S Anne Böttger

University of New Hampshire, Durham, NC, United States

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Publications (23)54.54 Total impact

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    ABSTRACT: Disseminated neoplasia, a diffuse tumor of the hemic system, is characterized in many bivalve mollusks by hemolymph containing 1–100% mitotic hemocytes. Little is known about the onset and chronic distribution of neoplasia in populations of Mya arenaria (Soft-shell Clam), though studies have reported episodic exposure to environmental contaminants or an infectious agent as a potential cause of this disease. Here we provide the first set of continuous data on neoplasia in Soft-shell Clams, from three sites in New England where sediments have been characterized regarding their granulometry, composition, contaminants, and clam densities. When correlating sediment characteristics to terminal neoplasia (76–100% neoplastic or rounded, unattached hemocytes), New Bedford Harbor, MA, which is the most contaminated site, had the highest frequency of treminal neoplasia (maximum of 9.49% ± 0.78 SE), and the most pristine site, Ogunquit, ME, displayed the lowest frequencies (maximum of 0.47% ± 0.05 SE). Correlations of frequency of neoplasia to known environmental contaminants also suggests that fully neoplastic individuals were found only at sites of increased levels of heavy metals, PCBs, and PAHs. In addition, we documented the highest frequency of clams with terminal neoplasia from New Bedford Harbor in December (9.49% ± 0.78 SE) when seawater temperatures were low, and the lowest frequency in July (1.08 ± 0.4 SE) when seawater temperatures were highest. These results may indicate vulnerability of neoplastic clams to seasonal increases in environmental temperature and resulting oxidative stress. Based on shell measurements and a theoretical mathematical age model (which correlates susceptibility to neoplasia with age and sexual maturity), we suggest that the Soft-shell Clam is most susceptible to this disease between one and two years of age (9.5% frequency at 1 year, 22.25% incidence at 1.5 years, and 57.14% incidence at 2 years).
    Northeastern Naturalist 01/2013; 20(3):430-440. · 0.36 Impact Factor
  • Norah G Taraska, S Anne Böttger
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    ABSTRACT: Disseminated neoplasia, a diffuse tumor of the hemolymph system, is one of the six most destructive diseases among bivalve mollusk populations, characterized by the development of abnormal, rounded blood cells that actively proliferate. Though the specific etiology of disseminated neoplasia in Mya arenaria remains undetermined, the involvement of viral pathogens and/or environmental pollutants has been suggested and considered. The current study used 5-bromodeoxyuridine (BrDU) known to induce the murine leukemia virus and filtered neoplastic hemolymph to initiate disseminated neoplasia in clams from different populations and size classes respectively. M. arenaria from three locations of different natural neoplasia occurrences were divided into a control and three experimental treatments and injected with 200μl of sterile filtered seawater or 50-200μg/ml BrDU respectively. In a concurrent experiment, animals from different size classes were injected with 2.5% total blood volume of 0.2μm filtered blood from a fully neoplastic animal. Animals were biopsied weekly and cell neoplasia development was counted and scored as 0-25, 26-50, 51-75 and 76-100% neoplastic hemocytes (stages 1-4) in 50μl samples. BrDU injection demonstrated that neoplasia development in M. arenaria was dose dependent on BrDU concentration. In addition, natural disease prevalence at the source location determined initiation of neoplasia induction, with animals from the area of the highest natural disease occurrence displaying fastest neoplasia development (p=0.0037). This could imply that depending on the natural disease occurrence, a potential infectious agent may remain dormant in normal (stage 1) individuals in higher concentrations until activated, i.e. through chemical injection or potentially stress. The size experiment demonstrated that only M. arenaria between 40 and 80mm developed 26-100% neoplastic hemocytes when injected with filtered neoplastic hemolymph, indicating that individuals smaller than 20mm or larger than 80mm were not or no longer susceptible to disease development. So far neoplasia studies have not considered natural disease prevalence or size involvement in neoplasia development and our results indicate that these should be future considerations in neoplasia examinations.
    Journal of Invertebrate Pathology 10/2012; · 2.67 Impact Factor
  • Erin E Gestl, S Anne Böttger
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    ABSTRACT: While it is known that cytoplasmic retention of p53 occurs in many solid tumors, the mechanisms responsible for this retention have not been positively identified. Since heatshock proteins like mortalin have been associated with p53 inactivation in other tumors, the current study sought to characterize this potential interaction in never before examined colorectal adenocarcinoma cell lines. Six cell lines, one with 3 different fractions, were examined to determine expression of p53 and mortalin and characterize their cellular localization. Most of these cell lines displayed punctate p53 and mortalin localization in the cell cytoplasm with the exception of HCT-8 and HCT116 379.2 cells, where p53 was not detected. Nuclear p53 was only observed in HCT-116 40-16, LS123, and HT-29 cell lines. Mortalin was only localized in the cytoplasm in all cell lines. Co-immunoprecipitation and immunohistochemistry revealed that p53 and mortalin were bound and co-localized in the cytoplasmic fraction of four cell lines, HCT-116 (40-16 and 386; parental and heterozygous fractions respectively of the same cell line), HT-29, LS123 and LoVo, implying that p53 nuclear function is limited in those cell lines by being restricted to the cytoplasm. Mortalin gene expression levels were higher than gene expression levels of p53 in all cell lines. Cell lines with cytoplasmic sequestration of p53, however, also displayed elevated p53 gene expression levels compared to cell lines without p53 sequestration. Our data reveal the characteristic cytoplasmic sequestration of p53 by the heat shock protein mortalin in human colorectal adenocarcinoma cell lines, as is the case for other cancers, such as glioblastomas and hepatocellular carcinomas.
    Biochemical and Biophysical Research Communications 06/2012; 423(2):411-6. · 2.41 Impact Factor
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    Mark C Pugliese, Stefanie Anne Böttger, Frank E Fish
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    ABSTRACT: Xenobalanus globicipitis, a unique type of small pseudo-stalked barnacle occurs on the appendages of cetaceans, including the common bottlenose dolphin Tursiops truncatus. In this study, we examined attachment structures of X. globicipitis and modifications to the skin of T. truncatus in areas of attachment compared to skin nearby an attachment site. Barnacles and their six calcareous footplates were measured for their length and width. There was a positive correlation of barnacle width and length to footplate width and length. The thickness of the stratum corneum increased significantly in areas of attachment compared to skin nearby a footplate. The mitotic stratum germinativum at the base of the dermal papillae did not change significantly in areas of attachment compared to skin nearby a footplate. The stratum germinativum lining the lateral walls of the dermal papillae was significantly thicker in areas of skin nearby a footplate compared to in areas of attachment. Skin of T. truncatus nearby a footplate, displayed dermal papillae extending from the dermis and pointing roughly perpendicular to the epidermal stratum corneum. At sites of X. globicipitis attachment, the dermal papillae were forced to extend laterally, parallel to the stratum corneum, and the dermal papillae length to width ratio at an attachment site was significantly higher than on skin near an attachment site. Our results show that attachment of X. globicipitis through production of footplates organized into calcareous rings, leads to a thickened stratum corneum of the epidermis, a thinner lateral mitotic stratum germinativum and displaced structures of the upper dermis. These resulting modifications to the epidermis and dermis of the host may add to securing barnacle attachment to its host.
    Journal of Morphology 04/2012; 273(4):453-9. · 1.60 Impact Factor
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    Charles W Walker, Ben Low, S Anne Böttger
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    ABSTRACT: In this chapter we demonstrate the remarkable sequence homology be-tween mortalin proteins from a broad array of invertebrate taxa, including the few species for which functional analyses have been conducted. We also discuss expres-sion and functional data for full-length and truncated clam homologs for human mortalin and their function in cytoplasmic sequestration in cancerous clam hemo-cytes. Both clam proteins have N-terminal mitochondrial targeting and p53 binding domains, though the truncated variant is missing exon 3 containing the N-terminal ATP/ADP binding and ATPase domains. Both variants are over-expressed and com-plexed with p53 and both may be responsible for cytoplasmic sequestration of p53 in cancerous clam hemocytes. Clam hemocyte cancer is the only animal model thus far investigated where cytoplasmically sequestered wild-type p53 can be reactivated both in vitro and in vivo using both genotoxic and non-genotoxic therapies. Our results suggest that mortalin-based cytoplasmic sequestration of wild-type p53 in cancerous clam hemocytes can be reversed by treatment with antineoplastic drugs also employed against similar human diseases and will result either in transcription-based apoptosis when the nucleus is accessible or non-transcription-based apoptosis when nuclear access is blocked., © Springer Science+Business Media B.V. 2012
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    ABSTRACT: All echinoderms have unique hydraulic structures called tube feet, known for their roles in light sensitivity, respiration, chemoreception and locomotion. In the green sea urchin, the most distal portion of these tube feet contain five ossicles arranged as a light collector with its concave surface facing towards the ambient light. These ossicles are perforated and lined with pigment cells that express a PAX6 protein that is universally involved in the development of eyes and sensory organs in other bilaterians. Polymerase chain reaction (PCR)-based sequencing and real time quantitative PCR (qPCR) also demonstrate the presence and differential expression of a rhabdomeric-like opsin within these tube feet. Morphologically, nerves that could serve to transmit information to the test innervate the tube feet, and the differential expression of opsin transcripts in the tube feet is inversely, and significantly, related to the amount of light that tube feet are exposed to depending on their location on the test. The expression of these genes, the differential expression of opsin based on light exposure and the unique morphological features at the distal portion of the tube foot strongly support the hypothesis that in addition to previously identified functional roles of tube feet they are also photosensory organs that detect and respond to changes in the underwater light field.
    Proceedings of the Royal Society B: Biological Sciences 03/2011; 278(1723):3371-9. · 5.68 Impact Factor
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    ABSTRACT: The human p53 tumour suppressor protein is inactivated in many cancers and is also a major player in apoptotic responses to cellular stress. The p53 protein and the two other members of this protein family (p63, p73) are encoded by distinct genes and their functions have been extensively documented for humans and some other vertebrates. The structure and relative expression levels for members of the p53 superfamily have also been reported for most major invertebrate taxa. The functions of homologous proteins have been investigated for only a few invertebrates (specifically, p53 in flies, nematodes and recently a sea anemone). These studies of classical model organisms all suggest that the gene family originally evolved to mediate apoptosis of damaged germ cells or to protect germ cells from genotoxic stress. Here, we have correlated data from a number of molluscan and other invertebrate sequencing projects to provide a framework for understanding p53 signalling pathways in marine bivalve cancer and stress biology. These data suggest that (a) the two identified p53 and p63/73-like proteins in soft shell clam (Mya arenaria), blue mussel (Mytilus edulis) and Northern European squid (Loligo forbesi) have identical core sequences and may be splice variants of a single gene, while some molluscs and most other invertebrates have two or more distinct genes expressing different p53 family members; (b) transcriptional activation domains (TADs) in bivalve p53 and p63/73-like protein sequences are 67-69% conserved with human p53, while those in ecdysozoan, cnidarian, placozoan and choanozoan eukaryotes are ≤33% conserved; (c) the Mdm2 binding site in the transcriptional activation domain is 100% conserved in all sequenced bivalve p53 proteins (e.g. Mya, Mytilus, Crassostrea and Spisula) but is not present in other non-deuterostome invertebrates; (d) an Mdm2 homologue has been cloned for Mytilus trossulus; (e) homologues for both human p53 upstream regulatory and transcriptional target genes exist in molluscan genomes (missing are ARF, CIP1 and BH3 only proteins) and (f) p53 is demonstrably involved in bivalve haemocyte and germinoma cancers. We usually do not know enough about the molecular biology of marine invertebrates to address molecular mechanisms that characterize particular diseases. Understanding the molecular basis of naturally occurring diseases in marine bivalves is a virtually unexplored aspect of toxicoproteomics and genomics and related drug discovery. Additionally, increases in coastal development and concomitant increases in aquatic pollutants have driven interest in developing models appropriate for evaluating potential hazardous compounds or conditions found in the aquatic environment. Data reviewed in this study are coupled with recent developments in our understanding the molecular biology of the marine bivalve p53 superfamily. Taken together, they suggest that both structurally and functionally, bivalve p53 family proteins are the most highly conserved members of this gene superfamily so far identified outside of higher vertebrates and invertebrate chordates. Marine bivalves provide some of the most relevant and best understood models currently available for experimental studies by biomedical and marine environmental researchers.
    Advances in Marine Biology 01/2011; 59:1-36. · 2.05 Impact Factor
  • Charles W Walker, S.A. Boettger, C.C. Eno
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    ABSTRACT: Here we provide methods for generating triploid green sea urchin embryos (Strongylocentrotus droebachiensis). These are the first triploid sea urchins of any species that have been produced at any life stage and represent an initial step in a larger effort to yield triploid adult green sea urchins for large-scale land-based and near-shore commercial aquaculture ventures. Following fertilization, triploid mollusks and fish result by preventing release of the second polar body. Adult triploids in all of these organisms are sterile and may be larger than diploids based on accelerated growth of somatic tissues. Similar methods are impossible for sea urchins since haploid (1n) ova are released from the ovary after completion of both meiotic divisions. We have produced triploid green sea urchin embryos (3n=63) by fusing two 1n ova (each n=21) and fertilizing the diploid products of these fusions with highly diluted spermatozoa (n=21). Mechanical, proteolytic and acidic treatments were employed to remove the jelly coat and vitelline membrane prior to fusion. Results indicate that acidic removal of the extracellular structures, subsequent fusion with poly(Arg) in 1.5 ml microcentrifuge tubes and fertilization with highly diluted sperm yield 60.64±1.39% triploid embryos that reach prism pluteus stage (with spicules). Triploidy was verified at all developmental stages by counting chromosomes. We discuss scaling up our laboratory methods for embryos to the hatchery environment and also issues unique to the reproductive biology of sea urchins that will be vital for commercial aquaculture ventures to consider if triploid adult green sea urchins can ultimately be produced. © 2011 Elsevier
    Aquaculture 01/2011; 318:199-206. · 2.01 Impact Factor
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    Carly C Ginter, S Anne Böttger, Frank E Fish
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    ABSTRACT: The unique pattern of small tubercles on the leading edge of the dorsal fins of harbor porpoises (Phocoena phocoena) has been widely noted in the literature, though their structure or function has never been conclusively identified. We examined external morphology and microanatomy of the tubercles for further understanding of the nature of the tubercles. Measurements were taken of height and peak-to-peak distance of the tubercles using scaled photographs. Mean tubercle height was standardized as a percentage of the dorsal fin height and ranged from 0.63 to 0.87%. Mean peak-to-peak distance ranged from 4.2 ± 2.0 to 5.6 ± 2.0 mm. The microstructure analysis of the dorsal fin leading edge, trailing edge and tubercles revealed an epidermal thickness of 0.7-2.7 mm with the thickest epidermis at the tubercular apex. The epidermis contained three distinct strata (=layers), including the stratum corneum, spinosum, and basale. The stratum corneum was significantly thickened in tubercles, over four times thicker than in the leading or trailing edge of the fin. The stratum spinosum, composed of lipokeratinocytes and lamellar oil bodies, was significantly thinner in the trailing edge than in the other two sites. There was no significant difference in the stratum basale among the three sites. Volume fraction of lipokeratinocytes was significantly higher at the sides of the leading edge and the apex of the tubercles, while volume fraction of lamellar oil bodies was significantly lower at the apex of the tubercles. Though the function of the tubercles is unknown, their position, hardened structure and increased epidermal stratum corneum suggest that they may have hydrodynamic importance.
    Journal of Morphology 01/2011; 272(1):27-33. · 1.60 Impact Factor
  • S Anne Boettger, James B McClintock
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    ABSTRACT: The common shallow-water sea urchin Lytechinus variegatus is capable of surviving inorganic phosphate exposures as high as 3.2 mg L(-1) and organic phosphate exposures of 1000 mg L(-1) . Nonetheless, chronic exposure to low, medium, and high-sublethal concentrations of organic phosphate inhibits the muscle enzyme acetyl cholinesterase (AChE), responsible for the break down of the neurotransmitter acetylcholine, as well as inhibiting contractions in the muscles associated with the Aristotle's lantern. AChE activity, measured in both a static enzyme assay and by vesicular staining, displayed concentration-dependent declines of activity in individuals maintained in organic phosphate for 4 weeks. The activity of AChE was not adversely affected by exposure to inorganic phosphate or seawater controls over the same time period. Maximum force of muscle contraction and rates of muscle contraction and relaxation also decreased with chronic exposure to increasing concentrations of organic phosphate. Chronic exposure to inorganic phosphates elicited no response except at the highest concentration, where the maximum force of muscular contraction increased compared to controls. These findings indicate that shallow-water populations of Lytechinus variegatus subjected to organic phosphate pollutants may display impaired muscular activity that is potentially related to the inhibition of the muscle relaxant enzyme AChE, and subsequently muscular overstimulation, and fatigue.
    Environmental Toxicology 07/2010; 27(4):193-201. · 2.71 Impact Factor
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    Celeste C Eno, Stefanie A Böttger, Charles W Walker
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    ABSTRACT: The purple sea urchin, Strongylocentrotus purpuratus, is the only non-chordate deuterostome model with a fully sequenced genome. Chromosomal localization of individual genes and resulting gene maps are unavailable for this or for any sea urchin. As a result, the purple sea urchin genome has not been mapped onto specific chromosomes and remains inaccessible to genome-wide approaches addressing questions that require positional information for particular genes. Here we describe the first successful methods for karyotyping and localizing specific gene loci on chromosomes of Strongylocentrotus purpuratus and those of the phylogenetically related Strongylocentrotus droebachiensis. Both species have 42 chromosomes in their diploid genomes (n = 21). There are 2 large, 8 medium, and 10 small pairs, plus one putative sex pair. In both species, bindin genes were localized to 2 pair of homologous chromosomes by fluorescent in situ hybridization. Fluorescently labeled bacterial artificial chromosome clones generated from S. purpuratus for the functionally related genes brachyury, foxa, and foxb were localized to different chromosomes. Our protocols provide previously unavailable tools for developing a gene map for the purple sea urchin genome.
    Biological Bulletin 12/2009; 217(3):306-12. · 1.23 Impact Factor
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    S Anne Böttger, James B McClintock
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    ABSTRACT: The sea urchin Lytechinus variegatus can survive chronic exposure to sodium phosphate (inorganic phosphate) concentrations as high as 3.2 mg L-1, and triethyl phosphate (organic phosphate) concentrations of 1000 mg L-1. However, chronic exposure to low (0.8 mg L-1 inorganic and 10 mg L-1 organic phosphate), medium (1.6 mg L-1 inorganic and 100 mg L-1 organic phosphate) or high (3.2 mg L-1 inorganic and 1000 mg L-1 organic phosphate) sublethal concentrations of these phosphates inhibit bactericidal clearance of the marine bacterium Vibrio sp. Bacteria were exposed to coelomic fluid collected from individuals maintained in either artificial seawater, or three concentrations of either inorganic phosphate or organic phosphate. Sterile marine broth, natural seawater and cell free coelomic fluid (cfCF) were employed as controls. Bacterial survival indices were measured at 0, 24 and 48 h periods once a week for four weeks. Bacteria were readily eliminated from the whole coelomic fluid (wCF) of individuals maintained in artificial seawater. Individuals maintained in inorganic phosphates were able to clear bacteria following a two week exposure period, while individuals maintained at even low concentrations of organic phosphates failed to clear all bacteria from their coelomic fluid. Exposure to phosphates represses antimicrobial defenses and may ultimately compromise survival of L. variegatus in the nearshore environment.
    Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology 08/2009; 150(1):39-44. · 2.71 Impact Factor
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    ABSTRACT: On the northeastern coast of the United States and Canada, Mya arenaria, the soft shell clam, develops a diffuse, hemopoetic tumor (a fatal leukemia-like cancer) resulting from inactivation of p53-like family member proteins.These malignant cells provide a model for an unrelated set of human cancer cells that are also characterized by mortalin-based cytoplasmic sequestration of wild-type p53 tumor suppressor protein (mortalin is the mitochondrial Hsp70 protein). Here we describe methods for mass culture and long-term storage of tumor cells from this cancer. These are the first successful efforts at maintaining malignant cells from any marine invertebrate in vitro. Following passage (subculture), these cultures undergo transition from primary cultures to non-immortalized cell lines that continue to proliferate and do not re-differentiate the normal hemocyte phenotype. We also characterize normal clam hemocytes and the pathology of cancerous clam hemocytes in vitro and in vivo using light and electron microscopy, cyto- and immunocytochemistry, molecular biology, and a phagocytosis assay. Our protocols provide biomedical and environmental researchers with ready access to this naturally occurring cancer model. We discuss the clam cancer model regarding (a) human health and disease; (b) animal health, disease, and aquaculture; (c) environmental health monitoring; and (d) future research directions.
    Biological Bulletin 03/2009; 216(1):23-39. · 1.23 Impact Factor
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    Charles W Walker, C.E. Eno, S.A. Boettger
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    ABSTRACT: Here we provide methods for generating triploid green sea urchin embryos (Strongylocentrotus droebachiensis). These are the first triploid sea urchins of any species that have been produced at any life stage and represent an initial step in a larger effort to yield triploid adult green sea urchins for large-scale land-based and near-shore commercial aquaculture ventures. Following fertilization, triploid mollusks and fish result by preventing release of the second polar body. Adult triploids in all of these organisms are sterile and may be larger than diploids based on accelerated growth of somatic tissues. Similar methods are impossible for sea urchins since haploid (1n) ova are released from the ovary after completion of both meiotic divisions. We have produced triploid green sea urchin embryos (3n=63) by fusing two 1n ova (each n=21) and fertilizing the diploid products of these fusions with highly diluted spermatozoa (n=21). Mechanical, proteolytic and acidic treatments were employed to remove the jelly coat and vitelline membrane prior to fusion. Results indicate that acidic removal of the extracellular structures, subsequent fusion with poly(Arg) in 1.5 ml microcentrifuge tubes and fertilization with highly diluted sperm yield 60.64±1.39% triploid embryos that reach prism pluteus stage (with spicules). Triploidy was verified at all developmental stages by counting chromosomes. We discuss scaling up our laboratory methods for embryos to the hatchery environment and also issues unique to the reproductive biology of sea urchins that will be vital for commercial aquaculture ventures to consider if triploid adult green sea urchins can ultimately be produced.
    Biological Bulletin 01/2009; 217:306-312. · 1.23 Impact Factor
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    Charles W Walker, Stefanie A Böttger
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    ABSTRACT: As Jessani et al.,(1) point out development of cell and animal models that accurately depict human tumorigenesis remains a major goal of cancer research. Clam cancer offers significant advantages over traditional models for genotoxic and non-genotoxic preclinical analysis of treatments for human cancers with a similar molecular basis. The naturally occurring clam model closely resembles an outbreeding, human clinical population and provides both in vitro and in vivo alternatives to those generated from inbred mouse strains or by intentional exposure to known tumor viruses. Fly and worm in vivo models for adult human somatic cell cancers do not exist because their adult somatic cells do not divide. Clam cancer is the best characterized, naturally occurring malignancy with a known molecular basis remarkably similar to those observed in several unrelated human cancers where both genotoxic and non-genotoxic strategies can restore the function of wild-type p53. To further emphasize this point of view, we here demonstrate a p53-induced, mitochondrial-directed mechanism for promoting apoptosis in the clam cancer model that is similar to one recently identified in mammals. Discerning the molecular basis for naturally occurring diseases in non-traditional models and correlating these with related molecular mechanisms responsible for human diseases is a virtually unexplored aspect of toxico-proteomics and genomics and related drug discovery.
    Cell cycle (Georgetown, Tex.) 09/2008; 7(15):2286-9. · 5.24 Impact Factor
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    Stefanie Böttger, Emily Jerszyk, Ben Low, Charles Walker
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    ABSTRACT: In nature, the soft shell clam, Mya arenaria, develops a fatal blood cancer in which a highly conserved homologue for wild-type human p53 protein is rendered nonfunctional by cytoplasmic sequestration. In untreated leukemic clam hemocytes, p53 is complexed throughout the cytoplasm with overexpressed variants for both clam homologues (full-length variant, 1,200-fold and truncated variant, 620-fold above normal clam hemocytes) of human mortalin, an Hsp70 family protein. In vitro treatment with etoposide only and in vivo treatment with either etoposide or mitoxantrone induces DNA damage, elevates expression (600-fold) and promotes nuclear translocation of p53, and results in apoptosis of leukemic clam hemocytes. Pretreatment with wheat germ agglutinin followed by etoposide treatment induces DNA damage and elevates p53 expression (893-fold) but does not overcome cytoplasmic sequestration of p53 or induce apoptosis. We show that leukemic clam hemocytes have an intact p53 pathway, and that maintenance of this tumor phenotype requires nuclear absence of p53, resulting from its localization in the cytoplasm of leukemic clam hemocytes. The effects of these topoisomerase II poisons may result as mortalin-based cytoplasmic tethering is overwhelmed by de novo expression of p53 protein after DNA damage induced by genotoxic stress. Soft shell clam leukemia provides excellent in vivo and in vitro models for developing genotoxic and nongenotoxic cancer therapies for reactivating p53 transcription in human and other animal cancers displaying mortalin-based cytoplasmic sequestration of the p53 tumor suppressor, such as colorectal cancers and primary and secondary glioblastomas, though not apparently leukemias or lymphomas.
    Cancer Research 03/2008; 68(3):777-82. · 8.65 Impact Factor
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    Charles Walker, Stefanie Böttger, Ben Low
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    ABSTRACT: In nature the soft shell clam Mya arenaria develops a fatal neoplasm that shares molecular similarity with an unrelated group of human cancers. In leukemic clam hemocytes, wild-type p53 and mortalin proteins co-localize in the cytoplasm. A similar phenotype, characterized by cytoplasmic sequestration of wild-type p53 protein, has been observed in several human cancers (undifferentiated neuroblastoma, retinoblastoma, colorectal and hepatocellular carcinomas, and glioblastoma). In some of these cancers p53 is tethered in the cytoplasm by mortalin when the latter protein is overexpressed. Using co-immunoprecipitation we have demonstrated that mortalin and p53 proteins are complexed in the cytoplasm of leukemic clam hemocytes (and not in normal hemocytes). In addition, treatment of leukemic clam hemocytes with MKT-077, a cationic inhibitor of mortalin, disrupts the interaction of mortalin and p53 proteins, resulting in translocation of some p53 to the nucleus. Based on these data, we introduce leukemic clam hemocytes as novel and easily accessible, in vivo and in vitro models for human cancers displaying a similar mortalin-based phenotype. Treatment of these models with novel chemotherapeutics may help reveal the molecular mechanism(s) involved in inactivating p53 by this form of cytoplasmic sequestration.
    American Journal Of Pathology 06/2006; 168(5):1526-30. · 4.52 Impact Factor
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    Charles W Walker, M.G. Devin, S.A. Boettger
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    ABSTRACT: and American markets and maximum US sales of $150M US dollars in 1996. Wild populations of sea urchins on all coasts of the US have been dramatically over-fished. Aquaculture of sea urchins in land-based facilities can help restore commercial populations and preserve this ecologically important herbivore. In this study, we used invariant summer photoperiod to prevent gametogenesis in the North American green sea urchin (Strongylocentrotus droebachiensis) maintained in a land-based aquaculture system and provided a commercially available formulated feed that promotes maximum growth of intra-gonadal somatic nutrient storage cells called nutritive phagocytes. Results were compared with individuals fed the same formulated feed under ambient photoperiod in cages in the ocean. Monthly samples of gonads from both treatments were evaluated for gonad index, volume fractions of cellular constituents of the germinal epithelium, oocyte diameters and taste. Over the 5 months of this study, gonad indices increased significantly ( pb0.001) in both treatments from 4.8%±0.9 (all values±SE) initially to 20.5%±2.1 under invariant and 23.2%±1.4 under ambient photoperiod with no significant difference between treatments ( p=0.55). Volume fractions of nutritive phagocytes increased to 80.3%±5.9 (initial 37.9%±7.1) in males and 71.0%±6.7 (initial 10.3%±4.0) in females ( pb0.001) only under invariant photoperiod. Nutritive phagocyte lengths increased under both photoperiod treatments, but the volume fraction containing nutrients was higher under invariant photoperiod. Volume fractions of gonial/gametogenic cells increased significantly ( pb0.001) only under ambient photoperiod from 20.4%±5.5 to 37.8%±1.8 in males and 0% to 22.6%±3.6 in females. The volume fraction of residual oocytes from last year's oogenesis increased under invariant photoperiod while that of both residual and new oocytes increased under ambient photoperiod. Residual oocyte diameters increased from 56.2 μm±2.2 initially to 93.5 μm±3.7 under invariant and those of residual and new oocytes to 126.0 μm±7.3 under ambient photoperiod. Invariant photoperiod yields gonads in both sexes of S. droebachiensis that do not initiate fall gametogenesis but attain large size as their nutritive phagocytes grow substantially in size. A Canadian study of wild-collected S. droebachiensis indicated that gonads taste best when they contain pre-dominantly nutritive phagocytes and not copious gametes, however gonad taste in our study was unsatisfactory suggesting that the only commercially available sea urchin diet requires modification to support commercial development of land-based aquaculture.
    Aquaculture 01/2006; 261:1422-1431. · 2.01 Impact Factor
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    S Anne Böttger, Charles W Walker, Tatsuya Unuma
    Methods in cell biology 02/2004; 74:17-38. · 1.44 Impact Factor

Publication Stats

127 Citations
54.54 Total Impact Points

Institutions

  • 2008–2011
    • University of New Hampshire
      • • Department of Molecular, Celluar and Biomedical Sciences
      • • Department of Zoology
      Durham, NC, United States
  • 2009–2010
    • West Chester University
      • Department of Biology
      West Chester, PA, United States
  • 2001–2002
    • University of Alabama at Birmingham
      • Department of Biology
      Birmingham, AL, United States