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.     

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.     

Effects of Methallibure (ICI 33,828) on the pars distalis of pituitary,testis and thumb pad of the green frog,Rana esculenta L.

Summary The influence of ICI compound 33,828 (1--methylallylthiocarbamoyl-2-methylthiocarbamoylhydrazine = Methallibure) on the pars distalis of pituitary, testes and thumb pads was investigated in the intact adult male green frog, Rana esculenta. Methallibure affected the gonadotropic basophils (particularly the B2 cells) of the pars distalis, which showed varying degrees of degranulation and underwent a notable decrease in their percentage. Within the testis this compound caused the arrest of spermatogenesis. The most uniform effect of Methallibure was observed in the thumb pads, which invariably showed regression of the epidermis and glandular epithelium. No histological changes occurred in the thyroid and adrenal glands and the B1 and A1 cells of the pars distalis remained unchanged cytologically. It is concluded, in concordance with the available data, that Methallibure is a non-steroidal antigonadotropic compound. The important question about its mode of action has been brought into discussion.Work supported by the National Research Council of Italy, and the Population Council (Grant No. M70.082 C) of New York.     

Morpho-functional changes in small intestine epithelium of frog Rana temporaria during hibernation

Structure and function of small intestinal epithelium were studied in overwintering frogs Rana temporaria at various stages of hibernation. In the process of testing of absorption of arginine vasotocin (AVT) in experiments in vitro it is established that at the period of hibernation there is preserved the capability of the epithelium for absorption of this nonapeptide without hydrolysis. However, as compared with October-December, in January-February and later, a decrease of the AVT absorption takes place, which is the most pronounced in March-April. Changes in epithelial structures appear by the middle of winter and are progressing by spring. In April-May, as compared with the beginning of hibernation, the height of enterocytes, the length of microvilli, and the number of microvilli decrease by 33 %, 40 %, and 57 %, respectively. The absence of features of destruction indicates an adaptive character of the observed changes. Dynamics of the studied parameters indicates morphological plasticity of the small intestine epithelium of R. temporaria at the period of hibernation.     

Cytological changes in the pars distalis of pituitary of the green frog,Rana esculenta L., during the reproductive cycle

Summary The cytological changes in the pars distalis of pituitary of male and female green frog, Rana esculenta during the reproductive cycle have been studied by examining the form, cytoplasmic granulation and numerical proportions of the different cell types. Some sex differences were recorded, particularly in relative total number of cells in a median sagittal section of the pars distalis. No extensive cytological changes occur during the hibernation period, but there are striking alterations at the time of spring awakening, during spawning months and during replenishment of the spent gonads i.e. during late summer and autumn months. Important changes are to be observed in the acidophils type one (A1) and basophils type two (B2) and three (B3). Changes in the A1 cells during breeding season have been speculated as being in support of the spawning activity of the animal. The B2 cells exhibit most profound changes which correlate precisely with the morphological changes in gonads, and these cells are considered as the FSH-gonadotrops. The functional significance of the B3 cells is discussed and it is suggested that they might be the source of an ICSH- or LH-like hormone.The work has been supported by grant from the National Research Council of Italy.The award of postdoctoral fellowship by the National Research Council of Italy is gratefully acknowledged.     

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.     

Pool desiccation and developmental thresholds in the common frog, Rana temporaria

The developmental threshold is the minimum size or condition that a developing organism must have reached in order for a life-history transition to occur. Although developmental thresholds have been observed for many organisms, inter-population variation among natural populations has not been examined. Since isolated populations can be subjected to strong divergent selection, population divergence in developmental thresholds can be predicted if environmental conditions favour fast or slow developmental time in different populations. Amphibian metamorphosis is a well-studied life-history transition, and using a common garden approach we compared the development time and the developmental threshold of metamorphosis in four island populations of the common frog Rana temporaria: two populations originating from islands with only temporary breeding pools and two from islands with permanent pools. As predicted, tadpoles from time-constrained temporary pools had a genetically shorter development time than those from permanent pools. Furthermore, the variation in development time among females from temporary pools was low, consistent with the action of selection on rapid development in this environment. However, there were no clear differences in the developmental thresholds between the populations, indicating that the main response to life in a temporary pool is to shorten the development time.     

Karyosphere of the oocytes of the common frog, Rana temporaria]

Karyospheres of ca. 200 mcm in diameter were isolated from the common frog oocytes of definitive size. An electron microscope study has revealed in the karyosphere fibrillar nucleoli and micronucleoli, modified synaptinemal complexes sometimes connected with chromatin and fibrillar material containing a great number of, mostly atypical, pore complexes resembling those of nuclear membrane and forming "pseudomembranes". An electrophoresis of the isolated karyosphere has revealed 12 distinct protein bands, of 3 which correspond to the protein triplet characteristic of the nuclear matrix and the rest 9 represent high molecular weight components with the molecular weight from 130 to 200,000 D.     

Seasonal changes in endocrine organs of the male common frog,Rana temporaria

Summary The annual cycle of the five morphological cell types in the pars distalis of the adenohypophysis of male common frogs,Rana temporaria, was studied in variously stained sections of the pituitary, collected at monthly intervals. All cell types showed a maximal development at the end of hibernation, a strong extrusion of secretory granules in the breeding season, and a more gradual release during later spring and early summer. A minimal development occurred in mid-summer, and a restoration followed in autumn and the hibernation period. During the phase of extrusion production was low, and vice-versa. These seasonal changes correspond with those of the testes and interrenal tissue of the same animals as well as with those of the thyroids and other endocrine tissues, and seems to be an important part of the biphasic annual life cycle of the species.The authors gratefully acknowledge the patient and skilful technical assistance of MissFemmy Brands. They thank Mr.H. van Kooten for making the photographs.     

Determination of growth and maturation in the common frog, Rana temporaria, by skeletochronology

Growth and maturation in a Swiss population of Rana temporaria were studied in 1983 and 1984 by means of skeletochronology. Resting line (growth ring) diameters were used to back-calculate individual body sizes in previous years; these permitted establishment of an average growth curve and determination of individual ages and sizes at first reproduction. Growth was rapid up to maturation, but continued thereafter at a decreased rate. Males were larger than females at age two but females grew faster thereafter, causing sexual dimorphism in adult body sizes. Body size distributions for both years and for frogs recaptured and first captured in 1984 were established. Growth in immatures was positively, but in adults negatively correlated with body size, with considerable variation at all sizes. Individual adult sizes were positively correlated with body sizes at the end of the first year. Average individual age at first reproduction was 2.8 years in males and 3.1 years in females (range in both sexes two to four years). There is no evidence for a two-year-cycle of reproduction.     

Effect of photostimulation on the wakefulness-sleep cycle in the common frog Rana temporaria

Effect of daily 30-min photostimulation in the 10 s light: 10 s pause (the total of 5 days) on the time structure of the wakefulness--protosleep cycle (WPC) was studied in the common frog Rana temporaria. Changes were analyzed of EEG wave components in three immobility forms of the type of catalepsy (P-1), catatonia (P-2), and cataplexy (P-3) that form protosleep. The first three photostimulations promoted a gradual increase of the P-1 state to 84.16 +/- 11.6% [the initial value (IV) 22.9 +/- 9.1%] and a decrease of representation of wakefulness to 4.86 +/- 2/1% (IV 13.8 +/- 7.8%), of P-2 to 11.1 +/- 5.3 (IV 53.3 +/- 13.3%), and of P-3 to 2.21 +/- 1.0% (IV 11.1 +/- 5.6%). After 4-5 photostimulations and especially after their complete cessation the percentage of P-1 in the WPC was restored to initial values, whereas the percentage of the frog WPC P-3 considered to be a precursor of the homoiothermal sleep rose to 20 +/- 8.3% after 5 photostimulations and to 38.5 +/- 6.7% the next day. Changes in the frog EEG spectra appeared only after one photostimulation and were characterized by a brief increase of power of alpha-like waves and by inhibition of slow 6-waves. In P-2 the power of the slow delta-waves gradually rose. In P-3 the EEG parameters did not change. In all experimental animals a decrease of the relative thymus and adrenal masses was revealed, which indicates the photostimulation regime used in the work induces stress. The obtained data allow thinking that a certain neurohormonal response to stress has already been formed at the amphibian level and that an important role in this response realization is played by a coordinated interaction of the hypothalamic sleep-regulating system providing protosleep manifestations and of the hypothalamic neurosecretory system triggering the stress-reaction hormonal cascade.