Validation and Application of Skeletochronology for Age Determination of the Ryukyu Ground Gecko,Goniurosaurus kuroiwae （Squamata： Eublepharidae）

Skeletochronology is a method commonly used for estimating the age of amphibians and reptiles in the wild. However, the number of lines of arrested growth （LAGs） does not necessarily reflect age in some species. We validated the applicability of this method to an endangered eublepharid gecko, Goniurosaurus kuroiwae, then inferred its longevity and age structures in wild populations. We classified young geckos into three groups using previously published data for early growth： Group 1 contained hatchlings before the first winter, Group 2 contained hatchlings after the first win- ter, and Group 3 included yearlings after the second winter. LAG numbers in these groups were then compared. All individuals in Group 1 possessed a single LAG, which was considered as a hatching line. Most individuals in Groups 2 and 3 possessed one and two additional LAGs, respectively （LAG1 and LAG2）, corroborating the notion that LAGs are formed annually. A few geckos exhibited fewer LAGs than expected. Analysis of variations in LAG and marrow cavity diameter demonstrated that in animals with fewer LAGs, endosteal resorption or fusion of hatching line and LAG1 had occurred. LAG2 was never lost by endosteal resorption and was identifiable by its diameter. Thus, the age of adult geckos could be determined by counting LAGs outward from LAG2. Application of this method to wild populations re- vealed that the longevity of this species is not less than 83 months, but that almost all individuals in fragmented habitats die before 50 months, suggesting lower population sustainability in such habitats.     

H-Y antigen in the study of sex determination and control of sex ratio

It has been proposed, on the basis of widespread phylogenetic conservation, that H-Y antigen is the inducer of primary sex, causing the undifferentiated XY gonad to become a testis in male heterogametic species such as the human and bovine. That proposition has withstood extensive testing in vivo and in vitro. Freemartin gonads are H-Y⁺, for example, and masculinization of the freemartin gonad has been attributed to soluble H-Y, borne and transmitted in the serum of the bull twin, and bound in ovarian receptors of the female. We have applied monoclonal H-Y antibodies to the identification of gender in embryos of the bovine. Our preliminary results imply presence of H-Y in bovine embryos of the morula and blastocyst stages recovered at about 6–12 days of gestation. Assignment of H-Y phenotype -- positive in males and negative in females -- allows selective implantation of male and female during embryo transfer. Thus in an early study, we correctly identified gender in 6 of 7 calves born healthy at term, after transfer of 8 blastocysts.     

Androgenic control of polysomal poly(A)-containing RNA in the cultured rat ventral prostate

Testosterone stimulated, at the concentration of 10-7 M and independently of other hormones, the accumulation of polysomal poly(A)-containing RNA (mRNA) in cultured explants of rat ventral prostate and concomitantly also protein synthesis. The hormone-induced accumulation of polysomal mRNA, which reached its maximum at 24 h after testosterone addition, paralleled the preferential labeling of high molecular weight RNA demonstrable with the electrophoretic analysis of the double-isotope labeled RNA after a short pulse (30 min). These findings are consistent with the idea that testosterone activated the synthesis of precursor mRNA leading to an increased amount of polysomal mRNA and eventually an activated protein synthesis. The synthesis and maturation of rRNA appeared to proceed even in the absence of testosterone, which is in contrast to the vivo findings on castrated rats. This partial uncoupling of RNA synthesis from androgenic control may account for the slow and less marked hormonal responses found in protein synthesis and glucose metabolism in cultured explants from normal animals. Because of the lack of uniformity in the suture, routine light microscopic control to assess the viability of cultured explants was found to be a prerequisite for successful biochemical work on prostate culture.     

External mouthpart morphology in the Tenuipalpidae (Tetranychoidea): Raoiella a case study

The use of low-temperature scanning electron microscopy (LTSEM) to study external mouthpart morphology in the Tenuipalpidae, in particular the genus Raoiella, has brought some aspects of the mechanics of feeding in this group into question. In addition, an LTSEM study on the specialized feeding behaviour of Raoiella indica Hirst (Tetranychoidea: Tenuipalpidae) revealed host plant use in this species could be affected by stomatal complex morphology.     

Karyotype, sex determination, and meiotic chromosome behavior in two pholcid (Araneomorphae, Pholcidae) spiders: implications for karyotype evolution

There are 1,111 species of pholcid spiders, of which less than 2% have published karyotypes. Our aim in this study was to determine the karyotypes and sex determination mechanisms of two species of pholcids: Physocyclus mexicanus (Banks, 1898) and Holocnemus pluchei (Scopoli, 1763), and to observe sex chromosome behavior during meiosis. We constructed karyotypes for P. mexicanus and H. pluchei using information from both living and fixed cells. We found that P. mexicanus has a chromosome number of 2n = 15 in males and 2n = 16 in females with X0-XX sex determination, like other members of the genus Physocyclus. H. pluchei has a chromosome number of 2n = 28 in males and 2n = 28 in females with XY-XX sex determination, which is substantially different from its closest relatives. These data contribute to our knowledge of the evolution of this large and geographically ubiquitous family, and are the first evidence of XY-XX sex determination in pholcids.     

When selection deceives phylogeographic interpretation: the case of the Mediterranean house gecko, Hemidactylus turcicus (Linnaeus, 1758)

A previous study on Hemidactylus turcicus based on mtDNA makers indicated that this gecko has a Middle-East origin, and that the current phylogeographic pattern is the result of a very rapid spread from the east to the west of the species’ range. The same study identified two distinct mitochondrial lineages with low differentiation and genetic diversity. Since H. turcicus is known to be closely associated to humanized environments, its present distribution range and phylogeography is frequently interpreted to be the result of recurrent human-mediated introductions. These conclusions used to be the same as those used to interpret the results obtained for the European populations of another gecko, Tarentola mauritanica. However, a recent study has revealed that the phylogeographic pattern of T. mauritanica is not solely the result of a recent colonization, but also of a mitochondrial selective sweep. Could the same be occurring in H. turcicus? To answer this question, two mitochondrial (12S rRNA and cytochrome b) and two nuclear genes (ACM4 and Rag2) were used in this study. From the mtDNA data we confirmed the existence of two distinct phylogeographic lineages; one occurring exclusively in the northern Mediterranean (Clade A), and another one more widespread that is the only lineage present in North Africa (Clade B). In light of these results, we could hypothesize that H. turcicus had its origin in Turkey, and from there Clade A moved to Europe and Clade B to North Africa spreading latter into Europe. However, Clade A presents significantly higher nucleotide diversity for the nuclear DNA compared to the mtDNA, and neutrality tests gave significant results for the mitochondrial data. These results suggest that the lack of mtDNA genetic diversity and structure in the European population of H. turcicus could also be due to a selective sweep, and not only because of a recent colonization. Together with the situation reported in T. mauritanica, the identification of a hitch-hiking process occurring in H. turcicus, represents two unprecedented cases of a selective sweep taking place in the same geographic area shaping the phylogeographic patterns of two unrelated genera of geckos.     

Methods for the study of amphipod swimming: behavior,morphology, and fluid dynamics

A thorough hydrodynamic approach to the study of swimming in amphipods demands a multipronged attack. A possible first step would be to gather swimming behavior data and determine the biomechanics and kinematics of pleopod beat. This requires careful observation of the swimming modes, swimming speeds, body positions and other aspects of behavior and limb motion that are crucial to swimming. Secondly, it is important to describe the morphology of the body and swimming appendages. Detailed drawings of body shape and design, skeletomusculature, condylic structure, and setal density and distribution on the pleopods and pereopods, are the tools required to ascribe hydrodynamic function to specific limb and body morphology. Finally, the information gathered from behavioral observations bolstered by functional morphology studies is applied to fluid dynamic calculations of drag, lift, and thrust. The theoretical calculations are then compared with empirical determinations of drag, wake generation, vortex shedding frequency, and flow patterns around an amphipod. The fluid dynamic facet of this research is the most challenging and requires an excellent grasp of the fundamental concepts of fluid flow and access to some highly technical equipment. The proposed tripartite approach for the study of amphipod swimming is by no means an exhaustive review of all the techniques that can be employed to quantify amphipod swimming. It will nevertheless permit a rigorous and systematic study of amphipod swimming.     

On the Live Coloration and Partial Mitochondrial DNA Sequences in the Topotypic Population of Goniurosaurus kuroiwae orientalis(Squamata: Eublepharidae), with Description of a New Subspecies from Tokashikijima Island,Ryukyu Archipelago,Japan

Careful investigations of live coloration and other characters in the eublepharid gecko Goniurosaurus kuroiwae orientalis from Tonakijima Island, the type locality of the subspecies in the Central Ryukyus, Japan, revealed its slight but consistent external differences from the consubspecific Tokashikijima population, whose phylogenetically divergent status against the former had been strongly suggested in a previous molecular study. A new subspecies, Goniurosaurus kuroiwae sengokui subsp. nov., is thus described for the Tokashikijima population. In life, Goniurosaurus k. sengokui subsp. nov. can be discriminated from G. k. orientalis in showing tint of reddish or pinkish, instead of yellowish, coloration in dorsal body pattern, and also usually darker and more or less reddish, instead of yellowish or grayish, iris in both adults and juveniles. Recalculation of sequence data at hand with relevant Gen Bank data confirmed substantial sequence divergences of G. k. sengokui from all other congeneric taxa of the Ryukyus described so far.     

A case study of primiparous maternal and infant gorilla (Gorilla gorilla gorilla) behavior

A primiparous mother and her infant were subjects of a longitudinal behavioral study at Lincoln Park Zoo in Chicago, Illinois. From November 1998 to November 1999, we collected a total of 100 hr of focal nonhuman animal instantaneous point sampling and all occurrence data on the mother and her infant. After 8 months, we introduced the 4-member focal group to an 8-member gorilla group, thus providing an opportunity to study the effect of the introduction on mother-infant behavior. Overall, time the pair spent physically apart was notably high with contact and proximate scores steadily decreasing over time. The infant was largely responsible for maintaining social proximity, rarely leaving the mother and frequently approaching her. In contrast, the mother left the infant frequently and approached the infant minimally. Maternal affiliative and nonaffiliative behaviors fluctuated throughout the study. Over time, both decreased. Despite a precarious maternal relationship, infant developmental trends were typical for captive gorillas. The results of this study suggest that allowing flexibility in judging maternal conduct can be of benefit to successful gorilla husbandry.     

Hygienic behavior of the honey bee (Apis mellifera) is independent of sucrose responsiveness and foraging ontogeny

Hygienic behavior in honey bees is a behavioral mechanism of disease resistance. Bees bred for hygienic behavior exhibit an increased olfactory sensitivity to odors of diseased brood, which is most likely differentially enhanced in the hygienic line by the modulatory effects of octopamine (OA), a noradrenaline-like neuromodulator. Here, we addressed whether the hygienic behavioral state is linked to other behavioral activities known to be modulated by OA. We specifically asked if, during learning trials, bees from hygienic colonies discriminate better between odors of diseased and healthy brood because of differences in sucrose (reward) response thresholds. This determination had to be tested because sucrose response thresholds are susceptible to OA modulation and may have influenced the honey bee's association of the conditioned stimulus (odor) with the unconditioned stimulus (i.e., the sucrose reward). Because the onset of first foraging is also modulated by OA, we also examined whether bees from hygienic colonies differentially forage at an earlier age compared to bees from non-hygienic colonies. Our study revealed that 1-day- and 15- to 20-day-old bees from the hygienic line do not have lower sucrose response thresholds compared to bees from the non-hygienic lines. In addition, hygienic bees did not forage at an earlier age or forage preferentially for pollen as compared to non-hygienic bees. These results support the idea that OA does not function in honey bees simply to enhance the detection of all chemical cues non-selectively or control related behaviors regardless of their environmental milieu. Our results indicate that the behavioral profile of the hygienic bee is sculpted by multiple factors including genetic, neural, social and environmental systems.     

Scale morphology and squamation patterns in cichlids (Teleostei, Perciformes): A comparative study

Problems in cichlid systematics call for new characteristics to be exploited. Scale surface morphology and squamation patterns could provide novel and useful information. A comparative study comprising 105 African, 10 American, and eight outgroup species was conducted to identify the most useful scale and squamation characters within the family and to clarify their phylogenetic significance. A great number of obviously genetically fixed characters were established. At least IS scale granulation types could be identified, proving to be ofparticular value for systematic purposes. Squamation patterns on the head, breast and fins were also found to be of interest in this context. Different character states are described and their distribution is reviewed, together with a discussion of their plesiomorphic status. From the results it can be concluded that for the Cichlidae scale and squamation studies can be valuable tools in investigating phylogenetic relationships.     

Origin of germ cells, sex determination, and sex inversion in medusae of the genus Clytia (Hydrozoa, leptomedusae): the influence of temperature

In Cnidaria, a separation between soma and germline remains unclear. In this work, we studied the origin of germinal cells and determination of the sexual phenotype in Clytia hemisphaerica and Clytia sp. Colonies of C. Hemisphaerica were cultivated and the medusae liberated by each colony raised until maturity. Two hermaphrodite colonies were obtained, liberating male and female medusae. These two colonies and their medusae were raised at 15 degrees C, 21 degrees C, or 24 degrees C. The medusae budded and cultured at 24 degrees C were mainly female (80%). In contrast, if the medusae were released at 15 degrees C, at whatever temperature they were raised later, they were mainly male (85%). The same occurred if, after release at 24 degrees C but before the formation of the gametes, they were kept at 15 degrees C for at least 24 hr. We suggest that there are two subpopulations of germ cells. The female line will be dominant at 24 degrees C but temperature sensitive, with inhibition of this line by a temperature drop to 15 degrees C, this inverting the population sex-ratio. The irreversible action of a temperature drop to 15 degrees C supports the view that the germ cells are isolated very early. In C. hemisphaerica, hermaphrodite medusae were never observed. On the contrary, in Clytia sp., probably a new species, we have found male, female, but also hermaphrodite specimens. This is the second definite example of hermaphroditism described in any hydromedusan. The transformation of female into hermaphrodite then into male specimens occurs at 13 degrees C. These results demonstrate the unstable character of genetic sex determination in cnidarians, at least in certain species.     

RNA binding protein sex-lethal (Sxl) and control of Drosophila sex determination and dosage compensation.

In the past two decades, scientists have elucidated the molecular mechanisms behind Drosophila sex determination and dosage compensation. These two processes are controlled essentially by two different sets of genes, which have in common a master regulatory gene, Sex-lethal (Sxl). Sxl encodes one of the best-characterized members of the family of RNA binding proteins. The analysis of different mechanisms involved in the regulation of the three identified Sxl target genes (Sex-lethal itself, transformer, and male specific lethal-2) has contributed to a better understanding of translation repression, as well as constitutive and alternative splicing. Studies using the Drosophila system have identified the features of the protein that contribute to its target specificity and regulatory functions. In this article, we review the existing data concerning Sxl protein, its biological functions, and the regulation of its target genes.     

Further studies on karyotypes and sex determination of phytoseiid mites (Acarina: Mesostigmata)

Chromosome studies were made on embryonic tissues of eggs of 15 species of predaceous mites of the family Phytoseiidae belonging to six genera: Amblyseius Berlese, Seiulus Berlese, Typhlodromus Scheuten, Phytoseiulus Evans, Phytoseius Ribaga and Paragigagnathus Amitai et Grinberg. In 14 species, the chromosome numbers n=4, 2n=8 were found. In cytological studies of post-embryonic stages based on these results, the haplo-diploid (arrhenotokous) mechanism was established. In the species Amblyseius deleoni Muma et Denmark, which was found to be thelytokous, only metaphases with 8 chromosomes were found.     

A behavior analysis of absolute pitch: sex, experience, and species

Absolute pitch (AP) perception refers to the ability to identify, classify, and memorize pitches without use of an external reference pitch. In tests of AP, several species were trained to sort contiguous tones into three or eight frequency ranges, based on correlations between responding to tones in each frequency range and reinforcement. Two songbird species, zebra finches and white-throated sparrows, and a parrot species, budgerigars had highly accurate AP, they discriminated both three and eight ranges with precision. Relative to normally reared songbirds, isolate reared songbirds had impaired AP. Two mammalian species, humans and rats, had equivalent and weak AP, they discriminated three frequency ranges to a lackluster standard and they acquired only a crude discrimination of the lowest and highest of eight frequency ranges. In comparisons with mammals even isolate songbirds had more accurate AP than humans and rats.     

Correlated evolution of sex and reproductive mode in corals (Anthozoa: Scleractinia)

Sexuality and reproductive mode are two fundamental life-history traits that exhibit largely unexplained macroevolutionary patterns among the major groups of multicellular organisms. For example, the cnidarian class Anthozoa (corals and anemones) is mainly comprised of gonochoric (separate sex) brooders or spawners, while one order, Scleractinia (skeleton-forming corals), appears to be mostly hermaphroditic spawners. Here, using the most complete phylogeny of scleractinians, we reconstruct how evolutionary transitions between sexual systems (gonochorism versus hermaphrodism) and reproductive modes (brooding versus spawning) have generated large-scale taxonomic patterns in these characters. Hermaphrodites have independently evolved in three large, distantly related lineages consisting of mostly reef-building species. Reproductive mode in corals has evolved at twice the rate of sexuality, while the evolution of sexuality has been heavily biased: gonochorism is over 100 times more likely to be lost than gained, and can only be acquired by brooders. This circuitous evolutionary pathway accounts for the prevalence of hermaphroditic spawners among reef-forming scleractinians, despite their ancient gonochoric heritage.