Cell and Developmental Biology Commons^™

Estructura Del Condrocráneo Y Esqueleto Visceral De Larvas De Pseudis Minuta (Anura, Pseudidae, E. O. Lavilla, Rafael O. De Sá

Biology Faculty Publications

The chondrocranium and visceral skeleton of Pseudis minuta tadpoles are described, based on a series of five larvae in stages 31 - 35 of Gosner (1960). Among their striking characters are the presence of peculiar articular surfaces between cornua trabeculae and suprarostral cartilage, the incomplete development of the orbital cartilage, the high fenestration of the floor of the cavum cranii, the fusion of posterior foramina, and the fusion of spicules 3 and 4 in the hyobranchial skeleton.

The Timing And Pattern Of Myogenesis In Hymenochirus Boettgeri, Matthew T. Smetanick, Rafael O. De Sá

Biology Faculty Publications

Differences in the relative timing of homologous developmental events among closely related species, known as heterochronies, may provide valuable clues in understanding evolutionary relationships (McKinney, 1988; McNamara, 1995). Examining the timing of myogenic events is a relatively easy and effective method for finding heterochronic events. For example, whether muscle proteins and myofibrils appear before or after multinucleation can be determined through histological techniques (Kielbowna, 1981). Simple observations of live specimens can pinpoint functional landmarks such as first twitch (spontaneous or due to external stimuli) and first heartbeat.

Development Of The Suprarostral Plate Of Pipoid Frogs, Rafael O. De Sá, Charles C. Swart

Biology Faculty Publications

The rostral region of nonpipoid tadpoles has two sets of cartilages, the cornua trabeculae and the suprarostral cartilages, whereas the rostral region in pipoid larvae is occupied by a single and continuous cartilage, the suprarostral plate. The homology of this region in pipoid and nonpipoids tadpoles has been controversial. We examined the early formation and development of the suprarostral plate using serially cross-sectioned specimens of Rhinophrynus, Xenopus, and Hymenochirus. We conclude that the cartilaginous structures present in the rostral area of pipoid and nonpipoid larvae are homologous. Furthermore, we found two different developmental patterns among pipoid larvae. The chondrocranium …

Osteological Analysis Of The Killifish Genus Cynolebias (Cyprinodontiformes: Rivulidae), Marcelo Loureiro, Rafael O. De Sá

Biology Faculty Publications

Relationships among the species of the annual fish Cynolebias are unclear. An analysis of the variation and utility of osteological characters for phylogenetic analysis was done using cleared and double-stained specimens representing 21 species of Cynolebias. This analysis showed that some of the characters previously used to diagnose this genus and some of the species are polymorphic. Osteologically, Cynolebias can be diagnosed by the following synapomorphies: (1) triangular-shaped parietal, (2) vomer positioned ventral to the parasphenoid, (3) long ventral process of the dentary, (4) teeth on fourth ceratobranchial, and (5) teeth on first epibranchial. In addition, characters that help …

Confocal Microscopy: A Powerful Tool For Biological Research, Amit Singh, K. P. Gopinathan

Biology Faculty Publications

Conventional light microscopy allows the observation of living as well as fixed cells and tissues to generate two-dimensional images. The out-of-focus information often obscures the ultrastructural details, especially in thick specimens with overlapping structures. The earliest available light microscopy visualized the objects in hydrated state in two-dimensions during their temporal development. The emergence of electron microscopy (EM) provided superb resolution of ultrastructural details, but it was applicable only for objects in the dehydrated state and thereby potentially introducing handling artifacts. The usefulness of optical methods, however, has been limited by the poor depth discrimination. Often, the fluorescence and reflectance images …

The Biology Of Xenopus By R. C. Tinsley And H. C. Kobel, Rafael O. De Sá

Biology Faculty Publications

The Biology of Xenopus presents a summary of current knowledge about a single genus resulting from a symposium held at the Zoological Society of London in September 1992. This approach to summarizing available information has also been taken for other taxa, such as Atelopus (Lotters, 1996). However, the task of compiling data for Xenopus is enormous relative to any other amphibian group, because Xenopus laevis has become a model system for molecular and development research (Cannatella and de Sa, 1993). Unfortunately, most of our knowledge of Xenopus is biased toward this single species. There are about 20 recognized species of …

Chondrocranial Morphology Of Leptodactylus Larvae (Leptodactylidae: Leptodactylinae): Its Utility In Phylogenetic Reconstruction, Peter M. Larson, Rafael O. De Sá

Biology Faculty Publications

Chondrocranial morphology of leptodactylid frogs is scarcely known and has not been completely described for any species of Leptodactylus. We describe the diversity of chondrocranial morphology in the genus Leptodactylus based on the analysis of 22 species, representing the four species groups: the fuscus Group, ocellatus Group, melanonotus Group, and pentadactylus Group. Furthermore, 26 characters are identified and used in a phylogenetic analysis. The phylogenetic analysis using Physalaemus, Crossodactylus, and Hylodes as outgroups suggests two monophyletic clades within Leptodactylus: the melanonotus-ocellatus clade and the pentadactylusfuscus clade. However, it does not support the monophyly of the species groups as currently recognized …

Altered Trafficking Of Mutant Connexin32, Suzanne M. Deschênes, Jessica L. Walcott, Tamara L. Wexler, Steven S. Scherer, Kenneth H. Fischbeck

Biology Faculty Publications

We examined the cellular localization of nine different connexin32 (Cx32) mutants associated with X-linked Charcot–Marie–Tooth disease (CMTX) in communication-incompetent mammalian cells. Cx32 mRNA was made, but little or no protein was detected in one class of mutants. In another class of mutants, Cx32 protein was detectable in the cytoplasm and at the cell surface, where it appeared as plaques and punctate staining. Cx32 immunoreactivity in a third class of mutants was restricted to the cytoplasm, where it often colocalized with the Golgi apparatus. Our studies suggest that CMTX mutations have a predominant effect on the trafficking of Cx32 protein, resulting …

Inhibition Of Axonal Growth By Brefeldin A In Hippocampal Neurons In Culture, Mark Jareb, Gary Banker

Biology Faculty Publications

The outgrowth of neuronal processes involves a great increase in the surface area of the cell. The supply of membrane material necessarily must be coordinated with the demands for neurite growth. The selective growth of only one or two neurites at any given time during the development of polarity raises the possibility that the production of materials by the soma is limiting for growth (Dotti and Banker, 1987; Dotti et al., 1988; Goslin and Banker, 1990). To examine the role of the availability of membrane components during the development of polarity and axonal elongation, we treated neurons with brefeldin A, …

Recovery Of Rna Polymerase Ii Synthesis Following Dna Damage In Mutants Of Saccharomyces Cerevisiae Defective In Nucleotide Excision Repair, Michael S. Reagan, Errol C. Friedberg

Biology Faculty Publications

We have measured the kinetics of the recovery of mRNA synthesis in the inducible GAL10 and RNR3 genes after exposure of yeast cells to ultraviolet (UV) radiation. Such recovery is abolished in mutant strains defective in nucleotide excision repair (NER) of DNA, including a rad23 mutant. Mutants defective in the RAD7 or RAD16 genes, which are required for the repair of the non-transcribed strand but not the transcribed strand of transcriptionally active genes, show slightly faster recovery of RNA synthesis than wild-type strains. A strain deleted of the RAD26 gene, which is known to be required for strand-specific NER in …

Analysis Of Gene Expression During Embryonic Development In Mulberry Silkworm Bombyx Mori, Amit Singh, K. P. Gopinathan

Biology Faculty Publications

We have developed a method for dechorionation and devitellinization of the silkworm eggs without damage, to facilitate the analysis of gene expression during embryonic development of Bombyx mori. Making use of antibodies available from heterologous systems, the spatio-temporal expression patterns of peroxidase and proliferating cell nuclear antigen have been directly visualized in whole mount embryos at various stages of development without the need for generating transformed ·lines carrying specific reporter constructs. The B. mori system, previously unamenable for such studies, could thus serve as an attractive model for molecular analysis of insect development.

The attention lavished on Drosophila melanogaster as …

The Tadpole Of Pseudis Minuta (Anura: Pseudidae), An Apparent Case Of Heterochrony, Rafael O. De Sá, Esteban O. Lavilla

Biology Faculty Publications

The external morphology, oral disc, and coloration pattern of the larva of Pseudis minuta are described. Tadpoles are mostly bottom dwellers that have a small, terminal oral disc with a labial tooth row formula 1(1-1)/(1-1)2. Internal oral anatomy is characterized using scanning electron microscopy, representing the only description available for the family Pseudidae. The coloration pattern of P. minuta tadpoles is compared with that of other pseudids. Pseudis minuta larvae do not exhibit the ontogenetic coloration change reported for P. paradoxa. Considering larval coloration, larval size, and reports of adult size, acceleration or hypermorphosis are suggested as possible heterochronic mechanisms …

Assignment Of Fragile Site 8e (Fra8e) To Human Chromosome Band 8q24.11 Adjacent To The Hereditary Multiple Exostoses 1 Gene And Two Overlapping Langer-Giedion Syndrome Deletion Endpoints, April L. Hill, Y. Harada, E. Takahashi, J. Hou, M. J. Wagner, D. E. Wells

Biology Faculty Publications

The distamycin A inducible fragile site, FRA8E, Fra(8) (q24.11) has been previously mapped to 8q24.11, proximal to the MYC gene, by fluorescent in situ hybridization (Takahasi et al., 1991). This fragile site has been shown to be present in about 1 out of every 140 healthy Japanese individuals (Takahasi et al., 1988). To more precisely map FRA8E, we have used cosmids isolated from the Langer-Giedion chromosomal region (LGCR) for FISH analysis. Langer-Giedion syndrome (LGS) is a contiguous gene syndrome characterized by chromosome deletions in Sq24.11 (Ludecke et al., 1991). One of the genes known to be involved in the etiology …

Connexin32 And X-Linked Charcot-Marie-Tooth Disease, Kenneth H. Fischbeck, Suzanne M. Deschênes, Linda Jo Bone, Steven S. Scherer

Biology Faculty Publications

This paper deals with the genetic defect responsible for the X-linked form of Charcot-Marie-Tooth disease.Mutations in the gap junction geneconnexin32(Cx32) cause the X-linked form of Charcot–Marie–Tooth disease, an inherited demyelinating neuropathy.

External Morphology Of The Chorion Of The Annual Fishes Cynolebias (Cyprinodontiformes: Rivulidae), Marcelo Loureiro, Rafael O. De Sá

Biology Faculty Publications

Members of the family Rivulidae (killifishes) inhabit temporary bodies of freshwater in South and Central America (one species is also found in North America). The most remarkable characteristic of the family Rivulidae is that species have an annual life cycle with a drought-resistant egg during the dry season. Parenti's (1981) analysis of the order Cyprinodontiformes considered a single genus, Cynolebias, whereas Cos- ta's (1990) phylogenetic analysis of the family Rivulidae separates Cynolebias from Cynopoecilus. One of Costa's synapomorphies to separate Cynopoecilus is the unique structure of their egg's chorion, which is shared with Leptolebias and Campellolebias.

Topographically Specific Effects Of Elf-1 On Retinal Axon Guidance In Vitro And Retinal Axon Mapping In Vivo, Masaru Nakamoto

Biology Faculty Publications

Topographic maps, which maintain the spatial order of neurons in the order of their axonal connections, are found throughout the nervous system. In the visual retinotectal projection, ELF-1, a ligand in the tectum, and its receptors in the retina show complementary gradients in expression and binding, indicating they may be positional labels for map development. Here we show that ELF-1 acts as a repellent axon guidance factor in vitro. In vivo, when the tectal ELF-1 pattern is modified by retroviral overexpression, retinal axons avoid ectopic ELF-1 patches and map to abnormally anterior positions. All these effects were seen on axons …

Características De La Osificación Craneal En Phyllomedusa Boliviana (Anura: Hylidae), Rafael O. De Sá, E. O. Lavilla

Biology Faculty Publications

RESUMEN. Dentro de la familia Hylidae disponemos de información sobre las secuencias de osificación craneal para menos del 4.0% de sus miembros. El presente trabajo describe la secuencia de osificación craneal de Phyllomedusa boliviana. El patrón general de osificación es similar a lo reportado para otras especies de la familia, pero existen algunas diferencias importantes tales como la osificación tardía de los próoticos y la osificación premetamórfica de neopalatino, vómer, cuadrado-yugal, pterigoides y columela. El lento desarrollo ulterior de la osificación resulta en juveniles con cráneos extensamente cartilaginosos.

ABSTRACT. Characteristics of the cranial ossification in Phyllomedusa …

Connexin32 Is A Myelin-Related Protein In The Pns And Cns, Steven S. Scherer, Suzanne M. Deschênes, Yi-Tian Xu, Judith B. Grinspan, Kenneth H. Fischbeck, David L. Paul

Biology Faculty Publications

We have examined the expression of a gap junction protein, connexin32 (Cx32), in Schwann cells and oligodendrocytes. In peripheral nerve, Cx32 is found in the paranodal myelin loops and Schmidt-Lanterman incisures of myelinating Schwann cells, and the levels of Cx32 protein and mRNA change in parallel with those of other myelin-related genes during development, Wallerian degeneration, and axonal regeneration. In the central nervous system, Cx32 is found in oligodendrocytes and their processes, but not in compact myelin, and the levels of Cx32 protein and mRNA increase during development in parallel with those of the other myelin genes. Thus, Cx32 is …

Enhancer Trap Technique: A Novel Tool For Identification And Developmental Characterization Of Genes Of Drosophila, Amit Singh

Biology Faculty Publications

The classical technique of mutational screen for identification of genes controlling early development has now approached saturation. A new era in genetic identification and developmental characterization of genes in Drosophila has commenced with the advent of the enhancer trap technique. This technique involves mobilization of a P-lacZ vector to diverse chromosomal locations in the fruit fly genome to bring it under the regulation of developmentally expressed genes or their enhancer elements. The technique offers a strikingly elegant method of gaining entry into fruit fly genes.

Isolation Of A Mouse Cdna Encoding Mtj1, A New Murine Member Of The Dnaj Family Of Proteins, Shannon E. Brightman, Gregory L. Blatch, Bruce R. Zetter

Biology Faculty Publications

We report the isolation and sequencing of MTJ1, a 1792-bp cDNA from an M27 murine lung carcinoma cell line. The largest ORF within MTJ1 encodes a 63,869-Da protein, containing a 73-amino-acid (aa) sequence (the J domain) that is conserved in proteins of the DnaJ family of chaperonins. The J domain of MTJ1 is bracketed by potential transmembrane domains in a similar configuration to the J domain of the yeast DnaJ-like protein, SEC63. Polyclonal antibodies raised against deduced aa sequences within MTJ1 recognized antigens of 62, 42 and 41 kDa that were enriched in the nuclear and heavy microsome subcellular fractions …

An Improved Method For Chemical Devitellinization Of X-Gal Stained Drosophila Embryos, Amit Singh, Madhuri Kango-Singh, P. Sinha

Biology Faculty Publications

In Drosophila developmental biological studies, X-gal staining is commonly employed to study the spatio-temporal expression of the lacZ reporter gene in the transformed flies or their embryos. Study of the lacZ pattern in embryos often suffers from the lack of an efficient and high yieldirrg technique for devitellinization of X-gal stained embryos. Devitellinization techniques employed during antibody staining, in situ hybridization or embryonic cuticular preparations generally do not give satisfactory results when used for similar purpose in X-gal stained embryos. This results in the flaky appearance of the blue stain. We present here an improved chemical devitellinization technique which gives …

Increased Stem Cell Factor Release By Hemangioma-Derived Endothelial Cells, C. J. Meininger, Shannon Brightman, K. A. Kelly, B. R. Zetter

Biology Faculty Publications

Background: Capillary hemangiomas, the most common tumors in young children, consist of proliferating capillary vessels and endothelial cells. These tumors also contain large numbers of mast cells, compared with the normal surrounding skin or tissue. We have recently shown that stem cell factor (SCF), the gene product of the murine steel locus, can act as a chemoattractant for mast cells. In this study, we investigated whether SCF might be involved in the recruitment and maintenance of mast cells in hemangiomas. Experimental Design: Cultured endothelial cells derived from a murine hemangioma were compared with normal vascular endothelial cells for the ability …

Complementary Gradients In Expression And Binding Of Elf-1 And Mek4 In Development Of The Topographic Retinotectal Projection Map, Masaru Nakamoto

Biology Faculty Publications

Topographic maps with a defined spatial ordering of neuronal connections are a key feature of brain organization. Such maps are believed to develop in response to complementary position-specific labels in presynaptic and postsynaptic fields. However, the complementary labeling molecules are not known. In the well studied visual map of retinal axons projecting to the tectum, the labels are hypothesized to be in gradients, without needing large numbers of cell-specific molecules. We recently cloned ELF-1 as a ligand for Eph family receptors. Here, RNA hybridization shows matching expression gradients for ELF-1 in the tectum and its receptor Mek4 in the retina. …

Mutations In The Non-Helical Linker Segment L1-2 Of Keratin 5 In Patients With Weber-Cockayne Epidermolysis Bullosa Simplex, Yiu-Mo Chan, Qian-Chun Yu, Janine M. Leblanc-Straceski, Angela Christiano, Lena Pulkkinen, Raju S. Kucherlapati, Jouni Uitto, Elaine Fuchs

Biology Faculty Publications

Keratins are the major structural proteins of the epidermis. Analyzing keratin gene sequences, appreciating the switch in keratin gene expression that takes place as epidermal cells commit to terminally differentiate, and elucidating how keratins assemble into 10 nm filaments, have provided the foundation that has led to the discoveries of the genetic bases of two major classes of human skin diseases, epidermolysis bullosa simplex (EBS) and epidermolytic hyperkeratosis (EH). These diseases involve point mutations in either the basal epidermal keratin pair, K5 and K14 (EBS), or the suprabasal pair, K1 and K10 (EH). In severe cases of EBS and EH, …

Development And Application Of A Thermistor Current Meter, Carl M. Way, Albert J. Burky, Christine Miller-Way

Biology Faculty Publications

This report provides details for the construction of a hot-bead thermistor current meter which is capable of measuring water velocities on a millimeter spatial scale and for the construction of a compact and accurate calibration system. Hot-bead thermistor current meters can be built with response times of 200 ms capable of measuring velocities between 0.1 and 80 cm s-1. The construction of a sturdy probe for application in lotic systems such as high gradient Hawaiian streams was achieved by the use of heavy-duty acrylic tubing, small stainless steel gas-chromatography tubing, and flexible Tygon spaghetti tubing. An acrylic handle anchors the …

Xenopus Laevis As A Model Organism, David C. Cannatella, Rafael O. De Sá

Biology Faculty Publications

Model organisms are often assumed to be representative of some more inclusive taxon of which the species is a part. This assumption leads to mistaken generalizations about the evolutionary and comparative significance of the data gathered. This paper reviews com? parative and evolutionary studies of Xenopus laevis and its relatives. Phylogenetic analysis of data from DNA sequences and morphology indicate that Xenopus is monophyletic and that Silurana is its sister group. The most basal lineages of Pipidae diverged prior to the breakup of Gondwana. The bizarre morphology of Xenopus is in part due to changes in the mode of meta? …

Ribosomal Dna And The Phylogeny Of Frogs, David M. Hills, Loren K. Ammerman, Michael T. Dixon, Rafael O. De Sá

Biology Faculty Publications

Phylogenetic analysis of 1656 aligned sites in the 28S ribosomal RNA gene of frogs supports some of the recently recognized higher groups of anurans but provides counter-support for others. The 28S rDNA data support the monophyly of the recently recognized Pipanura (me- sobatrachians plus neobatrachians), which in turn indicates paraphyly of archaeobatrachians. Me- sobatrachians (pelobatoids plus pipoids), which are either considered paraphyletic or weakly sup- ported as monophyletic in morphological analyses, also receive support as a monophyletic group from the 28S rDNA data. Hyloidea (=Bufonoidea), which is widely recognized but lacks morphological support, receives some molecular support as being monophyletic. …

A Retinoic Acid Responsive Gene, Mk, Produces A Secreted Protein A With Heparin Binding Activity, Masaru Nakamoto

Biology Faculty Publications

MK is a gene whose expression increases transiently during retinoic acid-induced differentiation of embryonal carcinoma cells. MK polypeptide was secreted by differentiating HM-1 embryonal carcinoma cells and by L-cells transfected with an MK cDNA under the control of the S-actin promoter and Rous sarcoma virus enhancer. MK polypeptide was found to have heparin binding activity. Conditioned medium of the transfected L-cells promoted growth of PC-12 pheochromocytoma cells. These findings support the view that MK polypeptide is a secreted factor involved in regulation of growth and differentiation.

Spermatogenesis And Histology Of The Testes Of The Caecilian, Chthonerpeton Indistinctum, Rafael O. De Sá, Nibia Berois

Biology Faculty Publications

Macroscopically, the testes of Chthonerpeton indistinctum consist of a series of oval lobes. The number and size of lobes is variable in Chthonerpeton indistinctum. Histologically, the testes are divided into structural units, locules separated by septa of connective tissue. Inside of each locule spermatogenesis takes place in cysts of germinal cells that divide synchronically. Chthonerpeton indistinctum is divided chronologically into four stages of development based on the shape of the nuclei of the spermatids and their arrangement in the cyst: (1) round, (2) kidney shaped, (3) attenuate, and (4) bouquet arrangement.

Seasonal Responses Of Filtration Rates To Temperature, Oxygen Availability, And Particle Concentration Of The Freshwater Clam, Musculium Partumeium (Say), Albert J. Burky, Richard B. Benjamin, Denis G. Conover, Jennifer Detrick

Biology Faculty Publications

Seasonal filtration rates of a pond population of the freshwater pisidiid clam, Musculium par­tumeium (Say), were assessed by measuring the clearance of 2.02 µ.m latex beads from suspension and expressed in terms of FR = a(AFDW}b where FR = filtration rate (ml H20 · hr- 1 ) , AFDW = mg ash-free dry weight of a whole clam and "a" and "b" are constants. The a-values ( = FR of a 1-mg AFDW clam} vary seasonally with the highest rates corresponding to periods of maximum growth and reproduction in the spring and fall. Changes in "b" reflect the influence of …