Effects of nitrate on the interactions of the tadpoles of two ranids (Rana clamitans and R. catesbeiana)

Nitrogen pollution as a result of agricultural runoff and atmospheric deposition is a major challenge to aquatic ecosystems, and is likely to increase in the future. Nitrogenous pollutants are potential stressors of amphibian larvae through their toxicological impacts on individuals; however, they may also increase primary productivity. We examined how such an anthropogenic stressor could influence the interactions between two potentially competing species of tadpoles (Rana clamitans and R. catesbeiana). In a 42 d mesocosm experiment, R. catesbeiana survival, but not final mass, was reduced by nitrate addition. Rana catesbeiana survival was lower when in competition with R. clamitans than when only experiencing intraspecific competition. For R. clamitans, survival was not affected by nitrate addition or competition type. Rana clamitans in nitrate addition mesocosms were heavier than tadpoles from no nitrate mesocosms, and were heavier in intraspecific than in interspecific mesocosms. Our results suggest that nitrate can influence the performance of amphibian larvae, and that its effects could have complex effects on aquatic ecosystems.     

Leukocyte surface markers in Rana catesbeiana,identified using mouse monoclonal antibodies

Monoclonal antibodies (mAbs), each reactive with thymocytes, neutrophils, and thrombocytes of the bullfrog, Rana catesbeiana, were raised and used for studies of hematopoiesis of metamorphic larvae. The mAbRc-T1, which immunoreacted exclusively with thymocytes in adults, was also strongly reactive to larval thymocytes. Thus the determinant Rc-T1 may provide an appropriate marker for thymocytes, although antigenic sharing is also seen in larval ovarian tissue. Neutrophils detectable by both mAbRc-N1 and Rc-N2 originated primarily in the mesonephros and showed up increasingly in peripheral blood of larvae. Thrombocytes detectable by mAbRc-P may originate in the larval spleen and/or liver but not in the mesonephros.     

Characterization of Rana catesbeiana HSP30 gene and its expression in the liver of this amphibian during both spontaneous and thyroid hormone-induced metamorphosis

During metamorphosis, the Rana catesbeiana tadpole undergoes developmental changes in almost every tissue/organ. These changes prepare the ammonotelic, swimming larva for its transition to a ureotelic, terrestrial adult, and involve dramatic remodeling. The postembryonic changes in this tadpole are initiated by the thyroid hormones (TH) and result in the extensive degradation of proteins and degeneration of tissues characteristic of the larval phenotype and in the de novo synthesis of proteins characteristic of the adult phenotype. We questioned whether the drastic nature and abruptness of the TH-dependent, postembryonic changes occurring in the tissues of this tadpole might be perceived by the cells in some tissues as stressful and, therefore, cause them to express heat shock and/or stress-like proteins. To address this question, we isolated and characterized a Rana catesbeiana hsp30 gene and used sequences from it to determine if mRNAs encoded from it, or other members of this gene family, are expressed in tissues of tadpoles undergoing metamorphosis. Our results demonstrate that the liver of metamorphosing Rana catesbeiana tadpoles accumulate hsp30 mRNAs and express the heat shock proteins they encode. The fact that the expression of these hsp30s in the liver of these tadpoles is coincidental with the TH-induced expression of genes encoding the liver-specific urea cycle enzymes suggests that TH may influence, directly or indirectly, the expression of these hsp30 genes and, moreover, implies that the presence of one or more of these heat shock proteins may be necessary for the developmental transitions occurring in this organ. © 1996 Wiley-Liss, Inc.     

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

Zusammenfassung Der Epiphysenkomplex von 32 Fröschen der Arten Rana temporaria, Rana esculenta und Rana catesbeiana wurde zuerst elektrophysiologisch, anschließend neuro-histologisch bzw. elektronenmikroskopisch untersucht. Auf Belichtung des Stirnorgans lassen sich vom Nervus pinealis (= Tractus frontalis) zwei Arten von Antworten ableiten. Antidrome elektrische Reizung des Tractus pinealis zeigt im Nervus pinealis fortgeleitete Aktionspotentiale und damit Nervenfasern, die vom Stirnorgan bis zum Mittelhirn durchgehen. Elektrophysiologisch wurden außerdem Fasern ermittelt, die die Epiphysis cerebri nicht erreichen. Weitere Fasern enden in der Epiphyse. Neben den vom Stirnorgan zum Gehirn ziehenden afferenten Fasern kann man im extracranialen Teil des Nervus pinealis auch efferente Fasern nachweisen.Histologisch ist in allen Fällen der Durchtritt des Nervus pinealis durch das Schädeldach zu beobachten. Der Durchmesser des Nerven nimmt jedoch wahrend seines Verlaufs kontinuierlich ab. Elektronenmikroskopisch konnte gesichert werden, daß diese Durchmesserabnahme auf einem Faserverlust beruht. Bei einer Rana catesbeiana wurden kurz vor Eintritt des Nervus pinealis in die Epiphyse — im Vergleich zum extracranialen Nervenabschnitt —nur noch die Hälfte der markhaltigen und 1/3 der marklosen Nervenfasern gezählt. Bei den verbleibenden markhaltigen Fasern war außerdem die Dicke der Markscheide stark reduziert. Der Nervus pinealis fächert sich in Epiphysennähe auf; seine Fasern treten dorsomedian einzeln durch die Basalmembran in das Parenchym des rostralen Epiphysenabschnittes ein. Gemeinsam mit den Neuriten der Epiphysennervenzellen bilden sie den Tractus pinealis (s. auch Paul u. Mitarb., 1971). Eine synaptische Umschaltung der Fasern des Nervus pinealis konnte in der Epiphyse nicht nachgewiesen werden. Bei einer Rana esculenta betrug der Faseranteil des Nervus pinealis am Tractus pinealis nur etwa 2%. Die Befunde werden im Zusammenhang mit der Entstehung und Leitung der chromatischen und achromatischen Antwort im Epiphysenkomplex diskutiert.

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Further experimental and neuroanatomical investigations of the nerve tracts of the pineal complex in anura

Summary The pineal systems of 32 frogs of the species Rana temporaria, Rana esculenta, and Rana catesbeiana were studied first electrophysiologically and then with histological and electron microscopical methods. The nervus pinealis (= tractus frontalis) showed two kinds of responses to photic stimulation. Uninterrupted conduction of nerve impulses from the frontal organ to the midbrain was demonstrated by antidromic electrical stimulation of the intracranial pineal tract and recording of action potentials from the extracranial pineal nerve. Beside the uninterrupted fibres from the frontal organ to the tractus pinealis, other fibres of the nervus pinealis either did not reach the pineal organ or ended in the pineal organ. The extracranial part of the nervus pinealis contained afferent as well as efferent fibres.Morphologically it was shown in 32 investigated animals that the nervus pinealis penetrates the skull and runs to the pineal organ. During its course, the diameter of the pineal nerve became smaller, which was related to a decrease of the number of myelinated and unmyelinated fibres as revealed by the electron microscope. In comparison with the extracranial portion of the pineal nerve, the intracranial portion of the nerve near the pineal organ contained only half of the myelinated and one third of the unmyelinated fibres counted in one specimen of Rana catesbeiana. The thickness of the myelin sheats of the residual myelinated fibres was much reduced. The nervus pinealis entered the rostral region of the pineal organ, and single nerve fibres were seen to penetrate the basal lamina of the pineal organ reaching its parenchyma dorsomedially. These fibres, and axons of the nerve cells of the pineal organ, constituted the pineal tract (see also Paul et al., 1971). In one specimen of Rana esculenta, only 2% of the nerve fibres of the tractus pinealis originated in the frontal organ. The results have been discussed in relation to the generation and conduction of the chromatic and achromatic responses in the pineal system.

Herrn Prof. Dr. Dr. h. c. W. Bargmann, Kiel, in Dankbarkeit gewidmet.     

The Presence of Intramitochondrial Yolk-crystals in Oocytes of Hypophysectomized Bullfrog Tadpoles*

Ovaries of hypophysectomized Rana catesbeiana tadpoles. weighing I to 14 g, were prepared for electron microscopic study. The oocytes are at the growth phase, ranging from 50 to 190 μm in diameter. The observation on these oocytes has revealed the presence of intramitochondrial yolk-crystals but not cytoplasmic yolk platelets. The crystalline structure, situated within the intracristal space, consists of a hexagonal array of dense particles about 50 Å in diameter and 72 Å in periodicity. Our data agree with those reported in oocytes of intact ranid species. According to literatures, crystals of intramitochondrial yolk and of cytoplasmic yolk platelets show similar ultrasturctures. The precursor of cytoplasmic yolk platelets in adult Xenopus oocytes is known to be synthesized in the estrogen-stimulated liver and incorporated via circulation into oocytes by gonadotropin-dependent micropinocytosis. The present finding suggests that the intramitochondrial yolk could be formed within oocytes, independently of the pituitary control.     

Blood oxygen dissociation curve of the frogsRana catesbeiana andRana brevipoda

Summary The blood O₂ dissociation curve was determined with a microphotometric reaction apparatus in two frog species,Rana catesbeiana andRana brevipoda, and formulated in terms of a modified Hill equation. The O₂ dissociation curve was in reasonable agreement with determinations of O₂ content, O₂ capacity, and pH (or ) in blood samples taken from various blood vessels and heart cavities. The blood O₂ affinity inRana catesbeiana (mean body weight 230 g, kept at 25°C) was low:P ₅₀=42 Torr (at 25°C and at ventricular blood pH=7.79). The blood of the smaller frog,Rana brevipoda (mean body weight 16 g), had even lower O₂ affinity,P ₅₀ being 52 Torr (at 25°C, pH=7.72). The decrease ofP ₅₀ with increasing body mass, resulting from comparison of these two frog species, is in accordance with literature data on mammals and birds.     

Light-Dependent Shift in Bullfrog Tadpole Magnetic Compass Orientation: Evidence for a Common Magnetoreception Mechanism in Anuran and Urodele Amphibians

Previous studies have demonstrated the presence of a light‐dependent magnetic compass in a urodele amphibian, the eastern red‐spotted newt Notophthalmus viridescens, mediated by extraocular photoreceptors located in or near the pineal organ. Newts tested under long‐wavelength (≥500 nm) light exhibited a 90° shift in the direction of orientation relative to newts tested under full spectrum (white) or short‐wavelength light. Here we report that bullfrog tadpoles Rana catesbeiana (an anuran amphibian) exhibit a 90° shift in the direction of magnetic compass orientation under long‐wavelength (≥500 nm) light similar to that observed in newts, suggesting that a common light‐dependent mechanism mediates these responses. These findings suggest that a light‐dependent magnetic compass may have been the ancestral state in this group of vertebrates.     

Diet composition of introduced bullfrog, <Emphasis Type="Italic">Rana catesbeiana</Emphasis>, in the Mizorogaike Pond of Kyoto,Japan

I analyzed stomach contents of the American bullfrog, Rana catesbeiana, in the Mizorogaike Pond of Kyoto, Japan. As a result, adult bullfrogs were found to feed predominantly on crayfish, and juveniles eat diverse arthropods. A wide variety of aquatic organisms occurred in the diet of both adult and juvenile bullfrogs.     

Über Kanälchenzellen und dunkle Zellen im Nephron von Anuren

Zusammenfassung Die elektronenmikroskopische und enzymcytochemische Untersuchung der Nieren von Rana cancrivora und R. esculenta, Hyla arborea und H. regilla, Bufo carens, B. mauretanicus und B. viridis ergibt, daß im Epithel ihrer Verbindungsstücke verschiedene Zelltypen vorkommen.Die Verbindungsstücke von Rana cancrivora und R. esculenta zeichnen sich — abgesehen von dem Vorhandensein heller Zellen — durch den Besitz sog. Kanälchenzellen aus, die nach Form und Struktur an die Belegzellen der Magendrüsen erinnern. Es handelt sich um sehr mitochondrienreiche, elektronendicht erscheinende Elemente, die durch teilweise verzweigte intrazelluläre Kanälchen mit der Lichtung des Verbindungsstückes kommunizieren. Aus der Oberfläche dieser Canaliculi ragen kurze Cytoplasmafortsätze in das Lumen. An der Basis der Kanälchenzellen bilden zahlreiche Cytoplasmafortsätze ein Labyrinth, das mit den seitlichen Interzellularräumen in Verbindung steht. Die Kanälchenzellen geben eine starke Reaktion auf Bernsteinsäuredehydrogenase und NADH-Diaphorase, die dem Reichtum der Zellen an Mitochondrien entspricht. Unterschiede in der Zahl der Kanälchenzellen bei dem marinen Frosch Rana cancrivora und bei R. esculenta scheinen nicht zu bestehen.Im Verbindungsstück der untersuchten Hyla- und Bufoarten fallen dunkle, d.h. elektronendichte Zellen auf, die zwar viele Mitochondrien enthalten und ein basales Labyrinth bilden, jedoch nicht kanalisiert sind. Auch diese dunklen Zellen geben eine intensive Reaktion auf Bernstein- säuredehydrogenase und NADH-Diaphorase.Die Kanälchenzellen weisen Zeichen einer sekretorischen Aktivität (Bildung von Mukosubstanzen) auf. Die Frage, ob sie außerdem resorptiv tätig sind und ob die dunklen Zellen sowohl sezernieren als auch Ionen aktiv transportieren, kann mit den angewandten Methoden nicht beantwortet werden. Unbekannt ist ferner, ob die Unterschiede im Aufbau der Verbindungsstücke bei den Rana-Arten auf der einen und den Hyla- und sowie Bufo-Arten auf der anderen Seite zu Verschiedenheiten der Harnbildung bei diesen Formen in Beziehung stehen.

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On canaliculi cells and dark cells in the nephron of anurans

Summary The electronmicroscopical and enzymecytochemical investigation of the kidneys of Rana cancrivora, R. esculenta, Hyla arborea, H. regilla, Bufo carens, B. mauretanicus and B. viridis reveals that the epithelium of the connecting tubule (Verbindungsstück) consists of different cell typs.The connecting tubules of Rana cancrivora and R. esculenta are characterized — apart from the occurrence of light cells — by the differentiation of so-called canaliculi cells, the shape and structure of which resemble those of the parietal cells of the gastric glands. These electron dense elements are rich in mitochondria and communicate by partly ramified intracellular canaliculi with the lumen of the connecting tubule. The plasma membrane bordering the canaliculi forms short microvilli-like processes. At the basis of the canaliculated cells numerous cytoplasmic processes form a labyrinth which is in connection with the lateral intercellular space. The canaliculi cells exhibit a strong activity of succinic dehydrogenase and NADH-diaphorase, which corresponds to the abundance of mitochondria in these cells. Differences in the number of canaliculi cells in the marine frog Rana cancrivora and the freshwater species Rana esculenta apparently do not exist.The connecting tubules of the Hyla and Bufo species investigated contain strikingly dark, i.e. electron dense cells, which contain numerous mitochondria, and which possess a basal labyrinth. Canaliculi, however, are lacking. These dark cells, too, give an intensive reaction for succinic dehydrogenase and NADH-diaphorase.The canaliculi cells show features of a secretory activity (production of mucous substances). The question, whether in addition they have a resorptive function and whether the dark cells are secretory and actively ion-transporting elements, cannot be answered by means of the methods applied. Further, it remains unknown whether the differences in the structure of the connecting tubule of the Rana species on the one hand and the Hyla-and Bufo species on the other, are related to differences in the mechanisms of urine production.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Light and Electron Microscopic Observations on the Intraerythrocytic Development of Babesiosoma stableri (Apicomplexa,Dactylosomatidae) in Frogs from Algonquin Park,Ontario1

ABSTRACT. The intraerythrocytic development and ultrastructure of Babesiosoma stableri Schmittner & McGhee, 1961 are described from Rana catesbeiana and Rana septentrionalis from Algonquin Park, Ontario. Morphometric and chronological observations on B. stableri in an experimentally infected Rana pipiens support the hypothesis that two successive types of merogonic cycles occur within the erythrocytes of infected frogs; the first cycle gives rise to the second and the second cycle produces merozoites destined to become gamonts. Merozoites, meronts, and gamonts are described by light and electron microscopy. Merozoites are typically coccidian and have a trilaminate pellicle with micropores, approximately 40 sub-pellicular microtubules, an apical and posterior polar ring, a conoid with two accessory rings and a pair of intra-conoidal microtubules, three rhoptries and numerous micronemes, and a nucleus with a nucleolus and a paranuclear Golgi body. The gamonts are larger than merogonic stages and are isogamous. They have approximately 55 sub-pellicular microtubules and large stores of amylopectin. These observations indicate that the genus Babesiosoma should be transferred from the Family Haemohormidiidae (Piroplasmida, Piroplasmia) to the Family Dactylosomatidae (Eucoccidiida, Coccidia).     

Do a threatened native amphibian and its invasive congener differ in response to human alteration of the landscape?

Anthropogenic changes to habitat are a global phenomenon and the impact of these changes may act in tandem to cause loss of biodiversity. One major global change is the introduction of invasive species. In order to determine whether other human impacts might correlate with populations of invaders, we examined the habitat correlates of distribution, persistence and reproduction of a global invader, the American bullfrog (Rana catesbeiana). We then compared these correlates with those of a threatened, native congener, the California red-legged frog (Rana draytonii). We found striking differences between the two species in response to habitat fragmentation and degradation. Our work suggests that human alteration of habitat, in particular the hydrology of freshwater sites and through building roads, favors this invasive species across the landscape.     

Development of arginine vasotocin innervation in two species of anuran amphibian:Rana catesbeiana andRana sylvatica

Arginine vasotocin (AVT) is a neurotransmitter in the amphibian central nervous system and is released from the neurohypophysis in the regulation of hydromineral balance and other homeostatic functions. Many amphibians experience drastic changes in habitat with respect to water availability during their transformation from aquatic larvae to terrestrial adults. To examine whether metamorphosis is accompanied by a reorganization of central vasotocinergic neurons, the developmental organization of vasotocin neurons and nerve fibers was studied with immunocytochemistry in the brains of bullfrogs (Rana catesbeiana) and woodfrogs (R. sylvatica). In bullfrogs, early limb-bud-stage tadpoles had AVT-immunoreactive neurons and nerve fibers in the lateral septal nucleus, amygdala, preoptic hypothalamus, suprachiasmatic nucleus, and posterodorsal tegmentum. Woodfrog larvae showed similar patterns of hypothalamic AVT immunoreactivity, although neuronal staining in the amygdala did not appear until metamorphic climax, and never appeared in septal neurons or in the posterodorsal tegmentum. Whereas the highly terrestrialR. sylvatica adults must adapt to an adult habitat with prolonged periods of dehydration,R. catesbeiana adults remain semiaquatic and, as such, need not develop extreme mechanisms for water retention. Nonetheless, vasotocinergic pathways showed developmental similarities in the two species. The early appearance of AVT innervation in bothRana suggests that AVT has neuroregulatory functions well before metamorphosis.     

Alpha IIb beta 3 integrin dissociation induced by EDTA results in morphological changes of the platelet surface-connected canalicular system with differential location of the two separate subunits

Treatment of human platelets by EDTA (5 mM at 37 degrees C and pH 7.4 for 30 min) induces ultrastructural morphological changes of the surface-connected canalicular system (SCCS). The first consists in dilations of some portions of the channels, whereas the second is represented by collapse of parts of the canaliculi. The collapsed elements of the EDTA treated SCCS are made up of two parallel limiting membranes and a central striated zone. Some of the EDTA treated platelets form microaggregates, the cohesion of which is apparently due to the appearance of pentalaminar interplatelet structures. EDTA treatment is known to induce an irreversible loss of platelet aggregability which is due to irreversible dissociation of the membrane GPIIb-IIIa complexes. In the present study, we looked for involvement of GPIIb-IIIa in the formation of these pentalaminar structures, and were able to demonstrate that the morphological changes described are in fact directly dependent on the EDTA induced dissociation of GPIIb- IIIa complexes. Indeed, we observed that these changes (a) cannot be induced in type I Glanzmann's thrombasthenia, where GPIIb-IIIa complexes are absent, (b) do not appear when human platelets are preincubated with monoclonal anti-GPIIb-IIIa complex-dependent (CD41a) antibodies, which protect the complex from EDTA induced dissociation, (c) appear only at alkaline pH and at 37 degrees C, which corresponds to the range of pH and temperature where EDTA can dissociate GPIIb-IIIa complexes, (d) are accompanied by the disappearance in fluorescence flow cytometry of the heterodimer complex-dependent epitopes, when using anti-CD41a antibodies and (e) do not appear in rat platelets, where GPIIb-IIIa does not dissociate after EDTA treatment. Furthermore, using gold-labeled mAbs concomitantly with the addition of EDTA, we observed that almost only GPIIb was present in the collapsed regions of the canaliculi. Using double labeling studies with polyclonal anti- GPIIb antibodies coupled to 10 nm gold particles and polyclonal anti- GPIIIa antibodies coupled to 20 nm gold particles, we observed that while both 10 and 20 nm particles were present in the dilated portions of the canaliculi almost only the small particles, coupled to the anti- GPIIb antibodies, labeled the collapsed portions of the SCCS. On Lowicryl thin sections, polyclonal antibodies against GPIIb labeled the central striated zone while both GPIIb and GPIIIa were found in the dilated portions of the SCCS. All these observations lead us to suggest that homopolymers of GPIIb could be responsible for "zipping" of the SCCS.     

The Frog Tongue: III. Histogenesis and Regeneration Following Complete and Partial Extirpations of Anlagen

Tongue anlagen from which the anterior, posterior, right or left lateral halves had been extirpated generally regenerated completely within 15–30 days in Rana catesbeiana and R. clamitans. Regeneration was most rapid and greatest in posterior and median regions. Removal of anterior-posterior and left-right middle thirds and of anterior, posterior, right or left dorsal or ventral quarter anlagen (R. catesbeiana) showed similar regenerative gradients. Regeneration never occurred when entire anlagen were removed. Extirpations of early and half-developed lingual cornua were made in metamorphosing and young adult R. catesbeiana, R. clamitans R. palustris and R. pipiens. Regeneration occurred where preoperative cornua did not exceed 1.5 mm, but never when they exceeded this length. It is concluded that anuran tongue anlagen, at the stages operated on, possess considerable reorganizing powers following partial extirpations.     

The mechanism of ultraplanktonic entrapment in anuran larvae

Tadpoles of several different genera were fed graded suspensions of uniform polystyrene particles to determine the lower size limit of particles that could be ingested. Certain tadpoles can extract suspended particles as small as 0.126 μ in diameter from the water. In terms of particle size, this is an efficiency comparable to the best mechanical sieves that can currently be produced by man. A mechanism for ultrasplanktonic entrapment is proposed on the basis of scanning electron micrographs of the secretory ridges in the branchial food traps of Rana catesbeiana before and after feeding. Xenopus tadpoles in yeast suspensions modify their clearance and buccal pumping rates in response to varying food concentrations. This may be an adaptation for maintaining a constant input of food mass to the tissues that extract the food from the water. Variability in the lower size limit of filterable particles among tadpoles of different genera correlates with the availability of suspended matter in the microhabitat where these tadpoles may be found.     

Relationship between steroidogenesis and spermiation in Rana catesbeiana and Leptodactylus ocellatus

The present study employs an in vitro system to analyse the role of steroid hormones in hCG-induced spermiation in two species of anuran amphibian: Rana catesbeiana and Leptodactylus ocellatus. In vitro spermiation was induced with 10 IU hCG and the effect of different steroid-biosynthesis inhibitors was analysed. Cyanoketone (10⁻⁵ M), an inhibitor of 3-oxo-4-ene steroid biosynthesis, did not block hCG-inducing activity even when biosynthesis of androgen was significantly reduced. These results clearly showed that, in both species, spermiation-inducing action of hCG does not depend on the biosynthesis of 3-oxo-4-ene steroids. Moreover, when combined inhibitors, aminoglutethimide (10⁻⁵ M) plus cyanoketone (10⁻⁵ M), were employed, spermiation evoked by hCG was not modified while hCG-induced androgen secretion significantly decreased. Additionally, none of the steroids used, progesterone, 17, 20α-dihydroxy-4-pregnen-3-one, testosterone and 5α-dihydrotestosterone, were able to induce spermiation in the absence of hCG, confirming that steroids are not involved in that process. In conclusion, as previously described in Bufo arenarum, in L. ocellatus and R. catesbeiana hCG-induced spermiation does not depend on steroid biosynthesis.     

Immunohistochemical localization of thyrotropin-releasing hormone (TRH) in skin of Rana pipiens

Summary Using immunofluorescent techniques thyrotropin releasing hormone (TRH) is demonstrated in skin of Rana pipiens and R. catesbeiana. The immunofluorescent-TRH is localized in all cell layers of the epidermis and in the epithelium lining the various cutaneous glands, but not in the dermal layer.We wish to thank Dr. Ronald DeLellis and Ms. Mary Blount for their expert advice and guidance in the immunohistochemical techniques.This investigation was supported by NIH National Research Service Award # 1F32 AMO6018-01 from the NIAMDD to Janice L. Bolaffi and NIH Grant AM 21863 to Ivor M.D. Jackson.     

The antibody response during amphibian ontogeny

The maturation of the anti DNP-response in adult and larval amphibians has been compared in two species of anurans,Rana catesbeiana andXenopus laevis. IgM responses were very similar in larvae and adults of both species. Specific IgG antibody synthesis, studied inRana catesbeiana only, was not obvious in larval primary response but was easily detectable in adults. Although metamorphosis results in profound changes in the lymphoid system associated with the apparent acquisition of self tolerance to new antigens, immunological memory of antigens injected during larval life ofRana andXenopus persists after the metamorphosis period.     

Light and electron microscopic investigation of ATPase activity in musculature during anuran tail resorption

Summary The histochemical activity of adenosine triphosphatase (ATPase) was studied at light and electron microscopic levels in larval tail musculature of Rana catesbeiana and Rana ornativentris during late metamorphic stages. The presence of low, moderate or dark reaction of K₂-EDTA-preincubated Ca⁺⁺-ATPase was correlated with the variable degree of degeneration of white fibres even at the late stage of tail resorption. The reasons for an increase in this ATPase activity in degenerating white muscle fibres are discussed. Irrespective of the degree of degeneration, all red fibres showed high ATPase reaction. During myocytolysis, it is shown that the SR vesicles accumulate electron dense amorphous material. The degree of myofibrillar disintegration correlated with decrease in ultrastructural reaction product for Mg⁺⁺-ATPase. Although grouped atrophy of muscle fibres (as seen in Xenopus laevis, den Hartog Jager et al., 1973, 1975) was absent in musculature of resorptive tails, ultrastructural characteristics including proliferation of SR and dilation of its vesicles represent alteration of the normal neural influence on the skeletal muscle fibres.     

INTRAMITOCHONDRIAL YOLK-CRYSTALS OF FROG OOCYTES : I. Formation of Yolk-Crystal Inclusions by Mitochondria during Bullfrog Oogenesis

Electron microscope examination of thin sections of bullfrog (Rana catesbeiana) ovarian oocytes has shown the presence of mitochondria containing yolk-crystal inclusions in oocytes of all sizes, from 160 to 1500 µ mean diameter. The hexagonally shaped yolk-crystals have major periodicities of 73.8 ± 10.7 A (n = 100). Several forms of modified mitochondria, observed in the smaller oocytes, may be arranged into a series of structurally intermediate forms between standard oocyte mitochondria and the typical mitochondria with yolk-crystal inclusions. The observation of such intermediate forms is consistent with proposals that the yolk-crystal inclusions arise within a limited portion of the oocyte chondriome by a complex process of mitochondrial differentiation.