Substance P-related peptides in the hypothalamus of Amphibia

Summary The occurrence of Substance P-(SP)-related peptides in the hypothalamus of three species of Amphibia (newt, clawed, toad, frog) was studied immunohistochemically employing the indirect immunofluorescence method or a double-step technique (indirect immunofluorescence followed by the peroxidase-antiperoxidase complex method). SP-like immunopositive fibers are seen throughout the hypothalamus. They are especially abundant in the preoptic area and in the outer zone of the median eminence, suggesting a role of SP-related peptides in the hypothalamo-hypophysial regulation in these animals. Some SP-like neurons are seen in the posterior hypothalamus and in the preoptic area. In the newt, such SP-like immunopositive neurons occur frequently in the preoptic periventricular grey.Work performed under the C.N.R. project Biologia della Riproduzione     

Immunohistochemical localisation of substances reactive to antisera against α-and β-endorphin and met-enkephalin in the brain of Rana temporaria L.

Summary In the brain of Rana temporaria, two distinct systems reactive with - and -endorphin antisera, respectively, and with a met-enkephalin antiserum, have been detected immunohistochemically.Neurons reacting with - and -endorphin antisera are located (1) in the preoptic nucleus, and (2) in the pars ventralis of the tuber cinereum. Immunoreactive nerve fibres of both groups of perikarya end in the infundibular floor near the capillaries and the preoptico-hypophysial tract. Control reactions have shown that the immunoreactivity is suppressed by the corresponding antigens, but also by -LPH. In view of these results the immunoreactive systems examined correspond to an /-endorphin system or a lipotropinergic system.Neurons reacting with the met-enkephalin antiserum are located in the paraventricular organ. Intense immunofluorescence was observed in the infundibular floor. Controls show that the labelling by met-enkephalin antiserum is exclusively suppressed by met-enkephalin.In the pituitary gland, on the other hand, - and -endorphin antisera reveal: 1) the MSH/ACTH-like cells of the pars intermedia and 2) the ACTH-like cells of the pars distalis.Supported by the D.G.R.S.T., Contrat no 77.7.0648     

Development and growth of long bones in European water frogs (Amphibia: Anura: Ranidae), with remarks on age determination

Differentiation and development of long bones were studied in European water frogs: Rana lessonae, R. ridibunda, and R. esculenta. The study included premetamorphic larvae (Gosner Stage 40) to frogs that were 5 years old. Femora, metatarsal bones, and proximal phalanges of the hindlimb exhibit the same pattern of periosteal bone differentiation and the same pattern of growth. Longitudinal and radial growth of these bones was studied by examination of the diaphyses and epiphyses, particularly where the edge of periosteal bone is inserted into the epiphysis. The periosteum seems to be responsible for both longitudinal and radial growth. Investigation of the formation, length, and arrangement of lines of arrested growth reveals that the first line is present only in the middle 25-35% of the length of the diaphysis of an adult bone; therefore, only the central portion of the diaphysis should be used for age estimation in skeletochronological studies. Comparison of the shapes and histological structures of epiphyses in the femur, metatarsal bones, and phalanges revealed that epiphyseal cartilages are composed of an inner and outer part. The inner metaphyseal cartilage has distinct zones and plugs the end of the periosteal bone cylinder; its role in longitudinal growth is questioned. The outer epiphyseal cartilage is composed of articular cartilages proper, in addition to lateral articular cartilages. Differences in the symmetry of the lateral articular cartilages of distal epiphyses of the femur and toes may reflect adaptations to different kinds of movements at the knee and in the foot.     

Genetic variation of an endangered Malagasy frog, <Emphasis Type="Italic">Mantella cowani</Emphasis>, and its phylogeographic relationship to the widespread <Emphasis Type="Italic">M. baroni</Emphasis>

We investigated the degree and distribution of the genetic variation, and phylogeography, of two species of Malagasy poison frogs, Mantella cowani and M. baroni. The former is critically endangered due to its restricted distribution, habitat destruction and overcollection for the pet trade. Analysis of 526 bp of mtDNA (cytochrome b) resulted in separate haplotype networks for the two species, and discovered hybridization at a single locality. The two networks confirm the status of M. baroni and M. cowani as separate evolutionary species and units for conservation. Within both mitochondrial haplotype networks, specimens from different localities shared numerous identical haplotypes, even those from the most distant sample sites of M. baroni. Most populations were characterized by high haplotype diversity and no haplotype clades exclusive to geographical regions were observed. Protection of a few large populations of these species is therefore likely to conserve much of the mtDNA genetic diversity found in the entire species. While M. baroni is widespread and occurs in many nature reserves, we recommend efficient legal protection of some M. cowani habitats to protect this species against extinction.     

A previously unrecognized radiation of ranid frogs in Southern Africa revealed by nuclear and mitochondrial DNA sequences

In sub-Saharan Africa, amphibians are represented by a large number of endemic frog genera and species of incompletely clarified phylogenetic relationships. This applies especially to African frogs of the family Ranidae. We provide a molecular phylogenetic hypothesis for ranids, including 11 of the 12 African endemic genera. Analysis of nuclear (rag-1, rag-2, and rhodopsin genes) and mitochondrial markers (12S and 16S ribosomal RNA genes) provide evidence for an endemic clade of African genera of high morphological and ecological diversity thus far assigned to up to five different subfamilies: Afrana, Cacosternum, Natalobatrachus, Pyxicephalus, Strongylopus, and Tomopterna. This clade has its highest species diversity in southern Africa, suggesting a possible biogeographic connection with the Cape Floral Region. Bayesian estimates of divergence times place the initial diversification of the southern African ranid clade at approximately 62-85 million years ago, concurrent with the onset of the radiation of Afrotherian mammals. These and other African ranids (Conraua, Petropedetes, Phrynobatrachus, and Ptychadena) are placed basally within the Ranoidae with respect to the Eurasian groups, which suggests an African origin for this whole epifamily.     

中国西北地区中国林蛙各居群的分类学研究(两栖纲:蛙科)

采用形态学比较和数值分类相结合,对中国西北部分布地中国林蛙Ｒａｎａ ｃｈｅｎｓｉｎｅｎｓｉｓ各居群间（包括分布于青藏庙的地区的原定种或亚种）的差差水平进行了探讨。结果认为：分布于中国西北部高原地区的各居群其形态特征和数值分析这〖其中包括原定名为Ｒａｎａ ａｍｕｒｅｎｓｉｓ ｋｕｋｕｎｏｒｉｓ（Ｎｉｋｏｌ ｓｋｉｉ,１９１８,青海湖）,Ｒ．ｗｅｉｇｏｌｉｄ（Ｖｏｇｔ,１９２４,四川甘孜和德格）,Ｒ．     

中国角蟾属一新种(两栖纲:锄足蟾科)

采自贵州省水城县凤凰乡的一种大形角蟾,因其外部形态与角蟾属Ｍｅｇｏｐｈｒｙｓ已知种有一些特异之处,尤其是鼓膜显露;上颌内缘无栉齿状突;头后部有由痣粒组成的“Ｖ”形棕褐色斑;背部痣纹呈“＊”形;有犁骨棱;雄性无声囊等特征与大花角蟾Ｍ．ｇｉｇａｎｔｉｃａ明显不同。据此认为该角蟾为一新种,定名水城角蟾Ｍｅｇｏｐｈｒｙｓ ｓｈｕｉｃｈｅｎｇｅｎｓｉｓ Ｔｉａｎ,Ｇｕ ｅｔ Ｓｕｎ,ｓｐ．ｎｏｖ．。     

Species diversity of Amphibia, Reptilia and Lipotyphla (Mammalia) at Ambolokopatrika, a rainforest between the Anjanaharibe-Sud and Marojejy massifs, NE Madagascar

The Réserve Spéciale d' Anjanaharibe-Sud and the Parc National de Marojejy are two important areas of biodiversity and endemicity in northeastern Madagascar. These reserves are separated by about 40 km, including the extensive Andapa Basin, and connected by a rather narrow mid-altitude montane ridge. Nothing was previously known about the biota of this corridor and its faunal relations with the two reserves. At this purpose, during 1997 the Ambolokopatrika rainforest (lying about midway between Anjanaharibe-Sud and Marojejy massifs) was surveyed for amphibians, reptiles, and for small mammals belonging to the order Lipotyphla. As a general rule these vertebrates may be important ecological indicators, while the herpetofauna (Amphibia, Reptilia) exhibits a high degree of habitat specialisation and endemicity. Furthermore, the herpetofauna and lipotyphlans of Ambolokopatrika Forest were compared to those known from the forests of Anjanaharibe-Sud, Marojejy, and Tsararano massifs, the latter site being a southern extension of the Anjanaharibe-Sud chain. These animals were surveyed by use of opportunistic searching and pitfall trapping during two seasonal periods, May–June (winter), and November–December (summer). Forty-two species of amphibians, 23 of reptiles, and nine of lipotyphlans were recorded at Ambolokopatrika Forest. The biodiversity of Ambolokopatrika is comparable to those of other analysed sites, and this stresses the value of this forest in assuring biotic exchange between Anjanaharibe-Sud and Marojejy reserves. Considerations are also provided on the faunal similarities and differences in terms of exclusivity and endemisms. It is therefore suggested that a certain degree of protection should be given to Ambolokopatrika rainforest, to assure a biological connection and exchange between the protected areas of Anjanaharibe-Sud and Marojejy.     

Genetic subdivision, glacial refugia and postglacial recolonization in the golden-striped salamander, Chioglossa lusitanica (Amphibia: urodela)

The golden-striped salamander (Chioglossa lusitanica) is an ecologically specialized species, endemic to north-western Iberia. Patterns of genetic variation were assessed at seven polymorphic enzyme loci and one mitochondrial DNA (mtDNA) marker (cytochrome b) in 17 populations across its range. Estimates of enzyme genetic diversity revealed a high degree of genetic subdivision (FST = 0.68), mainly attributable to the existence of two groups of populations. The groups were located, respectively, north and south of the Mondego River, indicating that this river coincided with a major historical barrier to gene flow. A significant decrease in genetic variability from the Mondego northwards was associated with the Douro and Minho rivers. mtDNA sequence variation revealed a congruent pattern of two haplotype groups (d = 2.2%), with a geographical distribution resembling that of allozymes. The pattern and depth of genetic variation is consistent with the following hypotheses: (i) subdivision of an ancestral range of the species prior to the middle Pleistocene; (ii) secondary contact between populations representing historical refugia; (iii) relatively recent range expansion giving rise to the northern part of the species range; and (iv) loss of genetic variation through founder effects during range expansion across major rivers.     

Blood-gas,acid-base and catecholamine responses to lactic acid infusion in larval and adult Ambystoma tigrinum

Larval and adult Ambystoma tigrinum were subjected to acidosis by infusing lactic acid (2 M·g^-1) into the peritoneal cavity. Blood was sampled at intervals to establish the time-course of the ensuing acidosis. Both larvae and adults became significantly acidotic after 1 h. The larval acidosis was more pronounced (-4 pH units versus-2 pH units) than adults due to greater extracellular buffering capacity (higher [HCO₃ ^-]) in adults. Both forms recovered in about 8 h. Larvae showed a typical increase in circulating norepinephrine during the initial stages of the acidosis; adults did not, having significantly lower norepinephrine titer than larvae during the acidosis. Both larvae and adults showed transient increases in PO₂ during the acidosis. The ₁ and ₂ antagonists, timolol and butoxamine respectively, (0.2 g·g^-1) were administered to separate groups of larvae. Butoxamine (₂) delayed the recovery from the acidosis by prolonging the increase in arterial PCO₂ and reversing the recovery of [HCO₃ ^-]. Timolol (₁) did not delay recovery. We conclude that ₂ receptors are involved in the catecholamine responses to acidosis in larvae. Catecholamines appear not to play the same role in adult acid-base disturbances as they seem to in larvae.Abbreviations RBC red blood cell