Ribonucleic acid (RNA) from calf spleen tissue was isolated and purified by a modified Kirby phenol procedure. The absorption maximum of the isolate occurred at 260 mµ indicating the presence of nucleic acids and the absorption minima recorded at 230 mµ and 280 mµ indicated the absence of peptides and proteins. Colorimetric analyses indicated the presence of RNA and the absence of peptide, protein, DNA and carbohydrate contamination. Chromatographic analysis indicated the absence of carbohydrate contamination only after the purification with 2-methoxyethanol. The spleen RNA prepared by the phenol method was undegraded and demonstrated three distinct molecular species when analysed with the ultracentrifuge; a 27S fraction, an 18S fraction and an 8S fraction. Competent early gastrula ectoderm and embryos of Xenopus laevis exposed to undegraded spleen RNA demonstrated no tissue-specific induction. However, in both the in vitro and in vivo experimental series an enhancement of development was observed. A possible explanation of this phenomena was discussed. / Science, Faculty of / Zoology, Department of / Graduate

The present study documents that baroreceptors located in the pulmocutaneous artery (PCA) are key components in control of lymph heart rate in amphibians. A negative feedback control loop exists between arterial pressure and lymphatic heart rate. The recurrent laryngeal nerve (rLN), which innervates the PCA baroreceptors, transmits information on arterial pressure to integration centers in the central nervous system. Lymphatic heart rate (LHR) is reduced as a result of increases in arterial pressure. This loop was determined using three experimental protocols. First, the correlation between LHR reduction and hormonally induced vasoconstriction was determined. Increases in arterial pressure due to pressor actions of angiotensin II and arginine vasotocin at high concentrations was negatively correlated to LHR. Second, lymphatic heart rate changes due to natural increases in arterial pressure were compared to rate changes due to increase in arterial pressure after bilateral denervation of the rLN. Post-denervation LHR was not affected by natural increase in arterial pressure prior to the establishment of a new resting arterial pressure. Increase in arterial pressure due to administration of vasoconstricting hormones was negatively correlated with LHR following denervation. Third, the effect on LHR due to direct stimulation of the rLN was studied. Stimulation of the rLN caused LHR to stop without increases in arterial pressure. Presumably, this negative feedback loop is present to limit fluid return to the cardiovascular system from the lymphatic system during periods of acute hypertension. Reduction in the return of lymph volume to the cardiovascular system could eliminate potential damage to pulmonary tissues due to high arterial pressures.

No description available.

The characteristics of the ion exchange mechanisms which regulate volume and pH in fish and amphibian erythrocytes were investigated and compared. Experiments were carried out under steady state conditions and also following adrenergic stimulation both in vivo and in vitro. Under steady state conditions, a decrease in extracellular pH caused an increase in the volume of rainbow trout erythrocytes, and a decrease in the intracellular pH. These pH-induced volume changes were mainly associated with movements of chloride across the chloride/bicarbonate exchange pathway. The sodium/proton exchange mechanism is quiescent at all pH's studied under steady state conditions. Beta adrenergic stimulation of rainbow trout erythrocytes promoted cell swelling and proton extrusion from the erythrocytes. Amiloride inhibited both the volume and pH changes associated with adrenergic stimulation indicating that this response is associated with an increase in the activity of the sodium/proton exchange mechanism on the erythrocyte membrane. The adrenergic swelling and pH responses are enhanced by a decrease in extracellular pH. An increase in bicarbonate concentration reduces the adrenergic pH response, but it is still significant even at 10 mM bicarbonate. DIDS markedly enhanced the beta adrenergic effect on the erythrocyte pH, but abolished the increase in erythrocyte volume. The adrenergic response was independent of temperature between 10 and 18°C. These results support a loosely coupled sodium/proton and chloride/bicarbonate exchange model for the adrenergic response in rainbow trout erythrocytes. The increases in erythrocyte pH and volume following adrenergic stimulation are associated with increases in the haemoglobin:oxygen affinity. The oxygen carrying capacity of the blood is, therefore, increased following adrenergic stimulation in rainbow trout. Carbon dioxide excretion, however, was not significantly affected by adrenergic stimulation. The functional significance of the adrenergic response of fish erythrocytes may be to offset the effects of the Root shift on the oxygen carrying capacity of the blood during acute changes in extracellular pH. In contrast to fish erythrocytes, the sodium/proton exchange mechanism in amphibian erythrocytes is active under steady state conditions. In the presence of bicarbonate movements, this exchange significantly affects the erythrocyte volume, but not the erythrocyte pH. Similar to fish erythrocytes, protons are passively distributed in amphibian erythrocytes under steady state conditions and in Donnan equilibrium with chloride ions. The erythrocyte volume also increases with decreases in extracellular pH as in fish erythrocytes, due to changes in the chloride distribution across the erythrocyte membrane. Adrenergic stimulation does not affect the volume or pH of amphibian erythrocytes either in vivo or in vitro. These animals, therefore, do not appear to regulate erythrocyte pH adrenergically. Amphibians are able to efficiently utilize oxygen stores via both central and peripheral shunting. In addition, the blood of these animals does not have a Root shift. Adrenergic regulation of erythrocyte pH in order to enhance oxygen transport during fluctuations in ambient and internal gas tensions, therefore, is probably less important than it would be in fish. / Science, Faculty of / Zoology, Department of / Graduate

The importance of hypovolemia in dehydrational death was assessed in two anuran species. Xenopus laeyis, a species which experiences a significant reduction in circulating plasma volume with dehydration, was used to evaluate the role of sympathetic reflex compensation in hypovolemia. Adrenergic blockade with propranolol or phenoxybenzamine produced no significant reduction in dehydration tolerance in this species, although β-blockade with propranolol appeared to have a minor effect. The role of hypovolemic shock in the terminal circulatory collapse that precedes death in dehydrating anurans was investigated in the toad, Bufo marinus. The activity of lysosomal proteinases (cathepsins) was used as an indicator of the shock state. An assay was developed for the determination of cathepsin activity in toad plasma and was used to demonstrate the presence of proteinases similar in pH dependence to mammalian cathepsins Bl and D. Plasma cathepsin D-type activity increased with dehydration in toads but was not significantly different from the activity in controls or in toads subjected to splanchnic artery occlusion. Toads appear to maintain plasma volume to 20% water loss at the expense of other extracellular fluids. These results do not support the hypothesis that hypovolemia is a direct cause of dehydrational death in anurans.

Two aspects of possible adaptation In cardiovascular performance caused by Increased plasma electrolytes were examined. Cells In anisotonic plasma may either act as osmometers or volume regulate. Blood flow rate Is dependent upon cell viscosity, which in turn is a consequence of cell volume and membrane deform-ability. Cell volume changes which Increase membrane deform-ability will thus potentially extend the limits of dehydration tolerance. It was found in R. catesbeiana and B. marinus that red blood cell Is maintain constant volume during dehydration. Cells in vitro Initially lose water, but then sodium, potassium and water move Into the cell. Cell viscosity within the physiologic range of hematocrits was higher In salt loaded non-regulating cell Is of B. marinus than In regulating isotonic cells.

This study attempted to selectively stimulate and record from either the amphibian or basilar papillae of Rana pipiens. Computer-generated, frequency-specific clicks were used to elicit BSER's from either amphibian or basilar papillae. Narrowband noise fatiguers were presented in the frequency region of which each papillae are tuned. It was expected that a threshold shift would be elicited in the papillae that received the acoustic trauma, and that no threshold shift would be observed from the collateral papilla. The results of this experiment indicated that there was no overall difference between the threshold shift of either papilla. Furthermore, the amount of AP threshold shift was relatively constant regardless of whether the fatiguer bandwidth was overloading the amphibian or basilar papillae. By contrast, the amount of BP threshold shift was greater when proceeded by a fatiguer with a bandwidth corresponding to the BP tuning region than by a fatiguer with a bandwidth corresponding to the AP tuning region. Additionally, curare maximized the amount of BP threshold shift following fatiguing noise presented with a bandwidth to which the AP is tuned.