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.     

The morphology of the juxtaglomerular apparatus in the toad,Bufo bufo

Summary In a light microscopic study the course of the tubule in the kidney of the toadBufo bufo was studied. The distal tubule returning to the glomerulus of its origin appears to enclose the afferent arteriole. In that area, from which a three dimensional graphic reconstruction is made, there is an intimate contact between tubular and vascular wall. The latter contains granulated media cells. In the part of the tubule adjacent to the afferent arteriole an accumulation of nuclei is present. It is suggested that this structure is similar to the macula densa of the mammalian juxtaglomerular apparatus. The functional significance of a stricture in the tubule distally from the macula densa-like structure is discussed.The authors wish to thank Mrs. Ineke van de Mee-Wienen and Miss Ans Rouwenhorst for their technical assistance and Mr. J. J. M. de Bekker for the realization of the graphic reconstruction.     

Egg production in the Common toad, Bufo bufo

The number and size of eggs produced by the Common toad (Bufo bufo) was investigated at a site in southern England. There was a trend within the population as a whole for egg weight, egg number and ovary weight to increase with female size, as defined by body length and body weight. However, with use of partial correlation analysis to remove the effects of female size from the data, some of these trends changed. Thus, for females of the same size, although egg number increased with ovary weight, no significant correlation was found between egg weight and ovary weight whilst egg weight became negatively correlated with egg number. The possible significance of variation in egg weight on tadpole survival and female reproductive success is discussed.     

Experimental study on the origin of pancreatic endocrine cells in the toad Bufo bufo L.

Summary Ablation, transplantation and culture experiments were used to determine the respective roles of the pancreatic dorsal and ventral anlagen in the formation of the endocrine cells. Three successive waves of endocrine formation occur in the pancreas of Bufo bufo at three developmental stages (III₆, IV₁ and IV₂). Each wave is derived from a different source: the first originates from the dorsal anlage, the second from the exocrine tissue of the cortex of the pancreas and the third from the pancreatic duct. Each generation of islets has a specific composition of different cell types. The first wave is only composed of insulin islets; the second wave gives rise to single insulin, glucagon and somatostatin cells; while the third wave generates single cells synthesizing one of the three hormones, homogeneous islets of insulin cells, rare glucagon islets and heterogeneous islets containing insulin cells in the centre and a few glucagon or somatostatin cells at the periphery.     

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.     

Breeding pond fidelity in the common toad, Bufo bufo

Two independent studies of adult common toad, Bufo bufo , movements between different spawning ponds/populations both within and between years were made in England and Sweden during the spring breeding periods of1987–1990. The results of the two studies were subsequently combined since they were found to be complementary. In addition, in England during 1984 and 1985, large numbers of 'toadlets' were marked so that they could be identified if recaptured as breeding adults.
The degree of relocation between ponds was negatively correlated to the distance between ponds. The proportion of both males and females that moved between ponds within a particular year was significantly less than the proportion relocating between years. In any year, between 79% and 96% of adults that survived to breed the following year, returned to the original pond. Similarly, of the toadlets marked during1984–85, 81% of the males that subsequently returned as breeding adults were captured in their pond of origin.
Isolation, in relation to both population dynamics and population genetics, is defined and its implications for the management of common toad populatins discussed.     

The effect of winter temperatures on the timing of breeding activity in the common toad Bufo bufo

A 19-year study of a common toad population in south Dorset, UK, was carried out between 1980 and 1998. The daily arrival of sexually mature male and female toads at a breeding pond was recorded each year. The timing of the main arrival of toads at the breeding pond was highly correlated with the mean daily temperatures over the 40 days immediately preceding the main arrival. When the temperatures were higher than average, breeding occurred significantly earlier in the year than if they were either average or lower than average. During the study, the toad breeding seasons were early (2–13 February) in 5 years (1989, 1990, 1993, 1995, 1998), late (16–23 March) in 2 years (1986, 1996) and average (25 February–8 March) during the remaining 12 years. Evidence was found suggesting that common toads have a daylength threshold of about 9 h, below which the migration to the breeding pond does not occur. Evidence was also found indicating that common toads in southern England have a threshold temperature for activity of about 6°C and that the onset of breeding activity is highly correlated with the number of days during the 40 days prior to the main arrival at the breeding pond that were at or above this temperature. Predicting the start of the main breeding migration to a pond in any year may be possible by comparing the pattern of the 40-day running mean daily temperatures from 21 December the preceding year with those from previous years when the start of breeding activity is known. Although all five of the earliest recorded toad breeding years occurred during the last 10 years, and were associated with the occurrence of particularly mild winters, a significant trend towards earlier breeding in recent years compared with previous years was not found. Received: 16 July 1998 / Accepted: 14 September 1998     

斯洛维尼亚池塘中大蟾蜍的交配模式

为了解大蟾蜍( Bufo bufo)的配对成功是否与身体大小有关,在蟾蜍产卵之前,我们在斯洛文尼亚捕捉和测量了2 224只成体样本。在这些捕获的个体中, 1 772只为雄性, 452只为雌性,其中355对处于抱对状态。雌性的体长(从吻端到泄殖腔的距离)一般比雄性稍长。无论雄性还是雌性,抱对者的体长均大于未抱对者。抱对的雄性和雌性的体长具有显著的正相关。我们所发现的大小匹配的交配模式,与从其它普通蟾蜍种群得到的结果一致[动物学报51 (3) : 513 -515 , 2005]。     

中华大蟾蜍斯钙素基因的组织表达及其在肾脏中的细胞定位

斯钙素(Stanniocalcin, STC)是一类首先在鱼类特有的内分泌腺--斯坦尼氏小体(Corpuscles of Stannius, CS)、随后又在人和哺乳动物中发现的同型二聚体糖蛋白激素,具有广泛的组织表达模式和多种生物学效应.为阐明两栖类动物是否存在STC1基因的表达及其表达模式,本研究基于部分已知鱼类和哺乳动物的STC1基因序列,从中华大蟾蜍(Bufo bufo gargarizans)卵巢获得了STC1基因的部分序列(GenBank注册号为EF586886).同源性分析显示,所获得的中华大蟾蜍STC1基因部分序列与鱼类STC1基因相应序列的同源性在40%-48%,而与小鼠和人STC1基因相应序列的同源性分别为41.89%和37.95%.RT-PCR分析显示STC1基因可在肾脏、性腺等多种组织中表达;原位杂交(in situ hybridization, ISH)技术表明中华大蟾蜍肾脏的近端小管、远端小管和集合管细胞内表达STC1 mRNA.这些结果首次证实两栖类动物中华大蟾蜍组织中存在STC1基因的表达     

Immunohistochemical demonstration of renin in the juxtaglomerular apparatus of three Bufo species

Summary The cellular localization of renin was examined in the kidneys of some amphibians of the genus Bufo by immunoperoxidase and immunofluorescence techniques with an antiserum to renin isolated from the submandibular gland of the mouse. Immunoreactivity could be demonstrated in the media cells of the afferent arterioles (juxtaglomerular cells) close to as well as at great distance from the glomeruli. Occasionally, media cells of larger arterial vessels were also stained. The immunohistochemical data seem to be in accordance with earlier results obtained with a modified silver impregnation technique (Movat's staining procedure) used for the visualization of juxtaglomerular cells in non-mammalian vertebrates. Mouse kidney tissue, studied for purposes of comparison, showed renin-immunoreactivity as described by earlier investigators, i.e., immunoreactive staining in the afferent arterioles near the glomeruli and in the proximal tubule cells.     

Male breeding behaviour and mate acquisition in the Common toad, Bufo bufo

The breeding behaviour of a Common toad ( Bufo bufo ) population was studied at a breeding pond on Portland, Dorset. Major toad movements towards the breeding pond occurred on warm, wet nights when the temperature did not fall below 6°C. Males arrived earlier than females resulting in an initially exaggerated sex ratio which then slowly reduced as the breeding season progressed. No size assortative pairing was found although large males were more successful at mating than small males. Although males demonstrated no ESS (evolutionary stable strategy) in searching for a female between sites within the pond evidence suggesting the existence of one between the pond and an undefined area surrounding it was found.     

A case of pulmonary cryptococcosis in a free-living toad (Bufo bufo)

Pulmonary cryptococcosis was observed in a free-living adult female common toad (Bufo bufo) that was killed by a vehicle. Both lungs had various eosinophilic, monomorphic, and spherical to elliptical organisms identified as Cryptoccocus spp. The yeasts were demonstrated by Grocott's silver method and the periodic acid-Schiff reaction and the capsule was positive for mucin with a mucicarmine stain. The agent was confirmed by immunohistochemistry, using the monoclonal antibody anti-Cryptococcus neoformans, and by a polymerase chain reaction-based method using a C. neoformans-specific primer. This report, to the best of our knowledge, represents the first case of cryptococcosis in a common toad.