Cell Biology Commons^™

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 …

Heat Shock Factor Gains Access To The Yeast Hsc82 Promoter Independently Of Other Sequence-Specific Factors And Antagonizes Nucleosomal Repression Of Basal And Induced Transcription, Alexander M. Erkine, C. C. Adams, T. Diken, D. S. Gross

Scholarship and Professional Work – COPHS

Transcription in eukaryotic cells occurs in the context of chromatin. Binding of sequence-specific regulatory factors must contend with the presence of nucleosomes for establishment of a committed preinitiation complex. Here we demonstrate that the high-affinity binding site for heat shock transcription factor (HSF) is occupied independently of other cis-regulatory elements and is critically required for preventing nucleosomal assembly over the yeast HSC82 core promoter under both noninducing (basal) and inducing conditions. Chromosomal mutation of this sequence, termed HSE1, erases the HSF footprint and abolishes both transcription and in vivo occupancy of the TATA box. Moreover, it dramatically reduces promoter chromatin …

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 …

Investigation Of The Substrate Recognition Characteristics And Kinetics Of Mammalian Mitochondrial Dna Topoisomerase I, Zeki Topcu

Theses and Dissertations in Biomedical Sciences

Topoisomerases are DNA-modifying enzymes found in prokaryotes, eukaryotes, viruses and organelles such as chloroplast and mitochondria. Information about these enzymes in eukaryotic systems is mostly limited to nuclear enzymes, although our laboratory has been characterizing the biochemical and biophysical properties of the mammalian mitochondrial topoisomerases. We have determined the polarity of the attachment of mitochondrial topoisomerase I to its substrate DNA. To study the substrate preference and kinetic parameters of mitochondrial topoisomerase I, selected regions of mammalian mitochondrial DNA (mtDNA) were inserted into pGEM plasmid vectors following a series of modification and optimization experiments of currently available methods for PCR-cloning. …

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 …

Analysis Of Human Sperm Chromatin Integrity, Silvina M. Bocca

Theses and Dissertations in Biomedical Sciences

Determining potential maternal or paternal sources of abnormal chromosomal constitution gives opportunity for preconception genetic counseling. The most direct determination is achieved by analyzing the nuclear constitution of the gametes.

The present study evaluated the integrity of human spermatozoal nuclear material in the two condensation stages of chromatin and chromosomes. Original semen samples (ORI) and their swim-up fractions (SW, selected for motility) from men of known (donors) and unknown (patients) fertility were analyzed. The extent of chromatin condensation was assessed by light microscopy and flow cytometry during the time course of a chemically-induced decondensation reaction.

Motile spermatozoa were used to …

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 …

The Cγ Subunit Is A Unique Isozyme Of The Camp-Dependent Protein Kinase, Stephen J. Beebe, Paul Salomonsky, Tore Jahnsen, Yixin Li

Bioelectrics Publications

There are at least three isozymes (Cα, Cβ, and Cγ) of the mammalian catalytic (C) subunit of cAMP-dependent protein kinase (PKA) (Beebe, S., Oyen, O., Sandberg, M., Froysa, A., Hansson, V., and Jahnsen, T. (1990) Mol. Endocrinol. 4, 465-475). To compare the Cγ and Cα isozymes, the respective cDNAs were expressed in permanently transformed Kin-8 PKA-deficient Y1 adrenal cells using the mouse metallothionein promoter. The recombinant C subunits were characterized as immunoreactive, zinc-inducible, cAMP-dependent kinase activities. In contrast to Cα, histone was a better substrate than Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) for Cγ. Furthermore, Cγ histone kinase activity was not inhibited by the …

Effects Of Chemical Aneuploidogens On Taxol Purified Drosophila And Mouse Brain Microtubules Polymerization And Depolymerization In Vitro, Anil Sehgal

Biological Sciences Theses & Dissertations

The effects of aneuploidogens (aneuploidy causing agents) on taxol-purified microtubules from Drosophila and mouse brain in vitro were studied by using a spectrophotometric assay and electron microscopy. Colchicine, acetonitrile, propionitrile, acrylonitrile, dimethyl sulfoxide (DMSO), griseofulvin and cadmium chloride inhibited microtubule polymerization whereas methoxyethyl acetate (MEA) and methyl mercuric chloride (MMC) did not. All aneuploidogens tested (at 50mM) resulted in reduced rate of elongation of mouse brain microtubules. MMC, cadmium chloride and DMSO resulted in increased rates of Drosophila microtubule elongation whereas the rest of the drugs resulted in decreases. The in vitro results from Drosophila correlate well with the previously …

A Simple Solid-Phase Electrophoretic Procedure For The Separation Of Plasmid Dna, Linda Ann Simurra

Biological Sciences Theses & Dissertations

A method was developed for extraction of plasmid DNA from bacterial cells embedded in agarose blocks. Cell - containing blocks were treated with various lysing reagents and inserted into the wells of an agarose gel. Upon electrophoresis the plasmid DNA migrated out of the embedding block and into the gel leaving intact chromosomal DNA in the well. This method was tested with various organisms and found to be effective for plasmid isolation. In comparison to "traditional" procedures, this new method is less tedious since chemical separation of plasmids is not required prior to electrophoresis. Also, a higher yield of plasmid …

Solid Phase Extraction Of Mammalian Cell Mitochondrial Dna And Its Electrophoretic Separation In Agarose Gels, Theresa Brick-Miller

Biological Sciences Theses & Dissertations

An improved and efficient electrophoretic procedure for the isolation of mitochondrial DNA (mtDNA) is described. The solid phase extraction procedure for the isolation of mtDNA involves embedding as few as 200,000 Ehrlich ascites tumor cells into a block of agarose, digestion of cellular membranes by detergent action and subsequent electrophoretic separation of nucleic acids in agarose gels. Genomic DNA remains in the original block of agarose while the mtDNA migrates in the separation gel to a position equivalent to that obtained for mtDNA isolated by traditional procedures.

The efficacy of a large number of detergents, agarose concentrations, embedding conditions, embedding …

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 …

Characterization Of The Dna And Chromosonal Analysis Of Embryonic Dermacentor Variabilis (Say) Cells In Culture, Stanley Nicholas Mason

Biological Sciences Theses & Dissertations

Cultured embryonic Dermacentor variabilis cells were examined by spectrophotometric techniques to determine total cellular DNA concentration and guanine-cytosine content of the DNA and by differential staining procedures to define the karyotype further. The cells were found to have a mean DNA content of 16.98 pg and a DNA guanine-cytosine content of 49%. Chromosome banding attempts resulted in the specific characterization of four chromosomes and differential grouping of the seven remaining chromosomes. A previously unreported submetacentric chromosome was identified.

Fine Structure Of The Nucleus And Cytoplasmic Feulgen-Positive Areas In The Developing Oocyte Of Argus (Persicargas) Radiatus, Bonnie J. Harding

Biological Sciences Theses & Dissertations

A transmission electron microscope analysis of the changes that occur in the nuclear chromatin and the Feulgen-positive areas of the oocyte cytoplasm as the female develops, feeds and mates is described. The description includes analysis of oocytes from three types of females: unfed, unmated; fed, unmated; and fed, mated. In all types, dense material passes through the extremely porous nuclear membrane where it accumulates in the cytoplasm.

Nuclei of previtellogenic oocytes appear the same in all females studied. Fibrillar bodies and up to four large nucleoli are seen. No typical chromatin is seen, but structures resembling nucleosomes appear throughout the …

Interpopulation Variation In Calcareous And Proteinaceous Shell Components In The Stream Limpet, Ferrissia Rivularis, Carl M. Way, Daniel J. Hornbach, Albert J. Burky

Biology Faculty Publications

Natural populations of the North American stream limpet, F. rivularis, were studied in upstate New York [USA], in a set of localities whose waters have a 15-fold range of dissolved Ca (4.6-67.6 mg/liter) and also range from oligotrophy to eutrophy. Shell component analyses (CaCO3, total organic C and total N) are reported as component mass-fractions (mg/g or .mu.g/g dry weight) and as values for a standard limpet shell of 35 mm aperture length (AL). More than 2-fold differences occur between populations in all 3 components, with relatively little variation occurring within each population. Expressed per standard limpet, CaCO3 values for …

Seasonal Metabolism Of The Sphaeriid Clam, Musculium Partumeium (Say), From A Permanent And A Temporary Pond, Carl M. Way, Daniel J. Hornbach, Albert J. Burky

Biology Faculty Publications

Seasonal metabolic rates were examined for the sphaeriid clam, Musculium partumeium (Say) from a permanent and a temporary pond. For the fall-born generation from the permanent pond and for the single generation from the temporary pond, metabolic rates peak during periods of greatest growth and reproduction. Metabolic rates were consistently higher for the permanent pond population. These rates for Musculium partumeium extend the available information on the family Sphaeriidae to the genus Musculium.

Growth Of Pisidium Casertanum (Poli) In West Central Ohio, Albert J. Burky, Daniel J. Hornbach, Carl M. Way

Biology Faculty Publications

Clarke (1973) states that Pisidium casertanum (Poli) is perhaps the most common species of Pisidium and is truly cosmopolitan, occurring throughout Eurasia, Africa, Australia, and South, Central and North America. Populations of these clams are reported in habitats ranging from ephemeral ponds to benthic zones of deep lakes. Our study describes Manuscript received 29 May 1979 and in revised form 25 July 1980 (#79-34). 2 Present address: Department of Biology, University of Virginia, Charlottesville, VA 22901. 3 Present address: Department of Zoology, Miami University, Oxford, OH 45056. 41 aspects of the life-cycle of P. casertanum in a spring-fed stream with …

Buoyancy Changes As Related To Respiratory Behavior In An Amphibious Snail, Pomacea Urceus (Müller), From Venezuela, Kathleen A. Burky, Albert J. Burky

Biology Faculty Publications

The ampullariid Pomacea urceus uses its ctenidium and lung in respiration. The snails reach the surface with their siphon and ventilate their lung by withdrawing their head-foot in a pumping action. The lung gas also serves to change the buoyancy of the snails. After a ventilation a snail does not necessarily return to the same buoyancy level. They may achieve overall specific gravities of <1 to > 1 after a ventilation. They may remain submerged (s.g. > 1) or float (s.g.

A Cytogenetic Study Of Xx/Xy Chimerism And Other Anomalies Of Bos Taurus In A Sampled Dairy Herd, Ferne M. Rogers

Masters Theses

The establishment of normal chromosomal patterns for a wide range of plants and animals has led to the recognition of abnormalities as well as their relationship to phenotypic irregularities. Research dealing with the expression of human karyotype is of prime interest; but, closely rivaling that is the genetic study of domestic animals which are of economic importance to man.

One observed anomaly which has come under scrutiny by cytogeneticists is that of the female member of a heterogeneous bovine twin pair, the freemartin. This is a sterile animal with internal morphology showing varying degrees of intersex. Cytogenetic studies reveal XX/XY …

Modification Of Ontogenic Expression Due To Relocation Of The Ry+ Gene In Drosophila Melanogaster, Allen A. Badgett Jr.

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The ry⁺ gene of Drosophila melanogaster and its associated enzyme, xanthine dehydrogenase, were employed in an analysis of the relationship between regulation and the location of a gene in the genome. Enzyme assays as an indication of gene activity were performed on genotypes containing zero, one and two doses of ry⁺ genes. Xanthine dehydrogenase activity of rosy genes in normal and relocated positions was determined during development.

The results indicate: 1) the gene ry⁺ is differentially active during ontogeny; 2) the activity during ontogeny is proportional to the number of ry⁺ genes present in the genome; …