Effect of osmotic gradient on ADH-induced intramembranous particle aggregates in toad bladder

Summary Paired toad urinary bladders were prepared without or with an osmotic gradient (175 mosm) across them, stimulated for 2.5 (n=6), 5 (n=6), 30 (n=6) or 60 (n=6) min with ADH (20 mU/ml), and studied by freeze-fracture electron microscopy. Water permeability at these times was assessed in additional bladders (n=6 for each case) after tissue fixation according to the technique of Eggena. After both 60 and 30 min of ADH stimulation, the presence of a gradient compared with the absence of one was associated with fewer aggregates (242±35vs. 382±14 ×235 m^–2 at 60 min,P<0.01; 279±36vs. 470±51 ×235 m^–2 at 30 min,P<0.01) and lower water permeability (8.4±1.1vs. 18.8±1.8g×min^–1×cm^–1 ×mosm ^–1 at60min,P<0.005; 9.2±1.0vs. 22.0±2.1 g ×min^–1×cm^–2×mosm ^–1 at 30 min,P<0.001). In addition, with a gradient both maximum water permeability and maximum aggregate frequency were reached nearly together; a similar correspondence occurred without a gradient. We conclude that in the presence of an osmotic gradient both the ADH-associated aggregates and the water permeability response to ADH are prevented from reaching the higher levels observed in bladders not exposed to a gradient.     

Types of supernumerary outgrowths produced after inverting the dorsoventral limb axis of the anuranBufo bufo

Summary Contralateral grafts were performed on the larval limb buds of the anuranBufo bufo. The dorsoventral axis of 80 buds at stages IV or V was inverted. Ten tadpoles were used as controls. Fifty-two supernumerary structures developed, all of them in dorsal or ventral locations on the host stump. The majority (32 out of the 44 outgrowths with more than 3 toes) were normal limbs of stump handedness. However, the following abnormal structures were also observed: 2 double-posterior, 3 mixed-symmetric, and 7 undetermined cases. These results are in agreement with the predictions of a hierarchical polar coordinate model for epimorphic regeneration.     

不同脊椎动物消化道内5-羟色胺免疫染色细胞的分布

用过氧化物酶——抗过氧化物酶（PAP）法,对乌鳢（Ophiccephalus argus）、中华大蟾蜍（Bufo bufo garaaarizans）、黄喉水龟（Clemmys mutica）、虎皮鹦鹉（Melopsittacus undulatus）和小白鼠（Mus musculus albula）五种脊椎动物消化道内的5-羟色胺（5-hydroxytryptamine,简称5-HT）免疫染色细胞的分布进行了研究。发现各种动物胃肠道（虎皮鹦鹉胃、乌鳢肠及胃贲门除外）均含有5-HT免疫染色细胞,并首次发现黄喉水龟和中华大蟾蜍食道内含有5-HT免疫染色细胞。一般地,各种动物胃内5-HT免疫染色细胞密度最高,十二指肠和大肠次之,小肠最低。5-HT免疫染色细胞位于粘膜上皮或腺上皮细胞间,常有一个或一个以上的细胞突起伸入固有层或肠腔面（或腺腔面）,有些细胞的一端突起伸入固有层,另一端突起伸入肠腔面,表明5-HT免疫染色细胞兼具内分泌或分泌功能。     

花背蟾蜍蝌蚪变态期角膜发育的研究

作者用光镜和电镜研究了花背蟾蜍蝌蚪变态期角膜的发育。在后肢发育晚期,内、外角膜在中央部位首先愈台,在完全变态期角膜完全愈合,此时角膜上皮细胞增殖,上皮基质变为Bowman’s膜,内、外角膜之间的成纤维细胞和由它分泌的胶原纤维形成角膜基质,内角膜细胞形成单层的角膜内皮,它与角膜基质间的Descemet’s膜最晚形成。     

Exocytosis of cortical granules from activated oocytes of the toad,Bufo arenarum

Summary Observation of the cortical region of oocytes of Bufo arenarum by transmission electron microscopy reveals modifications on their surface and in the contents of the cortical granules (CG) during activation. In non-activated oocytes only amorphous cortical granules (ACG) can be observed. Activated oocytes display ACG, intermediate cortical granules containing both amorphous and membranous material (ICG), and a third type containing only membranous material (MCG). During exocytosis, CG release their contents into the perivitelline space, where the amorphous and membranous materials are found. The three types of CG found during oocyte activation suggest transformation of ACG to MCG and indicate that the different components of the cortical granules, when released into the perivitelline space, might play different roles in prevention of polyspermy.Members of the Scientific Research Career of CONICET, R. Argentina.     

Phylogeography of the natterjack toad Bufo calamita in Britain: genetic differentiation of native and translocated populations

The natterjack toad Bufo calamita is rare in Britain, which is at the northwestern edge of its biogeographical range. We investigated the level of genetic differentiation amongst almost all (34 out of 38) of the surviving British populations of this species, and among six new populations established by translocations during the 1980s. For eight microsatellite loci, allele sizes and frequencies were analysed using samples from each of these populations. The populations clustered into three robustly differentiated groups, each of which corresponded with a geographical region (east/southeast England, Merseyside and Cumbria). The Cumbrian populations showed a further weak geographical substructuring into northern and southern clades. The populations in south Cumbria were genetically more diverse than those in any of the other regions, as judged by the mean numbers of alleles per locus and the mean heterozygosity estimates. The translocated populations clustered close to their founders and, with one exception, did not differ significantly with respect to mean allele numbers, heterozygosity or polymorphism level. However, significant genetic differentiation (as measured by unbiased R _ST) was found between all but one of the founder-translocation pairs. The implications of this phylogeographic study for the future conservation of B. calamita in Britain are discussed.     

Postmetamorphic dispersal and recruitment of first breeders in a Bufo calamita metapopulation

The migratory capacity of metamorphs and their later integration into local breeding populations was studied in a Bufo calamita metapopulation inhabiting the northern Rhineland, Germany (1991–1994). A new tagging method allowed unequivocal distinction among metamorphs (n = 1,709), which originated either from the spring-breeding cohort in 1991 and 1992 or from the summer-breeding cohort in 1991. The life history of batch-marked individuals was followed until adulthood, focusing on postmetamorphic dispersal, choice of breeding locality and timing of adult reproduction. Almost all metamorphs left the patches used for breeding within a few weeks after emergence. They were found at about the same density in all favourable microhabitats in a radius of up to 2 km around their native patch. This demonstrates that even small toadlets (6–11 mm snout-to-vent length) migrate distances of several hundred metres within a short time and that they significantly help prevent local extinction in sink populations (rescue effect). Recaptures of reproductive adults originating from tagged metamorphs were limited to males belonging to the 1997 spring and summer cohorts. The absence of recaptured females was probably due to their greater vagility and later sexual maturation. First-breeding males (2 years old) without exception joined the breeding cohort from which they originated: early breeders originated from spawn of the spring cohort, mid-summer breeders from spawn of the summer cohort. This result is in accordance with the hypothesis that the integration of first breeders to a specific breeding cohort is genetically controlled in central European natterjacks. The exchange of individuals between different cohorts in the same patch is considerably less than the migratory exchange between the same breeding cohorts among local populations, as predicted from the allozyme data. Received: 28 October1996 / Accepted: 4 June 1997     

Gene flow and functional connectivity in the natterjack toad

Functional connectivity is a key factor for the persistence of many specialist species in fragmented landscapes. However, connectivity estimates have rarely been validated by the observation of dispersal movements. In this study, we estimated functional connectivity of a real landscape by modelling dispersal for the endangered natterjack toad (Bufo calamita) using cost distance. Cost distance allows the evaluation of 'effective distances', which are distances corrected for the costs involved in moving between habitat patches in spatially explicit landscapes. We parameterized cost-distance models using the results of our previous experimental investigation of natterjack's movement behaviour. These model predictions (connectivity estimates from the GIS study) were then confronted to genetic-based dispersal rates between natterjack populations in the same landscape using Mantel tests. Dispersal rates between the populations were inferred from variation at six microsatellite loci. Based on these results, we conclude that matrix structure has a strong effect on dispersal rates. Moreover, we found that cost distances generated by habitat preferences explained dispersal rates better than did the Euclidian distances, or the connectivity estimate based on patch-specific resistances (patch viscosity). This study is a clear example of how landscape genetics can validate operational functional connectivity estimates.     

Parasites,info-disruption,and the ecology of fear

There is growing interest in the ecological consequences of fear, as evidenced by the numerous studies on the nonconsumptive, trait-mediated effects of predators. Parasitism, however, has yet to be fully integrated into research on the ecology of fear, despite it having direct negative and often lethal effects on hosts and being the most common life history strategy on the planet. This might at least be partly due to the traditional, but untested, assumption that anti-parasite responses are weak relative to anti-predator responses. To test this hypothesis, we quantified the activity and location responses of Bufo americanus tadpoles to one of six chemical cues: water; cercariae of Echinostoma trivolvis, a trematode which infects and can kill amphibians; a snail releasing E. trivolvis cercariae; an uninfected snail; food; or conspecific alarm chemicals signaling predation. There is also literature encouraging research on the context dependency and pollution-induced disruption of fear responses. Consequently, before quantifying responses to the chemical cues, half of the B. americanus were exposed to the herbicide atrazine (201 μg/l for 4 days), a reported inhibitor of fear responses in fish. Tadpoles were attracted to food, were indifferent to an uninfected snail, avoided alarm chemicals, and exhibited avoidance and elevated activity in response to a snail shedding cercariae and cercariae alone. Atrazine had no detectable effects on B. americanus’ responses to the tested cues despite the use of a higher concentration and longer exposure duration than has been repeatedly shown to inhibit chemical cue detection in fish. The magnitude of anti-parasite and anti-predator responses were qualitatively similar, suggesting that the fear of disease and its ecological consequences could be comparable to that of predation. Consequently, we call for a greater integration of parasites into research on the ecology of fear and trait-mediated indirect effects.