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Biomedical subjects

J Hanken

Publications and source records attributed to J Hanken.

27 records · Page 2Linked to original sources

Morphological integration in the cranium during anuran metamorphosis.

We examined the role of thyroid hormone in mediating morphological integration between cranial cartilage and bone during anuran metamorphosis. Exogenous T3 applied to premetamorphic tadpoles (Bombina orientalis) via intracranial implants of plastic micropellets precociously induced typical metamorphic changes in both tissues, but also dissociated the relative timing of developmental events between them. Morphological integration between the two primary cranial tissues is achieved in part by each tissue responding independently to endocrine factors and does not reflect a tight developmental coupling between them.

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Skull development during anuran metamorphosis: III. Role of thyroid hormone in chondrogenesis.

Metamorphosis of cranial cartilages in anuran amphibians constitutes one of the most dramatic and extensive ontogenetic transformations in vertebrates. We quantitatively examined the role of thyroid hormone (3,3',5-triiodo-L-thyronine; T3) in mediating gross aspects of this morphological repatterning in the skull of the Oriental fire-bellied toad, Bombina orientalis. T3 was administered via plastic (Elvax) micropellets in three treatment dosages (2.5, 0.25, and 0.025 microgram) and one control dosage (0 microgram) to tadpoles of three Gosner developmental stages--28/29, 30/31, and 32/33; tadpoles were recoved up to 8 d (treatment and control dosages) or 14 d (control dosage) later. Response of larval cartilages to exogenous T3 was dosage dependent but not implant-stage dependent; chondrogenic tissues that participate in metamorphic transformation are competent to respond to T3 well before they normally do. Metamorphic effects of T3 were visible within 2 d; in most treatment groups, the normal metamorphic sequence was two-thirds complete after 8 d. While T3 also induced precocious ossification, the normal temporal relation between bone formation and cartilage transformation was dissociated in experimental groups. Morphological integration between cartilage and bone during cranial metamorphosis is at least partly the result of each tissue responding independently to endocrine factors.

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Skull development during anuran metamorphosis: I. Early development of the first three bones to form--the exoccipital, the parasphenoid, and the frontoparietal.

In anuran amphibians, cranial bones typically first form at metamorphosis when they rapidly invest or replace the cartilaginous larval skull. We describe early development of the first three bones to form in the Oriental fire-bellied toad, Bombina orientalis--the parasphenoid, the frontoparietal, and the exoccipital--based on examination of serial sections. Each of these bones is fully differentiated by Gosner stage 31 (hindlimb in paddle stage) during premetamorphosis. This is at least six Gosner developmental stages before they are first visible in whole-mount preparations at the beginning of prometamorphosis. Thus, developmental events that precede and mediate the initial differentiation of these cranial osteogenic sites occur very early in metamorphosis--a period generally considered to lack significant morphological change. Subsequent development of these centers at later stages primarily reflects cell proliferation and calcified matrix deposition, possibly in response to increased circulating levels of thyroid hormone which are characteristic of later metamorphic stages. Interspecific differences in the timing of cranial ossification may reflect one or both of these phases of bone development. These results may qualify the use of whole-mount preparations for inferring the sequence and absolute timing of cranial ossification in amphibians.

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Skull development during anuran metamorphosis. II. Role of thyroid hormone in osteogenesis.

We examined the role of thyroid hormone (TH) in mediating cranial ossification during metamorphosis in the Oriental fire-bellied toad, Bombina orientalis. Exogenous T3 (3,3',5-triiodo-L-thyronine) was administered in three treatment dosages (0.025, 0.25, and 2.5 micrograms) plus a control dosage via plastic micropellets implanted within the dermis of tadpoles of three Gosner developmental stages: 28/29, 30/31, 32/33. Tadpoles were recovered after 2, 4, 6, and 8 d, and scored for the presence of three bones - median parasphenoid and paired frontoparietals and exoccipitals--as seen in cleared-and-stained, whole-mount preparations. T3 induced precocious ossification in both a stage-dependent and a dosage-dependent manner; stage dependence corresponded precisely with the degree of osteogenic differentiation at the time of hormone administration. Precocious ossification thus was due to the T3-promoted growth and calcified matrix deposition of these centers. Differential TH sensitivity among osteogenic sites may underlie both the temporal cranial ossification sequences characteristic of metamorphosing amphibians as well as sequence differences commonly observed among taxa.

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Native variant limb skeletal patterns in the red-backed salamander, Plethodon cinereus, are not regenerated.

Species of the salamander genus Plethodon have a characteristically uniform morphology. Morphological conservation at the level of interspecific comparisons, however, is not always reflected within species. Perhaps the most extreme example of intraspecific variation is the recent description of extensive variability in limb-skeletal patterning both within and between populations of the widespread species P. cinereus. We utilized limb regeneration following experimental amputation as a tool to examine whether naturally occurring variant skeletal patterns result from limb loss and regeneration in nature, and to assay the intrinsic (i.e., genetic) component of between-individual variation in mesopodial patterning. We observed the following. First, regenerate patterns are strikingly different from native patterns: interelement fusions in regenerates are typically between proximodistally adjacent cartilages, whereas interelement fusions in native variant limbs occur exclusively between laterally adjacent cartilages. Fusions also are over ten times more frequent in regenerates than in native limbs. Second, there is no strong correlation between native limb pattern (typical vs. variant) and the regenerate pattern. We conclude that variability in field-collected P. cinereus reflects extensive intrapopulation variation in limb-skeletal patterning during original limb development, rather than regeneration in nature, and that limb regeneration analysis provides no evidence of a strong genetic component to between-individual variation. Finally, unusual mesopodial patterns produced during limb regeneration may be related to the mechanical factors impinging on the regenerating limb in this terrestrial species.

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Morphological novelty in the limb skeleton accompanies miniaturization in salamanders.

Salamanders of the genus Thorius (Plethodontidae) are among the smallest tetrapods. Hypotheses of limb skeletal evolution in these vertebrates were evaluated on the basis of estimates of natural variation, comparisons of skeletal homology, and analysis of molecular phylogeny. Nine carpal arrangements occur in Thorius, more than in all twelve related genera of typically larger salamanders; six of these arrangements are unique. They represent a trend toward a decrease in the number of separate cartilages that is independent of locomotor and ecological specialization. Miniaturization may be an important source of morphological novelty, distinct from local adaptation, in vertebrates.

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Technique for revealing the three-dimensional architecture of whole preserved spiculated invertebrates.

Procedures for revealing the three-dimensional arrangement of calcareous sclerites, spicules, or ossicles embedded within connective tissue in formalin fixed invertebrates are described. Spicules are stained with alizarin red S following maceration of preserved animals or colonies with either trypsin or KOH solutions. Connective tissue is stained with alcian blue in different samples prior to maceration. Stained animals or colonies are cleared in glycerin. This method for revealing spicular structure and arrangement and the gross morphology of connective tissues offers several advantages over either scanning electron microscopy or reconstruction from serial sections.

Alcian Blue↗

Miniaturization and its effects on cranial morphology in plethodontid salamanders, genus Thorius (Amphibia, Plethodontidae): II. The fate of the brain and sense organs and their role in skull morphogenesis and evolution.

Relative size and arrangement of the brain and paired sense organs are examined in three species of Thorius, a genus of minute, terrestrial salamanders that are among the smallest extant tailed tetrapods. Analogous measurements of representative species of three related genera of larger tropical (Pseudoeurycea, Chiropterotriton) and temperate (Plethodon) salamanders are used to identify changes in gross morphology of the brain and sense organs that have accompanied the evolution of decreased head size in Thorius and their relation to associated changes in skull morphology. In adult Thorius, relative size (area measured in frontal plane, and length) of the eyes, otic capsules, and brain each is greater than in adults of all of the larger genera; relative size of the nasal capsules is unchanged or slightly smaller. Interspecific scaling phenomena--negative allometry of otic capsule, eye and brain size, isometry or slight positive allometry of nasal capsule size, all with respect to skull length--also are characteristic of intraspecific (ontogenetic) comparisons in both T. narisovalis and Pseudoeurycea goebeli. Predominance of the brain and eyes in Thorius results in greater contact and overlap among these structures and the nasal capsules in the anterior portion of the head. This is associated with anterior displacement of both the eyes and nasal capsules, which now protrude anterior to the skull proper; a change in eye shape; and medial deformation of anterior braincase walls. Posteriorly, predominance of the otic capsules has effected a reorientation of the jaw suspensorium to a fully vertical position that is correlated with the novel presence of a posteriorly directed squamosal process and shift in origin of the quadropectoralis muscle. Many of these changes in cranial morphology may be explained simply as results of mechanical (physical) interactions among the skeletal, nervous, and sensory components during head development at reduced size. This provides further evidence of the role of nervous, sensory, and other "soft" tissues in cranial skeletal morphogenesis, and reinforces the need to consider these tissues in analyses of skull evolution.

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Multiple paternity in Belding's ground squirrel litters.

Sexually receptive female Spermophilus beldingi (Rodentia: Sciuridae) usually mate with several different males. The paternity of 27 litters born in 1977 and 1978 was ascertained by combining field observations of mating with laboratory paternity exclusion analyses. Most of the litters (78 percent) were multiply sired, usually by two or three males. This may be the highest frequency of multiple paternity ever directly demonstrated in a natural population.

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