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The embryonic macrophage : scavenger or sculptor? /Cunningham, Michael Lawrence. January 1996 (has links)
Thesis (Ph. D.)--University of Washington, 1996. / Vita. Includes bibliographical references (leaves [77]-83).
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Embryonic stem cells for myocardial infarct repair /Nussbaum, Jeannette, January 2004 (has links)
Thesis (Ph. D.)--University of Washington, 2004. / Vita. Includes bibliographical references (leaves 92-105).
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Fonction des rétinoïdes dans les membranes foetales humaines. Action pro-cicatrisante médiée par LOXL4. / Function of retinoids in human fetal membranes. Wound-healing action mediated by LOXL4.Rouzaire, Marion 22 September 2016 (has links)
La vitamine A et ses dérivés actifs (les rétinoïdes) sont des molécules essentielles à la vie et ce dès la période embryonnaire. Molécules de choix dans de nombreuses applications thérapeutiques, notamment dans les domaines de la dermatologie et de l’ophtalmologie, elles sont le plus couramment utilisées pour leurs propriétés pro-cicatrisantes. Malgré cette utilisation courante en clinique, les mécanismes cellulaires et moléculaires permettant aux rétinoïdes de promouvoir un processus de cicatrisation restent encore mal connus. Afin de mieux appréhender ces mécanismes, mais aussi afin d’élargir leur utilisation clinique à la rupture prématurée des membranes (RPM), je me suis intéressée aux propriétés pro-cicatrisantes de l’atRA (l’un des dérivés actifs de la vitamine A) sur les membranes fœtales. Alors que l’amnios et le chorion sont physiologiquement incapables d’initier un processus de cicatrisation suite à une lésion, nos résultats démontrent un effet positif de l’atRA sur la migration des cellules épithéliales amniotiques (amniocytes primaires), conduisant ainsi à une potentialisation de la cicatrisation de plus de 80% in vitro. Ce travail, complété par une analyse transcriptomique réalisée sur des membranes fœtales et des amniocytes primaires traités ou non à l’atRA, a permis d’identifier de nombreux gènes régulés par l’atRA au sein de l’amnios et des amniocytes. Parmi ces gènes, je me suis intéressée à un membre de la famille des lysyl oxydases, LOXL4, qui joue un rôle clé dans la dynamique de la matrice extracellulaire en régulant la réticulation du collagène. Une étude de régulation transcriptionnelle associée à de la promotologie ont tout d’abord permis de montrer que l’atRA induisait l’expression du gène LOXL4 de manière directe. Puis, l’inhibition de la protéine LOXL4 lors de tests de blessure (par le β-aminopropionitrile et par un siARN spécifique) nous a permis de démontrer que l’effet pro-cicatrisant de l’atRA était médié, au moins en partie, par la rétino-induction deLOXL4. Ces nouveaux éléments apportés par ce travail dans la compréhension des mécanismes moléculaires utilisés par l’atRA afin de promouvoir la cicatrisation permettent d’envisager son utilisation future dans la prise en charge clinique des ruptures prématurées. / Vitamin A and its active derivates (retinoids) are essential molecules for life from the embryonic development to the adulthood. Because of these numerous functions and especially because of their pro-healing properties, they are commonly used in clinic in dermatology and ophthalmology in particular. Despite this routine clinical use, the molecular and cellular mechanisms used by retinoids to promote a healing process remain unclear. To better understand these mechanisms, but also to expand its clinical use to the premature rupture of fetal membranes (PROM), we looked to pro-healing properties of atRA (active derivative of vitamin A) on the human fetal membranes. While amnion and chorion are unable to initiate a physiological process of healing after injury, our results demonstrate a positive effect of atRA on the migration of primary amniocytes, leading to an healing improvement of up to 80% in vitro. This work, completed by a transcriptomic analysis performed on fetal membranes and primary amniocytes treated or not with atRA, allowed the identification of many genes regulated by atRA within the amnion and the amniocytes. Among these genes, we looked to a member of the lysyl oxidase family, LOXL4, which plays a key role in the dynamic of the extracellular matrix by regulating collagen crosslinking. First, transcription and promotology experiments have shown that atRA strongly induced the expression of LOXL4 in a direct manner. Then, the inhibition of the LOXL4 protein in scratch assay experiments (using β-aminopropionitrile or a specific siRNA) allowed us to demonstrate that the pro-healing effect of atRA was mediated, at least in part, by the retinoid-induction of this gene. Besides providing new elements to understand how atRA exerts their pro-healing properties, this work proposes atRA as a promising candidate to improve the clinical management of premature rupture of the fetal membranes by promoting re-epithelialization of the amnion.
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Efeitos da suplementação com semente de girassol em fêmeas bovinas na produção de oócitos e embriões in vitro /Baltazar, Angélica Leão. January 2016 (has links)
Orientador: Flávia Lombardi Lopes / Coorientadora: Claudia Maria Bertran Membrive / Banca:Calie Castilho Silvestre / Banca:Guilherme de Paula Nogueira / Resumo: A suplementação com compostos ricos em ácido linoleico, dentre os quais inclui-se a semente de girassol, promove aumento na taxa de concepção em fêmeas bovinas. Hipotetizou-se que a suplementação com semente de girassol em doadoras de oócitos aumenta o número e a qualidade de oócitos, incrementa as taxas de clivagem e determina um aumento no número e qualidade dos blastocistos produzidos in vitro. Assim, objetivou-se investigar o efeito de tal suplementação, no número e qualidade de oócitos cultivados in vitro, na taxa de clivagem e no número e qualidade de blastocistos produzidos in vitro. Para tanto, cinco vacas e vinte e cinco novilhas (n=30) Nelore foram divididas em dois grupos para receberem um dos seguintes tratamentos: 1,7 kg/dia de suplemento contendo 53% de farelo de soja com 44% de PB e 47% de milho (Grupo Controle - Grupo C; n= 15) ou 1,7 kg/dia de suplemento contendo 40% de farelo de soja com 44% de proteína bruta (PB) e 60% de semente de girassol (Grupo Girassol - Grupo G; n= 15), durante 57 dias. As fêmeas foram submetidas à aspiração folicular nos dias D0, D13, D29, D43, D56 e D77 (D0= início da suplementação; D56= término da suplementação). Os oócitos foram submetidos ao processo de produção de embriões in vitro. Os dados foram analisados pelo programa Mixed procedure (SAS) versão 9.2, pelo teste ANOVA modelo misto. Não se observou diferença entre os Grupos C e G no número de folículos visualizados (16,85 ± 1,32 vs. 16,12 ± 1,48); número... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Supplementation with compounds rich in linoleic acid, among which is included sunflower seed, promotes increase in conception rates in cows. It was hypothesized that sunflower seed supplementation in oocyte donors increases the number and quality of oocytes, increases the cleavage rates and determines an increase in the number and quality of blastocysts produced in vitro. Therefore the objective was to investigate the effect of such supplementation, in the number and quality of oocytes cultured in vitro on cleavage rate and in the number and quality of in vitro produced blastocysts. Therefore, cows five and twenty-five heifers (n = 30) Nellore were divided into two groups to receive one of the following treatments: 1.7 kg / day supplement containing 53% soybean meal 44% and 47% PB corn (Control Group - Group C, n = 15) or 1.7 kg / containing 40% add-on of soybean meal with 44% crude protein (CP) and 60% sunflower seed (Sunflower group - Group C; n = 15) for 57 days. Females underwent follicular aspiration on days D0, D13, D29, D43, D56 and D77 (D0 = start of supplementation; D56 = end of supplementation). The oocytes were subjected to in vitro embryo production process. Data were analyzed by the Mixed procedure (SAS) version 9.2, by ANOVA mixed model. There was no difference between Groups C and G on the number of displayed follicles (16.85 ± 1.32 vs. 16.12 ± 1.48); number of aspired oocytes (13.80 ± 1.27 vs. 13.05 ± 1.25); recovery rate (82 ± 1% vs. 80 ±... (Complete abstract click electronic access below) / Mestre
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Investigating a Role for the CCAAT/Enhancer-Binding Protein δ in the Developing ZebrafishBeirl, Alisha Jennifer 20 March 2014 (has links)
The CCAAT/enhancer-binding protein delta (C/EBPδ) is a highly conserved transcription factor capable of regulating numerous cell fate processes, such as cell growth, differentiation, proliferation and apoptosis. C/EBPδ is inducible during cellular stress responses, including inflammation and responses to growth factor deprivation or thermal stress. C/EBPδ is stress-inducible in a diversity of fishes, including the zebrafish Danio rerio; however, little is known about its role in fish development. Here I show that overexpression of C/EBPδ leads to severe developmental defects, including reduced body length, edema, liver malformation and retinal abnormalities. The proportion of individuals that display developmental abnormalities is significantly greater in C/EBPδ-overexpressing embryos compared to control embryos and overexpression significantly reduces survival of larvae over time. TUNEL analysis suggests C/EBPδ-overexpressing embryos exhibit a pattern of apoptotic cell death which is spatially distinct from control embryos. These data support a critical role for C/EBPδ in numerous developmental processes, including promoting programmed cell death during development. Mutations in C/EBPδ have been implicated in the progression of human tumors, including those of myeloid, hepatocellular and breast cancers. Therefore, the C/EBPδ-overexpressing zebrafish will serve as a valuable model for examining the role of this gene during development, as a part of the cellular response to stress and in pathological states such as tumor progression.
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Distribuição da proteína IMPACT em encéfalos de camundongos, ratos e sagüis / Distribution of the protein IMPACT in the mouse, marmoset brainBittencourt, Simone [UNIFESP] 26 November 2009 (has links) (PDF)
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Publico-11799m.pdf: 712324 bytes, checksum: 045cf67852c1de984973142eb424a0ec (MD5) / O controle da síntese protéica tem papel fundamental em diversas condições fisiológicas, mecanismos homeostáticos e diversos fenômenos de plasticidade neuronal. Um dos principais mecanismos de regulação da síntese protéica é a fosforilacão do fator de iniciação eIF2α. O aumento de fosforilação de eIF2α leva ao bloqueio do início de tradução geral da célula e tradução de genes específicos a exemplo do ATF4 (fator de ativação transcricional 4). Há 4 cinases específicas que fosforilam eIF2α, cada uma ativada sob condições distintas de estresse. A cinase GCN2, por exemplo, está presente em abundância no encéfalo, e faz parte do controle alimentar, memória e aprendizado de mamíferos. A proteína IMPACT, preferencialmente expressa no encéfalo, atua como uma inibidora da ativação de GCN2. Nesse contexto, torna-se relevante uma análise sistemática da distribuição de IMPACT no encéfalo e do possível envolvimento dessa proteína em condições fisiológicas e/ou patológicas. Um conjunto de resultados foi observado: (i) A proteína IMPACT é preferencialmente expressa em núcleos encefálicos associados ao ritmo circadiano, como também núcleos importantes na geração de ritmos, como o theta hipocampal; ii) A elevada intensidade e densidade de IMPACT no hipotálamo e hipocampo pode condizer com o envolvimento de IMPACT em mecanismos que envolvam homeostase corporal (função hipotalâmica) e memória (função hipocampal). (iii) Em tecido adulto, poucos grupos neuronais mostraram divergências na expressão de IMPACT entre roedores e primata; (iv) Há evidências de expressão diferencial (em uma mesma espécie) de IMPACT em grupos neuronais específicos (GABAérgicos); (v) Neurônios IMPACT-positivos são resistentes a vários tipos de estresse (leves, intensos, agudos ou crônicos, a exemplo de baixas temperaturas e indução de status epilepticus no modelo de epilepsia por pilocarpina); (vi) A proteína IMPACT é constitutiva, mostrou-se insensível a perturbações in vivo; (vii) IMPACT é detectada a partir do estágio embrionário E16. De acordo com os dados aqui dispostos sobre a expressão de IMPACT em neurônios e a evidência de sua expressão áreas encefálicas e em diversas condições, nós podemos especular sobre sua possível relevância na neurofisiologia. Assim, juntos esses dados sugerem que neurônios IMPACT-positivos tem função importante no encéfalo de mamíferos e podem estar envolvidos na geração e controle de ritmos encefálicos e na homeostase. / The control of protein synthesis plays a major role in several physiological conditions, homeostatic mechanisms and several phenomenon of neuronal plasticity. One of the most important mechanisms in regulation of protein synthesis is the phosphorylation of the initiation factor 2 (eIF2α). Increase in the eIF2α phosphorylation blocks general translation and enhance translation of specific genes such ATF4 (activating transcription factor 4) for example. In response to specific stress stimuli four specific kinases can phosphorylate eIF2α. The GCN2 kinase, for instance, is abundant in the brain, and is involved in feeding behavior control, learning and memory of mammalians. The IMPACT protein, preferentially expressed in the brain, acts as an inhibitor of the activation of GCN2. Therefore, a systematic analysis of the distribution of IMPACT in the brain and its possible involvement in physiological and/or pathological conditions are relevant. The following results were observed: (i) The IMPACT protein is preferentially expressed in brain nuclei involved in circadian rhythms, as well as in important nuclei involved in rhythm generation, such as the hippocampal theta rhythm; (ii) The high density and intensity of IMPACT labeling in the hypothalamus and hippocampus may reflect its involvement in homeostatic mechanisms (hypothalamic function) and memory (hippocampal function). (iii) In adult tissue, only a few neuronal groups showed divergence in IMPACT expression between rodents and primate; (iv) Within a single species there was differential expression of IMPACT in specific neuronal groups (e.g. GABAergic neurons); (v) IMPACT-positive neurons were resistant to several types of stress (weak, strong, acute or chronic), such as low temperature and status epilepticus induced by pilocarpine in a model of epilepsy; (vi) IMPACT is a constitutive protein, insensitive to perturbations in vivo; (vii) The detection of IMPACT starts at E16 embryonic stage. According to these results on the neurons expressing IMPACT and the evidence of its presence in some brain areas, we speculate on its possible relevance in neurophysiology. Thus, these data taken together suggest that IMPACT-positive neurons have important functions in the mammalian brains and may be involved in generation and control of brain rhythms and physiological homeostasis in general. / TEDE / BV UNIFESP: Teses e dissertações
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An Extra-Embryonic Wnt Signaling Event Controls Gastrulation in Mice: A DissertationTortelote, Giovane G. 06 November 2012 (has links)
The formation of the anterior-posterior axis requires a symmetry-breaking event that starts gastrulation. Ultimately, the morphogenetic movements of gastrulation reshape the embryo to its final tri-dimensional form. In mouse embryos, the identity of the molecule that breaks the bilateral symmetry and sets in motion gastrulation remains elusive. The Wnt signaling pathway plays a pivotal role during axial specification and gastrulation in metazoans. Loss-of-function experiments have demonstrated a requirement of Wnt3 for gastrulation in mice. But because Wnt3 is expressed sequentially in two tissues, the visceral endoderm and the epiblast, its tissue specific requirements remain uncertain. Here, we report that embryos lacking Wnt3 specifically in the visceral endoderm do not form a primitive streak, mesoderm, endoderm or any derivatives. Visceral endoderm-specific Wnt3 mutants also lack primordial germ cells. Moreover, we provide data demonstrating that Wnt3 carries out its actions in the epiblast via the canonical Wnt pathway. Together, these data suggest that the posterior visceral endoderm via Wnt3, regulates the development of mouse embryos in a similar fashion to the amphibian Nieuwkoop center. Next, we conditionally ablated Wnt3 locus in the epiblast to investigate whether Wnt3 expression is also required in that tissue. Embryos lacking Wnt3 expression in the epiblast, but retaining its expression in the visceral endoderm, show delayed but not absent gastrulation. We conclude that the expression of Wnt3 in the epiblast is required for maintenance but not initiation of gastrulation in mouse embryos. Furthermore, we used in vitro and in vivo approaches to demonstrate that the Wnt3-mediated activation of the canonical Wnt pathway leads to β-catenin occupancy followed by transcription of key loci, including the Wnt3 locus itself, during gastrulation in mice. Our data indicate the presence of an autoregulatory loop in which Wnt3 controls its own expression and orchestrates the process of gastrulation in the mouse embryo.
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Fatores que afetam a viabilidade e a proporção do sexo de embriões bovinos produzidos in vitro em programa de sexagem comercial /Alonso, Rodrigo Vitorio. January 2008 (has links)
Orientador: Silvia Helena Venturoli Perri / Banca: José Fernando Garcia / Banca: José Antônio Visintin / Resumo: O crescente avanço da produção in vitro de embriões bovinos intensificou a utilização de outras biotecnologias da reprodução tais como a micro-manipulação embrionária e o diagnóstico genético pré-implantacional, sendo a identificação do sexo embrionário utilizada na rotina comercial de laboratórios de produção in vitro. O objetivo deste trabalho foi avaliar as interações de diferentes fatores sobre a taxa de mortalidade embrionária e a proporção do sexo de embriões bovinos submetidos ao processo de sexagem. Foi realizado levantamento no banco de dados da Transfix - Transplante de Embriões Ltda, Patrocínio Paulista / Brasil, referente a 4.650 embriões produzidos in vitro e sexados entre 2005 e 2007. Os embriões foram submetidos à micro-manipulação pela técnica de micro-aspiração, e as biópsias à reação em cadeia pela polimerase (PCR). Somente as fêmeas foram transferidas para receptoras previamente sincronizadas. O diagnóstico de gestação e a determinação do sexo fetal foram realizados por ultra-sonografia. As variáveis foram classificadas de acordo com o sexo dos embriões (macho, fêmea e indeterminado), cinco laboratórios (A, B, C, D e E), seis raças bovinas (Nelore, Brahman, Girolando, Simental, Holandês e Jersey), estágio embrionário (MO, BI, BL, BX e BE), qualidade embrionária (1, 2 e 3) e qualidade da biópsia ("dentro do padrão" e "fora do padrão"). As análises estatísticas foram realizadas pelos testes 2 de associação, 2 de aderência para proporção 1:1 e pela análise de regressão logística com o método de Hosmer-Lameshow utilizando o procedimento logístico (PROC LOGISTIC) programa computacional SAS. A PCR apresentou eficiência de 93,3%, acurácia de 93,2% e taxa de machos e fêmeas de 52,9% e 47,1%, respectivamente. A taxa de mortalidade dos embriões... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Crescent progress of in vitro bovine embryo production has improved the use of other reproductive biotechnologies, as embryo micromanipulation and preimplantation genetic diagnosis, being embryo sexing used in commercial routine of in vitro embryo production laboratories. The present study aimed to evaluate the interactions among different factors on the mortality rate and sex ratio of in vitro produced bovine embryos. A survey was performed in the Transfix - Transplante de Embriões Ltda, Patrocínio Paulista / Brazil data base, referring to 4.650 in vitro produced bovine embryos sexed during years 2005/2007. Embryos were submited to the biopsy by the microaspiration technique, and biopsies to the polymerase chain reaction (PCR). Only female embryos were transferred to synchronized recipients. Pregnancy diagnosis and fetal sex determination were carried out by ultrasound. The variables were classified according embryo sex (male, female and indeterminate), five laboratories (A, B, C, D and E), six bovine breeds (Nellore, Brahman, Girolando, Simmental, Holstein and Jersey), embryo stage (MO, EB, BL, XB and HB), embryo quality (1, 2, and 3) and biopsy quality ("standard" and "non standard"). The statistical analysis was carried out by association 2 test, 2 for 1:1 ratio and logistic regression analysis with Hosmer- Lameshow method using logistic procedure (PROC LOGISTIC) of SAS package. The PCR showed 93.3% efficiency, 93.2% accuracy and male and female ratio of 52.9% and 47.1%, respectively. Mortality rate of biopsied embryos was 10.3% and pregnancy rate was 31.7%. Although no significant differences were observed between male and female ratio, indeterminate embryos possess greater possibility to die after micromanipulation. For quality 2 and 3 embryo mortality rate after biopsy was 3.19 and 11.37 fold higher, respectively, than for quality 1 embryo. For those whose biopsy... (Complete abstract click electronic access below) / Mestre
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Spag17 Deficiency Impairs Neuronal Cell Differentiation in Developing BrainChoi, Olivia J 01 January 2019 (has links)
The development of the nervous system is a multi-level, time-sensitive process that relies heavily on cell differentiation. However, the molecular mechanisms that control brain development remain poorly understood. We generated a knockout (KO) mouse for the cilia associated gene Spag17. These animals develop hydrocephalus and enlarged ventricles consistent with the role of Spag17 in the motility of ependymal cilia. However, other phenotypes that cannot be explained by this role were also present. Recently, a mutation in Spag17 has been associated with brain malformations and severe intellectual disability in humans. Therefore, we hypothesized that Spag17 plays a crucial role in nervous system development. To investigate this possibility, we first characterized the spatiotemporal expression of Spag17 in the developing brain by using Beta-galactosidase staining and immunohistochemistry. Results showed Spag17 expression in the spinal cord in embryonic E11. By E11.5-12.5 the expression extends to the rhombic lip from the developing hindbrain, as well as to the forebrain and midbrain regions. E14.5-15.5 embryos exhibit an intense expression in the developing ventricles as well as the cerebellum. From E17.5 to birth (P0), the gene is more broadly expressed. We then used a global Spag17 KO mouse model to characterize the function of Spag17 during brain development. Immunohistochemical studies performed in brain sections from E15.5 and P0 time points showed increased expression of the neural progenitor marker Nestin, and reduced expression of mature neuron marker NeuN, increasing positive trend with the young neuron marker Tuj1. Altogether, these findings reveal that Spag17 has a unique spatiotemporal distribution and may be critical for the maturation of neural progenitor cells.
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Regulation of δ-Aminolevulinic Acid Synthase and Heme Oxygenase in Cultured Chick Embryo Liver Cells: Synergistic Induction of Both Enzymes by Glutathimide and Iron and Repression of δ-Aminolevulinic Acid Synthase by Metalloporphyrins and Heme: A DissertationCable, Edward Earl 01 April 1993 (has links)
Primary chick embryo liver cells were used to explore the regulation of δ-aminolevulinic acid synthase and heme oxygenase, the enzymes that catalyze the rate-limiting reactions of heme anabolism and catabolism, respectively. The general focus of the work was the exploration of the novel observation in which glutethimide and iron synergistically induced both δ-aminolevulinic acid synthase and heme oxygenase, a phenomenon that would not be predicted a priori. The course of events appeared to be: first, that heme synthesis was increased after addition of the glutethimide and that iron potentiated heme synthesis; second, the heme induced heme oxygenase five to ten fold; and third, that heme oxygenase degraded the heme permitting an uncontrolled induction of δ-aminolevulinic acid synthase. This induction of δ-aminolevulinic acid synthase could be prevented by the addition of a metalloporphyrin inhibitor of heme oxygenase. Induced δ-aminolevulinic acid synthase activity could be dramatically reduced by the addition of nanomolar concentrations of a metalloporphyrin, inhibitory for heme oxygenase, and heme.
Specific observations related to the synergistic induction of heme oxygenase by glutethimide and iron was that the induction of heme oxygenase activity by glutethimide and iron occurred rapidly, with maximal increases occurring four to six hours after original treatment. Induction of heme oxygenase by glutethimide and iron was shown to be dependent on de novoheme synthesis since 4,6-dioxoheptanoic acid, a potent and specific inhibitor of heme biosynthesis, prevented the activity of heme oxygenase from increasing in the presence of glutethimide and iron. Induction of activity was associated with increases in heme oxygenase mRNA and protein; and, when induction was prevented by 4,6-dioxoheptanoic acid, no increase in either mRNA or immunoreactive protein was observed.
δ-Aminolevulinic acid synthase activity was also synergistically increased by glutethimide and iron; this increase occurred 4-6 hours after maximal heme oxygenase activity had been attained. The temporal relationship between the induction of δ-aminolevulinic acid synthase and heme oxygenase suggested that the oxygenase depleted a regulatory heme pool that would normally prevent uncontrolled induction of the synthase. When cultures were exposed to tin-mesoporphyrin, a potent inhibitor of heme oxygenase, induction of δ-aminolevulinic acid synthase, normally produced by glutethimide and iron, was prevented. Addition of tin-mesoporphyrin after δ-aminolevulinic acid synthase induction had already been established promptly halted any further induction. When heme or a combination of heme and tin-mesoporphyrin was added after induction of δ-aminolevulinic acid synthase was established, activity of the synthase was rapidly reduced.
Finally, experiments in primary chick embryo liver cells with tin-, zinc- and copper- chelated porphyrins were done to assess their effects on activities of δ-aminolevulinic acid synthase, induced by prior treatment of cells with glutethimide and iron. Nanomolar concentrations of zinc- or tin porphyrins reduced δ-aminolevulinic acid synthase activities, while copper-chelated porphyrins did not. When nanomolar concentrations of heme were added with zinc- or tin-porphyrins, δ-aminolevulinic acid synthase activity was further reduced. Effects of the non-heme metalloporphyrins on δ-aminolevulinic acid synthase were closely correlated with their abilities to inhibit heme oxygenase (r=0.78). The largest decrease of δ-aminolevulinic acid synthase (67%) was obtained with zinc-mesoporphyrin and heme. There was a rapid appearance of the cytosolic, precursor form of δ-aminolevulinic acid synthase in the presence of both 10 μM heme or 50 nM zinc-mesoporphyrin and 200 nM heme. Reduction of the half-life of the mRNA from 5.2 hours to 2.2-2.5 hours was observed in the presence of both 10 μM heme or 50 nM zinc-mesoporphyrin and 200 nM heme.
In summary, the chick embryo liver cell culture model treated with glutethimide and iron may serve as one experimental model for patients suffering from acute porphyrias, in whom uncontrolled induction of hepatic δ-aminolevulinic acid synthase plays a key role in pathogenesis of disease. The synergistic induction of δ-aminolevulinic acid synthase in the presence of glutethimide and iron may serve as an experimental paradigm for this disease. The reduction of δ-aminolevulinic acid synthase by low doses of zinc-mesoporphyrin and heme may help form the experimental foundation for eventual studies in patients suffering from acute porphyrias.
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