Vagal afferent projections in Rana pipiens, Rana catesbeiana, and Xenopus mülleri. With a note on lateral line and VIII nerve projection zones.
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Afferent Pathways↗
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Neurophysiological and behavioral evidence for selective reinnervation in skin-grafted Rana pipiens.
Rana pipiens with skin dorosoventrally reversed can respond to stimulation of the back with forelimb wipes to the belly and to stimulation of the belly with hindlimb wipes to the back. These "misdirected wiping responses" have been explained in terms of two alternative hypotheses of nerve regeneration: nerve respecification or selective reinnervation. Experimental behavioral and neurophysiological experiments reported here support the selective reinnervation hypothesis. Severing ventral nerves, which normally innervate the belly, greatly reduced the percentage of misdirected responses on stimulation of belly skin grafted to the back, while severing dorsal nerves, which normally innervate the back, increased the percentage of misdirected responses elicited under the same circumstances. Moreover, neurophysiological recordings of grafted animals showed three effects of skin grafting on nerve distributions: (i) termination of dorsal and ventral nerve receptive field at graft edges; (ii) overlap of nonadjacent ventral nerve receptive fields; and (iii) dorsal coursing of ventral nerves to reinnervate target belly skin displaced to the back. These neurophysiological observations, and particularly the third effect, also support selective reinnervation as the mechanism of nerve regeneration in skin-grafted Rana pipiens.
Action Potentials↗
Cell turnover in the beak of Rana pipiens.
Rana pipiens tadpoles were injected with 3H-thymidine at different stages to label basal cells of the horny beaks, the keratinized oral specializations of many anuran tadpoles. Cells in the column and sheath of the beak are derived from the basal layer. Movement of the labeled cells in beak columns can be traced, permitting calculation of the rate of cell addition into the columns. With the counts of the total number of column cells and cell addition rate, the rate of apical cell loss from a column throughout the larval stages can also be calculated. The rate of cell addition decreases steadily from stage III through stage XX, in all portions of both beaks. The rate of change of these column cell addition rates is similar among middle and lateral portions of both upper and lower beaks. There is a relatively constant rate of cell loss until stage XII or XIII. It then increases sharply, and the beaks are completely lost at stages XIX or XX. The longer, lower beak columns have a much faster cell loss rate than do the upper beak columns at late larval stages. Life spans for cells that move to the sheath may be about one-half those for cells that enter the beak columns, It is suggested that thyroid hormone accelerates the rate of column cell loss.
Age Factors↗
Central projection of optic tract from translocated eyes in the leopard frog (Rana pipiens).
In Rana pipiens embryos, eye anlagen were moved to the evacuated ear position, where they continued to differentiate and sent their optic nerve fibers into the hindbrain. Upon entering the medulla, the optic fibers turned caudally, penetrated the spinal cord, and traversed the dorsolateral white matter to the caudal end. We found this pattern of growth in every animal; the optic fibers did not enter the tecta. These results suggest the existence within the neural tube of a three-dimensional gradient system to which embryonic optic fibers are responsive and which may guide the normal development of the visual pathway.
Animals↗
The occurrence of diploid ova in Rana pipiens.
All 12 Rana pipiens females tested from three populations produced diploid ova. These were identified by the development of from 0.15 to 35 percent per clutch of normal diploid-like embryos among large numbers of haploid embryos following activation of R. pipiens eggs with irradiated R. clamitans sperm. Their diploid nuclear constitution was demonstrated by diploid cell size at Shumway Stage 19, and by the diploid number of chromosomes both as embryos and as mature frogs, and was confirmed by the occurrence of triploid embryos among normally fertilized progeny from the same female parents. Although the precise cytogenetic events leading to the origin of these diploid ova were not directly determined, we were led to conclude that, although diploid ova may result from polynucleate oocytes, the diploidy reported here was an expression of abnormal meiosis that occurred under genetic control. Such anomalies, at the orders of frequency we observed in R. pipiens, have important consequences for all studies using Anuran model systems.
Animals↗
Increased prevalence of mature females bearing pigmented oocytes from populations of Minnesota Rana pipiens.
Mature female leopard frogs, Rana pipiens, collected from several sites in Minnesota from 1967 through 1977, were examined for the presence of pigmented oocytes. An increase in the percent of mature females with pigmented oocytes was observed in the frogs captured in 1977 compared to those captured earlier. Coincident with the increase in prevalence of females with pigmented oocytes, there was a decline in abundance of the frogs.
Animals↗
The beta adrenergic receptors of chromatophores of the frog, Rana pipiens.
The isolated skin of Rana pipiens was found to be a suitable model for the quantitative study of chromatophore beta adrenergic receptors uninfluenced by prejunctional phenomena. Cumulative concentration-response curves for adrenergic agonists were obtained in preparations in which effective alpha adrenergic blockade had been produced with phenoxybenzamine. The beta adrenergic agonists darkened the preparation, as did melanocyte-stimulating hormone, but the maximum effects differed. The maximum of the l-isoproterenol cumulative concentration-response curve was approximately 50% less than that of melanocyte-stimulating hormone, while the maxima for l-epinephrine and l-norepinephrine were significantly less than that for isoproterenol. Microscopic examination revealed a qualitative difference: while maximal darkening produced by melanocyte-stimulating hormone was associated with maximal changes in both interspot melanophores and iridophores, maximal adrenergic-induced darkening was associated with maximal iridophore granule concentration only. No qualitative differences could be observed in the darkening caused by the three adrenergic agonists. The beta adrenergic potencies of l-norepinephrine and l-isoproterenol relative to l-epinephrine were determined by four-point bioassay. Isoproterenol was found to be 138 times as potent as epinephrine, while norepinephrine was 4 times as potent. Similarly, antagonism of isoproterenol-induced darkening of phenoxybenzamine-pretreated skin samples by the beta adrenergic blocking agents dl-propranolol, dl-sotalol, dl-practolol, l-butoxamine and d-butoxamine was studied, and their KB and pA2 values, respectively, were found to be: dl-propranolol (1.44 X 10(-8)M, 7.81); dl-sotalol (7.25 X 10(-8)M, 7.23); l-butoxamine (6.92 X 10(-6)M, 5.10); dl-practolol (1.91 X 10(-5)M, 4.96); d-butoxamine (no activity). Comparison of the potency ratios and pA2 values cited above with similar parameters obtained by other investigators in several mammalian tissues suggests that there is wide variation among beta adrenergic receptors.
Adrenergic beta-Agonists↗
Reduced prevalence of the Lucké renal adenocarcinoma in populations of Rana pipiens in Minnesota.
Northern leopard frogs (Rana pipiens) afflicted with the Lucké renal adenocarcinoma virtually disappeared from Minnesota in the autumn of 1977. Frogs from four sites in Minnesota counties (Polk, Otter Tail, Kandiyohi, and Scott) with a previously high prevalence of Lucké renal tumor were studied. In the past decade, prevalence averaged 4.2% in 29 collections (total, 1,870 frogs). No tumors were detected in 685 frogs autopsied in the autumn of 1977 by the method of previous studies. Frog collections, each comprised of 20 or more individuals, were compared for the presence or absence of tumor-bearing frogs. Significantly fewer collections contained tumor-bearing frogs in the autumn of 1977 than did collections of previous years.
Adenocarcinoma↗
The effects of four arthropod diets on the body and organ weights of the leopard frog, Rana pipiens, during vitellogenesis.
Wild-caught adult Rana pipiens females were captured in midsummer and fed diets of crickets, flies sowbugs or wax moth larvae during a three-month period of active vitellogenesis. The cricket diet supported the most extensive body weight gain during this time and promoted a prolonged period of weight increase in an additional long-term study. Synchronous growth of the oocytes occurred in all four groups, but the ovaries and oviducts of cricket-fed animals were significantly larger than those of frogs on the other three diets. The significantly higher liver weights of frogs fed wax moth larvae may have reflected an augmentation of hepatic energy stores. Fat body weights were also highest in this group of animals. Frogs fed crickets and wax moth larvae possessed larger fat bodies than did the midsummer control animals killed immediately after their arrival in the laboratory. In contrast, frogs fed flies and sowbugs had smaller fat bodies than did the initial controls, suggesting that animals on these diets had utilized fat body lipid during vitellogenesis. Gastrocnemius and final body weights were lowest in frogs fed wax moth larvae. These findings may have reflected the nutritional content of the diet or the reduction in appetite frequently noted in these animals during observations of feeding behavior.
Animals↗
Birefringence measurements of structural inhomogeneities in Rana pipiens rod outer segments.
The birefringence (deltan) of Rana pipiens rod outer segments (ROS) reveals microstructure inhomogeneities not seen with other techniques. In the basal 20-30-micron length of the ROS there is a nearly linear axial gradient in deltan of approximately equal to -2 x 10(-5)/micron. No consistent deltan gradients are found in the distal 20-30 micron of the ROS. Using glycerol imbibition to separate the intrinsic and form birefringence components, we found that the basal deltan gradient was principally due to a gradient of the intrinsic birefringence component. The disk membrane volume fraction decreases uniformly from the basal end to the distal end, while the disk membrane refractive index increases. The contributions of these changes to the form birefringence approximately cancel, so that the form component is fairly uniform along the ROS axis. Because its axial distance from the inner segment is a measure of the time since a disk membrane was formed, these gradients may reflect a disk membrane aging process. Occasionally a highly birefringent, 2-micron-wide band is seen at the basal end ot the ROS, possibly where the envelope membrane folds to form new disk membranes.
Animals↗
Environmental influence on ovulation and embryonic development in Rana pipiens.
Environmental effects on ovulation and embryogenesis in Rana pipiens were assessed using both freshly-captured fall animals and laboratory-conditioned females which had undergone vitellogenesis in the laboratory. Frogs in both categories were divided into two groups. Ovulation was hormonally induced in one group of females prior to cold exposure and in the second group of animals following an 8-week-period at 4 degrees C with an 8L 16D photoperiod. The incidence of both ovulation and normal embryonic development was increased following exposure of the animals to low temperatures and short daylength. Those animals which only partially ovulated prior to cold treatment did not respond to hormone injections following the period of cold exposure. Examination of the ovaries of these females revealed a much greater degree of oocyte resorption than was found in frogs whose initial ovulation was induced only after exposure to cold temperatures. The administration of ovulation-inducing hormones prior to artificial hibernation may thus have initiated a phase of oocyte resorption which progressed even at 4 degrees C. The incidence of ovulation was similar in wild-caught and laboratory-conditioned females, but eggs from the latter showed a much lower percentage of development to Shumway stage 20. This effect may have been related to differences in the environmental factors to whcih the two groups were exposed during oogenesis.
Animals↗
Amphibian embryo protease inhibitor. V. Effect of calcium on the distribution of amphibian trypsin inhibitor during fertilization and subsequent development of Rana pipiens.
In previous publications we have documented the existence in oocytes and embryos of a variety of forms (notably Rana pipiens), massive amounts of a powerful inhibitor of trypsin-like enzymes (ATI). The bulk of the inhibitor in Rana pipiens is localized in yolk platelets. We present evidence here that the distribution of the inhibitor between yolk platelets and cytosol changes and that this change is mediated by variations in the distribution of calcium ions. There is an inverse relationship between ATI and free calcium in the cytosol. Several workers have demonstrated a dramatic rise in free cytosol calcium immediately following fertilization. We confirm this observation, and demonstrate that there is a parallel and equally dramatic decrease in free cytosol ATI during this period. Experiments with purified yolk platelets indicate that calcium effects a release of sequestered inhibitor from these particles. Other experiments indicate that calcium mediates ATI-lippovitellin associations. A calcium mediated flux of ATI from cytosol to yolk is proposed as a device for controlling limited proteolysis in the cytoplasm. We offer this as a model for studying the unmasking of mRNA which follows fertilization.
Animals↗
Experimental studies on hemopoiesis in the pronephros of Rana pipiens.
Embryogenesis of hemopoietic cell populations in the pronephros of Rana pipiens was examined during embryonic and early larval development. Differential cell counts of Wright-Giemsa-stained cell suspensions demonstrated that granulopoiesis is the predominant hemopoietic activity in the pronephros, erythropoiesis accounts for a minor component of the hemopoietic activity (less than 10%), and lymphopoiesis within the organ is negligible. Microdensitometric analysis of Feulgen-DNA stained granulocyte populations in pronephroses from larvae that had received chromosomally labeled pronephric analgen transplants between 84 and 96 h of development demonstrated that hemopoiesis in this organ is dependent on colonization by an extrinsic hemopoietic stem cell. A similar analysis of pronephric hemopoiesis in larvae which had received chromosomally labeled, presumptive ventral blood island transplants between 62 and 67 h of development, indicates that granulopoietic cells are not derived from the embryonic blood islands. It is proposed that the pronephros may be the initial site of granulocyte differentiation during early embryogenesis. Although the embryonic origin of the hemopoietic stem cell is unknown, indirect evidence from this study indicates a dorsal stem cell compartment.
Animals↗
Some telencephalic connections in the frog, Rana pipiens.
Some afferent, efferent and intrinsic connections of the telencephalon of Rana pipiens were studied using a horseradish peroxidase method. Afferents to the telencephalon from thalamic and brain stem cell groups were demonstrated. These findings, taken together with the results of previous studies, indicate that separate thalamic cell groups project visual, auditory and somatosensory information onto the striatum. A separate thalamic cell group projects to the medial telencephalic wall and probably conveys visual and somatosensory information. These telencephalic afferent systems do not appear to be directly comparable to those of birds and reptiles. Additionally, some telencephalic afferents demonstrated in previous studies using anterograde degeneration techniques were confirmed, and some intratelencephalic connections were identified.
Amygdala↗
The germfree leopard frog (Rana pipiens): preliminary report.
Eggs and sperm were aseptically collected from leopard frogs (Rana pipiens) and passed into sterilized flexible film isolators. Egg fertilization was accomplished, and germfree tadpoles were produced. The germfree tadpoles were active and free-swimming for 3 weeks, but only 6.2% survived to 4 weeks of age. None survived after the 4th week.
Animals↗
Specificity of carbohydrate inhibition on leopard frog (Rana pipiens) erythrocyte agglutinating activity by concanavalin A and phytohemagglutinin.
1. Leopard frog (Rana pipiens) erythrocytes are agglutinated by the plant lectins concanavalin A and phytohemagglutinin. 2. Concanavalin A hemagglutination can be specifically inhibited by the sugars mannose, glucose, arabohexulose, alpha-methyl-D-mannopyranoside, N-acetyl-D-glucosamine, maltose, sucrose, trehalose and melezitose. 3. Sugars inhibiting concanavalin A hemagglutination are structurally composed of alpha-D-glucopyranosyl, alpha-D-mannopyranosyl or beta-D-fructofuranosyl residues with the same hydroxyl groups at carbons 3, 4 and 6. 4. Phytohemagglutinin hemagglutination can only be inhibited by the sugar N-acetyl-D-galactosamine. 5. Sugar inhibition of concanavalin A and phytohemagglutinin leopard frog erythrocyte agglutination is the same as that observed in many mammalian species.
Animals↗
Immunohistochemical localization of thyrotropin-releasing hormone (TRH) in skin of Rana pipiens.
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.
Animals↗
Continuity between the ventricular and subarachnoid cerebrospinal fluid in an amphibian, Rana pipiens.
Continuity between the ventricular and subarachnoid cerebrospinal fluid has been investigated in Rana pipiens. The structure of the posterior tela, a deficient membrane situated at the extreme caudal end of the roof of the fourth ventricle, has been studied using whole membrane mounts and by light microscopy of resin embedded tissue. The ependymal component consists of columnar and rounded cells which form a regular 'syncytium' enclosing round and oval fenestrations. Small fenestrations are covered on the subarachnoid side by elongated pial cells and thus do not give total continuity between the fourth ventricle and the subarachnoid space. Large fenestrations, on the other hand, are accompanied by equivalent pial fenestrations giving direct access between the fluid compartments. Towards the caudal end the fenestrations break up and the numbers of ependymal and pial cells decrease, the caudal end itself being characterised by a small remaining clump of ependyma and pia or of pia alone. Flow through the tela has been studied using fluorescein-labelled dextran placed in the intraventricular space. Infusion into the lateral ventricle and subsequent localisation by fluorescence microscopy shows the marker to be in the fourth ventricle, in the fenestrations of the posterior tela and in the subarachnoid space overlying the tela. Infusion of the marker followed by freezing and examination of the cut heads on a freezing microtome, shows fluorescence throughout the ventricular system, in the subarachnoid space adjacent to the posterior tela and also along the dorsal subarachnoid space of the spinal cord.
Animals↗