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.     

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.     

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.     

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.     

Cell junctions with funnels in the olfactory mucosa of frogs (Rana temporaria L.)

Summary A characteristic structure in the apical junctional belt of the olfactory epithelium in Rana temporaria is visible in freeze-fracture preparations. This structure is described as a funnel with channel across the junctional belt. It is supposed to represent a possible way for discarding used molecules after stimulation, and to allow the stimulation of free nerve endings in the depth of olfactory epithelia.I wish to thank Prof. C.F. Bardele and Mr. H. Schoppmann for their kind support and technical help     

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.     

Thymic vascular system of the European common frog,Rana temporaria: a scanning electron-microscopic study of vascular casts

Vascular corrosion casts of the thymus of adult individuals of the European common frog, Rana temporaria, were analysed by scanning electron microscopy. The main arterial vessel, which is derived either from the temporal artery or from the auricular ramus, approaches the central territory of the gland and branches into twigs that, on penetrating the parenchyma, give rise to capillaries. Most of these capillaries run vertically towards the surface of the gland; they either join the superficial capillary plexus or follow this plexus for a variable distance and then run back towards the medulla, forming capillary loops. The former capillaries link with the extensive venous plexus composed of irregular meshes, whereas the latter capillaries join the venules at the cortico-medullary boundary and finally escape into collecting veins on the gland surface. The venous twigs, which join together near the gland, form the main thymic vein, which empties into the external jugular vein. The details of the thymic vasculature of the anuran amphibian, R. temporaria, are compared with those described in mammalian species, viz. the mouse, rat and guinea pig.     

Directionality of phase locking in auditory nerve fibers of the leopard frog Rana pipiens pipiens

Summary A dorsal approach to the eighth nerve and free-field stimulation were used to investigate the effect of sound direction and intensity on phase locking in auditory nerve fibers of the leopard frog Rana pipiens pipiens.Tuning curves of 75 auditory neurons were analyzed (Fig. 2). Amphibian papillar neurons, but not basilar papillar neurons, exhibit significant phase locking to short tone bursts at the characteristic frequency (CF), the degree of phase locking (vector strength) decreasing with the neuron's CF (Figs. 3, 4 and 10E). Vector strength increases with sound pressure level to saturate about 20 dB above threshold, while the preferred firing phase is only slightly affected (Figs. 5 and 6).In contrast, sound direction hardly affects vector strength (Figs. 7, 8, 9A and 10A and C), but has a strong influence on the preferred firing phase (Figs. 7, 8, 9B and C, 10B and D): With respect to anterior tone presentation there are phase lags for ipsilateral and phase leads for posterior and contralateral presentation. Phase differences between both ears show a sinusoidal or cardioid/ovoidal directional characteristic; maximum differences are found with antero-lateral tone presentation (Fig. 11). The directionality of phase locking decreases with the neuron's CF (Fig. 10F) and only slightly changes with sound pressure level (Fig. 12). Thus, phase locking of amphibian papilla neurons can potentially provide intensity-independent information for sound localization.Abbreviations SPL sound pressure level - FTC frequency threshold curve - CF characteristic frequency - TF test frequency - VS vector strength - AP amphibian papilla - BP basilar papilla     

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.     

Die Ontogenese der aminhaltigen Neuronensysteme im Gehirn von Rana temporaria

Zusammenfassung Vorkommen und Verteilung biogener Amine im Gehirn von Rana temporaria-Kaulquappen wurden fluoreszenzmikroskopisch untersucht. Catecholaminhaltige Perikaryen erscheinen ab Stadium 20 im Nucleus reticularis mesencephali, im Tuber cinereum und im Bulbus olfactorius, ab Stadium 22 in den Flügelplatten der Medulla oblongata und in der Area praeoptica. Ab Entwicklungsstufe 20 zeigen sich ventrolateral in Medulla oblongata und Mittelhirn, lateral vom Organon vasculosum hypothalami, im Bereich des medialen Vorderhirnbündels und im Striatum catecholaminhaltige Faseranschwellungen, ab Stadium 22 außerdem in der Eminentia mediana und dem Hypophysenzwischenlappen, in der Commissura transversa (bis zur Stufe 26), in der Commissura anterior (bis zur Stufe 26) und in der Pars ventrolateralis nuclei lateralis septi. Im Striatum ist von dieser Entwicklungsstufe an ein zweites Areal mit grün fluoreszierenden Varikositäten nachweisbar. Ab Stadium 26 finden sich auch in der Pars dorsolateralis des lateralen Septumkerns catecholaminhaltige Faseranschwellungen.Ab Entwicklungsstufe 22 sind 5-HT-haltige, gelb fluoreszierende Perikaryen im Nucleus raphes und in seiner Umgebung zu beobachten, gelb fluoreszierende Varikositäten im Nucleus interpeduncularis und zwischen medialem und lateralem Septumkern.

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Ontogeny of the amine-containing nerve cell systems in the brain of Rana temporaria

Summary The occurrence and distribution of biogenic amines in the brain of Rana temporaria tadpoles have been investigated with the fluorescence-microscope. From the embryonic developmental stage 20 onwards catecholamine-containing cell bodies are shown to be present in the nucleus reticularis mesencephali, the tuber cinereum and the olfactory bulb, and from stage 22 onwards also within the dorsolateral areas of the medulla oblongata and within the preoptic area. Catecholamine-containing enlargements of nerve fibres occur in the ventrolateral parts of the medulla oblongata and the midbrain, in an area lateral to the hypothalamic organon vasculosum, within the region of the medial forebrain bundle and within the striatum, in all stages following stage 20. These enlargements also occur in the median eminence and the pars intermedia of the hypophysis, in the commissura transversa (up to stage 26), in the commissura anterior (also up to stage 26) and in the pars ventrolateralis nuclei lateralis septi in all stages after 22. From the same stage onwards a second area of green fluorescent varicosities can be demonstrated within the striatum. After stage 26 catecholamine-containing enlargements of nerve fibres additionally are to be found in the dorsolateral part of the lateral septal nucleus.After appearing at stage 22 5-HT-containing, yellow fluorescent perikarya are to be observed within the nucleus raphes and its neighbourhood, and yellow fluorescent varicosities in the interpeduncular nucleus and in an area between the medial and the lateral septal nucleus.

Herrn Professor Dr. med. W. Bargmann zum 65. Geburtstag gewidmet.     

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.     

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.     

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.     

Amiloride-sensitive Na+/H+ exchange stimulated by cellular acidification or shrinkage in red blood cells of the frog,Rana temporaria

Simultaneous net uptake of Na⁺ and net extrusion of H⁺, both inhibited by amiloride, could be stimulated in red blood cells of the frog, Rana temporaria, either by intracellular acidification or cellular shrinkage. Net transports of Na⁺ and H⁺ were transient, dying out after 10–20 min (20°C) when stimulated by intracellular acidification but developing more slowly and proceeding for more than 60 min (20°C) when stimulated by cellular shrinkage. Evidence is presented suggesting a coupling between the transports of Na⁺ and H⁺ with an exchange ratio of 1:1 Na⁺/H⁺ exchange, stimulated by intracellular acidification, was able to readjust intracellular pH also when operating in parallel to a fully working anion exchanger in CO₂/HCO ₃ ^- -buffered media. Inhibition of anion exchange resulted in reduced cellular net uptake of Na⁺.Abbreviations DIDS 4,4-diisothiocyanatostilbene-2,2-disulphonate - DMSO dimethylsulphoxide - IU international unit - pH _e extracellular pH - pH _i intracellular pH - RBC red blood cell     

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.     

Post-freeze recovery of peripheral nerve function in the freeze-tolerant wood frog,Rana sylvatica

We investigated the restoration of peripheral nerve function and simple neurobehavioral reflexes in the freeze-tolerant wood frog (Rana sylvatica). Thirty-two specimens, allowed to freeze for 39 h and ultimately cooled to-2.2°C, were sampled at various time intervals up to 60 h after thawing at 5°C was initiated. The sciatic nerves of treated frogs were initially unresponsive to stimulation, but usually regained excitability within 5 h. Except for a slight reduction in nerve excitability characteristics of the compound action potentials of treated frogs were indistinguishable from those of control frogs. Recovery times for the hindlimb retraction and righting reflexes were 8 h and 14 h, respectively. Concentrations of the cryoprotectant glucose increased 8.2-fold in the sciatic nerve and 10.5-fold in the underlying semimembranosis muscle of treated frogs, and remained elevated for at least 60 h after thawing was initiated. These organs lost 47.2% and 15.9%, respectively, of their water during freezing, but were rehydrated within 2 h of the onset of thawing. The accumulation of glucose and the withdrawal of tissue water apparently are cryoprotective responses which enable this species to survive freezing.     

Scanning electron microscopy of the wall of the third ventricle in the brain of Rana temporaria. Part IV.

Summary The surface specializations of the wall of the third cerebral ventricle of Rana temporaria were investigated with the scanning electron microscope. These specializations can be divided into three types: cilia, large bulbous protrusions, and microvillus-like protrusions.Most parts of the ventricular surface are densely ciliated. In contrast, other regions are either scantily ciliated or devoid of cilia. Four areas of the ventricular surface are studded with numerous large bulbous protrusions. These large protrusions can be divided into two types: One type consists of intraventricular end bulbs of dendrites of secretory neurons. The other type is represented by large cytoplasmic extensions of ependymal cells.In the third ventricle of Rana, microvillus-like surface specializations of ependymal cells are ubiquitous structures. Generally, filiform protrusions of varying length are the predominant type. The microvillus-like specializations are transient structures, the number of which varies according to different physiological states of the ependymal cells.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek