Depuis plusieurs dizaines d’années, la population des anguilles européennes a souffert d’un dramatique déclin et est classée parmi les espèces hors de leurs limites biologiques depuis 1998. Différents phénomènes, tels que la pêche, les obstacles aux migrations, ou le réchauffement climatique, en sont à l’origine. Cependant, les perturbations environnementales, telles que les contaminations métalliques et organiques ou les épisodes d’hypoxie, participent probablement à la vulnérabilité de cette espèce. Or, l’estuaire de la Gironde est soumis à des contaminations poly-métalliques historiques, ainsi qu’à des contaminations organiques de différentes origines, et à des épisodes hypoxiques réguliers. Les travaux menés au cours de cette thèse, se sont donc composés d’études de terrain visant à identifier les contaminants majeurs chez les anguilles jaunes de l’estuaire de la Gironde, ainsi que leur voie de bio-accumulation et leurs impacts au niveau physiologique, biochimique et moléculaire. Ces études ont nécessité la mise en place d’expériences préliminaires ayant pour objectifs de vérifier les impacts de certaines procédures, telles que l’anesthésie et la mise en cage des anguilles jaunes. D’autre part, les impacts des deux principaux contaminants métalliques et organiques des anguilles européennes de l’estuaire de la Gironde, ont été testés, de façon individuelle et combinée, ainsi que ceux de l'hypoxie sur des anguilles pré-contaminées ou non, lors d’une étude expérimentale.Ainsi, ces travaux ont mis en évidence que les anguilles européennes installées dans la zone avale de l’estuaire, étaient susceptibles d’être soumises à des contaminations poly-métalliques plus importantes, essentiellement d’origine trophique, responsables d’une croissance pondérale moins élevée et de perturbations transcriptionnelles hépatiques et cérébrales. De plus, bien que les contaminations métalliques de ces anguilles ne mettent pas ne danger la santé humaine, les contaminations en PCB sont, en revanche, supérieures aux normes de consommation. D’autre part, ces travaux ont également mis en évidence des dérèglements mitochondriaux ainsi qu’un stress oxydant, au niveau branchial et cérébral chez des anguilles contaminées au Cd, et au niveau cérébral, branchial, hépatique et rénal chez les anguilles contaminées aux PCB. Enfin, la concomitance de ces deux contaminants et/ou d’un épisode d’hypoxie, réduit et/ou retarde les capacités de réponses transcriptionnelles de ces anguilles. Il semble donc que les différentes perturbations chimiques subies par les anguilles européennes au stade jaune au sein de l’estuaire de la Gironde participent de façon non négligeable à la vulnérabilité de cette espèce. / For several decades, the European eel has been suffering from a dramatic decline and has been classified among species beyond their biological limit since 1998. Different phenomena, as fishing, migration barriers, or global warming, are to blame. Environmental perturbations, as metallic and organic contaminations, or hypoxic episodes, probably take part to the vulnerability of this species. The Gironde estuary has been submitted to historic poly-metallic contaminations, to organic contaminations from different origins, and to regular hypoxic episodes. This thesis work is composed of field studies aimed to identify the main contaminants in yellow eel living in the Gironde estuary, their major bioaccumulation way and impacts on physiological, biochemical and molecular parameters. These studies needed preliminary experiments aimed to verify whether some field and handling procedures are consistent with ecotoxicological analyses. Moreover, impacts of the two main contaminants in European eels from the Gironde estuary have been assessed individually or combined, with those of hypoxia on pre-contaminated eels.Thus, these studies have demonstrated that European eels installed in the downstream area of the estuary, were likely to be subject to largest poly-metallic contaminations, mostly by food web, and responsible for a less weight gain and high liver and brain transcriptional disturbances. Furthermore, although the metal contamination of eels does not endanger human health, PCB contaminations are higher than consumption standards. On the other hand, these studies have also revealed oxidative stress and mitochondrial disorders in gills and brain of Cd-contaminated eels, and in brain, gills, liver and kidneys of PCB-contaminated eels. The combination of these two contaminants and/or an episode of hypoxia, reduces and/or delays the transcriptional responses ability of these eels. It seems that the different chemical disturbances, suffered by the yellow eels in the estuary of the Gironde, participate significantly to this species vulnerability.

A luz ultravioleta (UV) bloqueia a replicação e transcrição devido à formação de lesões que distorcem o DNA. Descobrimos que a depleção da quinase ATR promove a indução precoce de apoptose após irradiação com luz UVB em fibroblastos humanos imortalizados com SV40 e que mesmo células proficientes em reparo de DNA e síntese translesão foram incapazes de alcançar a mitose após depleção de ATR. Essa quinase também representa um alvo promissor para sensibilizar tumores com mutações em p53 ao quimioterápico cisplatina e ao indutor de estresse oxidativo cloroquina. Além do bloqueio da replicação, danos no DNA bloqueiam a síntese de RNA. Utilizamos sequenciamento para mapear RNA nascentes e analisar a recuperação da transcrição em escala genômica. Genes mais longos são mais inibidos por luz UV, mas o nível de expressão gênica não contribui para a recuperação da transcrição. Além disso, o reparo de DNA é similar entre genes com recuperação da transcrição distinta e outras regulações, além da remoção de lesões no DNA, devem existir para que a síntese de RNA recomece. / Ultraviolet (UV) light stalls replication and transcription due to the formation of lesions that distort DNA. We found that ATR silencing promotes early induction of apoptosis after UVB light in human fibroblasts immortalized with SV40 and even cells proficient in DNA repair and translesion synthesis were unable to reach mitosis after ATR depletion. This kinase is also a promising target for sensitizing tumors with p53 mutations to chemotherapeutic that block replication, such as cisplatin, and the oxidative stress inducer chloroquine. In addition to blocking the replication, DNA damage arrest the synthesis of RNA. We used next-generation sequencing to map and analyze the nascent RNA transcription recovery genome-wide. We confirmed that longer genes are more inhibited following UV light, however, the level of gene expression does not contribute to the recovery of transcription. Moreover, DNA repair is similar among genes with different recovery of transcription and further regulation, besides DNA damage removal, must exist to promote resumption of RNA synthesis.

Changes in synaptic strength underlie synaptic plasticity, the cellular substrate for learning and memory. Disruptions in the mechanisms that regulate synaptic strength closely link to many developmental, neurodegenerative and neurological disorders. Release site probability (PAZ) and active zone number (N) are two important presynaptic determinants of synaptic strength; yet, little is known about the processes that establish the balance between N and PAZ at any synapse. Furthermore, it is not known how PAZ and N are rebalanced during synaptic homeostasis to accomplish circuit stability. To address this knowledge gap, we adapted a neurophysiological experimental system consisting of two functionally differentiated glutamatergic motor neurons (MNs) innervating the same target. Average PAZ varied between nerve terminals, motivating us to explore benefits for high and low PAZ, respectively. We speculated that high PAZ confers high-energy efficiency. To test the hypothesis, electrophysiological and ultrastructural measurements were made. The terminal with the highest PAZ released more neurotransmitter but it did so with the least total energetic cost. An analytical model was built to further explore functional and structural aspects in optimizing energy efficiency. The model supported that energy efficiency optimization requires high PAZ. However, terminals with low PAZ were better able to sustain neurotransmitter release. We suggest that tension between energy efficiency and stamina sets PAZ and thus determines synaptic strength. To test the hypothesis that nerve terminals regulate PAZ rather than N to maintain synaptic strength, we induced sustained synaptic homeostasis at the nerve terminals. Ca2+ imaging revealed that terminals of the MN innervating only one muscle fiber utilized greater Ca2+ influx to achieve compensatory neurotransmitter release. In contrast, morphological measurements revealed that terminals of the MN inner vating multiple postsynaptic targets utilized an increase in N to achieve compensatory neurotransmitter release, but this only occurred at the terminal of the affected postsynaptic target. In conclusion, this dissertation provides several novel insights into a prominent question in neuroscience: how is synaptic strength established and maintained. The work indicates that tension exists between energy efficiency and stamina in neurotransmitter release likely influences PAZ. Furthermore, PAZ and N are rebalanced differently between terminals during synaptic homeostasis. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2015. / FAU Electronic Theses and Dissertations Collection

Leung Wan-chi. / Thesis submitted in: November 2005. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 276-284). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese Version) --- p.v / Acknowledgements --- p.viii / Tables of Contents --- p.ix / List of Abbreviations --- p.xxx / List of Figures --- p.xxxiii / List of Tables --- p.xlii / Chapter Chapter 1 --- Literature review --- p.1 / Chapter 1.1 --- Peroxisome proliferator activator receptors --- p.1 / Chapter 1.2 --- Peroxisome proliferators --- p.6 / Chapter 1.2.1 --- Hepatomegaly --- p.9 / Chapter 1.2.2 --- Peroxisome proliferation --- p.11 / Chapter 1.2.3 --- Target genes regulation --- p.12 / Chapter 1.2.4 --- Hypolipidemic effect --- p.16 / Chapter 1.2.5 --- Hepatocarcinogenesis --- p.18 / Chapter 1.3 --- Mode of actions --- p.20 / Chapter 1.3.1 --- Oxidative stress --- p.21 / Chapter 1.3.2 --- Inhibition of apoptosis --- p.22 / Chapter 1.3.2 --- Increase in cell replication --- p.22 / Chapter 1.3.4 --- Alterations in cell cycle control --- p.23 / Chapter 1.4 --- Objectives --- p.23 / Chapter Chapter 2 --- Materials and Methods --- p.25 / Chapter 2.1 --- Animal tail-genotyping --- p.25 / Chapter 2.1.1 --- Materials --- p.25 / Chapter 2.1.2 --- Methods --- p.28 / Chapter 2.2 --- Animal treatment --- p.29 / Chapter 2.2.1 --- Materials --- p.29 / Chapter 2.2.2 --- Methods --- p.29 / Chapter 2.3 --- Serum cholesterol and tryiglyceride analysis --- p.30 / Chapter 2.3.1 --- Materials --- p.31 / Chapter 2.3.2 --- Methods --- p.31 / Chapter 2.3.2.1 --- Serum preparation --- p.31 / Chapter 2.3.2.2 --- Serum cholesterol analysis --- p.31 / Chapter 2.3.2.3 --- Serum triglyceride analysis --- p.32 / Chapter 2.4 --- Histological analysis --- p.32 / Chapter 2.4.1 --- Materials --- p.32 / Chapter 2.4.2 --- Methods --- p.33 / Chapter 2.5 --- Total RNA isolation --- p.34 / Chapter 2.5.1 --- Materials --- p.34 / Chapter 2.5.2 --- Methods --- p.34 / Chapter 2.6 --- DNase I treatment of total liver RNA --- p.37 / Chapter 2.6.1 --- Materials --- p.37 / Chapter 2.6.2 --- Methods --- p.37 / Chapter 2.7 --- Reverse transcription (RT) of mRNA and non- fluorescent PCR (non-fluoroDD PCR) --- p.38 / Chapter 2.7.1 --- Materials --- p.43 / Chapter 2.7.2 --- Methods --- p.43 / Chapter 2.8 --- Reverse transcription (RT) of mRNA and fluorescent PCR (fluoroDD PCR) --- p.44 / Chapter 2.8.1 --- Materials --- p.44 / Chapter 2.8.2 --- Method --- p.44 / Chapter 2.9 --- Fluorescent differential display (fluoroDD) --- p.45 / Chapter 2.9.1 --- Materials --- p.45 / Chapter 2.9.2 --- Methods --- p.45 / Chapter 2.9.2.1 --- FluoroDD gel preparation --- p.45 / Chapter 2.9.2.2 --- Sample preparation and electrophoresis --- p.45 / Chapter 2.10 --- Excision of differentially expressed cDNA fragments --- p.46 / Chapter 2.10.1 --- Materials --- p.46 / Chapter 2.10.2 --- Methods --- p.46 / Chapter 2.11 --- Reamplification of differentally expressed cDNA fragments --- p.48 / Chapter 2.11.1 --- Materials --- p.48 / Chapter 2.11.2 --- Methods --- p.50 / Chapter 2.12 --- Subcloning of reamplified cDNA fragmens --- p.50 / Chapter 2.12.1 --- Materials --- p.53 / Chapter 2.12.2 --- Methods --- p.53 / Chapter 2.12.2.1 --- Ligation --- p.53 / Chapter 2.12.2.2 --- Transformation --- p.53 / Chapter 2.12.2.3 --- Phenol-choloroform extraction --- p.54 / Chapter 2.12.2.4 --- Confirmation of insert size by EcoRI digestion --- p.54 / Chapter 2.12.2.5 --- Mini-preparation of plasmid DNA from recombinant clones --- p.55 / Chapter 2.13 --- Sequencing of subcloned cDNA fragments --- p.55 / Chapter 2.13.1 --- Materials --- p.56 / Chapter 2.13.2 --- Methods --- p.56 / Chapter 2.13.2.1 --- Sequencing of fluoroDD cDNA fragments --- p.56 / Chapter 2.13.2.2 --- Blast search against computer database --- p.57 / Chapter 2.14 --- Northern blot analysis of sequenced cDNA fragments --- p.57 / Chapter 2.14.1 --- Materials --- p.58 / Chapter 2.14.2 --- Methods --- p.58 / Chapter 2.14.2.1 --- Formaldehyde agarose gel electrophoresis of total RNA --- p.58 / Chapter 2.14.2.2 --- Preparation of DIG-labeled RNA probes for hybridization --- p.59 / Chapter 2.14.2.3 --- Preparation of PCR DIG-labeled cDNA probes for hybridization --- p.60 / Chapter 2.14.2.4 --- Hybridization and colour development --- p.60 / Chapter Chapter 3 --- Results --- p.62 / Chapter 3.1 --- Confirmation of genotypes by PCR --- p.62 / Chapter 3.2 --- Body weight changes --- p.62 / Chapter 3.3 --- Organ weight changes --- p.67 / Chapter 3.4 --- Serum cholesterol and triglyceride levels --- p.70 / Chapter 3.5 --- Liver histology --- p.78 / Chapter 3.6 --- Reverse transcription (RT) of mRNA and non-fluorescent PCR (non-flurroDD PCR) --- p.114 / Chapter 3.7 --- Reverse transcription (RT) of mRNA and fluorescent PCR (fluoroDD PCR) --- p.125 / Chapter 3.8 --- Reamplification of fluorescent differential display (FDD) fragments --- p.138 / Chapter 3.9 --- Subcloning of reamplifled FDD fragments --- p.162 / Chapter 3.10 --- Sequencing of subcloned cDNA fragments --- p.176 / Chapter 3.11 --- Northern blot analysis of sequenced cDNA fragments --- p.195 / Chapter Chapter 4 --- Discussion --- p.250 / Chapter 4.1 --- Body weight changes --- p.250 / Chapter 4.2 --- Organ weight changes --- p.251 / Chapter 4.3 --- Serum cholesterol and triglyceride levels --- p.253 / Chapter 4.4 --- Liver histology --- p.254 / Chapter 4.5 --- "Functions and roles of identified PPARa-dependent and Wy-14,643- responsive genes" --- p.255 / Chapter 4.6 --- Mechanism of PP-induced hepatocarcinogeneis --- p.270 / Chapter Chapter 5 --- Conclusions --- p.274 / References --- p.276 / Appendix A Tables of preparation of reaction mix --- p.285 / Table A1. Preparation of animal tail genotyping PCR reaction --- p.285 / Table A2. Preparation of DNase I treatment --- p.285 / Table A3. Preparation of reverse transcription of non-fluoroDD and fluoroDD --- p.285 / Table A4. Preparation of non-fluoroDD and fluoroDD RT-PCR --- p.286 / Table A5. Preparation of reamplification of differentially expressed cDNA fragments --- p.286 / Table A6. Preparation of PCR reaction for DNA sequencing --- p.286 / Table A7. Preparation of PCR reaction for RNA probe --- p.287 / Table A8. Preparation of PCR reaction for cDNA probe --- p.287 / Appendix B DNA sequences and sequencing alignments of FluoroDD Fragments --- p.288 / Chapter B 1.1: --- DNA sequence of cDNA subclone AA1#2 (AP1 & ARP2) using M13 forward (-20) primer --- p.288 / Chapter B 1.2: --- "Sequencing alignment of cDNA subclone AA1#2 with mouse peroxisomal delta 3, delta 2-enoyl-Coenzyme A isomerase (Peci) by BLAST searching against the National Center for Biotechnology Information database" --- p.288 / Chapter B 1.3: --- Summary of sequence alignment of cDNA subclone AA1#2 with mouse Peci --- p.288 / Chapter B 2.1: --- DNA sequence of cDNA subclone AA1#3 (AP1 & ARP2) using M13 forward (-20) primer --- p.289 / Chapter B 2.2: --- "Sequencing alignment of cDNA subclone AA1#3 with mouse peroxisomal delta 3, delta 2-enoyl-Coenzyme A isomerase (Peci) by BLAST searching against the National Center for Biotechnology Information database" --- p.289 / Chapter B 2.3: --- Summary of sequence alignment of cDNA subclone AA1#3 with mouse Peci --- p.289 / Chapter B 3.1: --- DNA sequence of cDNA subclone AA1#4 (AP 1 & ARP2) using Ml3 reverse primer --- p.290 / Chapter B 3.2: --- "Sequencing alignment of cDNA subclone AA1#4 with mouse peroxisomal delta 3, delta 2-enoyl-Coenzyme A isomerase (Peci) by BLAST searching against the National Center for Biotechnology Information database" --- p.290 / Chapter B 3.3: --- Summary of sequence alignment of cDNA subclone AA1#4 with mouse Peci --- p.290 / Chapter B 4.1: --- DNA sequence of cDNA subclone AA1#20 (AP 1 & ARP2) using Ml3 forward (-20) primer --- p.291 / Chapter B 4.2: --- "Sequencing alignment of cDNA subclone AA1#20 with mouse peroxisomal delta 3, delta 2- enoyl-Coenzyme A isomerase (Peci) by BLAST searching against the National Center for Biotechnology Information database" --- p.291 / Chapter B 4.3: --- Summary of sequence alignment of cDNA subclone AA1#20 with mouse Peci --- p.291 / Chapter B 5.1: --- DNA sequence of cDNA subclone AA4#1 (AP 1 & ARP2) using Ml3 forward (-20) primer --- p.292 / Chapter B 5.2: --- Sequencing alignment of cDNA subclone AA4#1 with mouse apolipoprotein A-V (Apoa5) by BLAST searching against the National Center for Biotechnology Information database --- p.292 / Chapter B 5.3: --- Summary of sequence alignment of cDNA subclone AA4#1 with mouse Apoa5 --- p.292 / Chapter B 6.1: --- DNA sequence of cDNA subclone AA4#9 (AP 1 & ARP2) using Ml3 reverse primer --- p.293 / Chapter B 6.2: --- Sequencing alignment of cDNA subclone AA4#9 with mouse apolipoprotein A-V (Apoa5) by BLAST searching against the National Center for Biotechnology Information database --- p.293 / Chapter B 6.3: --- Summary of sequence alignment of cDNA subclone AA4#9 with mouse Apoa5 --- p.293 / Chapter B 7.1: --- DNA sequence of cDNA subclone AA5#5 (AP 1 & ARP2) using Ml3 forward (-20) primer --- p.294 / Chapter B 7.2: --- Sequencing alignment of cDNA subclone AA5#5 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.294 / Chapter B 7.3: --- Summary of sequence alignment of cDNA subclone AA5#5 with mouse mitochondrion --- p.294 / Chapter B 8.1: --- DNA sequence of cDNA subclone AA6#1 (AP1 & ARP2) using Ml3 forward (-20) primer --- p.295 / Chapter B 8.2: --- Sequencing alignment of cDNA subclone AA6#1 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.295 / Chapter B 8.3: --- Summary of sequence alignment of cDNA subclone AA6#1 with mouse mitochondion --- p.295 / Chapter B 9.1: --- DNA sequence of cDNA subclone AA6#9 (AP 1 & ARP2) using Ml3 reverse primer --- p.296 / Chapter B 9.2: --- Sequencing alignment of cDNA subclone AA6#9 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.296 / Chapter B 9.3: --- Summary of sequence alignment of cDNA subclone AA6#9 with mouse mitochondrion --- p.296 / Chapter B 10.1: --- DNA sequence of cDNA subclone AA7#3 (AP 1 & ARP2) using Ml3 forward (-20) primer --- p.297 / Chapter B 10.2: --- Sequencing alignment of cDNA subclone AA7#3 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.297 / Chapter B 10.3: --- Summary of sequence alignment of cDNA subclone AA7#3 with mouse mitochondrion --- p.297 / Chapter B 11.1: --- DNA sequence of cDNA subclone AA7#5 (AP 1 & ARP2) using Ml3 reverse primer --- p.298 / Chapter B 11.2: --- Sequencing alignment of cDNA subclone AA7#5 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.298 / Chapter B 11.3: --- Summary of sequence alignment of cDNA subclone AA7#5 with mouse mitochondrion --- p.298 / Chapter B 12.1: --- DNA sequence of cDNA subclone AA10#1 (AP1 & ARP2) using M l3 forward (-20) primer --- p.299 / Chapter B 12.2: --- Sequencing alignment of cDNA subclone AA10#1 with mouse cysteine sulfinic acid decarboxylase (Csad) by BLAST searching against the National Center for Biotechnology Information database --- p.299 / Chapter B 12.3: --- Summary of sequence alignment of cDNA subclone AA10#1 with mouse Csad --- p.299 / Chapter B 13.1: --- DNA sequence of cDNA subclone AA10#1 (AP 1 & ARP2) using M13 reverse primer --- p.300 / Chapter B 13.2: --- Sequencing alignment of cDNA subclone AA10#1 with mouse cysteine sulfinic acid decarboxylase (Csad) by BLAST searching against the National Center for Biotechnology Information database --- p.300 / Chapter B 13.3: --- Summary of sequence alignment of cDNA subclone AA10#1 with mouse Csad --- p.300 / Chapter B 14.1: --- DNA sequence of cDNA subclone AA12#4 (AP1 & ARP2) using Ml3 forward (-20) primer --- p.301 / Chapter B 14.2: --- "Sequencing alignment of cDNA subclone AA12#4 with mouse acetyl-coenzyme A dehydrogenase, medium chain (MCAD) by BLAST searching against the National Center for Biotechnology Information database" --- p.301 / Chapter B 14.3: --- Summary of sequence alignment of cDNA subclone AA12#4 with mouse MCAD --- p.301 / Chapter B 15.1: --- DNA sequence of cDNA subclone AA12#4 (AP 1 & ARP2) using Ml3 reverse primer --- p.302 / Chapter B 15.2: --- "Sequencing alignment of cDNA subclone AA12#4 with mouse acetyl-coenzyme A dehydrogenase, medium chain (MCAD) by BLAST searching against the National Center for Biotechnology Information database" --- p.302 / Chapter B 15.3: --- Summary of sequence alignment of cDNA subclone AA12#4 with mouse MCAD --- p.302 / Chapter B 16.1: --- DNA sequence of cDNA subclone AB7#2 (AP3 & ARP3) using Ml3 forward (-20) primer --- p.303 / Chapter B 16.2: --- "Sequencing alignment of cDNA subclone AB7#2 with mouse UDP-glucuronosyltransferase 2 family, member 5 (UGT2b5) by BLAST searching against the National Center for Biotechnology Information database" --- p.303 / Chapter B 16.3: --- Summary of sequence alignment of cDNA subclone AB7#2 with mouse UGT2b5 --- p.303 / Chapter B 17.1: --- DNA sequence of cDNA subclone AB7#8 (AP3 & ARP3) using M13 reverse primer --- p.304 / Chapter B 17.2: --- "Sequencing alignment of cDNA subclone AB7#8 with mouse UDP-glucuronosyltransferase 2 family, member 5 (UGT2b5) by BLAST searching against the National Center for Biotechnology Information database" --- p.304 / Chapter B 17.3: --- Summary of sequence alignment of cDNA subclone AB7#8 with mouse UGT2b5 --- p.304 / Chapter B 18.1: --- DNA sequence of cDNA subclone AB17#16 (AP3 & ARP3) using M13 reverse primer --- p.305 / Chapter B 18.2: --- Sequencing alignment of cDNA subclone AB17#16 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.305 / Chapter B 18.3: --- Summary of sequence alignment of cDNA subclone AB17#16 with mouse mitochondrion --- p.305 / Chapter B 19.1: --- DNA sequence of cDNA subclone AB18#4 (AP3 & ARP3) using M13 forward (-20) primer --- p.306 / Chapter B 19.2: --- Sequencing alignment of cDNA subclone AB18#4 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.306 / Chapter B 20.1: --- DNA sequence of cDNA subclone AB18#4 (AP3 & ARP3) using M13 reverse primer --- p.307 / Chapter B 20.2: --- Sequencing alignment of cDNA subclone AB18#4 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.307 / Chapter B 20.3: --- Summary of sequence alignment of cDNA subclone AB 18#4 with mouse mitochondrion --- p.307 / Chapter B 21.1: --- DNA sequence of cDNA subclone AB19#2 (AP3 & ARP3) using M13 forward (-20) primer --- p.308 / Chapter B 21.2: --- Sequencing alignment of cDNA subclone AB 19#2 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.308 / Chapter B 21.3: --- Summary of sequence alignment of cDNA subclone AB19#2 with mouse mitochondrion --- p.308 / Chapter B 22.1: --- DNA sequence of cDNA subclone AB19#10 (AP3 & ARP3) using Ml3 reverse primer --- p.309 / Chapter B 22.2: --- Sequencing alignment of cDNA subclone AB 19#10 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.309 / Chapter B 22.3: --- Summary of sequence alignment of cDNA subclone AB19#10 with mouse mitochondrion --- p.309 / Chapter B 23.1: --- DNA sequence ofcDNA subclone AB22#9 (AP3 & ARP3) using M13 forward (-20) primer --- p.310 / Chapter B 23.2: --- Sequencing alignment of cDNA subclone AB22#9 with mouse peroxisome biogenesis factor 16 (Pexl6) by BLAST searching against the National Center for Biotechnology Information database --- p.310 / Chapter B 23.3: --- Summary of sequence alignment of cDNA subclone AB22#9 with mouse Pexl6 --- p.310 / Chapter B 24.1: --- DNA sequence of cDNA subclone AB22#9 (AP3 & ARP3) using Ml3 reverse primer --- p.311 / Chapter B 24.2: --- Sequencing alignment of cDNA subclone AB22#9 with mouse peroxisome biogenesis factor 16 (Pexl6) by BLAST searching against the National Center for Biotechnology Information database --- p.311 / Chapter B 24.3: --- Summary of sequence alignment of cDNA subclone AB22#9 with mouse Pexl6 --- p.311 / Chapter B 25.1: --- DNA sequence ofcDNA subclone AB24#9 (AP3 & ARP3) using Ml3 forward (-20) primer --- p.312 / Chapter B 25.2: --- Sequencing alignment of cDNA subclone AB24#9 with mouse Cyp4al4 by BLAST searching against the National Center for Biotechnology Information database --- p.312 / Chapter B 25.3: --- Summary of sequence alignment of cDNA subclone AB24#9 with mouse Cyp4al4 --- p.312 / Chapter B 26.1: --- DNA sequence of cDNA subclone AB24#9 (AP3 & ARP3) using M13 reverse primer --- p.313 / Chapter B 26.2: --- Sequencing alignment of cDNA subclone AB24#9 with mouse Cyp4al4 by BLAST searching against the National Center for Biotechnology Information database --- p.313 / Chapter B 26.3: --- Summary of sequence alignment of cDNA subclone AB24#9 with mouse Cyp4al4 --- p.313 / Chapter B 27.1: --- DNA sequence of cDNA subclone AB25#6 (AP3 & ARP3) using Ml3 forward (-20) primer --- p.314 / Chapter B 27.2: --- Sequencing alignment of cDNA subclone AB25#6 with mouse Cyp4a l4 by BLAST searching against the National Center for Biotechnology Information database --- p.314 / Chapter B 27.3: --- Summary of sequence alignment of cDNA subclone AB25#6 with mouse Cyp4al4 --- p.314 / Chapter B 28.1: --- DNA sequence of cDNA subclone AB26#17 (AP3 & ARP3) using Ml3 forward (-20) primer --- p.315 / Chapter B 28.2: --- Sequencing alignment of cDNA subclone AB26#17 with mouse Cyp4al4 by BLAST searching against the National Center for Biotechnology Information database --- p.315 / Chapter B 28.3: --- Summary of sequence alignment of cDNA subclone AB26#17 with mouse Cyp4al4 --- p.315 / Chapter B 29.1: --- DNA sequence of cDNA subclone AB26#3Q (AP3 & ARP3) using M13 reverse primer --- p.316 / Chapter B 29.2: --- Sequencing alignment of cDNA subclone AB26#30 with mouse Cyp4al4 by BLAST searching against the National Center for Biotechnology Information database --- p.316 / Chapter B 29.3: --- Summary of sequence alignment of cDNA subclone AB26#30 with mouse Cyp4al4 --- p.316 / Chapter B 30.1: --- DNA sequence of cDNA subclone AB29#7 (AP3 & ARP3) using Ml3 forward (-20) primer --- p.317 / Chapter B 30.2: --- Sequencing alignment of cDNA subclone AB29#7 with mouse catalase by BLAST searching against the National Center for Biotechnology Information database --- p.317 / Chapter B 30.3: --- Summary of sequence alignment of cDNA subclone AB29#7 with mouse catalase --- p.317 / Chapter B 31.1: --- DNA sequence of cDNA subclone AC1#1 (AP2 & ARP19) using Ml3 forward (-20) primer --- p.318 / Chapter B 31.2: --- Sequencing alignment of cDNA subclone AC1#1 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.318 / Chapter B 31.3: --- Summary of sequence alignment of cDNA subclone AC1#1 with mouse SPI --- p.318 / Chapter B 32.1: --- DNA sequence of cDNA subclone AC1#1 (AP2 & ARP 19) using Ml3 reverse primer --- p.319 / Chapter B 32.2: --- Sequencing alignment of cDNA subclone AC 1# 1 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.319 / Chapter B 32.3: --- Summary of sequence alignment of cDNA subclone AC1#1 with mouse SPI --- p.319 / Chapter B 33.1: --- DNA sequence of cDNA subclone AC1#2 (AP2& ARP 19) using M13 forward (-20) primer --- p.320 / Chapter B 33.2: --- Sequencing alignment of cDNA subclone AC 1#2 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.320 / Chapter B 33.3: --- Summary of sequence alignment of cDNA subclone AC1#2 with mouse SPI --- p.320 / Chapter B 34.1: --- DNA sequence of cDNA subclone AC1#2 (AP2& ARP 19) using M13 reverse primer --- p.321 / Chapter B 34.2: --- Sequencing alignment of cDNA subclone AC1#2 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.321 / Chapter B 34.3: --- Summary of sequence alignment of cDNA subclone AC1#2 with mouse SPI --- p.321 / Chapter B 35.1: --- DNA sequence ofcDNA subclone AC2#2 (AP2 & ARP19) using Ml3 reverse primer --- p.322 / Chapter B 35.2: --- Sequencing alignment of cDNA subclone AC2#2 with mouse bifunctional enzyme (PBFE) by BLAST searching against the National Center for Biotechnology Information database --- p.322 / Chapter B 35.3: --- Summary of sequence alignment of cDNA subclone AC2#2 with mouse PBFE --- p.322 / Chapter B 36.1: --- DNA sequence of cDNA subclone AC2#5 (AP2 & ARP19) using Ml3 reverse primer --- p.323 / Chapter B 36.2: --- Sequencing alignment of cDNA subclone AC2#5 with mouse catalase by BLAST searching against the National Center for Biotechnology Information database --- p.323 / Chapter B 36.3: --- Summary of sequence alignment of cDNA subclone AC2#5 with mouse catalase --- p.323 / Chapter B 37.1: --- DNA sequence of cDNA subclone AC2#6 (AP2 & ARP19) using Ml3 forward (-20) primer --- p.324 / Chapter B 37.2: --- Sequencing alignment of cDNA subclone AC2#6 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.324 / Chapter B 37.3: --- Summary of sequence alignment of cDNA subclone AC2#6 with mouse SPI --- p.324 / Chapter B 38.1: --- DNA sequence ofcDNA subclone AC4#3 (AP2 & ARP19) using Ml3 forward (-20) primer --- p.325 / Chapter B 38.2: --- Sequencing alignment of cDNA subclone AC4#3 with mouse Cyp2a5 by BLAST searching against the National Center for Biotechnology Information database --- p.325 / Chapter B 38.3: --- Summary of sequence alignment of cDNA subclone AC4#3 with mouse Cyp2a5 --- p.325 / Chapter B 39.1: --- DNA sequence ofcDNA subclone AC4#3 (AP2 & ARP 19) using M13 reverse primer --- p.326 / Chapter B 39.2: --- Sequencing alignment of cDNA subclone AC4#3 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.326 / Chapter B 39.3: --- Summary of sequence alignment of cDNA subclone AC4#3 with mouse SPI --- p.326 / Chapter B 40.1: --- DNA sequence of cDNA subclone AC7#5 (AP2& ARP 19) using M13 forward (-20) primer --- p.327 / Chapter B 40.2: --- Sequencing alignment of cDNA subclone AC7#5 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.327 / Chapter B 40.3: --- Summary of sequence alignment of cDNA subclone AC7#5 with mouse SPI --- p.327 / Chapter B 41.1: --- DNA sequence of cDNA subclone AD6#4 (AP2 & ARP 18) using Ml3 reverse primer --- p.328 / Chapter B 41.2: --- Sequencing alignment of cDNA subclone AD6#4 with mouse N-terminal Asn amidase (Ntanl) by BLAST searching against the National Center for Biotechnology Information database --- p.328 / Chapter B 41.3: --- Summary of sequence alignment of cDNA subclone AD6#4 with mouse Ntanl --- p.328 / Chapter B 42.1: --- DNA sequence of cDNA subclone AD6#10 (AP2 & ARP 18) using Ml3 forward (-20) primer --- p.329 / Chapter B 42.2: --- Sequencing alignment of cDNA subclone AD6#10 with mouse Cyp4al0 by BLAST searching against the National Center for Biotechnology Information database --- p.329 / Chapter B 42.3: --- Summary of sequence alignment of cDNA subclone AD6#10 with mouse Cvp4al0 --- p.329 / Chapter B 43.1: --- DNA sequence of cDNA subclone AD6#10 (AP2 & ARP18) using M13 reverse primer --- p.330 / Chapter B 43.2: --- Sequencing alignment of cDNA subclone AD6#10 with mouse Cyp4al0 by BLAST searching against the National Center for Biotechnology Information database --- p.330 / Chapter B 43.3: --- Summary of sequence alignment of cDNA subclone AD6#10 with mouse Cyp4al0 --- p.330 / Chapter B 44.1: --- DNA sequence of cDNA subclone AD8#2 (AP2 & ARP 18) using M13 forward (-20) primer --- p.331 / Chapter B 44.2: --- Sequencing alignment of cDNA subclone AD8#2with mouse Cyp4a l0 by BLAST searching against the National Center for Biotechnology Information database --- p.331 / Chapter B 44.3: --- Summary of sequence alignment of cDNA subclone AD8#2 with mouse Cvp4a10 --- p.331 / Chapter B 45.1: --- DNA sequence ofcDNA subclone AD8#7 (AP2 & ARP18) using Ml3 reverse primer --- p.332 / Chapter B 45.2: --- Sequencing alignment of cDNA subclone AD8#7 with mouse Cyp4al0 by BLAST searching against the National Center for Biotechnology Information database --- p.332 / Chapter B 45.3: --- Summary of sequence alignment of cDNA subclone AD8#7 with mouse Cyp4a10 --- p.332 / Chapter B 46.1: --- DNA sequence of cDNA subclone AD9#2 (AP2 & ARP 18) using Ml3 forward (-20) primer --- p.333 / Chapter B 46.2: --- Sequencing alignment of cDNA subclone AD9#2 with mouse Cyp4al0 by BLAST searching against the National Center for Biotechnology Information database --- p.333 / Chapter B 46.3: --- Summary of sequence alignment of cDNA subclone AD9#2 with mouse Cyp4al0 --- p.333 / Chapter B 47.1: --- DNA sequence of cDNA subclone AD9#3 (AP2 & ARP 18) using M13 reverse primer --- p.334 / Chapter B 47.2: --- Sequencing alignment of cDNA subclone AD9#3 with mouse Cyp4al0 by BLAST searching against the National Center for Biotechnology Information database --- p.334 / Chapter B 47.3: --- Summary of sequence alignment of cDNA subclone AD9#3 with mouse Cvp4a10 --- p.334 / Chapter B 48.1: --- DNA sequence ofcDNA subclone AF1#8 (AP10 & ARP13) using M13 forward (-20) primer --- p.335 / Chapter B 48.2: --- Sequencing alignment of cDNA subclone AF1#8 with mouse very-long-chain acyl-coA synthetase (VLACS) by BLAST searching against the National Center for Biotechnology Information database --- p.335 / Chapter B 48.3: --- Summary of sequence alignment of cDNA subclone AF1#8 with mouse VLACS --- p.335 / Chapter B 49.1: --- DNA sequence of cDNA subclone AF1#8 (AP 10 & ARP 13) using Ml3 reverse primer --- p.336 / Chapter B 49.2: --- Sequencing alignment of cDNA subclone AF1#8 with mouse very-long-chain acyl-coA synthetase (VLACS) by BLAST searching against the National Center for Biotechnology Information database --- p.336 / Chapter B 49.3: --- Summary of sequence alignment of cDNA subclone AF1#8 with mouse VLACS --- p.336 / Chapter B 50.1: --- DNA sequence of cDNA subclone AF21#5 (AP 10 & ARP 13) using M13 reverse primer --- p.337 / Chapter B 50.2: --- "Sequencing alignment ofcDNA subclone AF21#5 with mouse cell death-inducing DNA fragmentation factor, alpha subunit-like effector B (Cideb) by BLAST searching against the National Center for Biotechnology Information database" --- p.337 / Chapter B 50.3: --- Summary of sequence alignment of cDNA subclone AF21#5 with mouse Cideb --- p.337 / Chapter B 51.1: --- DNA sequence ofcDNA subclone AF25#6 (AP10 & ARP13) using M13 forward (-20) primer --- p.338 / Chapter B 51.2: --- Sequencing alignment of cDNA subclone AF25#6 with mouse major urinary protein 2 (MUPII) by BLAST searching against the National Center for Biotechnology Information database --- p.338 / Chapter B 51.3: --- Summary of sequence alignment of cDNA subclone AF25#6 with mouse MUP II --- p.338 / Chapter B 52.1: --- DNA sequence of cDNA subclone AF25#7 (AP 10 & ARP 13) using Ml3 reverse primer --- p.339 / Chapter B 52.2: --- Sequencing alignment of cDNA subclone AF25#7 with mouse major urinary protein 2 (MUP II) by BLAST searching against the National Center for Biotechnology Information database --- p.339 / Chapter B 52.3: --- Summary of sequence alignment of cDNA subclone AF25#7 with mouse MUPII --- p.339 / Chapter B 53.1: --- DNA sequence ofcDNA subclone AF30#4 (AP10 & ARP13) using M13 forward (-20) primer --- p.340 / Chapter B 53.2: --- Sequencing alignment of cDNA subclone AF30#4 with mouse mRNA for suppressor of actin mutations (SAC1 gene) by BLAST searching against the National Center for Biotechnology Information database --- p.340 / Chapter B 53.3: --- Summary of sequence alignment of cDNA subclone AF3Q#4 with mouse SAC1 --- p.340 / Chapter B 54.1: --- DNA sequence of cDNA subclone AF30#5 (AP 10 & ARP 13) using Ml3 reverse primer --- p.341 / Chapter B 54.2: --- Sequencing alignment of cDNA subclone AF30#5 with mouse mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.341 / Chapter B 54.3: --- Summary of sequence alignment of cDNA subclone AF30#5 with mouse mitochondrion --- p.341 / Chapter B 55.1: --- DNA sequence ofcDNA subclone AH1#6 (AP11 & ARP19) using M13 forward (-20) primer --- p.342 / Chapter B 55.2: --- Sequencing alignment of cDNA subclone AH1#6 with mouse EST by BLAST searching against the National Center for Biotechnology Information database --- p.342 / Chapter B 55.3: --- Summary of sequence alignment of cDNA subclone AH1#6 with mouse EST --- p.342 / Chapter B 56.1: --- DNA sequence of cDNA subclone AIl#5 (AP6 & ARP4) using Ml3 forward (-20) primer --- p.343 / Chapter B 56.2: --- Sequencing alignment of cDNA subclone AIl#5 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST searching against the National Center for Biotechnology Information database --- p.343 / Chapter B 56.3: --- Summary of sequence alignment of cDNA subclone All#5 with mouse SPI --- p.343 / Chapter B 57.1: --- DNA sequence of cDNA subclone AI1#5 (AP6 & ARP4) using Ml3 reverse primer --- p.344 / Chapter B 57.2: --- Sequencing alignment of cDNA subclone AIl#5 with mouse serine (or cysteine) proteinase inhibitor (SPI) by BLAST --- p.344 / Chapter B 57.3: --- Summary of sequence alignment of cDNA subclone AIl #5 with mouse SPI --- p.344 / Chapter B 58.1: --- DNA sequence of cDNA subclone AI18#6 (AP6 & ARP4) using Ml3 forward (-20) primer --- p.345 / Chapter B 58.2: --- Sequencing alignment of cDNA subclone AI18#6 with mouse argininosuccinate lyase (Asl) by BLAST searching against the National Center for Biotechnology Information database --- p.345 / Chapter B 58.3: --- Summary of sequence alignment of cDNA subclone AI18#6 with mouse Asl --- p.345 / Chapter B 59.1: --- DNA sequence of cDNA subclone AI18#6 (AP6 & ARP4) using M13 reverse primer --- p.346 / Chapter B 59.2: --- Sequencing alignment of cDNA subclone AI18#6 with mouse argininosuccinate lyase (Asl) by BLAST searching against the National Center for Biotechnology Information database --- p.346 / Chapter B 59.3: --- Summary of sequence alignment of cDNA subclone AI18#6 with mouse Asl --- p.346 / Chapter B 60.1: --- DNA sequence ofcDNA subclone AJ1#4 (AP6 & ARP14) using Ml3 forward (-20) primer --- p.347 / Chapter B 60.2: --- Sequencing alignment of cDNA subclone AJ1#4 with mouse carboxylesterase by BLAST searching against the National Center for Biotechnology Information database --- p.347 / Chapter B 60.3: --- Summary of sequence alignment of cDNA subclone AJ1#4 with mouse carboxylesterase --- p.347 / Chapter B 61.1: --- DNA sequence ofcDNA subclone AJ1#5 (AP6 & ARP14) using Ml3 reverse primer --- p.348 / Chapter B 61.2: --- Sequencing alignment of cDNA subclone AJ1#5 with mouse carboxylesterase by BLAST searching against the National Center for Biotechnology Information database --- p.348 / Chapter B 61.3: --- Summary of sequence alignment of cDNA subclone AJ1#5 with mouse carboxylesterase --- p.348 / Chapter B 62.1: --- DNA sequence ofcDNA subclone AJ2#10 (AP6 & ARP14) using M13 forward (-20) primer --- p.349 / Chapter B 62.2: --- Sequencing alignment of cDNA subclone AJ2#10 with peroxisomal acyl-coA oxidase (AOX) by BLAST searching against the National Center for Biotechnology Information database --- p.349 / Chapter B 62.3: --- Summary of sequence alignment of cDNA subclone AJ2#10 with mouse AOX --- p.349 / Chapter B 63.1: --- DNA sequence ofcDNA subclone AJ2#10 (AP6 & ARP14) using Ml3 reverse primer --- p.350 / Chapter B 63.2: --- Sequencing alignment of cDNA subclone AJ2#10 with peroxisomal acyl-coA oxidase (AOX) by BLAST searching against the National Center for Biotechnology Information database --- p.350 / Chapter B 63.3: --- Summary of sequence alignment of cDNA subclone AJ2#10 with mouse AOX --- p.350 / Chapter B 64.1: --- DNA sequence ofcDNA subclone AJ9#1 (AP6 & ARP 14) using Ml3 forward (-20) primer --- p.351 / Chapter B 64.2: --- Sequencing alignment of cDNA subclone AJ9#1 with mouse catalase by BLAST searching against the National Center for Biotechnology Information database --- p.351 / Chapter B 64.3: --- Summary of sequence alignment of cDNA subclone AJ9#1 with mouse catalase --- p.351 / Chapter B 65.1: --- DNA sequence ofcDNA subclone AJ9#1 (AP6 & ARP14) using Ml3 reverse primer --- p.352 / Chapter B 65.2: --- Sequencing alignment of cDNA subclone AJ9#1 with mouse suppressor of actin mutations (SAC1 gene) by BLAST searching against the National Center for Biotechnology Information database --- p.352 / Chapter B 65.3: --- Summary of sequence alignment of cDNA subclone AJ9#1 with mouse SAC1 --- p.352 / Chapter B 66.1: --- DNA sequence ofcDNA subclone AL2#8 (AP7 & ARP15) using M13 forward (-20) primer --- p.353 / Chapter B 66.2: --- Sequencing alignment of cDNA subclone AL2#8 with mouse hydroxy steroid (17-beta) dehydrogenase 11 (Hsdl7pil) by BLAST searching against the National Center for Biotechnology Information database --- p.353 / Chapter B 66.3: --- Summary of sequence alignment of cDNA subclone AL2#8 with mouse HSD17β11 --- p.353 / Chapter B 67.1: --- DNA sequence of cDNA subclone AL3#3 (AP7& ARP 15) using Ml3 forward (-20) primer --- p.354 / Chapter B 67.2: --- Sequencing alignment of cDNA subclone AL3#3 with mouse hydroxy steroid (17-beta) dehydrogenase 11 (Hsdl7pll) by BLAST searching against the National Center for Biotechnology Information database --- p.354 / Chapter B 67.3: --- Summary of sequence alignment of cDNA subclone AL3#3 with mouse HSD17β11 --- p.354 / Chapter B 68.1: --- DNA sequence of cDNA subclone AL3#3 (AP7& ARP 15) using M13 reverse primer --- p.355 / Chapter B 68.2: --- Sequencing alignment of cDNA subclone AL3#3 with mouse hydroxysteroid (17-beta) dehydrogenase 11 (Hsdl7β1l) by BLAST searching against the National Center for Biotechnology Information database --- p.355 / Chapter B 68.3: --- Summary of sequence alignment of cDNA subclone AL3#3 with mouse HSD17β11 --- p.355 / Chapter B 69.1: --- DNA sequence of cDNA subclone AO1#2 (AP5 & ARP 10) 356 using Ml3 forward (-20) primer --- p.356 / Chapter B 69.2: --- Sequencing alignment of cDNA subclone AO1#2 with mouse 356 adipose differentiation related protein (ADFP) by BLAST searching against the National Center for Biotechnology Information database --- p.356 / Chapter B 69.3: --- Summary of sequence alignment of cDNA subclone AO1 #2 with 356 mouse ADFP --- p.356 / Chapter B 70.1: --- DNA sequence ofcDNA subclone AO1#5 (AP5 & ARP10) 357 using M13 reverse primer --- p.357 / Chapter B 70.2: --- Sequencing alignment of cDNA subclone AO1#5 with mouse 357 carnitine O-octanoyltransferase (Crot) by BLAST searching against the National Center for Biotechnology Information database --- p.357 / Chapter B 70.3: --- Summary of sequence alignment of cDNA subclone AO1 #5 with 357 mouse Crot --- p.357 / Chapter B 71.1: --- DNA sequence ofcDNA subclone AO2#6 (AP5 & ARP10) 358 using Ml3 forward (-20) primer --- p.358 / Chapter B 71.2: --- Sequencing alignment of cDNA subclone A02#6 with mouse 358 RNase A family 4 (Rnase4) by BLAST searching against the National Center for Biotechnology Information database --- p.358 / Chapter B 71.3: --- Summary of sequence alignment of cDNA subclone AO2#6 358 with mouse Rnase4 --- p.358 / Chapter B 72.1: --- DNA sequence of cDNA subclone AO2#6 (AP5 & ARP 10) 359 using Ml3 reverse primer --- p.359 / Chapter B 72.2: --- Sequencing alignment of cDNA subclone A02#6 with mouse 359 RNase A family 4 (Rnase4) by BLAST searching against the National Center for Biotechnology Information database --- p.359 / Chapter B 72.3: --- Summary of sequence alignment of cDNA subclone A02#6 359 with mouse Rnase4 --- p.359 / Chapter B 73.1: --- DNA sequence ofcDNA subclone AO2#8 (AP5 & ARP10) 360 using Ml3 reverse primer --- p.360 / Chapter B 73.2: --- Sequencing alignment of cDNA subclone A02#8 with mouse 360 carnitine O-octanoyltransferase (Crot) by BLAST searching against the National Center for Biotechnology Information database --- p.360 / Chapter B 73.3: --- Summary of sequence alignment of cDNA subclone AO2#8 with 360 mouse Crot --- p.360 / Chapter B 74.1: --- DNA sequence ofcDNA subclone AO8#2 (AP5 & ARP10) 361 using M13 forward (-20) primer --- p.361 / Chapter B 74.2: --- Sequencing alignment of cDNA subclone A08#2 with mouse 361 RNase A family 4 (Rnase4) by BLAST searching against the National Center for Biotechnology Information database --- p.361 / Chapter B 74.3: --- Summary of sequence alignment of cDNA subclone AO8#2 with 361 mouse Rnase4 --- p.361 / Chapter B 75.1: --- DNA sequence of cDNA subclone AP4#4 (AP12 & ARP2) 362 using Ml3 forward (-20) primer --- p.362 / Chapter B 75.2: --- Sequencing alignment of cDNA subclone AP4#4 with mouse 362 mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.362 / Chapter B 75.3: --- Summary of sequence alignment of cDNA subclone AP4#4 with 362 mouse mitochondrion --- p.362 / Chapter B 76.1: --- DNA sequence ofcDNA subclone AP4#4 (AP12 & ARP2) 363 using Ml3 reverse primer --- p.363 / Chapter B 76.2: --- Sequencing alignment of cDNA subclone AP4#4 with mouse 363 mitochondrion by BLAST searching against the National Center for Biotechnology Information database --- p.363 / Chapter B 76.3: --- Summary of sequence alignment of cDNA subclone AP4#4 with 363 mouse mitochondrion --- p.363

Skin is the largest organ in the body that protects the organism from environmental, chemical and physical traumas of each passing day. The protective skin epidermal permeability barrier (EPB) is formed within the exterior layers of the epidermis, which are regularly sloughed off and repopulated by movement of inner cells. The epidermal permeability barrier is established during in utero development and maintained through lifetime. Impaired epidermal barrier formation is one of the major features of several dermatoses such as psoriasis and atopic dermatitis. Chicken ovalbumin upstream promoter transcription factor (COUP-TF)-interacting protein 2 (Ctip2), also known as Bcl11b, is a C���H��� zinc finger protein expressed in many organs and tissues. It has been shown to regulate the development of thymocyte, tooth and corticospinal motor neurons. Ctip2 is highly expressed in mouse epidermis during skin organogenesis and in adulthood. It is crucial for epidermal homeostasis and protective barrier formation in developing mouse embryos. Germline (Ctip2- null mice) and selective ablation of Ctip2 in mouse epidermis (Ctip2[superscript ep-/-] mice) leads to increased transepidermal water loss (TEWL), impaired epidermal proliferation and terminal differentiation as well as altered lipid distribution during embryogenesis. Sphingolipids account for ~50% of total skin lipids by weight and are crucial components of epidermal barrier. We have recently identified Ctip2 as a key regulator of skin lipid metabolism. Germline deletion of Ctip2 in mouse embryos leads to altered lipid composition in the developing mouse epidermis by modulating the expression levels of key enzymes involved in lipid metabolism (bio-synthesis and catabolism). We also demonstrated that Ctip2 is recruited to the promoter regions of several genes involved in the ceramide and sphingomyelin biosynthesis pathways and could directly regulate their expression. Thus, we have identified Ctip2 as a key regulator of several lipid metabolizing genes and hence epidermal sphingolipid biosynthesis during skin development. To study the role of Ctip2 in adult skin homeostasis, we have utilized Ctip2[superscript ep-/-] mouse model in which Ctip2 is selectively deleted in epidermal keratinocytes. We showed that keratinocytic ablation of Ctip2 leads to atopic dermatitis (AD)-like skin inflammation, characterized by alopecia, pruritus and scaling, as well as high infiltration of T lymphocytes and immune cells. We have also observed increased expression of Th2-type cytokines and chemokines in the mutant skin, as well as systemic immune responses that share similarity with human AD patients. Furthermore, we discovered that thymic stromal lymphopoietin (TSLP) expression is significantly upregulated in the mutant epidermis as early as postnatal day 1 and Ctip2 was recruited to the promoter region of the TSLP gene in mouse epidermal keratinocytes. The results suggest that upregulation of TSLP expression in the Ctip2[superscript ep-/-] epidermis could be due to a derepression of gene transcription in absence of Ctip2. Thus, our data demonstrated a cell-autonomous role of Ctip2 in barrier maintenance and epidermal homeostasis in adult skin, as well as a non-cell autonomous role of keratinocytic Ctip2 in suppressing skin inflammatory responses by regulating the expression of Th2-type cytokines in adult mouse skin. Present results establish an initiating role of epidermal TSLP in AD pathogenesis via a novel repressive regulatory mechanism mediated by Ctip2 in mouse epidermal keratinocytes. Altogether, our study indicates that Ctip2 could be involved in a diverse range of biological events in skin including barrier formation, maintenance and epidermal homeostasis. Ctip2 appears to be a master regulator in skin barrier functions by directly regulating the transcription of a subset of genes involved in lipid metabolism and inflammatory responses. / Graduation date: 2013

The fast-skeletal-muscle-fiber-specific expression of the troponin I(fast) (TnIfast) gene is driven by an Intronic Regulatory Element (IRE) located within the first intron of the gene. The IRE is a 148 bp transcriptional enhancer that contains several known and suspected cis-regulatory elements. These include the E-box, the closely-spaced MEF2 site and CACT box, the CACC site, and the CAGG element. Previous loss-of-function studies performed using the quail TnIfast IRE suggest that its activity depended on the MEF2 and CACT elements. The goal of my thesis research was to determine whether the MEF2 and CACT sites were not only necessary, but also sufficient, to support IRE activity. I prepared head-to-tail multimers of a 27-bp IRE segment that consisted largely of the near-adjacent MEF2 and CACT elements and did not contain any other known/suspected elements. These multimers were cloned upstream of a reporter gene consisting of the minimal promoter of the quail TnIfast gene linked to sequences encoding human placental alkaline phosphatase. The transcriptional capabilities of the constructs were assessed by gene transfer into the mouse soleus muscle in vivo by intramuscular injection/electroporation, and histochemical analysis of reporter enzyme plap expression including quantitative microdensitometry. I found that expression of these constructs was readily detectable and that it was markedly reduced by prior mutation of the CACT and, especially, of the MEF2 sites. These data indicate that the short DNA segment containing MEF2 and CACT elements is sufficient to drive expression in skeletal muscle and confirms the functional importance of these specific elements. / Although constructs containing the wild-type IRE 27-bp region were expressed, there was little preferential expression in fast fibers, in contrast to expression driven by the complete 148-bp IRE. Thus my results indicate that the MEF2 and CACT elements are not sufficient to drive fast fiber-type-specific expression, and suggest that additional elements outside of the 27-bp region tested are also necessary for fiber-type-specificity.

CYP17 is an ACTH/cAMP inducible gene in the human adrenal cortex encoding a cytochrome P450 enzyme with sterol 17α-hydroxylase activity and 17,20 lyase activity essential for biosynthesis of cortisol and androgens. Studies carried out during the past decade have shown that acclerated transcription of inducible eukaryotic genes involves sequential chromatin modifications by cooperative promoter-specific transcription factors and the class of proteins called transcriptional coregulators. In the present work, we aimed to first identify important chromatin modifications and chromatin modifying complexes at the CYP17 transcription start site and nearby steroidogenic factor-1 (SF-1) binding site. Then, we asked what modifications to SF-1 occur during the interaction of this nuclear receptor with the CYP17 promoter, and what their function may be. Finally, we asked how ACTH/cAMP signaling affects SF-1-containing chromatin-modifying complexes during the early phase of transcriptional induction of CYP17. Results from chromatin immunoprecipitation (ChIP) and mammalian two hybrid experiments identified complexes including one comprised of SF-1, steroid receptor coactivator-1 (SRC-1), and the histone acetyltransferase general control nonderepressed 5 (GCN5) as cAMP-inducible, but sensitive to the SF-1 antagonist sphingosine, and able to act in stimulating CYP17 transcription. Moreover, ATPases on the promoter coincided with manipulation of nucleosome histone H2 dimer content. Next, we found that SF-1 phosphorylation by glycogen synthase kinase 3beta (GSK3beta), reciprocal dephosphorylation by phosphatase(s), and acetylation by GCN5 at nearby sites at the ligand binding pocket opening were required for efficient CYP17 transcription. This leads us to propose that ligand binding to SF-1 is controlled by these post-translational modifications. Finally, we determined that the corepressors E1A C-terminal binding proteins (CtBP) 1 and 2 are protein kinase A (PKA) targets and are sensitive to PKA-dependent NADH accumulation. These effects of PKA activation by ACTH/cAMP in adrenal cortex cells enforce CYP17 transcription concomitant with dimerization of CtBP1 and CtBP2.

Lymphomas are neoplasms of the human immune system and can be divided into two categories, Hodgkin’s lymphoma (HL) and non-Hodgkin lymphoma (NHL). Anaplastic large cell lymphoma (ALCL) is a form of NHL that shares a common distinctive feature with HL, the overexpression CD30. The expression of cytokine receptor CD30 is restricted to proliferating B and T lymphocytes in healthy individuals while its overexpression is associated with several lymphoproliferative diseases such as ALCL and HL. The activation of CD30 via ligand or antibodies triggers various cellular responses ranging from apoptosis to cell proliferation and it is thought that the variable cellular response to CD30 activation may be due to cell surface levels of CD30. The human CD30 gene is regulated at the transcriptional level and previous studies characterising its promoter have identified several factors that regulate the expression this gene. However none of these identified factors explain for the high levels of CD30 observed in HL and ALCL. Therefore this study focused on the identification and functional analysis of transcriptionally active regions located up or downstream of the CD30 promoter region. The first aim for this study was to identify and characterise regions within the human CD30 gene that are involved in its transcriptional regulation. Phylogenetic footprinting identified several regions downstream of the CD30 promoter that displayed high levels of sequence homology indicating potential functional significance. Validation of these regions through two in vivo approaches, DNase 1 hypersensitivity assay and chromatin accessibility studies localised potential transcriptionally active regions to intron 1 of the CD30 gene.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Thesis (Ph. D.)--University of Missouri--Columbia, 1997. / Typescript. Vita. Includes bibliographical references (leaves : 144-159). Also available on the Internet.