Genetic variation and differentiation in the Japanese five-lined skink,Eumeces latiscutatus (Reptilia: Squamata)

The genetic variation in Eumeces latiscutatus from the main islands of Japan and the northern Ryukyus was investigated and compared with that of its close relatives (E. okadae and E. marginatus) using allozyme data. All three samples from the northern Tokara Island Group, currently identified as E. latiscutatus, were shown to belong to E. marginatus and not to E. latiscutatus. The non-monophyly of the northern Tokara samples and the great genetic differences within these samples may have resulted from colonization from more than one source population by northeastward overseas dispersal. The samples from the Izu Peninsula were genetically much closer to E. okadae than the other conspecific ones. This indicates that the samples from the Izu Peninsula and the other samples of E. latiscutatus should be treated as distinct species, and that E. latiscutatus from the Izu Peninsula and E. okadae from the Izu Island Group may be treated as conspecific. Samples from western Japan were genetically well differentiated from those of eastern Japan. Within the western group, the samples from the Osumi Island Group was genetically distinct from those from the other regions, by possessing unique alleles. Our phenograms also reveal a distant affinity between samples from the Danjo Island Group and the main islands of Japan. This may be the result of long geographic isolation of the Osumi and Danjo Island Groups from Kyushu. By contrast, samples from Sapporo and Aomori were poorly differentiated genetically in spite of the long separation of these two localities by the Tsugaru Strait. This suggests that overseas dispersal of E. latiscutatus occurred across this strait after its formation.     

Ontogeny of the extraembryonic membranes of the oviparous lizard, Eumeces fasciatus (Squamata: scincidae)

Oviposited eggs of Eumeces fasciatus contain embryos in the limb bud stage. Amniogenesis is complete and two yolk sac membranes, vascular trilaminar omphalopleure (choriovitelline membrane) and bilaminar omphalopleure, enclose the yolk vesicle. A small allantoic vesicle contacts the chorion. The choriovitelline membrane is the primary vascular system. Blood islands, sites of hematopoiesis, are associated with omphalomesenteric vessels of the choriovitelline membrane. The bilaminar omphalopleure, which contacts the eggshell over the abembryonic hemisphere of the egg, lies external to an isolated yolk mass and yolk cleft and is not vascularized. The definitive yolk sac (splanchnopleure) is formed when the extraembryonic coelom and allantoic vesicle intrude into the choriovitelline membrane. Omphalomesenteric vessels are retained with the yolk sac splanchnopleure and the associated hematopoietic sites are present throughout incubation. The chorioallantoic membrane reaches the equator of the egg, entirely supplanting the choriovitelline membrane, after 25% of incubation is completed. Further growth of the allantois is stalled until 65% of incubation is completed when rapid expansion of the allantoic vesicle, in conjunction with resorption of the isolated yolk mass, supplants the bilaminar omphalopleure. As a result, the chorioallantoic membrane completely envelopes the egg for the final 35% of incubation. This developmental event is coincident with published reports for the timing of increased growth and metabolism of embryos. As the isolated yolk mass regresses, intravitelline cells associated with the yolk cleft invade and resorb the yolk to form a large cavity. The wall of this cavity is a germinal epithelium that produces cells that fill the cavity. This structure appears to be a site of hematopoiesis previously undescribed in vertebrates.     

Ultrastructural studies of normal and alloxan-treated islet cells of the pancreas of the lizard, Eumeces fasciatus

Alpha and beta cells can be distinguished by differences in mitochondrial and secretion granule structure. Many mitochondria of alpha cells possess “tubular” or prismatic cristae oriented longitudinally and having triangular profiles in cross-section. The matrix is particulate, with the roughly spherical particles measuring about 100 A in diameter. Evidence is presented indicating that alpha and beta granules are sequestrated in association with Golgi elements. Fully-condensed beta granules, assumed to be insulin, appear homogeneously dense and crystalline. Recurrent profiles of crystalline beta granules suggest that they possess an octahedral configuration. Alpha cell granules also appear homogeneously dense but have round profiles. Many acinar cell nuclei in both normal and alloxan-treated pancreata display masses of moderately dense fibrils oriented roughly parallel to each other. These fibrils are about 200 A across and their terminal portions are rebranched and often appear to be continuous with the finely granular or filamentous component of the nucleoplasm. Not infrequently the fibrils show evidence of periodicity. Alloxan has a specific destructive effect on beta cells. An initial effect seems to be the disruption and coalescence of the bounding membranes of beta granules. Lysosome-like bodies are often seen in beta cell cytoplasm, which ultimately becomes degranulated and necrotic following prolonged administration of large doses of alloxan.     

中国石龙子成熟精子的超微结构

利用透射电镜观察中国石龙子附睾成熟精子的超微结构。顶体囊前部扁平、由皮质和髓质组成 ,穿孔器中度倾斜、顶端尖 ,穿孔器基板塞子状 ,细胞核长形 ,核内小管缺 ,核前电子透亮区小 ,核肩圆 ,核陷窝锥形。颈段具片层结构 ,近端中心粒和远端中心粒的长轴呈直角 ,9束外周致密纤维与远端中心粒相应的 9束三联微管相联 ,向后与轴丝相应的 9束双联微管相联 ,中央纤维与 2个中央单微管相联。中段短 ,多层膜结构缺 ,含有线状嵴的柱状线粒体 ,不规则卵状致密体组成不连续的环状结构 ,纤维鞘伸入中段 ,具终环。线粒体与环状结构的模式为 :rs1 /mi1 ,rs2 /mi2 ,rs3/mi3,rs4 /mi4。主段前面部分具薄的细胞质颗粒区。纤维 3和 8至主段前端消失。轴丝呈“9 2”型。中国石龙子精子超微结构具有塞子状的穿孔器基板、致密体形成不连续的环状结构和纤维鞘始于ms2等特征与巨石龙子群和蜓蜥 -胎生群不同。没有发现石龙子科精子的独征     

Karyotype of the lizard Eumeces taeniolatus]

Using bone marrow cell preparations, the karyotype of Eumeces taeniolatus Blyth was examined. The diploid chromosome set of E. taeniolatus is found equal to 28 (24 meta- and 4 submetacentrics), NF=56. Sex chromosomes do not differ morphologically.     

Mitochondrial phylogeography of the Dalmatian wall lizard, Podarcis melisellensis (Lacertidae)

A 903 bp section of the mitochondrial cytochrome b gene was sequenced from 73 specimens of Podarcis melisellensis collected at 52 localities distributed over the major part of the species’ range. In addition, parts of the 12S (about 470 bp) and 16S rRNA (about 500 bp) genes were analysed for 11 representative samples leading to a congruent phylogeny. Our study includes representatives of all 20 subspecies recognized today. The phylogenetic analysis of the sequence data revealed three main clades: mainland with nearby islands, Vis archipelago, and Lastovo archipelago. The degree of mitochondrial DNA divergence among these clades suggests a separation of the respective population groups during the earliest Pleistocene. The phylogenetic pattern observed within the species is in sharp contrast to the actual taxonomic division into subspecies. A correlation between genetic diversity of P. melisellensis populations and paleogeography of the regions they inhabit is discussed.     

Cladogenesis and phylogeography of the lizard Phrynocephalus vlangalii (Agamidae) on the Tibetan plateau

Phrynocephalus vlangalii is restricted to dry sand or Gobi desert highlands between major mountain ranges in the Qinghai (Tibetan) Plateau. Mitochondrial DNA (mtDNA) sequence (partial ND2, tRNA(Trp) and partial tRNA(Ala)) was obtained from 293 Phrynocephalus sampled from 34 sites across the plateau. Partitioned Bayesian and maximum parsimony phylogenetic analyses revealed that P. vlangalii and two other proposed species (P. erythrus and P. putjatia) together form a monophyletic mtDNA clade which, in contrast with previous studies, does not include P. theobaldi and P. zetangensis. The main P. vlangalli clade comprises seven well-supported lineages that correspond to distinct geographical areas with little or no overlap, and share a most recent common ancestor at 5.06 +/- 0.68 million years ago (mya). This is much older than intraspecific lineages in other Tibetan animal groups. Analyses of molecular variance indicated that most of the observed genetic variation occurred among populations/regions implying long-term interruption of maternal gene flow. A combined approach based on tests of population expansion, estimation of node dates, and significance tests on clade areas indicated that phylogeographical structuring has been primarily shaped by three main periods of plateau uplift during the Pliocene and Pleistocene, specifically 3.4 mya, 2.5 mya and 1.7 mya. These periods corresponded to the appearance of several mountain ranges that formed physical barriers between lineages. Populations from the Qaidam Basin are shown to have undergone major demographic and range expansions in the early Pleistocene, consistent with colonization of areas previously covered by the huge Qaidam palaeolake, which desiccated at this time. The study represents one of the most detailed phylogeographical analyses of the Qinghai Plateau to date and shows how geological events have shaped current patterns of diversity.     

Induction of sexual receptivity in the female broad-headed skink, Eumeces laticeps, by estradiol-17 beta

Sexual receptivity in the female scincid lizard Eumeces laticeps occurs naturally only during the spring breeding season, which is also when maximal follicular development occurs. The presumption that high estrogen levels are coincidentally present and the need for a reliable method of inducing sexual receptivity for behavioral studies prompted tests of the hypothesis that estrogen induces sexual behavior. A series of experiments established that estradiol-17 beta induces sexual behavior. A series of experiments established that estradiol-17 beta induces sexual receptivity within 4 days when injected every other day at 2.0 micrograms in 20 microliters peanut oil in intact or ovariectomized females. In behavioral tests conducted during August, all control females (intact or ovariectomized injected with vehicle only) rejected courtship whereas all females receiving estrogen copulated. Estrogen injections also induced a statistically significant change from rejection to receptivity within individuals. Initial attempts to implant estradiol-17 beta in Silastic tubes killed all females so treated.     

Deferred agonistic behavior in a long-lived scincid lizard Eumeces laticeps

Summary A laboratory experiment with the broad-headed skink (Eumeces laticeps) involving staged agonistic encounters demonstrates that larger males have an advantage over smaller ones in agonistic bouts. Field data on head wounds produced by intraspecific fighting during the breeding season show a much higher frequency of new wounds among males over 100 mm in snout-vent-length than in smaller males. The significant difference in new-wound frequency strongly suggests avoidance of fights by the small males, which is corroborated by laboratory and field observations. Access by males to reproductively active females depends on the ability to defeat other males in aggressive contests virtually always involving head biting if the males are of nearly equal size. Because the probability of winning agonistic encounters increases with size, young males avoid fights with older males. Aggressive contests with larger males and reproductive attempts other than courtship in the absence of larger males are deferred.Aggressive behavior in E. laticeps may be employed in direct defense of females, but might also be expressed in defense of specific sites and/or territories. In the laboratory, males in their home cages were significantly more likely to win encounters with males of similar size than were males fighting in the home cages of opponents. This suggests that encounter site could be important in determining encounter outcome and that field study of possible site defense or territoriality is needed.     

Influence of food type on specific dynamic action of the Chinese skink Eumeces chinensis

We used the Chinese skink (Eumeces chinensis) as an experimental model to study influence of food type on specific dynamic action (SDA) of feeding. Thirty-three adult males collected from a natural population were divided equally into three (one control and two experimental) groups. We starved all skinks at 30 degrees C for 3 days and then provided the experimental skinks with a single meal consisting of either mealworms or meat [the flesh of the bullfrog (Rana catesbeiana)]. Food ingested by skinks of the two experimental groups differed in lipid content and lean dry mass but not in total dry mass and energy. Defecation following feeding occurred slightly earlier in skinks ingesting mealworms (mean=41.7 h) than in those ingesting meat (mean=47.7 h), but the difference was not significant. Analyses of variance (ANOVAs) with repeated measures showed that temporal variation in oxygen consumption over 72 h after feeding was evident in the experimental skinks but not in the control ones. Oxygen consumption was higher in the experimental skinks than in the control ones during the time interval between 4.5 and 36 h after feeding. The peak metabolic rate was greater but occurred later in skinks ingesting meat than in those ingesting mealworms. The estimated amounts of oxygen consumed by mealworm-fed, meat-fed and unfed skinks at 30 degrees C over 72 h after feeding were 356.5, 393.8 and 295.2 mL, respectively. Our results provide a support for the previous prediction that SDA is affected by types of food ingested by animals as skinks ingesting mealworms and meat differed in the time to reach a peak metabolic rate, the level of the peak metabolic rate and the magnitude of the SDA effect.     

Tail loss affects fecundity but not offspring traits in the Chinese skink Eumeces chinensis

We used the Chinese skink Eumeces chinensis as a model animal to study the effects of tail loss on reproductive investment and offspring traits.A total of 147 wild-captured adult females were divided into four groups according to their tail conditions.Tail breaks occurred most frequently in the proximal portion of the tail and least frequently in the distal portion,with the middle portion in between.This finding suggests that tail breaks occurring in nature often entails substantial energetic costs in E.chinensis where the tail is a major site of energy storage.The proportion of females that laid eggs was higher in females with intact or completely regenerated tails than in those with broken tails.Following whole-tail autotomy,the clutch size was reduced by 17％,and the clutch mass was reduced by 14％.Females undergoing substantial tail autotomy reduced reproductive investment,and they did so by reducing the number but not the size of eggs produced.None of the egg and hatchling traits was affected by tail loss.Comparing our data with those reported for other oviparous and viviparous skinks allows us to draw two general conclusions:one is that fecundity (clutch or litter size) is affected by tail loss in all species so far studied,whereas offspring size is affected by tail loss in some species,but not in others; the other is that the reduction in fecundity following tail loss is more evident in species lacking abdominal fat bodies [Current Zoology 58 (2):228-235,2012].     

Demography of an island population of the lizard,Eumeces okadae,on Miyake-jima,Izu Islands

The demography of the lizard Eumeces okadae was studied for 4 years on Miyake-jima, in the Izu Islands, central Japan. Most males matured at 3 years of age with a mean snout-vent length of 77.5 mm. Females matured at 3–4 years with a mean SVL of 77.8 mm, and reproduced biennially thereafter. Growth continued after maturity with a decreasing rate. In females, annual growth was negligible in reproductive years, but resumed markedly in non-reproductive years. Clutch size increased with SVL and was used to construct a fecundity schedule. Population density (exclusive of hatchlings) was estimated to be ca. 4000 individuals/ha in August, an exceptionally high density for lizards. Before introduction of the weasel onto Miyake-jima, low predation pressure had allowed E. okadae to maintain a high population density. Estimated annual survival rates were 36% for hatchlings, 56% for yearlings, 80% for 2-year-olds, 63% for adult males and 76% for adult females. The cohort generation time (6.95 years) was so long that the E. okadae population can be characterized by a combination of low natality and slow turnover.     

Glacial refugia in a maritime temperate climate: Cicada (Kikihia subalpina) mtDNA phylogeography in New Zealand

Understanding the biological significance of Pleistocene glaciations requires knowledge of the nature and extent of habitat refugia during glacial maxima. An opportunity to examine evidence of glacial forest refugia in a maritime, Southern Hemisphere setting is found in New Zealand, where the extent of Pleistocene forests remains controversial. We used the mitochondrial phylogeography of a forest-edge cicada ( Kikihia subalpina ) to test the hypothesis that populations of this species survived throughout South Island during the Last Glacial Maximum. We also compared mitochondrial DNA phylogeographic patterns with male song patterns that suggest allopatric divergence across Cook Strait. Cytochrome oxidase I and II sequences were analyzed using network analysis, maximum-likelihood phylogenetic estimation, Bayesian dating and Bayesian skyline plots. K. subalpina haplotypes from North Island and South Island form monophyletic clades that are concordant with song patterns. Song divergence corresponds to approximately 2% genetic divergence, and Bayesian dating suggests that the North Island and South Island population-lineages became isolated around 761 000 years bp . Almost all South Island genetic variation is found in the north of the island, consistent with refugia in Marlborough Sounds, central Nelson and northwest Nelson. All central and southern South Island and Stewart Island haplotypes are extremely similar to northern South Island haplotypes, a 'northern richness/southern purity' pattern that mirrors genetic patterns observed in many Northern Hemisphere taxa. Proposed southern South Island forest habitat fragments may have been too small to sustain populations of K. subalpina , and/or they may have harboured ecological communities with no modern-day analogues.     

A SINE of restricted gene flow across the Alpine Fault: phylogeography of the New Zealand common skink (Oligosoma nigriplantare polychroma)

New Zealand has experienced a complex climatic and geological history since the Pliocene. Thus, identifying the processes most important in having driven the evolution of New Zealand's biota has proven difficult. Here we examine the phylogeography of the New Zealand common skink ( Oligosoma nigriplantare polychroma ) which is distributed throughout much of New Zealand and crosses many putative biogeographical boundaries. Using mitochondrial DNA sequence data, we revealed five geographically distinct lineages that are highly differentiated (pairwise Φ_ST 0.54–0.80). The phylogeographical pattern and inferred age of the lineages suggests Pliocene mountain building along active fault lines promoted their divergence 3.98–5.45 million years ago. A short interspersed nuclear element (SINE) polymorphism in the myosin gene intron ( MYH-2 ) confirmed a pattern of restricted gene flow between lineages on either side of the mountain ranges associated with the Alpine Fault that runs southwest to northeast across the South Island of New Zealand. An analysis of molecular variance confirmed that ~40% of the genetic differentiation in O. n. polychroma is distributed across this major fault line. The straits between the main islands of New Zealand accounted for much less of the variation found within O. n. polychroma , most likely due to the repeated existence of landbridges between islands during periods of the Pleistocene that allowed migration. Overall, our findings reveal the relative roles of different climatic and geological processes, and in particular, demonstrate the importance of the Alpine Fault in the evolution of New Zealand's biota.     

Molecular phylogeny of the Greek legless skink Ophiomorus punctatissimus (Squamata: Scincidae): the impact of the mid-Aegean trench in its phylogeography

Sequence data derived from three mitochondrial markers (cytochrome b, 16S rRNA and 12S rRNA genes) were used to infer the evolutionary history of several insular and mainland populations of the Greek legless skink (Ophiomorus punctatissimus), covering most of its distributional range. All phylogenetic analyses produced topologically identical trees that revealed a well-resolved phylogeny. These trees support two O. punctatissimus clades, which are geographically separated (west and east of the mid-Aegean trench). The assumption of a clock-like evolution could not be rejected, and thus a local clock was calibrated for the O. punctatissimus lineages. The non-overlapping geographic distributions of the major clades suggest a spatial and temporal sequence of diversification that coincides with paleogeographic separations during the geological history of the Aegean region. It seems that O. punctatissimus is an old eastern Mediterranean species that has been differentiating in this region at least from middle Miocene. It is possible that the ancestral form of O. punctatissimus invaded the Aegean region from Anatolia before the complete formation of the mid-Aegean trench, when the Aegean was still a uniform landmass, while other vicariant events have led to its present distribution.     

Comparative phylogeography of three skink species (Oligosoma moco, O. smithi, O. suteri; Reptilia: Scincidae) in northeastern New Zealand

Sea-level fluctuations during the Pliocene and Pleistocene have shaped the landscape of the Northland region of New Zealand. We examined the comparative phylogeography of three skink species (Oligosoma moco, O. smithi, O. suteri) in northeastern New Zealand in order to investigate the impact of the historical processes that have prevailed since the Pliocene on the Northland fauna. O. moco, O. smithi and O. suteri have similar distributions across northeastern New Zealand, frequently occurring in sympatry. We obtained sequence data from across the entire range of each species, targeting the ND2 mitochondrial gene. Using Neighbor-Joining, Maximum likelihood and Bayesian methods, our analysis revealed contrasting phylogeographic patterns in each species. We found substantial phylogeographic structure within O. moco, with three distinct clades identified. Similarly, deep phylogeographic divergence was evident within O. smithi, with three distinct clades present. Clade 1 included O. smithi populations from the Three Kings Islands and the western coastline of Northland, while Clade 2 encompassed the remainder of the range. However, since Clade 3 corresponded to a described species (O. microlepis), O. smithi might represent a species complex. In both O. moco and O. smithi, divergences among clades are estimated to have occurred in the Pliocene, with divergences within clades occurring during the Pleistocene. In contrast, genetic divergence among O. suteri populations was extremely limited and indicative of more recent divergences during the Pleistocene. The lack of phylogeographic structure in O. suteri might be a consequence of its oviparous reproductive mode, which restricted its distribution to warm northern refugia during glacial maxima. Differences in the ecology and biology of each species might have produced contrasting responses to the same historical processes, and ultimately diverse phylogeographic patterns. Our study reveals an absence of consistent and concordant phylogeographic patterns in the Northland biota, even within the same taxonomic group.