Journal Articles and Book Chapters: 1999 - 1990

Hanken, J., 1999. Larvae in amphibian development and evolution. In The Origin and Evolution of Larval Forms. San Diego. San Diego: Academic Press, pp. 61-108. PDF
Carl, T.F., et al., 1999. Inhibition of neural crest migration in Xenopus using anti-sense-slug RNA. Developmental Biology , 213 , pp. 101-115.Abstract

Based primarily on studies in the chick, it has been assumed that the zinc finger transcription factor Slug is required for neural crest migration. In the mouse, however, Slug is not expressed in the premigratory neural crest, which forms normally in Slug -/- animals. To study the role of Slug in Xenopus laevis, we used the injection of XSlug antisense RNA and tissue transplantation. Injection of Slug antisense RNA did not suppress the early expression of the related gene XSnail, but led to reduced expression of both XSlug and XSnail in later stage embryos, whereas the expression of another neural crest marker, XTwist, was not affected. Down-regulation of XSlug and XSnail was associated with the inhibition of neural crest cell migration and the reduction or loss of many neural crest derivatives. In particular, the formation of rostral cartilages was often highly aberrant, whereas the posterior cartilages were less frequently affected. The effects of Slug antisense RNA on neural crest migration and cartilage formation were rescued by the injection of either XSlug or XSnail mRNA. These studies indicate that XSlug is required for neural crest migration, that XSlug and XSnail may be functionally redundant, and that both genes are required to maintain each other's expression in the neural crest development of xenopus laevis. (C) 1999 Academic Press.

Hanken, J., 1999. Why are there so many new amphibian species when amphibians are declining?. Trends in Ecology & Evolution , 14 , pp. 7-8. PDF
Hanken, J. & Wake, D.B., 1998. Biology of tiny animals: Systematics of the minute salamanders (Thorius : Plethodontidae) from Veracruz and Puebla, Mexico, with descriptions of five new species. Copeia , pp. 312-345.Abstract

Minute plethodontid salamanders, genus Thorius, are far more diverse taxonomically than has been recognized previously. Populations of these salamanders from the Mexican states of Veracruz and Puebla are assigned to 10 species, five of which are described as new. Combinations of morphological and allozymic characters are used to sort the species and to make initial assessments of relationships. Valid existing names include Thorius pennatulus, T., troglodytes, T. dubitus, and T. schmidti. Thorius narismagnus; from the Sierra de Los Tuxtlas, which previously was considered to be a disjunct subspecies of T., pennatulus, is elevated to species rank. Thorius maxillabrochus is treated as a subjective junior synonym of the sympatric T.:schmidti. New taxa include Thorius lunaris, T. magnipes, T. minydemus, T. munificus, and T. spilogaster: All 10 species can be distinguished by morphological characters, but the distinctiveness of the taxa is bolstered by allozymic characters and by extensive sympatry. As many as three, and possibly four, species occur in sympatry, with some evidence of segregation by microhabitat (arboreal vs terrestrial). Adult body sizes span the range known for the genus, from very small in T., pennatulus (maturing at < 16 mm standard length) to large in T. lunaris (adults reaching > 31 mm). Collectively these species display a Hide elevational distribution, from less than 1000 m (T. pennatulus, T. narismagnus) to more than 3000 m (T. lunaris, T., spilogaster). Discovery of these new species adds to the rich herpetological diversity of east-central Mexico and underscores its importance as a principal center of radiation of tropical plethodontid salamanders.

Richardson, M.K., et al., 1998. Limb development and evolution: a frog embryo with no apical ectodermal ridge (AER). Journal of Anatomy , 192 , pp. 379-390.Abstract

The treefrog Eleutlrerodactylus coqui is a direct developer-it has no tadpole stage. The limb buds develop earlier than in metamorphosing species (indirect developers, such as Xenopus laevis). Previous molecular studies suggest that at least some mechanisms of limb development in E. coqui are similar to those of other vertebrates and we wished to see how limb morphogenesis in this species compares with that in other vertebrates. We found that the hind limb buds are larger and more advanced than the forelimbs at all stages examined, thus differing from the typical amniote pattern. The limb buds were also small compared to those in the chick. Scanning and transmission electron microscopy showed that although the apical ectoderm is thickened, there was no apical ectodermal ridge (AER). In addition, the limb buds lacked the dorsoventral flattening seen in many amniotes. These findings could suggest a mechanical function for the AER in maintaining dorsoventral flattening, although not all data are consistent with this view. Removal of distal ectoderm from E. coqui hindlimb buds does not stop outgrowth, although it does produce anterior defects in the skeletal pattern. The defects are less severe when the excisions are performed earlier. These results contrast with the chick, in which AER excision leads to loss of distal structures. We suggest that an AER was present in the common ancestor of anurans and amniotes and has been lost in at least some direct developers including E. coqui.

Jennings, D.H. & Hanken, J., 1998. Mechanistic basis of life history evolution in anuran amphibians: Thyroid gland development in the direct-developing frog, Eleutherodactylus coqui. General and Comparative Endocrinology , 111 , pp. 225-232.Abstract

Direct development is a widespread, alternative life history in Recent amphibians. There is no free-living, aquatic larva; adult features form in the embryo and are present at hatching. The mechanistic bases of direct development remain relatively unexplored. The current study describes the embryonic ontogeny of the thyroid gland in the direct-developing frog Eleutherodactylus coqui (Leptodactylidae) and quantifies histological changes that occur in the gland after its initial appearance. The thyroid gland of E. coqui is first apparent at Townsend-Stewart stage 10, approximately two-thirds of the way through embryogenesis. Soon after this the thyroid begins to accumulate follicular colloid. Quantitative analyses of thyroid histology reveal embryonic peaks in two measures, follicle number and follicle volume, which are followed by declines in these measures prior to hatching. These peaks in thyroid activity in E. coqui are correlated with morphological changes that are directly comparable to metamorphic changes in frogs that retain the ancestral, biphasic life history. In metamorphic taxa, a histologically identifiable thyroid gland does not form until the larval period, well after hatching. Nevertheless, measures of thyroid histology observed in E. coqui follow the pattern reported for metamorphosing amphibians. The present results support the hypothesis that the evolution of direct development in anurans is associated with precocious development and activity of the thyroid axis. (C) 1998 Academic Press.

Richardson, M.K., et al., 1998. Somite number and vertebrate evolution. Development , 125 , pp. 151-160.Abstract

Variation in segment number is an important but neglected feature of vertebrate evolution, Some vertebrates have as few as six trunk vertebrae, while others have hundreds. We examine this phenomenon in relation to recent models of evolution and development. Surprisingly, differences in vertebral number are foreshadowed by different somite counts at the tailbud stage, thought to be a highly conserved (phylotypic) stage, Somite number therefore violates the 'developmental hourglass' model, We argue that this is because somitogenesis shows uncoupling or dissociation from the conserved positional field encoded by genes of the zootype.Several other systems show this kind of dissociation, including limbs and feathers. Bmp-7 expression patterns demonstrate dissociation in the chick pharyngeal arches, This makes it difficult to recognise a common stage of pharyngeal development or 'pharyngula' in ail species, Rhombomere number is more stable during evolution than somite number, possibly because segmentation and positional specification in the hindbrain are relatively interdependent, Although developmental mechanisms are strongly conserved, dissociation allows at least some major evolutionary changes to be generated in phylotypic stages.

Hanken, J., et al., 1997. Jaw muscle development as evidence for embryonic repatterning in direct-developing frogs. Proceedings of the Royal Society B-Biological Sciences , 264 , pp. 1349-1354.Abstract

The Puerto Rican direct-developing frog Eleutherodactylus coqui (Leptodactylidae) displays a novel mode of jaw muscle development for anuran amphibians. Unlike metamorphosing species, several larval-specific features never form in E. coqui; embryonic muscle primordia initially assume an abbreviated, mid-metamorphic configuration that is soon remodelled to form the adult morphology before hatching. Also lacking are both the distinct population of larval myofibres and the conspicuous, larval-to-adult myofibre turnover that are characteristic of muscle development in metamorphosing species. These modifications are part of a comprehensive alteration in embryonic cranial patterning that has accompanied life history evolution in this highly speciose lineage. Embryonic 'repatterning' in Eleutherodactylus may reflect underlying developmental mechanisms that mediate the integrated evolution of complex structures. Such mechanisms may also facilitate, in organisms with a primitively complex life cycle, the evolutionary dissociation of embryonic, larval, and adult features.

Hanken, J., Jennings, D.H. & Olsson, L., 1997. Mechanistic basis of life-history evolution in anuran amphibians: Direct development. American Zoologist , 37 , pp. 160-171.Abstract

The primitive, or ancestral reproductive mode for Recent amphibians involves a complex, biphasic life history, Yet evolutionarily derived, alternate modes are seen in all three living orders and predominate in some clades, Analysis of the consequences and mechanistic bases of one such mode-direct development-can provide insights into the evolutionary opportunities and constraints conferred by the ancestral metamorphic ontogeny, Direct development in the anuran genus Eleutherodactylus involves fundamental alterations to many features of embryonic and posthatching development, At hatching, young emerge as fully formed, albeit tiny versions of the adult; most larval features are absent, Pervasive changes in ontogenetic timing, in particular the precocious (embryonic) formation of many adult structures, appear to be correlated with early development of the thyroid axis, although responsiveness to exogenous thyroid hormone is diminished or even lacking in at least some peripheral tissues, Changes in cranial patterning are likely mediated by the embryonic neural crest, although many gross features of crest biology are highly conserved, Laboratory-based analyses of direct development and other derived reproductive modes in amphibians, using contemporary methods developed for more standard, ''model'' organisms, may contribute important insights into life-history evolution that complement those derived from analyses of morphology, ecology and phylogeny.

Richardson, M.K., et al., 1997. There is no highly conserved embryonic stage in the vertebrates: Implications for current theories of evolution and development. Anatomy and Embryology , 196 , pp. 91-106.Abstract

Embryos of different species of vertebrate share a common organisation and often look similar. Adult differences among species become more apparent through divergence at later stages. Some authors have suggested that members of most or all vertebrate clades pass through a virtually identical, conserved stage. This idea was promoted by Haeckel, and has recently been revived in the context of claims regarding the universality of developmental mechanisms. Thus embryonic resemblance at the tailbud stage has been linked with a conserved pattern of developmental gene expression - the zootype. Haeckel's drawings of the external morphology of various vertebrates remain the most comprehensive comparative data purporting to show a conserved stage. However, their accuracy has been questioned and only a narrow range of species was illustrated. In view of the current widespread interest in evolutionary developmental tal biology, and especially in the conservation of developmental mechanisms, re-examination of the extent of variation in vertebrate embryos is long overdue, We present here the first review of the external morphology of tailbud embryos, illustrated with original specimens from a wide range of vertebrate groups, We find that embryos at the tailbud stage - thought to correspond to a conserved stage - show variations in form due to allometry, heterochrony, and differences in body plan and somite number. These variations foreshadow important differences in adult body form. Contrary to recent claims that all vertebrate embryos pass through a stage when they are the same size, we find a greater than 10-fold variation in greatest length at the tailbud stage. Our survey seriously undermines the credibility of Haeckel's drawings, which depict not a conserved stage for vertebrates, but a stylised amniote embryo. In fact, the taxonomic level of greatest resemblance among vertebrate embryos is below the subphylum. The wide variation in morphology among vertebrate embryos is difficult to reconcile with the idea of a phyogenetically-conserved tailbud stage, and suggests that at least some developmental mechanisms are not highly constrained by the zootype, Our study also highlights the dangers of drawing general conclusions about vertebrate development from studies of gene expression in a small number of laboratory species.

Olsson, L. & Hanken, J., 1996. Cranial neural crest migration and chondrogenic fate in the oriental fire-bellied toad Bombina orientalis: Defining the ancestral pattern of head development in anuran amphibians. Journal of Morphology , 229 , pp. 105-120.Abstract

We assess cranial neural-crest cell migration and contributions to the larval chondrocranium in the phylogenetically basal and morphologically generalized anuran Bombina orientalis (Bombinatoridae). Methods used include microdissection, scanning electron microscopy, and vital dye labeling, in conjunction with confocal and fluorescence microscopy. Cranial neural-crest cells begin migrating before neural-fold closure and soon form three primary streams. These streams contribute to all cranial cartilages except two medial components of the hyobranchial skeleton (basihyal and basibranchial cartilages), the posterior portion of the trabecular plate, and the otic capsule, the embryonic origin of which is unknown. Chondrogenic fate is regionalized within the cranial neural folds, with the anterior regions contributing to anterior cartilages and the posterior regions to posterior cartilages. A neural-crest contribution also was consistently observed in several cranial nerves and the connective tissue component of many cranial muscles. Notwithstanding minor differences among species in the initial configuration of migratory streams, cranial neural-crest migration and chondrogenic potential in metamorphosing anurans seem to be highly stereotyped and evolutionarily conservative. This includes a primary role for the neural crest in the evolutionary origin of the paired suprarostral and infrarostral cartilages, two prominent caenogenetic features of the rostral skull unique to anuran larvae. Our results provide a model of the ancestral pattern of embryonic head development in anuran amphibians. This model can serve as a basis for examining the ontogenetic mechanisms that underlie the diversity of cranial morphology and development displayed by living frogs, as well as the evolutionary consequences of this diversity. (C) 1996 Wiley-Liss, Inc.

Wake, D.B. & Hanken, J., 1996. Direct development in the lungless salamanders: What are the consequences for developmental biology, evolution and phylogenesis?. International Journal of Developmental Biology , 40 , pp. 859-869.Abstract

Direct development is a widespread alternate reproductive mode in living amphibians that is characterized by evolutionary loss of the free-living, aquatic larval stage. Courtship, mating, and oviposition occur on land, and the terrestrial egg hatches as a fully formed, miniature adult. While it is the most common reproductive mode in urodeles, development outside the reproductive tract of the female that proceeds directly to a terrestrial hatchling occurs in only a single lineage, the lungless salamanders of the family Plethodontidae. Evolution of direct development in plethodontids has contributed importantly to the extraordinary evolutionary success of this speciose, geographically widespread, and morphologically and ecologically diverse taxon. Developmental consequences and correlates include increased egg size and embryonic development time, loss of larval structures and ontogenetic repatterning, and altered pattern formation in organogenesis. Evolutionary and phylogenetic consequences and correlates include the loss of larval constraints and origin of morphological novelty, and frequent homoplasy. Analysis of direct development in an evolutionary context illustrates the complex interplay between processes of phylogenetic divergence and developmental biology, and substantiates the prominent role of developmental processes in both constraining phenotypic variation and promoting phenotypic diversity. Despite the proven suitability of direct-developing plethodontid salamanders for laboratory and field study, knowledge of basic features of their developmental biology remains far below that available for many other urodeles. Examination of such features of these ''non-model'' organisms is an appropriate and deserving goal of future research.

Packard, M.J., Jennings, D.H. & Hanken, J., 1996. Growth and uptake of mineral by embryos of the direct-developing frog Eleutherodactylus coqui. Comp. Physiol. Biochem. , 113A , pp. 343-349.Abstract

Embryos of the direct-developing frog Elutherodactylus coqui cake up small quantities of yolk and yolk mineral early in incubation but increase their uptake of yolk reserves at later stages of development. Growth and accumulation of calcium and magnesium by embryos also occur slowly at first and at a higher rate later. Accumulation of calcium and magnesium by embryos is largely a function of variation in size of embryos, but uptake of phosphorus is unrelated to size. Although patterns of growth and uptake of mineral by embryonic coquis resemble those for embryos of oviparous amniotes, embryonic coquis do not deplete the yolk of its nutrients to the same degree. Thus, residual yolk of coqui hatchlings contains a high percentage of the nutrient reserves originally present in the egg. This difference between embryonic coquis and embryos of oviparous amniotes may indicate that transfer of nutrients from yolk to embryo becomes limiting during the growth phase. Alternatively, some aspects of the neurologic system are so poorly developed at hatching that coqui may not be able to find prey effectively. A large nutrient reserve could sustain hatchlings while the neurologic system continues to mature.

Hanken, J., 1995. Development and evolution in amphibians. In M. Slatkin, ed. Exploring Evolutionary Biology: Readings from American Scientist. Sunderland. Sunderland: Sinauer Assoc., Inc. pp. 224-231.
Moury, J.D. & Hanken, J., 1995. Early cranial neural crest migration in the direct-developing frog, Eleutherodactylus coqui. Acta Anatomica , 153 , pp. 243-253.Abstract

Direct development is a common reproductive mode in Living amphibians characterized by absence of the free-living, aquatic larval stage. In Eleutherodactylus, a species-rich genus of New World frogs, evolution of direct development from the ancestral biphasic ontogeny is correlated with a comprehensive modification in embryonic cranial patterning, including the loss of many larval-specific components and the precocious formation of many adult (postmetamorphic) structures, We use scanning electron microscopy (SEM) to examine the emergence and early migration of cranial neural crest cells in Eleutherodactylus coqui to begin to assess the possible role of the neural crest in mediating these evolutionary changes. As in metamorphosing frogs, cranial crest cells emerge prior to neural fold closure and assemble into three streams: rostral, rostral otic, and caudal otic. These streams contribute to the face and first visceral (mandibular) arch, to the second (hyoid) arch, and to posterior (branchial) arches, respectively. Rostrocaudal position, morphology, and/or migration patterns distinguish subpopulations of cells within the rostral stream and caudal otic stream. With the possible exception of the small size of the rostral otic and caudal otic streams, evolution of direct development in E. coqui has not altered basic patterns of neural crest emergence or early migration as assessed by SEM. Lf observed evolutionary changes in embryonic cranial patterning are mediated by the neural crest, then they likely involve later aspects of crest migration or more subtle features related to pattern formation such as cell behavior and commitment, or gene expression.

Hanken, J. & Wake, D.B., 1994. Five New Species of Minute Salamanders, Genus Thorius (Caudata, Plethodonitdaee, from Northern Oaxaca, Mexico. Copeia , pp. 573-590.Abstract

Five new species of diminutive salamanders of the endemic Mexican genus Thorius (Plethodontidae) are described from the Sierra de Juarez in northern Oaxaca. The species are diagnosed by adult body size, external proportions, dentition, osteology and coloration. The three species that have been studied using protein electrophoresis are genetically unique; all differ from T. macdougalli, the only species of the genus previously known from these mountains. Each of the six species studied has distinct geographic and elevational ranges, and there is a complex pattern of geographic overlap and replacement. As many as three species co-occur locally at elevations up to 2955 m on Cerro Pelon, and each species is sympatric with at least one other. One species descends to approximately 800 m, which is the lowest known elevational record for the genus. The new taxa include the full size range of the genus, with two large and three small species.

Seufert, D.W., Hanken, J. & Klymkowsky, M.W., 1994. Type II Collagen Distribution during Cranial Development in Xenopus Laevis. Anatomy and Embryology , 189 , pp. 81-89.Abstract

Epithelially expressed type II collagen is thought to play a prominent role in the embryonic patterning and differentiation of the vertebrate skull, primarily on the basis of data derived from amniotes. We describe the spatiotemporal distribution of type II collagen in the embryonic head of the African clawed frog, Xenopus laevis, using whole-mount and serial-section immunohistochemical analysis. We studied embryos spanning Nieuwkoop and Faber (1967) stages 21-39, a period including cranial neural crest cell migration and ending immediately before the onset of neurocranial chondrogenesis. Xenopus displays a transient expression of type II collagen beginning at least as early as stage 21; staining is most intense and widespread at stages 33/34 and 35/36 and subsequently diminishes. Collagen-positive areas include the ventrolateral surface of the brain, sensory vesicles, notochord, oropharynx, and integument. This expression pattern is similar, but not identical, to that reported for the mouse and two bird species (Japanese quail, domestic fowl); thus epithelially expressed type II collagen appears to be a phylogenetically widespread feature of vertebrate cranial development. Consistent with the proposed role of type II collagen in mediating neurocranial differentiation, most collagen-positive areas lie adjacent to subsequent sites gf chondrogenesis in the neurocranium but not the visceral skeleton. However, much of the collagen is expressed after the migration of cranial neural crest, including presumptive chondrogenic crest, seemingly too late to pattern the neurocranium by entrapment of these migrating cells.

Hanken, J., 1993. Adaptation of bone growth to miniaturization of body size. In B. K. Hall, ed. Bone, Volume 7: Bone Growth - B. Boca Raton. Boca Raton: CRC Press, pp. 79-104. PDF
Hall, B.K. & Hanken, J., 1993. Bibliography of skull development: 1937-1989. In J. Hanken & B. K. Hall, ed. The Skull: Vol. 1, Development. Chicago. Chicago: University of Chicago Press, pp. 378-577.
Hanken, J. & Hall, B.K., 1993. Mechanisms of skull diversity and evolution. In J. Hanken & B. K. Hall, ed. The Skull: Vol. 3, Functional and Evolutionary Mechanisms. Chicago. Chicago: University of Chicago Press, pp. 1-36. PDF