The effect of vasotocin, mesotocin and adrenaline on the interrenal gland in the frog Rana temporaria under in-vitro conditions]

Morphometric studies indicate that vasotocin increases the activity of the adrenal gland in frogs, whereas mesotocin and epinephrine do not essentially affect it. Combination of epinephrine with nonapeptides in cultural medium showed that the former may abolish the effect of vasotocin on the adrenal gland.     

Morphofunctional research on the interrenal gland of the frog Rana temporaria following arginine vasotocin administration

Studies have been made of the effect of injections of hypothalamic nonapeptide neurohormone, arginine vasotocin, on functional condition of the interrenal gland in mature frogs. In unoperated, sham-operated and in animals 10 days after hypophysectomy, single and especially three subsequent injections of arginine vasotocin (5 x 10(-9) M per 1 kg of the body weight) result in evident activation of glandular cells of the interrenal gland which is manifested in the increase of the volume of their nuclei and cytoplasmic area, as well as in the dilatation of the blood vessels. Activation of the interrenal gland in hypophysectomized frogs, which lack endogenous ACTH, indicate the direct para-adenohypophyseal influences of nonapeptide hypothalamic hormones on the activity of glandular cells in the peripheral endocrine glands, in particular, the interrenal gland of the grass frog.     

The skin sensitivity of the frog Rana temporaria to chlorides]

Analysis of total impulse reactions evoked in skin branches of the spinal nerves by application of 0.1-1.0 mol/l solutions of KCl, NaCl, NH4Cl, and CaCl2 to the skin in the grass frog showed that the reaction of the peripheral nervous elements depends not only on the concentration of a test-solution, but on the nature of the cation as well. The problem of specificity of acidic and saline sensitivity of the skin in amphibians is discussed.     

The temperature hysteresis of water-salt metabolism in the frog Rana temporaria]

Frogs acclimated to 4 degrees C were transported to a medium with temperature 20 degrees C, which caused polyuria; recovery of normal diuresis took about 24 h. During this period, hypernatremia was observed together with the increase in natriuresis, the rate of renal excretion of potassium ions with urea remaining constant. Water content of skeletal muscles decreased. Transportation of frogs acclimated to 20 degrees C into a medium with a temperature 4 degrees C decreased their diuresis. Renal excretion of sodium, calcium and magnesium ions remained unchanged, whereas that of potassium ions significantly decreased. The content of potassium and magnesium in the blood serum increased, that of sodium--decreased. Hydratation of muscled and kidneys was accompanied by the decrease of calcium, potassium and magnesium ions calculated per wet weight of the tissues, the level of sodium remaining unaffected. The data obtained indicate significant changes in the pattern of water and salt metabolism in frogs during temperature hysteresis.     

The effect of cyclic nucleotides on the rhythmogenicity of the lymphatic center in the frog Rana temporaria]

Studies have been made of the effect of cAMP on the spontaneous impulse activity of lymphatic pacemaker neurons in the frog. It was shown that inhibition of phosphodiesterases by papaverine and 3-isobutyl-1-methylxanthin resulted in the increase in rhythm of bursting impulse activity of the lymphatic centre turning it into a continuous one. The same effect was produced by dibutyryl-cAMP, and to a lower extent--by cAMP. CGMP blocked rhythmic activity of the centre. Possible role of cyclic nucleotides in rhythmic activity of pacemaker cells in the spinal centre of the lymphatic hearts in the frog is discussed.     

Effect of the epiphysis and hypophysis on the circadian organization of the wakefulness-primary sleep cycle in the frog Rana temporaria

Using continuous non-contact registration of the heart rate and motor activity in unrestrained frogs, studies have been made of the effects of hypophysectomy and epiphysectomy on functional state of the organism. It was shown that removal of the epiphysis alleviates the circadian physiological rhythm of natural changes of functional states in wakefulness-primary sleep cycle, whereas removal of the hypophysis results in additional cyclic rhythm of two functional states in the diurnal periodicity of the behaviour. The latter effect is considered to be the ultradian rhythm, which in the intact animals is masked by the circadian one modulated by natural diurnal illumination. It is suggested that removal of the epiphysis reveals endogenous rhythm of the activity typical of periodic regimes of the spontaneous excitation at early stages of ontogenesis.     

The effect of metopirone and aldactone on the interrenal tissue of intact and pars distalis-ectomized common frogs Rana temporaria

Summary The effects of metopirone (SU-4885) and aldactone (SC-9420) on the histology and histochemistry of the interrenal tissue of male common frogs, Rana temporaria, have been studied. From the changes in nuclear and cellular size, mitotic rate, and lipid and cholesterol content it has been concluded that in intact animals the administration of metopirone as well as of aldactone resulted in an increase of the interrenal secretory activity. The effects were very similar to that of ACTH. Ablation of the pars distalis prevented the interrenal activation by metopirone. The effect of aldactone was markedly reduced in operated frogs, but some stimulation remained. It is suggested that both substances act by way of enhancing pituitary ACTH ouput. The dependence of the secretion of corticosteroids on pituitary ACTH is discussed.I wish to thank Dr. W. J. van Dongen for his cooperation and criticism and for enabling me to do this work in his laboratory. I am endebted to Prof. Dr. P. G. W. J. van Oordt for his valuable and constructive suggestions. Thanks are also due to Miss Mieke Roelofsen and Miss Ineke Wienen for their conscientious technical assistance.     

Morphological changes in the anterior lobe of the hypophysis in allotransplantation]

Under study were histological changes resulting from rejection of adenohypophysis allotransplantat without special influence upon the recipient immunity. The allotransplantat rejection was established to be accompanied by pronounced morphological changes in the adenohypophysis. Delta-basophils were the first to loose their granules, and beta-basophils showed more moderate degranulation. Acidophils were preserved for a longer period. The chromophobes were more resistant cellular elements. Simultaneously the cells were dying at all stages of their conversion. The fibrous connective tissue alone was observed at the site of allotransplantat 32-35 days after transplantation.     

Role of various regulation factors of interrenal gland activity in response to stress in the green frog Rana esculenta L]

The effects of an ether stress on the corticosterone levels were studied in variously experimented frogs. The removal of the distal lobe of the pituitary gland, with preservation of the intermediate lobe which contains also ACTH, suppresses, as total hypophysectomy, the increase of corticosterone level observed in normal frogs. The administration of dexamethasone reduces but not suppresses this increase. A second ether stress 20 minutes after the first is effective. The injection of frog plasma treated or not with an corticosteroid adsorbant do not alter the response to stress. These results suggest that modifications of corticosterone level, in some limits, do not disturb the effects of a stress.     

The structure and vascularization of the pars tuberalis of the hypophysis of Rana temporaria

Summary The pars tuberalis of the hypophysis of Rana temporaria presents the general structural and the cytological characteristics of an endocrine gland. It is composed of elongated cells with long, branching processes ending on the external basement membrane of the pericapillary space. The pars tuberalis cells produce secretory granules which are accumulated in the pericapillary endings of the processes.Corresponding to its separate localization, the pars tuberalis of Rana temporaria has a separate vascularization of which the efferent capillaries anastomose with the capillary plexus of the median eminence. The general direction of the blood flow of the pars tuberalis is towards the capillaries of the median eminence. Also, the secretory products of the pars tuberalis pass into the blood stream of the hypophysial portal system.Several characteristics of the pars tuberalis show that its function must be different from that of the pars distalis of the hypophysis. Moreover, in contrast with the pars distalis, the activity of the pars tuberalis is not regulated by neurohumoral factors.The results show that a role of the pars tuberalis in the regulation of the activity of the pars distalis of the hypophysis is not excluded.     

The potential-dependent incoming ionic currents of myoblasts from the frog Rana temporaria developing in culture]

Voltage-dependent inward ionic currents in 1-6-day cultured skeletal myoblasts have been studied using whole-cell patch clamp technique. Sodium (INa) and two types of calcium (ICa) currents were recorded at all stages. INa did not differ from that described in frog striated muscle fibres. Both types of ICa were found to be DHP-sensitive. They differ in their activation time. It is suggested that two types of ICa correspond to ICa of twitch and tonic muscle fibres.     

Daily form of rest in the catfish Ictalurus nebulosus and the frog Rana temporaria]

By means of recording the cardiac rhythm, respiration rate and motor activity in unrestrained animals, studies have been made on the dynamics of formation of cataleptic-like immobilization in the teleost I. nebulosus. Rhythmic illumination at a rate 3/min. (10 sec light on - 10 sec light off) in the daytime gradually caused the onset of immobilization in the fish. This immobilization is characterized by the specific heart rate, type of respiration and the degree of plastic tone. Under the same experimental conditions, the formation of photogenic catalepsy in the frog Rana temporaria was confirmed. Typical heart rate, respiration pattern and the degree of plastic tone were determined as well. The role of illumination rhythm in the onset of all conditions which are characteristic for the "awakefulness-rest" cycle is discussed.     

2 unusual responses of the myocardium in the frog Rana temporaria to acetylcholine

When studying seasonal variations in acetylcholine sensitivity of isolated frog ventricle, two forms of unusual responses have been observed: 1) tonic response induced by supermaximal concentration of acetylcholine, and 2) double response to the increasing acetylcholine concentrations with two different threshold and maximum doses (KAch values differ by 1-2 orders). The tonic response is blocked by mytolone. Both effects are of a seasonal nature. Similar pattern of these reactions in frogs and molluscs suggests a possibility of restoration of evolutionary more ancient form of cholinergic response, hormonal influences being involved into this process.     

The structure and activity of the hypophysis of Rana temporaria in normal and in experimental conditions

Summary Volumetric, karyometric and histological studies were done on the pars distalis of the hypophysis of Rana temporaria, in normal and in experimental conditions.In the normal pars distalis, a gradient of activity of the P.A.S.-positive cells is described which shows a close relation with the blood supply. This relation strongly suggests the presence of a gradient of humoral factors which would influence the activity of the pars distalis. This gradient is not dependent on the influence of A.F.-positive neurosecretory fibres. In the P.A.S.-negative cells (O.G.-cells) a similar gradient could not be found. In transplants of pars distalis the gradient of activity is absent.There exists a striking difference between autotransplants and homotransplants of pars distalis in the eye, and also between autotransplants on the iris and autotransplants in the cornea.A parallelism between autotransplants of pars distalis on the iris and the pars distalis of animals deprived of their median eminence could be established. In both groups the pars distalis shows a similar decrease in volume. The glandular cells are well differentiated, but show a low activity. This low activity is most pronounced in the P.A.S.-cells. The normal variability in staining of the P.A.S.-cells has diminished: in general, the great majority of the P.A.S.-cells are P.A.S.-purples. The gradient of activity of the pars distalis has disappeared. The seasonal development of the gonads and of the secundary sexual characteristics is absent. This absence is probably not caused by a decrease of the total number of P.A.S.-cells of the pars distalis. It is probably due to the low activity of the P.A.S.-cells, which cannot be ascribed to the absence of the neurosecretory influence of the magnocellular preoptic nuclei.A preliminary note on the gradient of activity of the pars distalis has been published (Dierickx 1963c).