Cerebellar afferents in the frogs,Rana esculenta and Rana temporaria

Summary Afferents to the cerebellum in frogs (Rana esculenta, Rana temporaria) were studied by use of retrograde transport of horseradish peroxidase. Following injections restricted to the molecular layer of the cerebellum cell labelling was found in the contralateral inferior olive and the ventral portion of the caudal medullary raphe. Injections involving the granular layer resulted in labelling in the ventral horn of the cervical spinal cord, the caudal spinal trigeminal nucleus, the nucleus caudalis and the medial portion of the nucleus ventralis of the vestibular nerve, the inferior reticular nucleus and the nucleus of the fasciculus longitudinalis medialis. Following larger injections, which may have spread significantly into the cerebellar, secondary gustatory, trigeminal or vestibular nuclei, labelled cell bodies were also found in the nucleus ruber, nucleus solitarius, the rostral spinal trigeminal nucleus and the rostral rhombencephalic reticular formation. It is unclear whether the fibers from these latter areas innervate the cerebellum of the frog, as they do in mammals, or only reach the underlying areas. This situation emphasizes a limitation of the HRP technique when applied to small structures as is often the case in lower vertebrates.Supported by Grant Gr 276 to U. G.-C. from the Deutsche Forschungsgemeinschaft.     

Intra-individual length heterogeneity of Rana esculenta mitochondrial DNA

Mitochondrial DNA extracted from Rana esculenta oocytes appears heterogeneous in size. The length of these molecules varies continuously from 18,700 bp to 19,700 bp. Each animal is heteroplasmic and can be characterized by the range of the variation (400-700 bp) and the extreme sizes of the various molecules it carries. The variable region of the genome has been localized between the coding region and the replication origin area.     

Seasonal variations of Rana esculenta L. skin tyrosinase

Various enzymes are known to be involved in melanin biosynthesis, but the key role appertains to tyrosinase. In amphibians this enzyme displays peculiar characteristics: i) it requires an activation process; ii) its level of enzymatic activity in the animal skin changes depending on the season. In this work, by using chymotrypsin, subtilisin and SDS as putative activators, we studied the activation process of the skin pro-tyrosinase of Rana esculenta L. in different seasons over a period of two years. We found that chymotrypsin and subtilisin were able to yield an active enzyme, but not SDS. The maximum levels of tyrosinase activity were recorded in winter and the minimum in summer. We detected tyrosinase activity in the melanosomal fraction, where the enzyme form was least sensitive to proteolytic activation, probably corresponding to a "mature" tyrosinase. The enzyme forms found in the microsomal and soluble fractions were more sensitive to proteolytic activation, probably corresponding to "immature" tyrosinase. On SDS-PAGE, the tyrosinase activity assays showed a dopa-positive band at 200 kDa and a second aggregated band with a still higher molecular mass. The significance of these results in frog melanogenesis regulation is discussed.     

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

Zusammenfassung Die 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.

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Fluorescence microscopic studies of biogenic amines in the pineal organ of Rana esculenta and Rana pipiens

Summary The 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.

Finanzierung eines Studienaufenthaltes an der Universität Lund durch die Deutsche Forschungsgemeinschaft (Forschungsvorhaben — Ok 1/15 — von Prof. Dr. A. Oksche u.a.).     

Glucocorticoid receptor of frog (Rana esculenta) liver

The presence of a glucocorticoid soluble receptor is demonstrated in frog liver cytosol. The kinetic characterization of frog liver cytosolic receptor for glucocorticoids is reported and its steroid specificity assessed. Results indicate a gross similarity between frog liver and mammalian glucocorticoid receptor, being a major difference the reduced binding capacity.     

Male vocalization and female choice in the hybridogenetic Rana lessonae/Rana esculenta complex

In many species, females can improve their fitness by preferring particular males over others. In Palaearctic water frogs of the Rana lessonae/R. esculenta complex the consequences of such mate choice are particularly pronounced. To produce viable offspring, the hybrid R. esculenta (genotype RL) must mate with the parental species R. lessonae (LL); but R. lessonae should avoid mating with R. esculenta, because the resulting hybrid offspring will eliminate the L genome from the germline (hybridogenesis). Hence, there exists a conflict between the sexual parasite (RL) and its sexual host (LL) over the best mating partner. Previous studies have shown a preference for LL males in LL and RL females; but they have also shown that females cannot usually realize their choice when in close proximity to males, because the males forcefully and indiscriminately amplex them. We tested whether females use male vocalizations as a long-distance signal to increase their chances of mating with the preferred LL males. We exposed female R. lessonae and R. esculenta to playbacks of single LL and RL mating calls (experiment 1) and to choruses with a 3:1 excess of LL and RL calls, respectively (experiment 2). In experiment 1, both female types were attracted more by the LL than by the RL calls. In experiment 2, no discrimination between LL- and RL-dominated choruses was observed. The results suggest that females do not use distant male vocalization to approach preferentially ponds or arenas within a pond that hold an excess of LL males. But once they have arrived in a chorus, mating calls from nearby males can direct them to the preferred LL mates. We discuss possible reasons for the failure to discriminate between choruses and the chances for successful choice between individuals within choruses. Copyright 2000 The Association for the Study of Animal Behaviour.     

Organization of efferent extraspinal sympathetic pathways in Rana esculenta

Origin and distribution of pre- and postsynaptic fibres in the sympathetic trunk of Rana esculenta (from ganglion 3 to ganglion 10) have been investigated by means of extracellular recordings. Two systems conducting efferent information appear to exist: 1) a system made of faster conducting fibres (B group pre- and postsynaptic fibres); presynaptic fibres originating from a very high monosegmental source (4th spinal root); postsynaptic fibres leave the sympathetic chain plurisegmentally (rami communicantes 5-10); 2) a system made of slower conducting fibres (C group pre- and postsynaptic fibres) originating plurisegmentally from spinal roots 5-8. Intracellular recordings have shown that: 1) integrative processes take place in the amphibian sympathetic trunk, as in the mammalian one, but are quantitatively lesser. Homonomous (B-B) and heteronomous (B-C) convergence has been observed in B neurons, and also the convergence of a collateral of a C postsynaptic axon on B neurons. 2) posthumous depolarizations are present: these are events modulating the activity of sympathetic neurons. In B neurons posthumous depolarization follows orthodromic responses, and a late posthumous depolarization can be seen in B and C neurons following either ortho- or antidromic stimulation.     

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.