Catalase-like crystals in parathyroid gland cells of Rana temporaria L.

Summary Crystalline inclusions in parathyroid gland cell nuclei of Rana temporaria were studied by electron microscopy using a specimen tilting stage. Images were analysed by optical diffraction. Results were compared with X-ray and electron microscopic data of trigonal bovine liver catalase to which a striking resemblance of the inclusions was found.We are grateful to Professor R. Mosebach (Giessen) for discussions, to the Deutsche Forschungsgemeinschaft for a grant (La 229/4) and instruments and to Messrs. Spindler & Hoyer, Göttingen and Messrs. Rank Precision Instruments, Nürnberg for putting apparatus at our disposal and performing diffraction photographs.     

Cyclical mitotic activity in the adeno-hypophysis of Rana temporaria

Summary In the pars distalis, the pars intermedia and the pars tuberalis of the hypophysis of Rana temporaria, an annual mitotic cycle is described.In the pars distalis there exists a gradient of mitotic activity, corresponding to a gradient of distribution of the PAS positive cells.In the pars distalis, the rate of cell degeneration and of cell renewal of the PAS positive cells seems to be faster than of the orange G positive cells.The replacement of degenerated PAS positive cells of the pars distalis seems to occur mainly during the months immediately following on the period of maximal activity of the PAS positive cells.     

Lipidkugeln im Plexusepithel von Rana temporaria L.

Zusammenfassung Das Plexusepithel von Rana temporaria L. enthält zwei verschiedene Typen von Lipidkugeln, die einen Durchmesser bis zu 8 erreichen können. Diese Einschlüsse haben keine Zytomembran. Die Struktur der Einschlußkörper wird mit Hinweisen auf die Fixierung und den Stoffwechsel diskutiert.

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Lipid droplets in the epithelium of the choroid plexus of Rana temporaria L.

Summary The epithelium of the choroid plexus of Rana temporaria L. contains two different types of spherical lipid inclusions up to 8 in diameter. The droplets do not have a limiting membrane. The preservation and metabolism of the lipid bodies are discussed.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Influence of light and darkness on the hypothalamo-hypophysial system of Rana temporaria L.

Summary The influence of a long daily photoperiod and continuous darkness was studied on the hypothalamo-hypophysial system of Rana temporaria. After six weeks of treatment (December-January 1968/1969) the secretory activity of the hypothalamo-hypophysial system of the light-treated animals was enhanced.The amount of aldehyde-fuchsin positive material in the preoptic nucleus increased as a result of the light treatment, and decreased in the median eminence, whereas in the posterior lobe no difference could be observed between the light- and dark-treated animals.The karyometric results showed an increase of the nuclear volume of the cells of the preoptic nucleus after light treatment; small differences were noticed between distinct areas of the preoptic nucleus.Autoradiographic data especially showed that light influenced the hypothalamo-hypophysial system to a high degree. Half an hour after the injection of ³⁵S-cysteine, the incorporation of the isotope in the preoptic nucleus of the light-treated animals was five times as much as in the dark-treated animals. In the median eminence and in the posterior lobe, the autoradiographic results confirmed the results found with regard to the amount of aldehyde-fuchsin positive material. Labelled material is released only from the median eminence and not from the posterior lobe.The author is indebted to Prof. Dr. J. C. van de Kamer and Dr. F. C. G. van de Veerdonk for their interest and helpful encouragement. Thanks are also due to Ir. J. J. Bezem for his assistance in solving the statistical problems and to Mr. H. van Kooten and his staff for making the illustrations. The co-operation of the student Mr. E. Schenk is gratefully acknowledged. Dr. L. Boomgaart was so kind to check and amend the English writing.     

Arterial supply of the choriocapillaris of anuran amphibians (Rana temporaria,Rana esculenta)

Summary The pattern of the vascular supply to the choroid of the frog eye was studied in toto with the use of the injection-replication-SEM technique. The choroid of anuran amphibians is composed mainly of the choriocapillaris. In both species studied (Rana temporaria, Rana esculenta), an independent arterial supply to the choriocapillaris supplemented that from the ciliary arteries. This additional vascular route arises from the optic artery, a separate branch of the arteria infundibularis superficialis. The optic artery, accompanied by its vein within the vascular sheath of the optic nerve, joins the rich arterial capillary network of the choriocapillaris and supplies the posterior pole of the ocular bulb. The superficial capillary network displays a dense collar around the entrance of the optic nerve into the eye and is composed of a circular meshwork of small capillaries, several layers deep. More peripherally, however, it becomes single layered. This capillary network, as a whole, establishes numerous connections with the adjacent choriocapillaris at the posterior pole of the ocular bulb. In anuran amphibians the complex arrangement of both arterial systems supporting the choriocapillaris may be regarded as a more complete equivalent of the short posterior ciliary arteries of mammals.     

Elektronenmikroskopische Untersuchungen an spezifischen Organellen von Endothelzellen des Frosches (Rana temporaria)

Zusammenfassung Endothelzellen der Wirbeltiere enthalten spezifische Organellen, deren Funktion unbekannt ist. Diese Organellen werden beim Frosch (Rana temporaria) nach unterschiedlichen Fixierungen elektronenmikroskopisch untersucht. Die Organellen sind walzenförmig mit mannigfachen Abweichungen, bis zu 2 lang und 0,1 bis 0,5 dick. Ihre oft unterbrochene Außenmembran ist dicker als zytoplasmatische Membranen. Das Innere der Organellen besteht aus Tubuli, die in eine elektronendichte Matrix eingebettet sind. Die Dichte dieser Matrix zeigt deutliche Abstufungen. Die Tubuli sind möglicherweise aus einer spiralförmigen Molekülkette aufgebaut. Das Verteilungsmuster der Organellen wird mit stereologischen Methoden untersucht. Die größte Volumendichte weisen die Aortae thoracicae mit 8% auf. Die Volumendichte der Organellen im Zytoplasma der Endothelzellen scheint mehr von der Entfernung der betreffenden Gefäßstrecke zum Herzen abzuhängen als von der Gefäßgröße. Es werden Verbindungen der Organellen zu zytoplasmatischen Membransystemen aufgezeigt. Auf Besonderheiten des Endothels, darunter Aggregationen von Ribosomen, wird hingewiesen.

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Electron microscopic studies on specific organelles of endothelial cells in the frog (Rana temporaria)

Summary Endothelial cells of vertebrates contain specific organelles of unknown function. These organelles are studied by electron microscopy with different fixations. The organelles are rod-shaped with many variations, up to 2 in length and 0.1 to 0.5 in thickness. Their outer membrane, which is often discontinuous, is thicker than cytoplasmic membranes. The density of the matrix shows distinct gradations. The organelles contain tubules, possibly built up by a spiral molecular chain. The distribution of the organelles is investigated with stereological methods. Their volume density in endothelial cell cytoplasm appears to depend more on the distance from the heart than on vessel size, the thoracic aortae showing the highest organellae content of 8%. Connections between organelles and cytoplasmic membranes are demonstrated. Particularities of endothelium, among them aggregations of ribosomes, are pointed out.

Herrn Prof. Dr. med. F. Hammersen und Fräulein Dr. med. M. Lewerenz danke ich herzlich für Anregungen, Unterstützung und Kritik. Die Arbeit entstand während eines vom Deutschen Akademischen Austauschdienst dankenswerterweise gewährten Stipendiums; sie wurde unterstützt vom Schweizerischen Nationalfonds zur Förderung der wissenschaftlichen Forschung, Kredit-Nr. 3952.     

The superficial vascular hyaloid system in the eye of the frogs,Rana temporaria and Rana esculenta

Summary The angioarchitecture of the superficial vascular hyaloid system (membrana vasculosa retinae) of the frog eye was studied by means of scanning electron microscopy of vascular corrosion casts. The terminal vessels form a single-layered sheath intimately adjacent to the vitreal side of the avascular retina. The hyaloid system is subdivided by the ventral venous trunk into three central areas: the dorsal, the temporo-ventral, and the naso-ventral area. Toward the ora serrata, the hyaloid system is bordered by an arterial ring, and by nasal and temporal venous branches forming more or less complete hemicircles. A vascular zone composed of several tongue-like sectors establishes an inter-connection between the peripheral vascular rings and the central areas of the fundus. The arterial blood is supplied from the arterial ring. The drainage of the hyaloid system is provided via two routes: (1) the Y-shaped ventral trunk collects blood from the central areas, (2) the two peripheral venous branches drain the tongue-like sectors. The vessels within the dorsal area follow preferentially a dorso-ventral meridional direction. This densely capillarized territory corresponds in localization to the area centralis retinae. The ultrastructure of microvessels of the hyaloid system is characterized by features typical for capillaries of the central nervous system.     

Electron-microscopic investigation of the innervation of the pars distalis of the hypophysis in adult Rana temporaria L.

Summary In the pars distalis of the hypophysis of adult Rana temporaria, three types of nerve-fiber profiles were found at two distinct sites, in both lateral parts of the bordering regions of the anterior lobe with the intermediate lobe of the hypophysis. The first type of nerve-fiber profile consists of bundles of very fine axonal elements (diameter: <0.7 m). The second type is formed by larger nerve fibers (diameter up to 4 m) containing a few neurosecretory granules of approximately 100 nm. The third type of nervefiber profile resembles the second type but these nerve fibers make synaptoid contacts on at least two different types of glandular cells. The possible functional significance of these nerve fibers in the pars distalis is discussed.No nerve fibers were found (1) in the central part of the bordering region of the pars distalis with the intermediate lobe, (2) at the bordering region with the median eminence and (3) with the neurohypophysial stalk, and (4) in all other parts of the pars distalis.     

The structure and vascularization of the pars tuberalis of the hypophysis of Rana temporaria

Summary The pars tuberalis of the hypophysis of Rana temporaria presents the general structural and the cytological characteristics of an endocrine gland. It is composed of elongated cells with long, branching processes ending on the external basement membrane of the pericapillary space. The pars tuberalis cells produce secretory granules which are accumulated in the pericapillary endings of the processes.Corresponding to its separate localization, the pars tuberalis of Rana temporaria has a separate vascularization of which the efferent capillaries anastomose with the capillary plexus of the median eminence. The general direction of the blood flow of the pars tuberalis is towards the capillaries of the median eminence. Also, the secretory products of the pars tuberalis pass into the blood stream of the hypophysial portal system.Several characteristics of the pars tuberalis show that its function must be different from that of the pars distalis of the hypophysis. Moreover, in contrast with the pars distalis, the activity of the pars tuberalis is not regulated by neurohumoral factors.The results show that a role of the pars tuberalis in the regulation of the activity of the pars distalis of the hypophysis is not excluded.     

Seasonal changes of the ultrastructure of the pars tuberalis of the hypophysis of Rana temporaria

Summary In the pars tuberalis of the hypophysis of Rana temporaria, which shows the ultrastructural characteristics of a polypeptide hormone secreting endocrine gland, seasonal changes of the ultrastructure are described. In accordance with the literature, these seasonal changes of ultrastructure are interpreted as the morphological expression of seasonal changes of endocrine activity of the pars tuberalis.     

The hypothalamo-hypophysial system of the frog Rana temporaria

Summary The median eminence (ME) of the adult frog, Rana temporaria, was studied by means of electron microscopy including quantitative electron-microscopic autoradiography. In frogs captured in May and June numerous peptidergic neurosecretory fibres extending via the internal zone to the pars nervosa display large swellings containing few granules, mitochondria, neurotubules and cisternae of the smooth endoplasmic reticulum. In addition, few secretory globules up to 1.5 m in diameter occur in these varicosities. In animals collected during the autumn period many of these neurosecretory swellings filled with neurosecretory granules and polymorphic inclusions resemble Herring bodies. Three types of granule-containing neurosecretory fibres were observed in the external zone (EZ) of the ME of adult R. temporaria. Peptidergic A₁- and A₂-type fibres are characterized by granules 150–220 nm and 100–160 nm in diameter, respectively. Monoaminergic fibres of type B with granules approximately 100 nm in diameter represent 50% of all neurosecretory elements in the EZ of the frog ME; 12% of the total number of granule-bearing axons in the EZ actively taking up radiolabelled 5-hydroxytryptophan are thought to be serotoninergic terminals. Neurosecretory terminals of all types and glial vascular endfeet establish direct contacts with the perivascular space of the primary portal capillaries. Some neurosecretory terminals are separated from the lumen of the third ventricle by a thin cytoplasmic lamella of tanycytes. The possible physiological significance of this structural pattern is discussed.     

The optic tracts of Rana temporaria and a possible retino-preoptic pathway

Summary The optic tracts and centres of optic terminals of Rana temporaria have been investigated with silver impregnation techniques after unilaterally cutting the optic nerve and autoradiographically after injection of a mixture of tritiated amino acids into the vitreous body of the left eye.The observations on the course of the optic tracts and on the optic terminals in the thalamus and optic tectum are to a great extent in agreement with those of other authors. The probability of a retino-preoptic pathway is supported by the detection of fibres running in dorsal direction in front of the place where the optic nerve penetrates the brain. In horizontal sections these fibres can be seen deviating from the optic tract, covering a short distance in frontal direction and then turning upward under an angle of about 90 degrees. They disappear between the ventral aldehyde-fuchsin positive cells of the preoptic nucleus. The ventral and median parts of the ipsilateral preoptic nucleus contain fragments of degenerated fibres.Autoradiographic data are also in favour of the presence of an ipsilateral retino-preoptic tract. After carefully counting the number of grains over left and right preoptic nucleus, it appeared that over the ventral and median parts of the left preoptic nucleus more grains occur than over the identical areas of the right one.It is a pleasure to acknowledge Prof. Dr. J. C. van de Kamer and Dr. F. C. G. van de Veerdonk for their helpful encouragement and constructive suggestions, Dr. L. Boomgaart for checking and amending the English writing. Thanks are also due to Miss Ans de Groot and Miss Thera Verstappen for their technical assistance and to the staff of the photographic department for making the illustrations.     

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

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

Sound and vibration sensitivity of VIIIth nerve fibers in the grassfrog, Rana temporaria

We have studied the sound and vibration sensitivity of 164 amphibian papilla fibers in the VIIIth nerve of the grassfrog, Rana temporaria. The VIIIth nerve was exposed using a dorsal approach. The frogs were placed in a natural sitting posture and stimulated by free-field sound. Furthermore, the animals were stimulated with dorso-ventral vibrations, and the sound-induced vertical vibrations in the setup could be canceled by emitting vibrations in antiphase from the vibration exciter. All low-frequency fibers responded to both sound and vibration with sound thresholds from 23 dB SPL and vibration thresholds from 0.02 cm/s². The sound and vibration sensitivity was compared for each fiber using the offset between the rate-level curves for sound and vibration stimulation as a measure of relative vibration sensitivity. When measured in this way relative vibration sensitivity decreases with frequency from 42 dB at 100 Hz to 25 dB at 400 Hz. Since sound thresholds decrease from 72 dB SPL at 100 Hz to 50 dB SPL at 400 Hz the decrease in relative vibration sensitivity reflects an increase in sound sensitivity with frequency, probably due to enhanced tympanic sensitivity at higher frequencies. In contrast, absolute vibration sensitivity is constant in most of the frequency range studied. Only small effects result from the cancellation of sound-induced vibrations. The reason for this probably is that the maximal induced vibrations in the present setup are 6–10 dB below the fibers' vibration threshold at the threshold for sound. However, these results are only valid for the present physical configuration of the setup and the high vibration-sensitivities of the fibers warrant caution whenever the auditory fibers are stimulated with free-field sound. Thus, the experiments suggest that the low-frequency sound sensitivity is not caused by sound-induced vertical vibrations. Instead, the low-frequency sound sensitivity is either tympanic or mediated through bone conduction or sound-induced pulsations of the lungs.Abbreviations AP amphibian papilla - BF best frequency - PST peristimulus time     

Weight loss,reproductive output,and the cost of reproduction in the common frog,Rana temporaria

Summary The consequences of reproduction for body weight, growth and survival were studied in a Swiss population of the explosive breeder, Rana temporaria. Males and females continuously loss weight in the range of 0.5% of total body weight per day from the breeding migration throughout May. Females also lost about 33% (1983) and 29% (1984) due to spawning. In addition to this significant year-to-year variation, there was also considerable individual variation in reproductive output. Skeletochronological techniques indicated that breeding male or female frogs experienced a growth reduction of several millimeters relative to non-breeding frogs of the same body size. There was no relationship between an individual female's reproductive output in consecutive years or with her subsequent growth or survival. It was concluded that weight loss is caused by a seasonally elevated metabolism in combination with a lack of feeding and represents a basic energetic cost of reproduction, resulting in lowered growth. Individual variation in relative reproductive output is mostly environmentally induced and is not an expression of different reproductive strategies. This may explain the lack of trade-offs that are predicted by the cost-of-reproduction-hypothesis.     

Reaktionsmuster verschiedener Mittel-und Zwischenhirnzentren von Rana temporaria L. nach Unterbrechung der Nervenbahnen des Pinealkomplexes

Zusammenfassung Der Einfluß des lichtempfindlichen Pinealkomplexes (Stirnorgan und Epiphysis cerebri) von Rana temporaria auf dorsale diencephale und mesencephale Hirnzentren breitet sich über ein größeres Gebiet aus als man bisher angenommen hat. Nach operativer Unterbrechung des Nervus- und Tractus pinealis wurde mit Punktzählverfahren die Dichte der Zell- und Zellkernvolumina sowie der Kapillaroberfläche im dorsalen Thalamus, mesencephalen (postthalamischen) zentralen Grau, Ganglion habenulae, Tectum opticum, in der Area praetectalis und im Subcommissuralorgan (Sco) bestimmt. Die morphometrischen Meßergebnisse lassen sich wie folgt interpretieren: 1. Der Pinealkomplex wirkt auf das Sco, das zentrale Grau und die Area praetectalis ein (vgl. Paul u. Mitarb., 1971); das Sco wird in erster Linie über das Stirnorgan beeinflußt. 2. Weitere Kerngebiete des Zwischenhirns (Ganglion habenulae und dorsaler Thalamus) gehören funktionell zu dem vom Pinealkomplex kontrollierten System. 3. Die von Kemali und Braitenberg (1970) beschriebene Asymmetrie der Ganglia habenularum zeigt sich auch im funktionellen Zusammenspiel dieser Kerngebiete mit dem Pinealkomplex, dem dorsalen Thalamus und dem dorsalen Abschnitt des zentralen Graus. Die in Stirnorgan und Epiphysis cerebri transformierten Lichtreize werden in Hirnabschnitte weitergeleitet, die olfaktorische und retinale Signalmuster verarbeiten. Über die biologische Bedeutung der nervösen Beziehungen des Stirnorgans zum sekretorischen Sco können noch keine Angaben gemacht werden.

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Study of mesencephalic and diencephalic centers after transection of the centripetal nervous pathways of the pineal complex in Rana temporaria L.

Summary Nervous influences of the pineal complex of the frog, Rana temporaria, exerted upon mesencephalic and diencephalic centers were investigated using morphometric methods after the transection of the nervus or the tractus pinealis. Densities of the cell- and nuclear volumes and of the capillaries were measured by a point-counting method in the dorsal thalamus, the mesencephalic (postthalamic) central gray, the habenular ganglion, the optic tectum, the pretectal area, and the subcommissural organ (sco). The morphometric measurements showed: 1. A functional influence of the pineal complex on the sco, the central gray, and the pretectal area (see Paul et al., 1971); the frontal organ seems to be primarily related to the sco. 2. The habenular ganglia and the dorsal thalamus belong to a system which is controlled by the pineal complex. 3. The morphological asymmetry of the habenular ganglia (Kemali and Braitenberg, 1970) has an influence on the functional interactions between the pineal complex, the dorsal thalamus, and the central gray. The impulses arising in the light-sensitive pineal complex of the frog are conducted to brain regions which are known to integrate olfactory and retinal sensory inputs. The functional significance of the nervous connexion of the frontal organ with the secretory sco is still open to discussion.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft (Arbeitskreis Prof. Dr. A. Oksche).     

Patterns of cellular proliferation and migration in the developing tectum mesencephali of the frog Rana temporaria and the salamander Pleurodeles waltl

The development of the tectum mesencephali was studied in the frog Rana temporaria and the salamander Pleurodeles waltl by means of nuclear staining and by labeling of cells with bromodeoxyuridine (BrdU). The general spatial and temporal pattern of cell proliferation and cell migration is the same in both species, despite drastic differences in overall tectal morphology. However, the salamander species differs from the frog species by (1) a generally lower cell proliferation rate, (2) a reduction in the activity of the lateral proliferation zone, and (3) a reduction in the formation of superficial cellular layers. Because point (3) affects processes that occur late in ontogeny, our experiments provide evidence that the simple morphology of the tectum of Pleurodeles waltl, compared with the multilayered tectum of Rana, is a consequence of a paedomorphic alteration of the ancestral developmental pattern of the amphibian tectum.