Access to abstract permanently restricted to Ball State community only. / Access to thesis permanently restricted to Ball State community only. / Department of Physiology and Health Science

Access to abstract permanently restricted to Ball State community only. / Access to thesis permanently restricted to Ball State community only. / Department of Physiology and Health Science

Mouse models (that develop or can be stimulated to develop lymphomas) are used to examine cancer-related processes. Mouse models can be effective tools used to identify new, early, and pre-malignant markers of lymphomas. Signal Transducer and Activator of Transcription (STAT) 6 is a transcription factor activated through the Jak-Stat pathway. Transgenic mice expressing a constantly activated Stat6 (Stat6VT) were previously generated and characterized to have altered lymphocyte homeostasis. Some of these Stat6VT mice developed a lymphoproliferative disorder (LPD). LPD, including lymphomas, develops when lymphocytes are overproduced or act abnormally. These Stat6VT mice may serve as a model for examining lymphoma development. In order to characterize the altered lymphocytes and determine if LPD observed in the Stat6VT mice is characteristic of lymphoma, RT-PCR analysis and Western analysis were done to examine if the presence of Stat6VT alters the expression of the cell cycle genes Gfi-1 and Gfi-1b and if these genes differ in LPD Stat6VT verses control mice. / Department of Biology

No description available.

Brain deposition of the $ beta$-amyloid protein is an early marker of Alzheimer's disease (AD). AD is a neurodegenerative disorder characterized by learning and memory impairments. Here, mice (B6C3, 8 and 20 months old) transgenic for a human $ beta$-amyloid fragment were compared to normal litter mates in spatial and non-spatial learning tasks in the Morris water maze, according to standard procedures. Four measures of learning and performance were analyzed statistically: latency, total distance swam, mean distance to a platform, and number of trials correct in reaching a platform. Transgenic mice were impaired relative to their litter mates in spatial learning and performed better in the non-spatial task than in the spatial task in the first three measures. An age effect for transgenics was observed in the total distance measure. The results suggest that expression of the human $ beta$-amyloid protein may produce a selective learning deficit in mice.

We investigated the role of two intracellular second messengers, extracellular signal-regulated protein kinase (ERK), and protein kinase C (PKC) in a model of persistent pain, using intrathecal (i.t) (R,S )-DHPG to induce spontaneous nociceptive behaviours (SNBs). SNBs were measured in animals that were treated with an ERK inhibitor (PD 98059), and a PKC inhibitor (GF 109203X) compared with controls. Mechanical allodynia, was measured using paw withdrawal thresholds in the von Frey test, and thermal hyperalgesia was measured using response latencies in the plantar test. In study 1, it was shown that spinal administration of PD 98059, dose-dependently decreased SNBs, and reduced mechanical allodynia and thermal hyperalgesia. In study 2, it was shown that i.t. pretreatment with the GF 109203X, reduced SNBs and thermal hyperalgesia, but not mechanical allodynia. These results suggest that both ERK and PKC are involved in SNBs and the concomitant and thermal hyperalgesia and possibly mechanical allodynia.

Zebrafish (Brachydanio rerio) are small, freshwater teleost fishes in the family Cyprinidae, the true minnows. They are native to the tropical latitudes of India, but have become widespread through their use as aquarium fish and as models for several branches of biological research. Their ease of rearing, short generation time, year-around egg laying potential, brief developmental period, and embryo transparency have made them especially desirable as models for developmental biology, genetics, and neurobiology. Because of their popularity, they were also the first small aquarium fish to be used as test organisms for carcinogenesis in the early 1960's. For reasons that have never been stated, their use as a model for carcinogenesis research did not continue. Due to the number of positive characteristics that this species has, the goal of this research effort was to systematically evaluate the potential of zebrafish for use as an environmental monitor, to evaluate the toxicology and carcinogenesis of surface and/or ground waters. The overall project was multidisciplinary in nature, but the focus of this thesis research was on the whole animal, dose-response to a number of well-known carcinogens, administered by multiple exposure routes, and the pathological description of the resulting lesions. Exposure to N-nitrosodiethylamine (DEN) and N-nitrosodimethylamine (DMN) in the diet was ineffective, but static water bath exposure of fry and embryos to these nitrosamines resulted in neoplasms, primarily in the liver. Embryo exposure to DEN resulted in a low response of neoplasms in several other organs as well. Dietary exposure of zebrafish to aflatoxin B₁ resulted in few hepatic neoplasms, revealing a marked resistance to this carcinogen. Dietary exposure to methylazoxymethanol acetate (MAM-Ac) produced mostly liver tumors, as did both fry and embryo water bath exposures. Each water bath exposure also produced neoplasms at other tissue and organ sites, but the embryo stage produced the greatest variety. These results demonstrate a relative resistance to neoplastic development compared to the well-known rainbow trout model. But in one comparative trial, zebrafish were similar to Japanese medaka in their response to dietary MAM-Ac. The major limitation of this species, that will prevent its use as a model for environmental monitoring, however, is its narrow range of temperature tolerance. Temperatures below 15°C produce marked sluggishness, and below 10-12°C cause anesthesia and death. Therefore, this research indicates that this species is not as versatile as some other small fish species for laboratory and especially field monitoring of environmental carcinogenic hazards. / Graduation date: 1997

Thesis (Ph. D.)--University of Hawaii at Manoa, 2004. / Includes bibliographical references (leaves 92-105). / Also available by subscription via World Wide Web / xv, 105 leaves, bound ill. 29 cm

Transcription factors GATA-1, Fli-1 and FOG-1 are essential proteins for normal megakaryopoiesis, however, the detailed analyses of their functions within developmental stages of megakaryopoiesis are lacking. In my thesis, over expression of gene in target cells was adopted as the main strategy to study the biological functions of these proteins, therefore, an efficient gene delivery method was first developed by using retrovirus.This approach was then utilized to over express GATA-1, Fli-1 and FOG-1 in murine leukemia M1 cells and mouse hematopoietic stem cells (HSCs), and their effects on different developmental stages of megakaryopoiesis were investigated. In the transduced M1 cells, enforced expression of GATA-1 and Fli-1 was found to induce the megakaryocytic development, which was associated with the formation of megakaryocyte (Mk) and the increased expression of Mk specific genes c-Mpl and GPIX. In the transduced mouse HSCs, it was found that the expression of endogenous GATA-1, Fli-1 and FOG-1 was up-regulated throughout Mk differentiation; enforced expression of these transcription factors led to the significantly enhanced Mk development. Megakaryocytes over expressing GATA-1, Fli-1 and FOG-1 were characterized by the increased expression of various Mk-specific genes including GPIX, c-Mpl, platelet factor 4 (PF4), acetylcholinesterase (AChE) and NF-E2, an important transcription factor for terminal megakaryopoiesis; however, GATA-1, Fli-1 and FOG-1 displayed the different abilities in promoting the proliferation of hematopoietic cells and MK differentiation, as well as regulating other transcription factors involved in hematopoiesis. To further elucidate the role of the functional domains of Fli-1, various mutants of Fli-1 were also over expressed in mouse HSCs. The results demonstrated that first, the combination of the activation domain of Fli-1 and its Ets domain is required for early megakaryopoiesis but not sufficient for terminal megakaryopoiesis; second, DNA binding of Fli-1 was not the only requirement for early Mk enhancement, moreover, the interaction between Fli-1 and GATA-1 through the Ets domain and the resultant transcriptional synergy was the essential determinant for Fli?1 ability in Mk development. Taken together, the studies presented in this thesis provided strong in vitro evidence that GATA-1, Fli-1 and FOG-1 indeed play the critical roles in normal megakaryopoiesis.

Idiopathic Membranous nephropathy (MN) is a common cause of nephrotic syndrome in humans, and many patients progress to end-stage kidney disease. The best available animal model of MN is active Heymann nephritis (HN) in which rats are immunized with renal tubular antigen (RTA) in complete Freund's adjuvant (CFA). Rats develop heavy proteinuria, a key measure of glomerular damage, and the disease is histologically identical to human MN. It has been thought that HN is mediated by antibody-based mechanisms. More recent evidence demonstrates a critical role for cytotoxic T cells. This thesis aims to further examine the role of T cell responses in active HN. First, the effect of the anti-CD3 monocIonal antibody (mAb) G4.18 was investigated. Anti-CD3 given 4 weeks after immunization prevented the development of proteinuria, delayed anti-RTA antibody responses, and reduced glomerular infiltration of CD8+ T cells and macrophages, but did not affect glomerular deposition of IgG or complement. Increased mRNA expression of the Th2 cytokines IL-4 and IL-5 was detected in draining lymph nodes. These findings suggest that immune deviation to a Th2 response reduces glomerular injury in HN. Second, the role of CD4+ T cells in immune tolerance was examined. Rats were given RTA in incomplete Freund's adjnvant (lFA) to induce tolerance to RTA, and three weeks later were immunized with RTA in CFA. Anti-CD4 mAb therapy at the time of RTA1IFA treatment had no effect on subsequent proteinuria or anti-RTA autibodies. Third, the role of IL-4 in this model of immune tolerance was examined. Anti-IL-4 mAb therapy blocked the induction of tolerance, and led to the development of proteinuria. Finally, the effect of treatment with IL-4 and IL-5 was examined. Treatment with these cytokines separately or together after immunization blocked the development of proteinuria, without a consistent effect on anti-RTA antibodies. These results demonstrate a central role for T cell regulation in HN, and show that immune deviation to a Th2 response is protective against glomerular injury. The findings may have implications in the future for focused therapeutic intervention in human idiopathic MN.