Character Displacement in Frogs

Additional evidence has accumulated over the past 20 years regardingthe significance of the anuran mating call and regarding reinforcementof this premating isolating mechanism when hybridization occursin zones of symnatry. No two sympatric species have the samecall. That the mating call is a home-specific mate attractanthas been demonstrated in five major anuran families. Discriminationagainst hybrid calls versus homospecific calls has been demonstratedin Bufo and Hyla. Principal examples of presumed reinforcementcontinue to be in Gastrophryne, Acris, and the Australian Hylaewingi group. Reinforcement may be the basis for striking differentiationof some populations of Scaphiopus bombifrons. Sympatric speciespairs of anurans show greater differentiation in call than doallopatric pairs in the same group. One hybridizing situationin Bufo appears to have changed over 30 years to discrete populationswithout intermediate calls. Effective call differences can evolvewithout sympatry, and there are sympatric situations in whichno reinforcement is evident.     

Opsin shift in an aldolase antibody.

An antibody-retinal assembly that mimics the opsin shift (OS) of the naturally occurring visual pigments is reported. Both experiments and calculations show that the aldolase antibody 33F12 covalently binds all-trans retinal via a protonated Schiff base with a lysine residue. This chromophore, which exhibits a remarkable opsin red shift (140 nm), represents a useful model system for studying the factors that contribute to the OS.     

Opsin oligomerization in a heterologous cell system

Using bioluminescence resonance energy transfer (BRET) we studied opsin oligomerization in heterologous expression systems and quantitatively assessed its oligomerization state. BRET2 saturation and competition experiments were performed with live COS-7 cells expressing Rluc-and GFP2-tagged receptor constructs. BRET2 saturation curves obtained were hyperbolic, and the calculated oligomerization state (N = 1 for dimers) suggested that opsin (N = 1.34 +/- 0.25) forms higher oligomers. Very high BRET2 values obtained for the opsin homo-dimer pair indicated a large energy transfer efficiency (E) and for cases where E > 0.1 a modified saturation curve was proposed. The existence of homo-dimer complexes was additionally supported by competition assay results and was also observed in HEK-293 cells. Furthermore, evidence was provided for homo-and hetero-dimerization of family A (beta2-adrenergic) and B (gastric inhibitory polypeptide, GIP) receptors. In summary, these experiments demonstrate homo-and hetero-dimerization for opsin, beta 2-adrenergic, and GIP receptors.     

Opsin Oligomerization in a Heterologous Cell System

Using bioluminescence resonance energy transfer (BRET) we studied opsin oligomerization in heterologous expression systems and quantitatively assessed its oligomerization state. BRET² saturation and competition experiments were performed with live COS-7 cells expressing Rluc-and GFP²-tagged receptor constructs. BRET² saturation curves obtained were hyperbolic, and the calculated oligomerization state (N = 1 for dimers) suggested that opsin (N = 1.34 ± 0.25) forms higher oligomers. Very high BRET² values obtained for the opsin homo-dimer pair indicated a large energy transfer efficiency (E) and for cases where E ? 0.1 a modified saturation curve was proposed. The existence of homo-dimer complexes was additionally supported by competition assay results and was also observed in HEK-293 cells. Furthermore, evidence was provided for homo-and hetero-dimerization of family A (β₂-adrenergic) and B (gastric inhibitory polypeptide, GIP) receptors. In summary, these experiments demonstrate homo-and hetero-dimerization for opsin, β ₂-adrenergic, and GIP receptors.     

Linkage of Retinal to Opsin

RHODOPSIN, the visual pigment of vertebrate rods, has been shown to consist of a chromophore (11-cis retinal) bound to a protein (opsin)^1–2. It has been proposed that the linkage is a Schiff base between phosphatidyl ethanolaniine (PE) and retinal and that when exposed to light, the retinal migrates from PE to the ε-amino-group of a lysine residue in opsin^3–7. Most of the support for this theory comes from the observation that N-retinylidenephosphatidylethanolamine (N-RPE) can be extracted in the dark from rod outer segments (ROS)^3,4. Furthermore, N-retinylphosphatidylethanolamine (N-RH₂PE) has been extracted from ROS preparations after treating the visual pigment with acid and NaBH₄—conditions which are assumed fix retinal to its “native” binding site through a secondary amine linkage⁷. All these studies, however, were carried out on crude extracts of ROS in various detergents. These crude extracts contain large amounts of phospholipid and retinal which is not bound to opsin. Thus, the isolation of N-RPE from crude ROS extracts does not necessarily point to its involvement in the binding of retinal to opsin. In contradiction to these reports are findings that purified visual pigment contains no phospholipid^9,10 and that the molar concentration of N-RPE in bovine ROS is less than that of rhodopsin¹¹. We have taken advantage of the observation that visual pigment in the outer segment disks is continually being renewed¹² to label the rhodopsin with ³H-retinal and to show in yet another way that N-RPE does not exist in purified visual pigment.     

Opsin is a phospholipid flippase

Polar lipids must flip-flop rapidly across biological membranes to sustain cellular life [1, 2], but flipping is energetically costly [3] and its intrinsic rate is low. To overcome this problem, cells have membrane proteins that function as lipid transporters (flippases) to accelerate flipping to a physiologically relevant rate. Flippases that operate at the plasma membrane of eukaryotes, coupling ATP hydrolysis to unidirectional lipid flipping, have been defined at a molecular level [2]. On the other hand, ATP-independent bidirectional flippases that translocate lipids in biogenic compartments, e.g., the endoplasmic reticulum, and specialized membranes, e.g., photoreceptor discs [4, 5], have not been identified even though their activity has been recognized for more than 30 years [1]. Here, we demonstrate that opsin is the ATP-independent phospholipid flippase of photoreceptor discs. We show that reconstitution of opsin into large unilamellar vesicles promotes rapid (τ<10 s) flipping of phospholipid probes across the vesicle membrane. This is the first molecular identification of an ATP-independent phospholipid flippase in any system. It reveals an unexpected activity for opsin and, in conjunction with recently available structural information on this G protein-coupled receptor [6, 7], significantly advances our understanding of the mechanism of ATP-independent lipid flip-flop.     

Opsin gene duplication and divergence in ray-finned fish

Opsin gene sequences were first reported in the 1980s. The goal of that research was to test the hypothesis that human opsins were members of a single gene family and that variation in human color vision was mediated by mutations in these genes. While the new data supported both hypotheses, the greatest contribution of this work was, arguably, that it provided the data necessary for PCR-based surveys in a diversity of other species. Such studies, and recent whole genome sequencing projects, have uncovered exceptionally large opsin gene repertoires in ray-finned fishes (taxon, Actinopterygii). Guppies and zebrafish, for example, have 10 visual opsin genes each. Here we review the duplication and divergence events that have generated these gene collections. Phylogenetic analyses revealed that large opsin gene repertories in fish have been generated by gene duplication and divergence events that span the age of the ray-finned fishes. Data from whole genome sequencing projects and from large-insert clones show that tandem duplication is the primary mode of opsin gene family expansion in fishes. In some instances gene conversion between tandem duplicates has obscured evolutionary relationships among genes and generated unique key-site haplotypes. We mapped amino acid substitutions at so-called key-sites onto phylogenies and this exposed many examples of convergence. We found that dN/dS values were higher on the branches of our trees that followed gene duplication than on branches that followed speciation events, suggesting that duplication relaxes constraints on opsin sequence evolution. Though the focus of the review is opsin sequence evolution, we also note that there are few clear connections between opsin gene repertoires and variation in spectral environment, morphological traits, or life history traits.     

蛙类趋声行为的研究进展

蛙类是典型的依赖声音通讯的物种,具有季节性繁殖特点。繁殖季节雄性大多通过广告鸣叫竞争领地和吸引配偶,雌性通过雄性鸣声来评价个体特征及其所占有的资源,并以趋声行为完成配偶选择。趋声过程的行为学参数,通常用于检验动物对声音信号的偏爱和行为响应的特征。本文通过对蛙类趋声行为相关研究的回顾,总结其产生机制、评价方式及影响因素,并结合当前发展趋势对蛙类趋声行为未来的研究方向进行了展望。     

Ultrastructure of Cytotoxic Cells in Frogs

Our previous work gave evidence that leopard frogs ( Rana pipiens ) possess inducible killer (IK) cells (CTL-like) and spontaneous killer (SK) cells (NK-like) that can destroy allogeneic RBC. To further characterize these cytotoxic cells we have examined their ultrastructural morphology and effector cell-target cell membrane contacts. We also studied the sensitivity of these two effector cells to low temperature (4°C). Our results showed that SK effector cells are lymphocytes (10 μm) which contain numerous vacuoles and osmiophilic granules, while IK effectors are another lymphocyte population (up to 12 μm) with a small amount of cytoplasm and few mitochondria. Both IK- and SK-mediated lytic responses were suppressed significantly by low temperature. These observations allow us to propose that frog SK and IK cells may be homologous to mammalian NK and CTL.     

林蛙在中国的分布

作者收集有关我国林蛙分布的资料绘制了林蛙分布图,其中图1标明了主要分布于中国北方的中国林蛙种绀（Rana chensinensis species-group）和黑龙江林蛙种组（R．amurensis species-group）的分布,图2标明出了在亲缘关系上与日本林蛙很相近（R．jaonica）的分布于南方的长肢林蛙种组（R．longicrus species-group）的分布,图3重点标明了环渤海区域已知6种林蛙的分布。最后,作者提出环渤海区域是林蛙重要的分布中心,并对镇海林蛙（R．zhenhaiensis）和中国林蛙（R．chensinensis）的一些可疑或有争议的分布问题进行了简要讨论。     

Opsin Genes and Visual Ecology in a Nocturnal Folivorous Lemur

Primate color vision has traditionally been examined in the context of diurnal activity, but recent genetic and ecological studies suggest that color vision plays a role in nocturnal primate behavior and ecology as well. In this study, we united molecular analyses of cone visual pigment (opsin) genes with visual modeling analyses of food items to explore the evolution of color vision in the folivorous woolly lemur (genus Avahi). Previous studies have shown that leaf quality, e.g., protein content, leaf toughness, and protein/toughness ratio, is significantly correlated with green-red and blue-yellow chromatic differences, suggesting a potential role of color in leaf discrimination in Avahi, and, consequently, a potential adaptive advantage to color vision in this taxon. Phylogenetic selection tests determined that the strength of selection on the SWS1 opsin gene to retain blue-sensitive SWS cones did not significantly differ in Avahi compared to day-active primates. Genotyping of the M/LWS opsin gene in 60 individuals from nine species found that the 558-nm-sensitive (red-sensitive) allele is conserved across all Avahi. Finally, we measured spectral reflectance from five species of young leaves consumed by Avahi and background foliage in Ranomafana National Park and modeled performance of possible S and M/L pigment pairs for detecting these food items under different nocturnal illuminations (e.g. twilight, moonlight). We found that the observed cone pigment pair in Avahi was optimally tuned for color-based detection of young green leaves in all nocturnal light environments, suggesting a potential adaptive role of nocturnal color vision in selection for dichromacy in this genus.     

Opsin gene polymorphism predicts trichromacy in a cathemeral lemur

Recent research has identified polymorphic trichromacy in three diurnal strepsirrhines: Coquerel's sifaka (Propithecus coquereli), black and white ruffed lemurs (Varecia variegata), and red ruffed lemurs (V. rubra). Current hypotheses suggest that the transitions to diurnality experienced by Propithecus and Varecia were necessary precursors to their independent acquisitions of trichromacy. Accordingly, cathemeral lemurs are thought to lack the M/L opsin gene polymorphism necessary for trichromacy. In this study, the M/L opsin gene was sequenced in ten cathemeral blue-eyed black lemurs (Eulemur macaco flavifrons). This analysis identified a polymorphism identical to that of other trichromatic strepsirrhines at the critical amino acid position 285 in exon 5 of the M/L opsin gene. Thus, polymorphic trichromacy is likely present in at least one cathemeral Eulemur species, suggesting that strict diurnality is not necessary for trichromacy. The presence of trichromacy in E. m. flavifrons suggests that a re-evaluation of current hypotheses regarding the evolution of strepsirrhine trichromacy may be necessary. Although the M/L opsin polymorphism may have been independently acquired three times in the lemurid-indriid clade, the distribution of opsin alleles in lemurids and indriids may also be consistent with a common origin of trichromacy in the last common ancestor of either the lemurids or the lemurid-indriid clade.     

Opsin expression predicts male nuptial color in threespine stickleback

Theoretical models of sexual selection suggest that male courtship signals can evolve through the build‐up of genetic correlations between the male signal and female preference. When preference is mediated via increased sensitivity of the signal characteristics, correlations between male signal and perception/sensitivity are expected. When signal expression is limited to males, we would expect to find signal‐sensitivity correlations in males. Here, we document such a correlation within a breeding population of threespine stickleback mediated by differences in opsin expression. Males with redder nuptial coloration express more long‐wavelength‐sensitive (LWS) opsin, making them more sensitive to orange and red. This correlation is not an artifact of shared tuning to the optical microhabitat. Such correlations are an essential feature of many models of sexual selection, and our results highlight the potential importance of opsin expression variation as a substrate for signal‐preference evolution. Finally, these results suggest a potential sensory mechanism that could drive negative frequency‐dependent selection via male–male competition and thus maintain variation in male nuptial color.     

Relative Rates of Nucleotide Substitution in Frogs

Accurate estimation of relative mutation rates of mitochondrial DNA (mtDNA) and single-copy nuclear DNA (scnDNA) within lineages contributes to a general understanding of molecular evolutionary processes and facilitates making demographic inferences from population genetic data. The rate of divergence at synonymous sites (K_s) may be used as a surrogate for mutation rate. Such data are available for few organisms and no amphibians. Relative to mammals and birds, amphibian mtDNA is thought to evolve slowly, and the K_s ratio of mtDNA to scnDNA would be expected to be low as well. Relative K_s was estimated from a mitochondrial gene, ND2, and a nuclear gene, c-myc, using both approximate and likelihood methods. Three lineages of congeneric frogs were studied and this ratio was found to be approximately 16, the highest of previously reported ratios. No evidence of a low K_s in the nuclear gene was found: c-myc codon usage was not biased, the K_s was double the intron divergence rate, and the absolute K_s was similar to estimates obtained here for other genes from other frog species. A high K_s in mitochondrial vs. nuclear genes was unexpected in light of previous reports of a slow rate of mtDNA evolution in amphibians. These results highlight the need for further investigation of the effects of life history on mutation rates. Current address (Andrew J. Crawford): Smithsonian Tropical Research Institute, Apartado 2072, Balboa, Ancon, Republic of Panama     

Survivorship in Wild Frogs Infected with Chytridiomycosis

Chytridiomycosis is an emerging infectious disease that has been implicated as the causative agent of many recent amphibian population declines and extinctions that have taken place in relatively pristine locations worldwide. While there exists a growing body of literature regarding the effect of the fungus on experimentally infected frogs, few studies have examined the effect of the fungus on apparently healthy wild frogs from nondeclining, infected populations. We examined the temporal pattern of chytrid infection in individually marked Stony Creek Frogs (Litoria wilcoxii) at a lowland site in southeast Queensland, Australia. We provide the first evidence that wild frogs are capable of both acquiring chytridiomycosis as adults, and also of clearing their infections entirely. Changes in disease status in individual frogs largely tracked changing climatic conditions, with infections tending to appear in cooler months and disappearing in warmer months. Though 27.2% of the adult frogs we sampled were infected at some point in the study, we found no evidence that chytridiomycosis was negatively affecting adult survivorship, suggesting either: (1) chytrid-induced mortality in this population is generally restricted to metamorphs and juveniles; (2) this population was not exposed to conditions which favored lethal disease outbreaks; or (3) this population has evolved sufficient resistance to the disease to persist relatively unaffected.     

蛙类声音通讯研究进展

声音通讯包含鸣声的产生、传播及对鸣声的感知与行为响应。对大多数无尾两栖类而言,雄性个体间的竞争(即雄雄竞争)、雌性配偶识别与选择几乎完全依赖声音通讯,因此准确及时的声音信息传递与接收对蛙类的生存和繁殖起着决定性作用。本文总结了蛙类鸣叫特征及其产生机制,归纳了声音通讯在蛙类性选择中的功能及协同进化,探讨了鸣声感知的神经机制及声音通讯的内分泌机制。最后对蛙类声音通讯研究方向进行了展望,并提出了可能的解决方案。     

青草池塘处处蛙

海南岛气候温暖潮湿,是两栖动物和爬行动物的最佳栖息环境。蛙和蛇是这里两个很重要的类群。有关海南脊椎动物鸟和兽的资料还能找到一些,而有关两栖爬行动物的却非常少。为此,我专门请教了上海复旦大学动物学教授黄正一先生。黄先生告诉我,海南已经记录到39种两栖类和104种爬行动物。其中海南疣螈、细刺蛙、脆皮蛙、海南湍蛙、小湍蛙,海南溪树蛙、鳞皮厚蹼蟾、眼斑小树蛙、海南拟髭蟾等9种两栖类和海南蛇蜥、海南脊蛇、粉链蛇和费氏花龟等4种爬行动物为海南岛独有。海南蛇的种类极多,蟒蛇、眼镜蛇、眼镜王蛇、金环蛇、水蛇、滑鼠蛇等大量栖息在海南岛中西部山区的热带丛林中。近年海南岛的两栖爬行动物新种和新     

Comparison of Gastrulation in Frogs and Fish

SYNOPSIS. Comparative embryological studies of frogs and fishprovide valuable information about the mechanisms and evolutionof vertebrate development. First, by mapping developmental datafrom a range of species onto a cladogram, one can distinguishgeneral features of a ground plan from variation within it.Two studies illustrate this: comparison of gastrulation mechanismsin sturgeon and Xenopus, and morphogenesis of the dorsal mesodermin five species of anurans. Second, phylogenetic analysis ofdevelopmental data makes it possible to identify radical departuresfrom the ground plan among related groups. Teleost gastrulationis a highly derived process that appears to have little in commonwith the ancestral version. However, teleost gastrulation mayhave evolved as a result of two specific developmental changes:loss of bottle cells in the surface layer, and changes in theyolk. The phylogenetic distribution of developmental charactersforms the basis for mechanistic hypotheses about the originsof major evolutionary changes in development