Relationships between feeding, digestion and water balance in a carnivorous vertebrate, the toad Bufo bufo L.

1. Toads fed meal worms retained water, amounting to about the mass of the meal. 2. Secretion of gastric juice in the initial phase of digestion of a large meal would deplete the extracellular fluid of about half of the chloride ions. 3. Ingestion and rapid digestion of large meals in toads and other carnivorous vertebrates imply tolerance of large variations in anion composition and acidity of the extracellular fluid.     

Radial astrocytes and ependymocytes in the spinal cord of the adult toad (Bufo bufo L.)

Summary Immunohistochemical and ultrastructural techniques have been used to demonstrate glial fibrillary acidic protein (GFAP) immuno-positive cells in the adult toad spinal cord. Two types of GFAP-immunoreactive cells were observed: ependymocytes and radial astrocytes. GFAP-positive ependymocytes were scarce and contained the immunoreactive product in their processes. They showed intermediate filaments in the basal pole and in their processes when studied with the electron microscope. These immuno-positive ependymocytes represent the tanycytic form of ependymal cells because their processes ended at the subpial zone. The radial astrocytes showed a more intensive immunoreactive product in somata and processes when they were located far away from the ependymal layer. Cell bodies and processes were also associated with blood vessels, but most of the processes ended at the subpial zone forming a continuous subpial glia limitans. The GFAP-positive processes, which form this subpial glia limitans in the toad spinal cord, belong to both tanycytic ependymocytes and radial astrocytes, whose somata are located in the grey matter. These findings lead us to suggest that both types of GFAP-immunopositive cells might be the functional equivalents of mammalian astrocytes.     

Water economy in a terrestrial toad (Bufo bufo), with special reference to cutaneous drinking and urinary bladder function

Undisturbed toads, acclimated to a simulated terrestrial habitat with access to water, generally visited the water resource for cutaneous drinking before evaporative water losses had resulted in dehydration of the body, and often the bladder still contained ample amounts of urine. The toads did not urinate when they stayed out of water, but exposure to water in the terrestrially-acclimated state facilitated urination, even when the bladder contained only insignificant amounts of urine. Daily emptying of the bladder often resulted in substantial water deficits prior to drinking, but the severity and frequency of the deficits declined with time, concurrently with an increase in the frequency of cutaneous drinking. Volumes of urine stored in the bladder when the toads suspended cutaneous drinking varied from negligible to large, corresponding to up to 20% of the body mass. Daily emptying of the bladder tended to increase the volume of urine stored at the end of drinking episodes. It is concluded that toads and other terrestrial amphibians primarily maintain normal water balance by anticipatory cutaneous drinking; emergency drinking in response to dehydration plays a secondary role.     

Organogenesis and differentiation of the pancreas in the toad Bufo bufo L.

Summary In Bufo bufo at stage III₆ the first endocrine islets appear in the part of the pancreas corresponding to the dorsal anlage. At stage IV₂, 5 days later, the pancreatic duct develops and new islets arise by budding off from the ductal epithelium. The ultrastructural study of the secretory granules morphology of endocrine cells has distinguished four different cell types: B-cells (stage III₉), A-cells (stage IV₃), D-cells (stage IV₃) and a fourth type not yet identified (stage IV₃). By immunocytology insulin and corticotropin-releasing factor (CRF) cells have been demonstrated at stage III₉, and glucagon and somatostatin cells at stage IV₁. Lastly, endocrine islets can be homogeneous (predominantly containing insulin cells, rarely glucagon cells) or heterogeneous (insulin cells at the centre and glucagon or somatostatin cells at the periphery). Hypotheses are put forward for the origin and the constitution of the different generations of endocrine islets and isolated cells.     

Cell kinetics in the epidermis of the toad Bufo bufo bufo (L.) after partial hypophysectomy followed by corticotrophin treatment (an autoradiographic study)

1. Cell kinetic parameters in the epidermis of partially hypophysectomized, ACTH-treated toads whose moulting frequency was increased threefold were studied using labelled thymidine and the autoradiographic technique. 2. The labelling index, length of S-phase and cycle time of stratum germinativum cells changed slowly as a function of the number of ACTH injections, and so did the migration of cells from stratum germinativum to stratum corneum. 3. A cell production in stratum germinativum of normal toads in excess of cell loss, as demonstrated by Levi & Nielsen (1982) should make it possible to maintain a status quo over a limited period of time following ablation of pars distalis and ACTH treatment.     

Vascularization of the pars intermedia of the hypophysis in the toad,Bufo bufo (L.) (Amphibia,Anura)

The vascularization of the pars intermedia of the hypophysis of the toad, Bufo bufo (L.) was studied by injection of a mixture of India-ink and gelatine into the circulatory system of the head via the arteria carotis communis. Further methyl-methacrylate corrosion casts of brains were made and the hypophysial region of the corrosion casts was examined with the scanning electron microscope. The results showed that the vascularization of the pars intermedia of the toad hypophysis consists of a single-layered vascular network, which is located on the ventral surface of the pars intermedia. The network is formed by capillaries, which primarily run caudally in a fan-like manner and which show only a few cross-connections. In the rostral region of the pars intermedia this network lies rather superficially, while in the caudal region it slightly penetrates the parenchyma. The vascular network originates from vessels of the neural stalk and from wide capillaries of the rostro-ventral region of the neurointermediate junction. The venous drainage of the pars intermedia is exerted by veins, which leave the caudal region and drain into the veins leaving the venous pole of the pars distalis. The flat, wide meshed vascular net on the ventral side of the pars intermedia, demonstrated in this study, fits into the concept that the pars intermedia of the anuran hypophysis is under the control of nerve fibers coming from the hypothalamus.     

Vascularization of the pars distalis of the hypophysis in the toad,Bufo bufo (L.) (Amphibia,Anura)

Summary The vascularization of the pars distalis of the hypophysis of the toad, Bufo bufo (L.), was studied by the traditional method of injecting a mixture of India-ink and gelatine into the circulatory system of the head via the arteria carotis communis. Further, methyl-methacrylate corrosion casts of the brains were made; the hypothalamo-adenohypophysial region of these corrosion casts was studied with the scanning electron microscope. The results showed that the portal vessels which arise from the median eminence do not supply distinct areas in the pars distalis as is supposed by the point-to-point-hypothesis. The portal vessels enter the ventro-median region of the pars distalis and branch off into a three-dimensional network of the secondary capillary plexus of the pars distalis. The plexus is made up mostly by four- to six-sided meshes. This angioarchitecture guarantees an optimal supply of the glandular cells of the pars distalis with nutritional factors and releasing hormones, on the one hand, and facilitates the removal of the hormones which are released by these cells, on the other hand. The venous drainage of the pars distalis is exerted mainly by two large veins, which bilaterally leave the dorso-lateral region (venous pole) of the pars distalis and by a few small veins, which drain into the wide, sinus-like vessel, which curves around the dorso-caudal region of the pars distalis and joins bilaterally the vena hypophysea transversa.This work was supported by the Fonds zur Förderung der wissenschaftlichen Forschung (Projekt Nr. 2183/N 39) and by the Stiftungs- und Förderungsgesellschaft der Paris-Lodron-Universität SalzburgThe authors wish to thank Mag. Ursula Albrecht for excellent technical assistance and Mr. Gerhard Sulzer for photographic work     

Specific features of the melanophore system in different color morphs of larvae of the common toad (Bufo bufo L.)

In a natural pond among usual black larvae of the common toad (Bufo bufo L.), a few unusual individuals of red-olive coloring were found out. In both morphs we investigated the melanophores of skin using different methods. The ESR-spectrometric analysis has shown the absence of distinctions between morphs by the amount of melanin. Analysis of total preparations of skin has shown the presence of various kinds of melanophore cells both in the derma and in the epidermis. Among typical melanophores, essentially differing cells appeared (atypical cells). In black morph tadpoles, the number of all kinds of melanophores is significantly greater than in red-olive morphs. It is shown that dark coloring is connected with a considerable number of atypical cells in the epidermis imposed on a dense layer of typical dermal melanophores with dispersed melanin.     

Angiotensin II elicits water seeking behavior and the water absorption response in the toad Bufo bufo

Fully hydrated toads, Bufo bufo were acclimated to a simulated terrestrial habitat, with access to shelters and water. To get from the shelters to the water, the toads had to walk across the pan of an Ohaus balance and the body weights were recorded on a computer. Toads were placed inside shelters immediately following injection of human angiotensin II (A II), Thr(8)-saralasin, or Ringer's in the dorsal lymph sac, and their behavior was recorded continuously by video surveillance. The injection doses were 1-100 microg/100 g body weight A II and 100 microg/100 g body weight saralasin dissolved in 0.1 ml Ringer's; control animals received the same volume of Ringer's. The latency from injection to the initiation of water absorption behavior (WR) was significantly shorter in both A-II- and saralasin-injected toads, compared to controls. A-II- and saralasin-injected toads also spent significantly more time in the water than controls. The bladder depots when WR was terminated were significantly larger in A-II- or saralasin-injected toads than in controls. The stimulatory action of Thr(8)-saralasin, an antagonist of A II in mammals, on WR behavior in B. bufo suggests differences in receptor structure and/or receptor distribution between amphibians and mammals.     

Histochemistry of the juxtaglomerular apparatus in the toad Bufo bufo

Summary An investigation regarding the question of whether there exists a macula densa as part of the juxtaglomerular apparatus in the kidney of amphibians has been carried out. With the aid of a histochemical reaction for glucose-6-phosphate dehydrogenase activity, the presence of a macula densa zone as a specialized part of the distal tubule in the toad Bufo bufo was demonstrated. The functional significance of the high glucose-6-phosphate dehydrogenase activity in the macula densa cells is discussed.