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.     

Plasma immunoreactive calcitonin in the frog (Rana pipiens)

1. Plasma immunoreactive calcitonin (iCT) and ionic calcium [( Ca]i) were measured in intact frogs (Rana pipiens) within complete 24 hr light-dark cycles over an 18 month period. 2. Plasma iCT exhibits an annual periodicity about the annual mean of 10.0 ng/ml, with an amplitude of 5.4 ng/ml that peaks in October. 3. Within an annual cycle, a significant inverse association exists between the basal monthly levels of plasma iCT and [Ca]i for animals maintained in freshwater control conditions. 4. When subjected to a high calcium environment during the latter half of the year, plasma [Ca]i and iCT were elevated above control levels but exhibited independent cyclic patterns. 5. A distinct seasonal response of increased iCT in a high calcium environment may be related to the secretory activity of the ultimobranchial glands and physiological responsiveness to other calcemic hormones; e.g. parathyroid hormone and vitamin D.     

Tyrosine aminotransferase activity of frog (Rana esculenta) liver

1. The presence of tyrosine aminotransferase is reported both in particulate and soluble fractions of frog liver. 2. The activity of the soluble enzyme of frog liver was investigated with regard to its dose and time dependence, its substrate specificity and concentration dependence, its thermal sensitivity as well as pH and temperature dependence. 3. It appears that the properties of the soluble tyrosine aminotransferase of frog liver are in close agreement with those reported for the mammalian liver enzyme.     

Nutritional composition of frog (Rana esculanta) waste meal

In the present study, the waste obtained from the frozen frog leg industry was used for the production of frog waste meal, and its proximate, amino acids, fatty acids, mineral and vitamin compositions were evaluated to determine the nutritional quality. In addition, the total bacterial count, Salmonella, total volatile basic nitrogen (TVB-N, mgN/100g) and thiobarbituric acid (TBA, mg malonaldehyde/kg) were also measured to determine the microbiological and chemical quality of frog waste meal (FWM). The crude protein, fat and ash content of FWM on a dry weight basis were 68.6%, 17.0% and 13.2%, respectively. The amino acid profiles were found to be fairly close to those of fish meal in terms of protein sources and rich in the glutamic acid, glycine, proline, arginine, and methionine. The proportions of fatty acid composition in FWM were analysed and findings were 26.7% for total saturated fatty acid (SFA), 42.5% for total monounsaturated fatty acid (MUFA), 17.0% for total n - 6 and 3.3% for n - 3 ratio. The major SFA, MUFA and PUFA in FWM were palmitic acid (19.1%), oleic acid (26.0%) and linoleic acid (16.7%), respectively. FWM was found to be high in mineral content, especially Zn, K, Cu, Mn, and Mg and high level of some vitamins such as folic acids and thiamin. The total bacterial count was found to be 2.9x10(4) CFU/g, and Salmonella was not observed. TVB-N and TBA in FWM was determined to be 157.4+/-5.8 mg N/100g and 1.2+/-0.1 mg malonaldehyde/kg, respectively.     

NAG infection of grass frog (Rana temporaria) tadpoles]

Rana temporaria tadpoles (2250) were contaminated with three types of NAG vibrio cultures. Clinical, bacteriological and morphological examinations showed the larvae to be suffering from an acute infection during the first 2 days after the contamination. Then the vibrios persisted in the tadpole organism for a long time and were excreted in the feces.     

Ultrastructure of venom glands in the frog (Rana esculenta)

Electron microscopic study of skin venom glands in the frog, Rana esculenta, revealed the syncytial structure of the inner (secretory) wall which presents two distinct zones: a basal (juxtamuscular) one, which contains nuclei and major cytoplasmic organelles, and an apical one where large electron-dense granules form and accumulate. Granules are seen to arise inside clusters of tightly packed smooth endoplasmic reticulum (SER) elements, which suggests that the SER system is mainly involved in synthesis of this material. A high glutaraldehyde concentration (5%) also reveals a poorly defined material filling the intergranular cytoplasm. No apical limits to the syncytium could be traced, which suggests massive holocrine secretion. Nerves insinuate between the muscle cells and occur all along the internal face of the muscular layer, sometimes in close contact with the syncytium. The gland duct, the wall of which consists of epidermal cells, is blocked, in contact with the gland, by an epidermal bud linked externally to the muscle layer surrounding the gland. Thus, only strong muscle tension such as to expel all or part of the epidermal bud can trigger granule release. This phenomenon can be induced by the subcutaneous injection of epinephrine, but the high and distressing dose needed to provoke appreciable changes in venom glands renders unlikely any natural intense venom release triggered by epinephrine in the frog.     

Individual voice recognition in a territorial frog (Rana catesbeiana)

Some territorial animals display low levels of aggression towards a familiar territorial neighbour in its usual territory, but exhibit high levels of aggression towards neighbours in novel locations and unfamiliar individuals. Here, we report results from a field playback study that investigated whether territorial males of the North American bullfrog (Rana catesbeiana) could discriminate between the acoustic signals of simulated neighbours and strangers in the absence of contextual cues associated with a specific location. Following repeated exposures to synthetic bullfrog calls from a particular location, subjects responded significantly less aggressively to a familiar call, compared with an unfamiliar one, when both calls were broadcast from familiar and novel locations, indicating that bullfrogs could recognize a neighbour's calls independently of the contextual cues provided by the direction of the neighbour's territory. Subjects responded equally aggressively to unfamiliar calls broadcast from either a familiar or a novel location, which indicates that they could perceive unfamiliar calls as those of a stranger, regardless of where the stranger was encountered. Together, these two results provide evidence that a frog possesses a capacity for individual voice recognition.     

Circannual variations in mevalonate utilization in frog (Rana esculenta)

1. The fate of mevalonate, the product of HMGCoA reductase, was studied in male and female frogs (Rana esculenta) in order to explain the circannual variations of enzyme activity. 2. The incorporation of 2-14C MVA into unsaponifiable lipids, cholesterol and dolichol in liver, plasma and eggs was followed. 3. Labeled MVA shows a different utilization depending on season and sex. In spring and summer cholesterol synthesis is related to hepatic reserve storage in both sexes, while the peak of enzyme activity, present only in females in fall, seems committed to cholesterol export into the blood and uptake by the oocytes. 4. The presence of a MVA-derived protein identifiable with vitellogenin and labeled on the lipid moiety, suggests that HMGCoA reductase activity in fall is committed to the lipidation of this protein essential for oocyte maturation.     

The adrenergic receptor subtypes present in frog (Rana esculenta) skin

Frog skin transports ions and water under hormonal control. In spite of the fundamental role played by adrenergic stimulation in maintaining the water balance of the organism, the receptor subtype(s) present in the skin have not been identified yet. We measured the increase in short-circuit current (ISC, an estimate of ion transport) induced by cirazoline, clonidine, xamoterol, formoterol, or BRL 37344, in order to verify the presence of alpha1, alpha2, beta1, beta2, or beta3 receptor subtypes, respectively. Only after treatment with formoterol, BRL 37344 and, to a lesser extent, cirazoline was measured a significant increase in ISC (57%, 33.2%, and 4.7%, respectively). The formoterol and BRL 37344 concentrations producing half-maximal effect (EC50) were 1.12 and 70.1 nM, respectively. Moreover, the formoterol effect was inhibited by treatment with ICI 118551 (antagonist of beta2 receptors) while SR 59230A (antagonist of beta3 receptors) had no effect; opposite findings were obtained when the BRL 37344 stimulation was investigated. Finally, by measuring the transepithelial fluxes of 22Na+ and 36Cl-, we demonstrated that Na+ absorption is increased by activation of beta2 and beta3 and is cAMP-sensitive, whereas the Cl- secretion is only increased by activation of beta2 receptors and is cAMP- and calmodulin-sensitive.     

Nociceptin binding sites in frog (Rana esculenta) brain membranes.

The recently discovered natural heptadecapeptide nociceptin (orphanin FQ) shares some homology with the opioid peptides but it binds to a distinct receptor type, termed nociceptin receptor. This study demonstrates the presence of specific nociceptin recognition sites in brain membrane fractions of an amphibian, Rana esculenta. Para-iodo-Phe(1)-nociceptin-amide was radiolabelled by catalytic dehalotritiation, resulting in p[(3)H]Phe(1)-nociceptin-amide of 25 Ci/mmol specific radioactivity. Specific binding of [(3)H]nociceptin-amide to frog brain membranes was found to be saturable and of high affinity with equilibrium K(d) values in the low nanomolar range. A single set of binding sites with about 180 fmol/mg protein maximal binding capacity was obtained in saturation and competition experiments. [(3)H]Nociceptin-amide binding could easily be inhibited by synthetic nociceptin compounds but not by opioid ligands. Both sodium ions and 5'-guanylylimidodiphosphate decreased the binding of the radioligand by transferring the receptor to a lower affinity state. Nociceptin dose-dependently stimulated the binding of the nonhydrolysable, radiolabeled GTP-analogue guanosine-5'-O-(3-thio)triphosphate ([(35)S]GTPgammaS) to G-proteins in frog brain membranes. Addition of 1 microM naloxone caused no significant change in the curves, indicating that nociceptin-mediated activation of G-proteins occurred through nonopioid mechanism.     

Genetic linkage groups in the Japanese brown frog (Rana japonica)

To elucidate the genetic linkage groups of the Japanese brown frog (Rana japonica) and compare them with those of other amphibians, we analyzed the inheritance of alleles at 15 enzyme and blood protein loci and one pigment locus (Blk) in 3,298 offspring derived from 37 crosses using 28 males heterozygous at these loci. Of 63 pairs of loci tested for linkage, 55 pairs showed independent assortment in all crosses examined. In another six pairs, that is, between FUM/ME-1, alpha-GDH/MDH-2, MDH-1/ME-1, MDH-2/ME-2, MDH-2/PEP-C, and ME-2/PEP-C, all the offspring analyzed were parental, and none were recombinant. In the other two pairs, that is, between GPI/PEP-D and AAT-1/Blk, most of the offspring were parental, and some were recombinants with recombination rates ranging from 4.8% to 8.0%. Thus the following four linkage groups comprising 11 loci were established in R. japonica. The first group included the loci for alpha-GDH, MDH-2, ME-2, and PEP-C; the second group included the loci for MDH-1, ME-1, and FUM; the third group included the loci for GPI and PEP-D; and the fourth group included the loci for AAT-1 and Blk. No linkage between the other five loci--ADA, MPI, PEP-A, PGM, and Alb--was observed in the present study.     

Morphology of epiphyseal apparatus of a ranid frog (Rana Esculenta)

Morphological, histochemical and ultrastructural investigations on epiphyseal apparatus of Rana Esculenta were made. The most important findings were the following: 1) metaphyseal cartilage is localized inside proximal diaphyseal compact bone as a plug; 2) metaphyseal cartilage do not reduce in thickness during ageing; 3) metaphyseal cartilage do not show vascular invasion and do not mineralize in degenerative zone; 4) trabecular bone was not at all evident in this animal; 5) external periosteum is well vascularized and proliferates in correspondence to marginal epiphyseal end of the diaphyseal. From these results the hypothesis that the ranid frog bone growth is not due to metaphyseal metabolism (as in avian and mammals) but to bone periosteal marginal mineralization is reached.     

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.     

Twitch potentiation of frog (Rana japonica) skeletal muscle by antipyrine

1. The effects of antipyrine on single muscle fibres from the frog Rana japonica were examined. Antipyrine potentiated the twitch tension, but did not affect the tetanus response or induce any contracture by itself. 2. The duration of the action potential was prolonged in a dose-dependent manner. 3. Antipyrine did not affect the mechanical threshold, the uptake of Ca2+, or the Ca2+ induced release of Ca2+ from the sarcoplasmic reticulum. 4. Ca2+-channel blockers did not affect the twitch potentiation by antipyrine.