibular recess (IR) at rostral levels, (ii) the lateral extensions of the IR, and (iii) the floor of the premammillary recess appear to represent the same type of tanycyte ependyma (1 tanycytes). In the entire width of the rostral and postinfundibular palisade regions, as well as in the lateral palisade region of the preinfundibular ME, the processes of the 1 tanycytes form a continuous cuff. This cuff separates the nerve endings from the blood vessels and the pars tuberalis. At this level, synaptoid contacts between neurosecretory axons and the ependymal cuff can be observed. The ultrastructural characteristics of the 1 tanycytes are described and their ependymal endings tentatively classified into three types. In the lateral regions of the ME, the Golgi study revealed the presence of two fiber systems: (i) one possessing a latero-medial trajectory and distributed in the subependymal region; (ii) the other formed by a loose longitudinal tract originating from neurons of the arcuate nucleus. Some functional implications of the cellular organization of the rat ME are discussed.Supported by Grants from PLAMIRH (92.171.2.77) and from the Dirección de Investigaciones, Universidad Austral (S-77-28)The authors wish to thank Miss Rosario Andrade, Mrs. Elizabeth Santibáñez and Mr. Armando Bilbao for their assistance     

6-Methylpurine-induced inhibition of sclerotia morphogenesis in Sclerotium rolfsii and its reversal by adenosine

In liquid synthetic medium inoculated with Sclerotium rolfsii (SR), addition of 6-methylpurine (MP, 50g/ml) immediately after inoculation led to approximately 100% reduction in sclerotia production. Adenosine, and to a lesser extent guanosine, each at final concentration of 100g/ml significantly reduced inhibition of sclerotia formation by SR in presence of 50g/ml MP. Uridine and cytidine each at 100g/ml had no such effect. The inhibition of sclerotia morphogenesis could be prevented by addition of 800g/ml of adenosine together with 50g/ml MP. Reversal by adenosine of MP-induced inhibition of sclerotia development was concentration dependent.     

Das Cytostom von Lankesterella garnhami

Zusammenfassung Die Nahrungsaufnahme der Merozoiten von Lankesterella garnhami in den Milzzellen von Hamburger Hausspatzen (Passer d. domesticus) wurde elektronenmikroskopisch untersucht. Teile des Wirtszytoplasmas gelangen durch Abschnürung zahlreicher, etwa 20–30 m. großer Pinozytosevesikel in das Lumen der periparasitären Vakuole. Von dem Inhalt der periparasitären Vakuole werden Teile mit Hilfe mehrerer Cytostomstrukturen in Nahrungskanäle aufgenommen, von denen sich 100–200 m große Nahrungsvakuolen abschnüren, in denen die weitere Verdauung stattfindet. Das Cytostom ist durch zwei intensiv kontrastierte, konzentrische Zylinder charakterisiert, die sich von den beiden Pelliculamembranen ableiten. Der innere Durchmesser beträgt ca. 80m, der äußere Durchmesser ca. 150 m und die Tiefe im Ruhestadium mindestens 50–100 m. Die Bedeutung des Cytostoms als taxonomisches Merkmal wird diskutiert.

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The cytostome of Lankesterella garnhami

Summary The feeding mechanism of merozoites of Lankesterella garnhami in the cells of the spleen of young sparrows (Passer d. domesticus), which were captured in Hamburg, was studied by means of electron microscopy. The cytoplasm of the host cell permeates the membrane of the periparasitic vacuole by means of pinocytosis of vesicles with a diameter of 20–30 m. The parasite takes up parts of the content of the periparasitic vacuole with several cytostome structures. This structure consists of two electron dense, concentric cylinders, which originate from the two membranes of the pellicle. The cytostome is 80 m in the inner diameter and 150 m in the outer diameter. The depth of the inactive cytostomal cavity is about 50–100 m, whereas it is much prolonged in the stage of active feeding. In this stage the cytostome opens into a channel of 400–800 m and more, from which food vacuoles of 100–200 m are pinched off. Digestion occurs in the food vacuole. These findings are compared with observations of the cytostome and the micropyle in other sporozoa. Sometimes long, V-shaped invaginations of unknown signification are noticed.

     

Ultrastructural changes in adrenaline- and SGC-cells after morphine coincide with alterations of adrenaline and dopamine levels

Summary The effects of morphine on chromaffin cell ultrastructure and catecholamine contents were studied using the adrenal glands from male mice (ICR strain). After 2 h adrenaline was increased 25% from 8.1 to 11.6 mol/g tissue, followed by a 50% decrease to 5.2 mol/g between 8–24 h and low values persisting at 72 h. Dopamine increased initially, reaching peak values of 0.5 mol/g between 8–24 h, but had returned towards control values of 0.29 mol/g by 72 h. Noradrenaline remained unchanged at 2.5 mol/gram. Naloxone prevented alterations in adrenaline and dopamine levels.Ultrastructural examination revealed several types of catecholamine-storing cells. Of these the adrenaline and small-granule chromaffin (SGC) cells were more affected by morphine than noradrenaline cells. While the initial elevation of adrenaline 2 h after morphine was not accompanied by significant ultrastructural changes, the decrease after 8 and 24 h was paralleled by a significant (p<0.001) loss of adrenaline granules. Signs of active membrane turnover included an increase in the number of vacuoles, and the appearance of numerous coated omega profiles and coated (77.7±0.6 nm) vesicles. Clusters of synaptic-like vesicles (59.8±8.2 nm), slightly larger than neuronal vesicles (45.4±6.4), increased in the SGC-cells. After 72 h, the chromaffin granules in adrenaline cells remained low in numbers and were heterogeneous in electron density. Many synapticlike vesicles were aligned along the SGC-cell membranes where only few chromaffin granules (109.3±20 nm) remained. Thus, continuous morphine exposure for 8–72 h increases the turnover of storage granules in adrenaline and SGC-cells with less effect on the noradrenaline cells which maintain catecholamine levels as indicated by biochemical analysis.     

Stimulation of ethylene production by catecholamines and phenylethylamine in potato cell suspension cultures

The catecholamines (50 M dopamine, 50 M norepinephrine and 100 M epinephrine) and phenylethylamine (200 M) were found to stimulate ethylene production in potato suspension cultures. When 100 M amino-oxyacetic acid was added together with epinephrine, ethylene release returned to control levels. The endogenous 1-aminocyclopropane-1-carboxylic acid levels were increased in parallel with the release of ethylene, suggesting that the observed effect probably occurs via regulation of aCC synthase. Our results suggest that there is a link between these naturally occurring monoamines and ethylene in plants.Abbreviations AOA amino-oxyacetic acid - ACC 1-aminocyclopropane-1-carboxylic acid - DA dopamine - NE norepinephrine - E epinephrine - CA catecholamines - PEA phenylethylamine     

Fusion of photoreceptor membrane vesicles

Summary Then-alkyl bromides with 6 to 10 carbons induce formation of vesicles of 5 to 100 m diameter from the small vesicles (0.1 m average diameter) produced by disruption of the discs from frog rod photoreceptors. Then-alkanes,n-alkyl iodides andn-alkyl chlorides are relatively ineffective. The formation of large vesicles is independent of calcium concentration and is distinguished from fusion processes previously reported by the large number of vesicles involved. The results reported here together with others suggest the occurrence of multiple fusion (and/or rupture-resealing) events between vesicles, induced by then-alkyl bromides.     

Nerve-purkinje fiber relationship in the moderator band of bovine and caprine heart

Summary The moderator band in the heart of the ox and goat contains bundles of Purkinje fibers and nerve fibers separated by connective tissue. The axons are mostly unmyelinated and embedded in the cytoplasm of Schwann cells.Small bundles of axons run close to the Purkinje fibers. The axons dilate into varicosities 0.5 to 1.6 in diameter (mean 0.95 ), containing three types of vesicles: 1) agranular vesicles with a diameter of 400–500 Å, 2) large dense-cored vesicles with a diameter of 800–1200 Å, 3) small dense-cored vesicles with a diameter of 500 Å. Most varicosities contain agranular vesicles together with a few large dense-cored vesicles.The gap between the varicosities and the nearest Purkinje fiber is unusually wide and normally varies between 0.3 and 0.8 . No intimate nerve-Purkinje fiber contacts, with a cleft of 200 Å, were observed.     

Secretory capacity of the lachrymal salt gland of hatchling sea turtles,Chelonia mydas

Summary The lachrymal salt glands ofChelonia mydas were functional when hatchlings emerged from the nest. Osmotic concentrations up to 720 mosmol kg^–1 were recorded in spontaneously produced tears (salt gland secretions). When injected with a Na⁺ load (1500–2700 mol (100 g)^–1) newly emerged hatchlings produced tears ranging in osmotic concentration from 1000–1900 mosmol kg^–1 with Na⁺ secretion rates from single glands of 200–475 mol (100 g·h)^–1. In these circumstances the rate of sodium excretion, via the salt glands, was equivalent to the sodium content of 0.2 to 0.5 ml of sea water per hour. Since the apparent drinking rate of hatchlings within the first two days of entering sea water was approximately 0.5 to 1.7 ml per day, the excretion of Na⁺ imbibed by drinking is well within the secretory capacity of the lachrymal salt glands.In feeding hatchlings extraordinarily high Na⁺ secretion rates were induced by Na⁺ loading. Hatchlings which were loaded with Na⁺ by injection (1500–5400 mol (100 g)^–1) produced tears having osmotic concentrations between 1500 and >2000 mosmol kg^–1. The Na⁺ secretion rates from single glands were 750–4185 mol (100 g·h)^–1 with extremely high short term rates of 10700 mol (100 g·h)^–1 (50 mol min^–1 for 28 g hatchlings).In terms of gland mass the highest long term secretion rate translates into 21 mmol of Na⁺ per gram of salt gland per hour and is the highest secretion rate yet recorded for a reptilian salt gland. This rate is almost three times the highest rate recorded for sea snakes (8 mmol g·h^–1) and is similar to rates commonly observed in avian salt glands (25 mmol g·h^–1).Secretion by the lachrymal salt glands was initiated by increased blood concentrations of Na⁺ or K⁺, K⁺ being as effective as Na⁺ but with the composition of the teras being virtually unchanged compared to tears from Na⁺ stimulated hatchlings. Preliminary experiments indicated that secretion was not initiated by increased Cl^– concentration in the blood or by increased volume or osmotic concentration of the blood.Abbreviation O.P. osmotic pressure     

Zur Spaltbarkeit verschiedener Naphthol-Carbonsäureester durch Esterasen im Tanycytenependym des III. Ventrikels der Wistarratte

Zusammenfassung Im Tanycytenependym des III. Ventrikels kommen bei der Wistarratte unspezifische, E 600-empfindliche und gegenüber p-Chlormercuribenzoat resistente Esterasen vor. Um diese Enzyme histochemisch weiter zu charakterisieren, wurde ihre Fähigkeit geprüft, verschiedene Naphthol-Carbonsäureester bei Anwesenheit von Lösungsvermittlern zu spalten. Es zeigte sich hierbei ein unterschiedliches Verhalten der Esterasen in Ganglienzellen und Tanycyten. In letzteren weist von verschiedenen homologen Carbonsäureestern des -Naphthols der Buttersäureester die höchste Spaltungsrate auf; Acetatester der Naphthol-AS-Reihe ergeben unterschiedliche, aber stets schwächere Reaktionen als -Naphthylacetat. Wie in früheren Untersuchungen wurden Unterschiede im Verhalten der Esterasen-Aktivität in Perikaryen und Fortsätzen der Tanycyten beobachtet. — Auf Grund der Befunde wird die Möglichkeit einer Einschaltung der Tanycytenesterasen in die Erregungsübertragung diskutiert.

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Splitting of various naphthol carbonic acid esters by esterases in the tanycyte ependyma of the III ventricle of the Wistar rat

Summary In the Wistar rat, the tanycytes of the IIIrd ventricle contain unspecific esterases which are E 600 sensitive and are not inhibited by p-chlormercuribenzoate. To further investigate the character of these enzymes, their ability to split various naphtholcarbonic acid esters in the presence of organic solvents was tested. It was hereby established that the esterases of the tanycytes behave differently to those of the diencephalic nerve cells. In the tanycytes among various homologous carbonic acid esters of -naphthol applied, -naphthyl butyrate shows the highest splitting ratio; with acetate esters of AS-naphtholes differing but always weaker reactions are obtained than with -naphthyl-acetate. As in a former study, behavioral divergencies of esterase activity in the tanycyte perikaria and processes were observed. — On the basis of these findings the possibility is discussed that the esterases of the tanycytes might play a role in synaptic transmission.

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Herrn Prof. Dr. med. O. Veit in Verehrung zum 85. Geburtstag gewidmet.     

Aluminum-induced secretion of both citrate and malate in rye

Aluminum (Al)-resistant mechanisms responsible for Al-induced secretion of organic acids are poorly understood. In this study, we characterized the Al-induced secretion of both citrate and malate from rye (Secale cereale L. cv. King). Secretion of organic acids increased with increasing concentration (10, 30 and 50 M) and duration of Al treatments. Neither phosphorous (P) deficiency up to 15 days nor addition of 50M lanthanum, 50 M lead, 10 M cadmium, or 200 M manganese caused secretion of organic acids, suggesting that this secretion was a specific response to Al stress. Aluminum activated citrate synthase, the main enzyme for the synthesis of citrate, but its activation occurred only in the root tip. The elongation of roots of an Al-sensitive cultivar of wheat (Tritium aestivum L. cv. Scout 66) was not inhibited by 50 M Al in the presence of externally applied 50 M citrate or 400 M malate. The secretion of citrate and malate from intact rye roots exposed to 50 M Al corresponded to 31.3 ± 1.7 M and 11.5 ± 2.5 M, respectively, in the rhizosphere based on an assumption of a 2 mm thick unstirred layer around root tips. This result indicated that Al-resistance in rye was achieved by the Al-induced synthesis of citrate in root apices followed by Al-induced specific secretion of citrate from root tips.     

Angiogenesis and myogenesis as two facets of inflammatory post-ischemic tissue regeneration

Recently Hippophae rhamnoides has been reported to render chromatin compaction and significantly inhibit radiation induced DNA strand breaks. To investigate the mechanism of action of RH-3, a preparation of Hippophae rhamnoides, in this connection, present study was undertaken. Chromatin compaction induced by RH-3 (100 g/ml or more) was maximum at alkaline pH but was completely negated by acidic pH (< 6) or presence of free radical scavengers like glycerol, DMSO etc. In a concentration dependent manner, RH-3 inhibited the intercalation of ethidium ions from Et Br into calf thymus DNA and also increased the precipitation of DNA-protein cross-links (DPC) in thymocytes. Chromatin compaction caused by RH-3 treatment did not permit the separation of proteins from DNA even after treatment with 2 M NaCl solution. SDS-PAGE profiles also revealed that RH-3 in a dose dependent manner compacted the chromatin organization, induced DPC and inhibited the extraction of both histone and non-histone matrix proteins from chromatin maximally at 80 g/ml. More than 80 g/ml of RH-3, though extracted low molecular weight histones but did not separate non-histone proteins. The RH-3 mediated DPCs were resistant even to 1% SDS, 4 M NaCl and 3.8 M hydroxyl amine hydrochloride but were prone to both urea (8 M) and guanidine hydrochloride (6 M) indicating covalent bonding between DNA and proteins (serine/threonine). RH-3 in a concentration dependent manner induced superoxide anions and the phenomenon was dependent upon nature of medium, presence of metal ions and pH. RH-3 at concentrations up to 100 g/ml in presence of 50 M copper sulfate inflicted significant damage to extraneously added 2-deoxyribose molecules and maximum TBARS were formed at a concentration of 100 g/ml. Higher concentrations of RH-3 more than 100 g/ml quenched free radicals and inhibited 2-deoxyribose degradation. RH-3 also induced strand breaks in plasmid DNA at concentrations lower than 100 g/ml but completely inhibited at concentrations higher than 250 g/ml, indicating bimodal function. Strand breaks induced by lower concentrations of RH-3 (up to 100 g/ml) were inhibited by antioxidants like GSH, DFR etc. RH-3, in a concentration dependent mode also inhibited the relaxation of supercoiled plasmid DNA (PBR322) by topoisomerase I.Present study indicated that RH-3 caused compaction of reversible (< 100 g/ml) and irreversible (> 100 g/ml) nature which was related to the magnitude of DNA-protein cross-links formed. Maintenance of chromatin organization, induction of hypoxia, hydrogen atom donation, free radical scavenging and blocking of cell cycle at G2-M phase by interfering with topoisomerase I activity seem to contribute towards the radioprotective efficacy of RH-3.     

Ependymokrinie und Rezeptoren in der Wand des Recessus infundibularis der Maus und ihre Beziehung zum kleinzelligen Hypothalamus

Zusammenfassung Im Gegensatz zu den neurosekretbildenden Zellen des großzelligen Hypothalamus enthalten die Neurone des kleinzelligen Hypothalamus im Perikaryon und proximalen Neuronabschnitt nur eine verschwindend geringe Anzahl von Elementargranula. Dagegen finden sich in den Nervenendigungen der Zona externa gehäuft Vesikel (ø 900 bis 1500 Å), synaptische Vesikel und Elementargranula (ø 850 Å). Die vorgelegten Ergebnisse lassen nach Vergleich mit lichtmikroskopischen Befunden auf eine Synthese von Neurohormonen und neurohumoralen Substanzen schließen, die im distalen Neuronabschnitt stattfindet.Axone des Tractus tuberohypophyseus bilden in der Zona externa des Infundibulum synapsenähnliche Kontakte mit den Fortsätzen der ependymalen Tanyzyten. Im Bereich dieser Kontaktstellen kommt es zu Substanzübertritten aus den Nervenendigungen in die Tanyzytenfasern. Ansammlungen heller Vesikel von unterschiedlicher Größe finden sich im Verlauf des Tanyzytenfortsatzes bis zum Perikaryon.An der freien Oberfläche der Ependymzellen sammeln sich die Vesikel, vom Golgiapparat ausgehend, zu multivesikulären Protrusionen und werden in den Liquor ausgestoßen. Somit ist eine mikroapokrine Sekretion des Tanyzytenependyms am Boden des Recessus infundibularis festzustellen. Ob das Sekret ependymspezifisch ist oder ob es sich um Substanzen handelt, die aus den Nervenendigungen der Zona externa in die Tanyzytenfortsätze gelangten und verändert oder unverändert an den Ventrikel abgegeben werden, bleibt zu klären.Im Recessus infundibularis finden sich nahe der Ependymoberfläche markarme Axone, die — ihrem Bau nach zu urteilen — den Ganglienzellen des Nucleus infundibularis anzugehören scheinen und eine afferente Funktion besitzen könnten. Sekretorisch tätige Tanyzyten und intraventrikulär gelegene afferente Endigungen von Nervenzellen könnten Glieder in einem feedback-Mechanismus zwischen den Nervenendigungen in der Zona externa und den ihr zugeordneten Ganglienzellen sein.

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Ependymal secretion and receptors in the wall of the 3rd ventricle of the mouse

Summary In contrast to the cells of the large-celled hypothalamus, where neurosecretory substance is produced, the neurones of the small-celled hypothalamus contain in their cell bodies, and in the proximal part of the cell only a very small number of elementary granules. On the other hand, the nerve endings in the zona externa (outer layer of the median eminence) contain vesicle aggregations (ø 900–1,500 Å), synaptic vesicles and elementary granules (ø 850 Å). Comparing the results of the present investigation with lightmicroscopical findings, the conclusion seems to be justified that a synthesis of neurohormones and neurohumoral substances takes place in the distal part of the neuron.Axons of the tractus tuberohypophyseus form, in the zona externa of the infundibulum, synapse-like contacts with the processes of the ependymal tanycytes. In the region of these contiguities, substance is perhaps transferred from the neural endings into the fibres of the tanycytes. Accumulations of pale vesicles of various sizes are to be found in the process of the tanycyte, extending into the perikaryon.The vesicles spread from the Golgi complex to the free surface of the ependymal cells where they become multivesicular protrusions, and are subsequently expelled into the liquor. Hence a micro-apocrine secretion of the tanycyte-ependyma on the floor of the recessus infundibularis is demonstrated. It remains to be clarified whether this secretion is ependymaspecific or whether we are dealing with substances, which leaving the nerve endings of the zona externa (outer layer) and being transported into the tanycyte processes, are eventually, either in changed or unchanged state, discharged into the ventricle.In the recessus infundibularis axons are to be found, which lie near the surface of the ependyma, and which possess a thin myelin sheath. They appear to belong to the ganglion cells of the nucleus infundibularis and presumably have an afferent function. Both tanycytes engaged in secretion and intraventricularly-lying, afferent neuronal endings could be part of a feedback mechanism acting between the nerve endings in the zona externa and the ganglion cells to which they belong.