CIGARETTE SMOKE-INDUCED HUMAN FETAL GROWTH RESTRICTION
Frederick Naftolin, Director Reprod. Biol. Res.
New York University School Of Medicine, 550 1st Ave, New York, Ny 10016
Grant 5R01HD047003-06 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: Studies have shown that maternal exposure to cigarette smoke during pregnancy correlates significantly with adverse conceptus development, especially fetal growth restriction (FGR). In addition to obstetrical complications, FGR has been implicated in long-term effects on adult health, morbidity and even mortality. To elucidate the mechanisms of cigarette smoke-induced FGR we will use samples of placentae, maternal blood and cord blood from smoking and non-smoking pregnancies whose degree of FGR are known. Our hypothesis is that alteration of human growth factor genes, specifically insulin-like growth factor-1 (IGF-1), can be induced by environmental exposure to cigarette smoke toxins and produce FGR when the expressed abnormal gene products fail to properly regulate growth and development. Smoking-related gene alterations (polymorphisms) can result from the failure of enzymes required for detoxification of toxins coupled with enzymatic activation to reactive products that bind with DNA (adduct formation). This leads to DNA polymorphism and expression of abnormal gene products. The primary focus of our studies will be IGF-1. Placental IGF-1 gene will be assessed for polymorphism and alteration of nucleic acid sequences. IGF-1 gene expression products in maternal and cord blood will be determined and characterized. Adduct formation between activated toxins and the placental IGF-1 gene will be assessed. Since the process of detoxification versus activation involves specific enzymes, these will be measured in affected and non-affected pregnancies from smoking and non-smoking mothers (controlled by serum nicotine and cotinine assays) by assessing detoxification or activation of polynuclear aromatic hydrocarbons (PAH) into reactive products that bind with DNA. Similar studies will be performed on cultured chorionic villus sample cells from first trimester (CVS). Finally, by genotyping of the IGF-1 gene in the first trimester (CVS) and post delivery placentas during the same pregnancy we will determine both early and late effects of smoking, and identify pregnancies with high risk of smoking-induced FGR. If successful, such testing could be of considerable clinical utility
Keywords: 2-Pyrrolidi, 1-methyl-5-(3-pyridinyl)-, (S)-; 21+ years old; Adult; Adverse Late Effects; Affect; Aromatic Polycyclic Hydrocarbons; Assay; Base Sequence; Binding; Binding (Molecular Function); Bioassay; Biologic Assays; Biological Assay; Biopsy, Chorionic Villi; Blood; Blood Serum; Blood, Cord; Body Tissues; CVS; Cells; Characteristics; Chorionic Villi Sampling; Chorionic villi; Cigarette; Clinical; Conceptus; Cotinine; Coupled; DNA; DNA Adduct Formation; DNA Adduction; DNA Alteration; DNA mutation; Deoxyribonucleic Acid; Detoxification Process; Development; Drug Metabolic Detoxication; Dysfunction; Early Placental Phase; Effects, Longterm; Embryonic Tissue, Placenta; Environmental Exposure; Enzymes; Equilibrium; Exposure to; FLR; Failure (biologic function); Fetal Growth; Fetal Growth Restriction; Fetal Growth Retardation; First Pregnancy Trimester; Functional disorder; Gene Abnormality; Gene Alteration; Gene Arrangement; Gene Expression; Gene Mutation; Gene Order; Gene Position; Generations; Genes; Genetic Polymorphism; Genetic mutation; Genotype; Gestation; Growth Factor Gene; Growth and Development; Growth and Development function; Growth, Fetal; Health; Human; Human, Adult; Human, General; IGF; IGF-1; IGF-I; IGF-I-SmC; IGF1; IUGR; Incidence; Individual; Insulin-Like Growth Factor 1; Insulin-Like Growth Factor I; Insulin-Like Growth Factors; Insulin-Like Somatomedin Peptide I; Intrauterine Growth Retardation; Late Effects; Lead; Localized; Long-Term Effects; Man (Taxonomy); Man, Modern; Maternal Exposure; Measures; Metabolic Detoxication, Drug; Metabolic Detoxification, Drug; Metabolic Drug Detoxications; Metabolism of Toxic Agents; Molecular; Molecular Biology, Nucleic Acid Sequencing; Molecular Interaction; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Mothers; Nicotine; Nucleic acid sequencing; Nucleotide Sequence; Outcome; PAH; Pb element; Physiopathology; Placenta; Placenta-Tissue, Cells; Placental Villi; Placentoma, Normal; Placentome; Polycyclic Hydrocarbons, Aromatic; Polymorphism (Genetics); Polymorphism, Genetic; Polynuclear Aromatic Hydrocarbons; Pregnancy; Pregnancy Outcome; Pregnancy Trimester, First; Programs (PT); Programs [Publication Type]; Proteins; Proto-Oncogene, Growth Factor; Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-; Radiolabeled; Regulation; Relative; Relative (related person); Research Specimen; Reticuloendothelial System, Blood; Risk; Risk Factors; Role; Sampling; Scotine; Sequence Alteration; Serum; Severities; Smoke; Smoker; Smoking; Somatomedin C; Somatomedins; Specimen; Sulfation Factor; Testing; Tissues; Toxic effect; Toxicities; Toxin; Trimester, First; Umbilical Cord Blood; Variant; Variation; Villus; adduct; adult human (21+); balance; balance function; cigarette smoke; cigarette smoke-induced; cigarette smoking; detoxification; enzyme activity; failure; fetal; fetal cord blood; gene product; heavy metal Pb; heavy metal lead; insulinlike growth factor; intrauterine growth; intrauterine growth restriction; non-smoking; nucleic acid sequence; pathophysiology; polymorphism; polynuclear aromatic hydrocarbon; prenatal growth disorder; programs; radiolabel; radiotracer; smoke cigarette; smoke of cigarettes; social role
Project start date: 2004-09-30
Project end date: 2010-07-31
Budget start date: 1-AUG-2008
Budget end date: 31-JUL-2010
PFA/PA: RFA-HD-03-018
5R01HD047003-06 (2008): $0
Sponsored Links Excellgen http://Excellgen.com
Cigarette Smoke-induced Human Fetal Growth Restriction
Frederick Naftolin
New York University School Of Medicine New York, Ny 10016
Grant 5R01HD047003-05 from National Institute Of Child Health And Human Development IRG: ZHD1
Abstract: Studies have shown that maternal exposure to cigarette smoke during pregnancy correlates significantly with adverse conceptus development, especially fetal growth restriction (FGR). In addition to obstetrical complications, FGR has been implicated in long-term effects on adult health, morbidity and even mortality. To elucidate the mechanisms of cigarette smoke-induced FGR we will use samples of placentae, maternal blood and cord blood from smoking and non-smoking pregnancies whose degree of FGR are known. Our hypothesis is that alteration of human growth factor genes, specifically insulin-like growth factor-1 (IGF-1), can be induced by environmental exposure to cigarette smoke toxins and produce FGR when the expressed abnormal gene products fail to properly regulate growth and development. Smoking-related gene alterations (polymorphisms) can result from the failure of enzymes required for detoxification of toxins coupled with enzymatic activation to reactive products that bind with DNA (adduct formation). This leads to DNA polymorphism and expression of abnormal gene products. The primary focus of our studies will be IGF-1. Placental IGF-1 gene will be assessed for polymorphism and alteration of nucleic acid sequences. IGF-1 gene expression products in maternal and cord blood will be determined and characterized. Adduct formation between activated toxins and the placental IGF-1 gene will be assessed. Since the process of detoxification versus activation involves specific enzymes, these will be measured in affected and non-affected pregnancies from smoking and non-smoking mothers (controlled by serum nicotine and cotinine assays) by assessing detoxification or activation of polynuclear aromatic hydrocarbons (PAH) into reactive products that bind with DNA. Similar studies will be performed on cultured chorionic villus sample cells from first trimester (CVS). Finally, by genotyping of the IGF-1 gene in the first trimester (CVS) and post delivery placentas during the same pregnancy we will determine both early and late effects of smoking, and identify pregnancies with high risk of smoking-induced FGR. If successful, such testing could be of considerable clinical utility.
Keywords: embryo /fetus toxicology, gene environment interaction, genetic polymorphism, insulinlike growth factor, placental transfer, prenatal growth disorder, tobacco abuse, disease /disorder proneness /risk, environmental exposure, enzyme activity, gene expression, outcomes research, placenta, pregnancy, clinical research, cord blood, embryo /fetus, female, human pregnant subject, young adult human (21-34)
Project start date: 2004-09-30
Project end date: 2009-07-31
5R01HD047003-05 (2007): $316561
5R01HD047003-04 (2006): $326015
5R01HD047003-02 (2005): $322994
Grants awarded to Frederick Naftolin
ESTRADIOL-INDUCED SIALYATION OF NCAM PREVENTS LEUCOCYTE:ENDOTHELIAL ADHESION
Frederick Naftolin
New York University School Of Medicine, New York, Ny 10016
Grant 5RC1HL100769-02 from National Heart, Lung, And Blood Institute
Abstract: This proposal addresses broad Challenge Area (04) Clinical Research and specific Challenge topic, 04- HL-103 Assess the role of leukocyte interaction with platelets, erythrocytes, and endothelium in the pathogenesis of heart, lung, and blood diseases. The cadherin Neural Cell Adhesion Molecule (NCAM) is key in leukocyte adhesion to endothelial cells. Upon signaling by chemokines such as MCP1 leukocytes are drawn toward endothelial cells. Appositional interactions ("zippering") between NCAM molecules from the leukocyte and endothelial cell forms a tether between them and allows the leukocyte´s cell surface antigens to interact with those of the endothelial cell. This is followed by cellcell interactions such as diapedesis. The resulting sub-endothelial inflammatory processes can result in atheroma formation and atherosclerosis. However, this sequence may be blocked by the insertion of the NCAM by sialic acid decorated isoforms termed polysialated NCAM (PSA-NCAM). These have poly hydrated ¿2,8 sialic acid chains in the extracellular domain that interfere with the zipper adhesion to NCAM. We have documented the presence of PSA-NCAM in the glycocalyx of human vascular endothelial cells in arteries, veins and lymphatics of all organs (10 organ systems) studied. We also have shown that expression of the two required polysialyltransferases (ST8Sia II/STX and ST8Sia IV/PST) is estrogen receptor-mediated. Thus, the enzymatic replacement of NCAM with PSA-NCAM depletes the parent NCAM molecules, so that the adhesion (NCAM)non- adhesion (PSA-NCAM) balance is highly leveraged. In the case of the vascular endothelium, the result may be minimization of leukocyte adhesion and sub endothelial inflammation, which would diminish the formation of atheromas and atherosclerosis. In previous studies we have shown that estradiol induces PST and STX in rat brain tissue and increases PSA- NCAM at the expense of NCAM in human umbilical vein endothelial cells (HUVEC). The estrogen receptor blocker fulvestrant had the opposite effect. In this proposal, using cardiovascular-relevant cells [human leukocytes (THP1) and human arterial endothelial cells (HCAEC)] we will examine estrogen-regulated leukocyte adhesion We will determine the NCAMPSA-NCAM ratio induced by estradiol and fulvestrant, their effects on sialylation enzymes and their effect on leukocyte-endothelial adhesion. We will compare the results with effects of the Ras pathway inducer valproic acid which is known to induce PSA-NCAM. Further, the effects of siRNA inhibition of STX and PST on the NCAMPSA-NCAM balance and on leukocyte adhesion to endothelial cells will also be investigated. In this manner, we will test whether estrogen´s cardioprotective effects could be due to increased NCAM sialylation, leading to decreased leukocyte adhesion and sub-endothelial inflammation. Impact Heart disease is the single greatest cause of death and results in staggering healthcare expenditures and personal suffering. Finding new ways to protect against atherosclerosis is imperative for the health of the nation. Because of its importance to atherogenesis, understanding of the regulation of leukocyte-endothelial cell interactions is of the greatest clinical importance. This proposal will test the role of estrogen in preventing leukocyte adhesion that is a premonitory step toward atheroma formation. Such action would be consistent with the well- documented cardio-protective effects of estrogen in experimental and clinical studies. If this proposal is successful in exposing the mechanism by which estrogen affects leukocyte adhesion to endothelial cells it may be possible to alter already available treatments to avoid atherosclerosis and develop novel preventative treatments specifically aimed at regulating NCAM sialylation in clinical cardioprotection. Arteriosclerotic heart disease, the number one killer in the USA, starts from blood vessel inflammation that ultimately puts the individual at risk of a heart attack. This proposal will explore whether estrogen protects against atherosclerosis through production of specific, estrogen-regulated enzymes that inhibit inflammatory cells from entering into blood vessels. This proposal may improve the application of anti-atherosclerosis agents and lead to development of new agents to protect blood vessels from atherosclerosis and its complications. Arteriosclerotic heart disease, the number one killer in the USA, starts from blood vessel inflammation that ultimately puts the individual at risk of a heart attack. This proposal will explore whether estrogen protects against atherosclerosis through production of specific, estrogen-regulated enzymes that inhibit inflammatory cells from entering into blood vessels. This proposal may improve the application of anti-atherosclerosis agents and lead to development of new agents to protect blood vessels from atherosclerosis and its complications
Keywords: 2-Propylpentanoic Acid; 7-(9-(4, 4, 5, 5, 5-pentafluoropentylsulfinyl)nonyl)estra-1, 3, 5(10)-triene-3, 17-diol; 7a-[9-[(4, 4, 5, 5, 5, -Pentafluoropentyl)sulphinyl]nonyl]-estra-1, 3, 5(10)-triene-3, 17b-diol; Address; Adhesions; Affect; Aquadiol; Area; Arterial Fatty Streak; Arteries; Assay; Atheroma; Atheromatous; Atheromatous degeneration; Atheromatous plaque; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Binding; Binding (Molecular Function); Bioassay; Biologic Assays; Biological Assay; Bizzozero`s corpuscle/cell; Blood Diseases; Blood Platelets; Blood Vessels; Blood erythrocyte; Blood leukocyte; Blood monocyte; Blood normocyte; Blotting, Western; CCL2; CCL2 gene; CD56; Cadherins; Cardiac Diseases; Cardiac Disorders; Cardiac infarction; Cardiovascular; Cardiovascular Body System; Cardiovascular system; Cardiovascular system (all sites); Care, Health; Carotid Artery Ulcerating Plaque; Cause of Death; Cell Coat; Cell Communication; Cell Communication and Signaling; Cell Interaction; Cell Line; Cell Lines, Strains; Cell Signaling; Cell Surface Antigens; Cell-to-Cell Interaction; CellLine; Cells; Change of Life, Female; Clinical; Clinical Research; Clinical Study; Common Rat Strains; Cytokines, Chemotactic; Deetjeen`s body; Depakote; Depakote ER; Development; Dimenformon; Diogyn; Diogynets; Divalproex; Documentation; Endothelial Cells; Endothelium; Enzymes; Equilibrium; Erythrocytes; Erythrocytic; Estra-1, 3, 5(10)-triene-3, 17-diol (17beta)-; Estrace; Estradiol; Estradiol-17 beta; Estradiol-17beta; Estraldine; Estrogen Receptors; Estrogenic Agents; Estrogenic Compounds; Estrogens; External Domain; Extracellular Domain; Foam Cells; Fulvestrant; Fulvestrant (Faslodex); GDCF-2; GDCF-2 HC11; Glycocalyx; Goals; HC11; Hayem`s elementary corpuscle; Health; Healthcare; Heart; Heart Diseases; Hematologic Diseases; Hematological Disease; Hematological Disorder; Homologous Chemotactic Cytokines; Hormonal; Human; Human, General; INFLM; Individual; Inflammation; Inflammatory; Intercrines; Intracellular Communication and Signaling; Isoforms; Lead; Leukocytes; Life; Liver Cell Adhesion Molecules; Lung; Lymphatic; MCAF; MCP-1; MCP1; MGC9434; Mammals, Primates; Mammals, Rats; Man (Taxonomy); Man, Modern; Marrow erythrocyte; Marrow leukocyte; Marrow monocyte; Marrow platelet; Mediating; Menopause; Methods; Modeling; Molecular Interaction; Myocardial Infarct; Myocardial Infarction; N-Acetylneuraminic Acids; NCAM; NCAM1; NCAM1 gene; NKH-1; Neural Cell Adhesion Molecules; New Agents; Organ; Organ System; Organ System, Cardiovascular; Ovarian; Ovocyclin; Ovocylin; Parents; Pathogenesis; Pathway interactions; Pb element; Pentanoic acid, 2-propyl-; Personal Expenditure; Platelets; Primates; Process; Production; Progynon; Protein Isoforms; RNA, Small Interfering; RT-PCR; RTPCR; Rat; Rattus; Reaction; Red Blood Cells; Red Cell; Red blood corpuscule; Red cell of marrow; Regulation; Research Design; Respiratory System, Lung; Reticuloendothelial System, Erythrocytes; Reticuloendothelial System, Leukocytes; Reticuloendothelial System, Platelets; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; SCYA2; SIS cytokines; SMC-CF; ST8Sia II; Sialic Acids; Sialyltransferases; Signal Transduction; Signal Transduction Systems; Signaling; Small Interfering RNA; Streaks, Arterial Fatty; Study Type; Surface Antigens; Surface Markers, Immunologic; Surface Markers, Immunological; Testing; Therapeutic Estradiol; Therapeutic Estrogen; Thrombocytes; Time; Ulcerating Plaque, Carotid Artery; Ulcers, Carotid; Umbilical vein; Valproic Acid; Vascular Endothelial Cell; Vascular Endothelium; Vascular, Heart; Veins; Western Blotting; Western Blottings; Western Immunoblotting; White Blood Cells; White Cell; Woman; ing; analog; atherogenesis; atheromatosis; atherosclerosis plaque; atherosclerotic lesions; atherosclerotic plaque; atherosclerotic vascular disease; balance; balance function; base; biological signal transduction; blood corpuscles; blood disorder; body system; brain tissue; cardiac infarct; chemoattractant cytokine; chemokine; circulatory system; clinical practice; coronary attack; coronary infarct; coronary infarction; cultured cell line; design; designing; eroded vascular plaques; heart attack; heart disorder; heart infarct; heart infarction; heavy metal Pb; heavy metal lead; immunocytochemistry; improved; liver cell adhesion molecule; macrophage; menopausal; monocyte; novel; pathway; prevent; preventing; protective effect; protein blotting; pulmonary; reverse transcriptase PCR; siRNA; sialylation; social role; study design; thrombocyte/platelet; vascular; vascular corrosion plaques; vascular erosion plaques; vascular inflammation; vascular plaque; vulnerable plaque; white blood cell; white blood corpuscle
Relevance: Arteriosclerotic heart disease, the number one killer in the USA, starts from blood vessel inflammation that ultimately puts the individual at risk of a heart attack . This project will explore whether estrogen protects against atherosclerosis through production of specific, estrogen-regulated enzymes that inhibit inflammatory cells from entering into blood vessels. This project may improve the application of anti-atherosclerosis agents and lead to development of new agents to protect blood vessels from atherosclerosis and its complications
Project start date: 2009-09-30
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-003
5RC1HL100769-02 (2010): $452126
1RC1HL100769-01 (2009): $496482
ESTROGEN AND PRIMATE BRAIN CELLS REGULATING GONADOTROPIN
Frederick Naftolin
Obstetrics Gynecology & Reprod Scisyale University
47 College Street, Ste 203
new Haven, Ct 065208047
Grant 3R01NS036111-04S1 from National Institute Of Neurological Disorders And Stroke IRG: REB
Abstract: This application will test the hypothesis that, in addition to its effects on neurotransmitter production and receptors, estrogen affects gonadotropin secretion in primates by regulating the synaptic connections between elements of the GnRH delivery system. The investigator proposes that by the time of the preovulatory gonadotropin surge, the ratio of the number of stimulatory and inhibitory synapses is higher on GnRH neurons than at any other time during the menstrual cycle. To test this hypothesis, the investigator will determine the synaptic connection and activity of GnRH neurons in the monkey brain during estrogen-regulated control of gonadotropins. During the project period, the investigator will study 1) experimentally stimulated positive feedback induced by administered estradiol 2) negative and positive feedback during the normal ovarian cycle; 3) feedback in normal males; 4) effects of ovariectomy with or without hormone replacement; and 5) naturally occurring menopause. Three major inhibitory and three major stimulator neurotransmitter systems will be studied in this project
Keywords: estradiol, gonadotropin, hormone regulation /control mechanism, menstrual cycle, secretion autocrine, gonadotropin releasing factor, hormone therapy, menopause, neural transmission, neurotransmitter, neurotransmitter receptor, ovulation, synapse Primate, female, ovariectomy
Project start date: 1996-09-30
Project end date: 2002-08-31
3R01NS036111-04S1 (2000): $82500
PHILIPS TECNAI 12 ELECTRON MICROSCOPE
Frederick Naftolin
Obstetrics Gynecology & Reprod Scisyale University
47 College Street, Ste 203
new Haven, Ct 065208047
Grant 1S10RR015731-01 from National Center For Research Resources IRG: ZRG1
Abstract: This application responds to NIH program announcement PAR-99-031 (NCRR shared instrumentation grant) and requests funding to purchase a replacement transmission electron microscope for the Department of Obstetrics and Gynecology, Yale University School of Medicine. For the last three decades, under NIH support, the reproductive neuroscience group in our department has been studying basic central and peripheral mechanisms that underlie normal reproduction and pathological conditions, primarily using morphological methods. We have also been collaborating with other departments. The transmission electron microscope is used for funded projects in the department (80%) plus collaborative projects (20%). The 12 projects listed in the application use ultrastructural analyses by transmission electron microscopy. Through a similar NIH initiative, we purchased a Philips CM 10 electron microscope 15 years ago. The Department has paid for the construction and maintenance of the electron microscopic suite and supporting facilities during this period, and will continue to do so. Although there are other electron microscopes in other departments of the Medical School and the University, because all the projects of the PI and those of Drs. Csaba Leranth and Tamas Horvath utilize electron microscopy as a primary tool of investigation our group has the need for the equipment almost every hour of the working day and on weekends. Thus, it is impossible to resolve our need of an electron microscope on a shared basis with other departments. Through this initiative, we are applying to purchase a new electron microscope Our 15 year old Philips CM 10 transmission electron microscope contributed observations that have been published in hundreds of peer reviewed research articles, but it is now completely broken and out of service. The heavy use of the machine resulted in increasing numbers of breakdowns during the past year. Because this model has been long discontinued, it is difficult to find the appropriate replacement parts to rebuild the microscope. It has been abandoned as a viable instrument and we are temporarily using a "mothballed" obsolete machine. But, this one cannot withstand the load and will soon break down for good. Twelve government-supported projects are listed (see text) that in the aggregate require constant use of an electron microscope. Without the possibility of purchasing a new electron microscope, these projects are in jeopardy! It is the conclusion of all participants that the most user-friendly and durable microscope currently available is the Philips Tecnai 10, which is the model that replaced the popular Philips CM 100 model. We describe the Philips Tecnai 10 in the text. We are requesting funds urgently to purchase this machine and an image analysis package in order to support our NIH-funded projects
Keywords: biomedical equipment purchase, female reproductive system, transmission electron microscopy bioimaging /biomedical imaging
Project start date: 2001-04-01
Project end date: 2002-09-30
1S10RR015731-01 (2001): $387862
HORMONE ACTION ON GABA/GLUTAMATE BALANCE OF GNRH CELLS
Frederick Naftolin
Obstetrics Gynecology & Reprod Scisyale University
47 College Street, Ste 203
new Haven, Ct 065208047
Grant 5R03TW000933-03 from Fogarty International Center IRG: ICP
Abstract: The parent grant will use morphometry, immunohistochemistry, quantitative synaptology and in situ hybridization to investigate the effect of estrogen on the input to GnRH neurons of primates. The present application is an extension of the same aims, with the use of pre-embedding double labeled light and EM immunocytochemistry for GnRH and one of six transmitters that are either inhibitory (GABA, catecholamines, beta-endorphin) or excitatory (galanin, substance P, neuropeptide Y) to GnRH release. In addition, postembedding immunocytochemistry for GABA and glutamate will be used to quantitate their respective inputs to GnRH neurons. The micrographs will be quantified for stained and unstained boutons contacting GnRH positive and negative cells, using a stereological method for quantification in 5 experimental animals per group. The groups will be comprised of primates in endogenous or simulated estrogen surge of mid menstrual cycle, and surgical or naturally menopausal animals, some replaced with estrogen. Similar experiments will be performed on ovariectomized rats, treated with estradiol or control. In general, the treatments and tissue procurements in primates will be performed at the US lab, and rat experiments at both labs. The immunocytochemistry and analysis will be performed at the foreign site
Keywords: estrogen, gamma aminobutyrate, glutamate, gonadotropin releasing factor, hormone regulation /control mechanism, menstrual cycle, neuron catecholamine, endorphin, estradiol, galanin, menopause, messenger RNA, neuropeptide Y, neurotransmitter transport, substance P, synapse Primate, electron microscopy, immunocytochemistry, in situ hybridization, laboratory rat, light microscopy, ovariectomy, tissue /cell culture
Project start date: 1998-03-01
Project end date: 2001-02-28
5R03TW000933-03 (2000): $31500
1R03TW000933-01 (1998): $25200
BRAIN MACROPHAGES AND REPRODUCTIVE AGING
Frederick Naftolin
Obstetrics Gynecology & Reprod Scisyale University
47 College Street, Ste 203
new Haven, Ct 065208047
Grant 5R01AG015457-03 from National Institute On Aging IRG: REN
Abstract: The aging female rat´s reproductive life ends when she can no longer produce a surge of gonadotrophin in response to the midcycle surge of estrogen (positve feedback). Normally, this hypothalamic aging is gradual, marked by peroxidase accumulation in astroglia in the hypothalamus. We have shown a relationship between estrogen-induced synaptic retraction, elaboration of glial processes and positive feedback. To explain this aging, we propose that in the cycling rat, resident brain macrophages in the arcuate nucleus are repeatedly activated by estrogen and produce free radicals that, in turn, disables hypothalamic function. In testing this hypothesis 1) In cycling female rats, we will characterize the physiological mid cycle activation of arcuate nucleus macrophages and determine the morphological relationship between synaptic plasticity of arcuate nucleus synapses/neurons, astrocytes and brain macrophages using light and electron microscopic immunolabeling for the neuronal marker microtubule-associated protein 2 (MAP 2), the astroglia marker glial fibrillary acidic protein (GFAP), the macrophage marker OX42, and, the appearance of a cell adhesion molecule 1 (I-CAM-1) to mark activated macrophages. 2) We propose to delay the onset of reproductive senescence in normally aging rats with the administration of vitamin E, an anti-oxidant that previously has been shown to suppress free radical production in the central nervous system. The reproductive cycles of normally aging animals will be monitored by vaginal smears and blood LH measurements. At three distinct aging milestones (3 months old, 11 months old and 15 months), control and vitamin E-treated females will be studied by light- and electron microscopic immunocytochemistry for brain macrophage markers in combination with quantitative synaptology will be carried out. The failure of reproduction is normally evident by eleven months and completed by 15 months. Thus, we will characterize aging functionally and morphologically and determine the protective effect of vitamin E. 3) To determine in vitro the cellular basis of activation of brain macrophages by estrogen, we will treat primary cultures of microglia, astrocytes, and, neuronal cell lines that express alpha, beta or alpha/beta estrogen receptors, alone and in combination, assess the effects of estrogen, alone or with vitamin E or superoxide dismutase on these. We will assess production of reactive oxygen species in these cultures by in-situ chemiluminescence and histochemistry
Keywords: aging, estrus, hypothalamus, leukocyte activation /transformation, macrophage, tocopherol antioxidant, astrocyte, biomarker, estrogen, estrogen receptor, free radical oxygen, hormone regulation /control mechanism, hypothalamic pituitary axis, microglia, neural plasticity, neuron, oxidative stress, pituitary gonadal axis, synapse female, flow cytometry, immunocytochemistry, laboratory rat, tissue /cell culture
Project start date: 1999-07-01
Project end date: 2003-06-30
5R01AG015457-03 (2001): $270074
5R01AG015457-02 (2000): $255582
Cigarette Smoke-induced Human Fetal Growth Restriction
Frederick Naftolin
New York University School Of Medicine New York, Ny 10016
Grant 7R01HD047003-03 from National Institute Of Child Health And Human Development IRG: ZHD1
Abstract: Studies have shown that maternal exposure to cigarette smoke during pregnancy correlates significantly with adverse conceptus development, especially fetal growth restriction (FGR). In addition to obstetrical complications, FGR has been implicated in long-term effects on adult health, morbidity and even mortality. To elucidate the mechanisms of cigarette smoke-induced FGR we will use samples of placentae, maternal blood and cord blood from smoking and non-smoking pregnancies whose degree of FGR are known. Our hypothesis is that alteration of human growth factor genes, specifically insulin-like growth factor-1 (IGF-1), can be induced by environmental exposure to cigarette smoke toxins and produce FGR when the expressed abnormal gene products fail to properly regulate growth and development. Smoking-related gene alterations (polymorphisms) can result from the failure of enzymes required for detoxification of toxins coupled with enzymatic activation to reactive products that bind with DNA (adduct formation). This leads to DNA polymorphism and expression of abnormal gene products. The primary focus of our studies will be IGF-1. Placental IGF-1 gene will be assessed for polymorphism and alteration of nucleic acid sequences. IGF-1 gene expression products in maternal and cord blood will be determined and characterized. Adduct formation between activated toxins and the placental IGF-1 gene will be assessed. Since the process of detoxification versus activation involves specific enzymes, these will be measured in affected and non-affected pregnancies from smoking and non-smoking mothers (controlled by serum nicotine and cotinine assays) by assessing detoxification or activation of polynuclear aromatic hydrocarbons (PAH) into reactive products that bind with DNA. Similar studies will be performed on cultured chorionic villus sample cells from first trimester (CVS). Finally, by genotyping of the IGF-1 gene in the first trimester (CVS) and post delivery placentas during the same pregnancy we will determine both early and late effects of smoking, and identify pregnancies with high risk of smoking-induced FGR. If successful, such testing could be of considerable clinical utility.
Keywords: embryo /fetus toxicology, gene environment interaction, genetic polymorphism, insulinlike growth factor, placental transfer, prenatal growth disorder, tobacco abuse, disease /disorder proneness /risk, environmental exposure, enzyme activity, gene expression, outcomes research, placenta, pregnancy, clinical research, cord blood, embryo /fetus, female, human pregnant subject, young adult human (21-34)
Project start date: 2004-09-30
Project end date: 2009-07-31
1R01HD047003-01 (2004): $322994
Sponsored Links Excellgen http://Excellgen.com
ESTROGEN AND PRIMATE BRAIN CELLS REGULATING GONADOTROPIN
Frederick Naftolin
Yale University 47 College Street, Ste 203 New Haven, Ct 065208047
Grant 5R01NS036111-04 from National Institute Of Neurological Disorders And Stroke IRG: REB
Abstract: Adapted from the Investigator s ) This application will test the hypothesis that, in addition to its effects on neurotransmitter production and receptors, estrogen affects gonadotropin secretion in primates by regulating the synaptic connections between elements of the GnRH delivery system. The investigator proposes that by the time of the preovulatory gonadotropin surge, the ratio of the number of stimulatory and inhibitory synapses is higher on GnRH neurons than at any other time during the menstrual cycle. To test this hypothesis, the investigator will determine the synaptic connection and activity of GnRH neurons in the monkey brain during estrogen-regulated control of gonadotropins. During the project period, the investigator will study 1) experimentally stimulated positive feedback induced by administered estradiol 2) negative and positive feedback during the normal ovarian cycle; 3) feedback in normal males; 4) effects of ovariectomy with or without hormone replacement; and 5) naturally occurring menopause. Three major inhibitory and three major stimulator neurotransmitter systems will be studied in this project.
Keywords: estradiol, gonadotropin, hormone regulation /control mechanism, menstrual cycle, secretion, autocrine, gonadotropin releasing factor, hormone therapy, menopause, neural transmission, neurotransmitter, neurotransmitter receptor, ovulation, synapse, Primate, female, ovariectomy
Project start date: 1996-09-30
Project end date: 2002-08-31
5R01NS036111-04 (1999): $371036
5R01NS036111-03 (1998): $283691
5R01NS036111-02 (1997): $283270
METABOLISM AND MECHANISM OF ACTION OF ESTROGENS
Frederick Naftolin
Yale University 47 College Street, Ste 203 New Haven, Ct 065208047
Grant 5R01HD013587-12 from National Institute Of Child Health And Human Development IRG: BCE
Abstract: Adapted from applicant s ) This application proposes to identify which synapses are involved in phased synaptic remodeling. By quantitative ultra-structural morphometry and synapse counts, the effects of ovariectomy (low estrogen), progesterone, and ring A-reduced androgens will be studied. Freeze-fracture analysis will be used to determine whether intramembranous protein particle(IMP) deletion regularly precedes synaptic changes and whether there is a critical level of IMP density in postsynaptic membranes below which it is not possible to maintain cyclic circuitry. Neuronal proliferation and differentiation in fetal hypothalamic cell cultures will be studied utilizing cell- and molecular biology methods to probe the appearance of oncogenes and intracellular/membrane proteins.
Keywords: estrogen, hormone regulation /control mechanism, neural plasticity, synapse, synaptogenesis, aging, castration, developmental neurobiology, estradiol, estrogen inhibitor, estrogen receptor, hormone binding protein, hypothalamus, membrane protein, membrane structure, neuroanatomy, neuroendocrine system, neurotransmitter metabolism, ovariectomy, pinocytosis, progesterone, sex differentiation, electron microscopy, embryo /fetus cell /tissue, freeze etching, immunocytochemistry, laboratory rat, radioimmunoassay, radiotracer, sex difference, tissue /cell preparation
Project start date: 1980-09-30
Project end date: 1995-07-31
5R01HD013587-12 (1992): $287629
ESTROGEN AND PRIMATE BRAIN CELLS REGULATING GONADOTROPIN
Frederick Naftolin
Yale University 47 College Street, Ste 203 New Haven, Ct 065208047
Grant 1R01NS036111-01 from National Institute Of Neurological Disorders And Stroke IRG: REB
Project start date: 1996-09-30
Project end date: 2000-08-31
1R01NS036111-01 (1996): $277345
METABOLISM AND MECHANISM OF ACTION OF ESTROGENS
Frederick Naftolin
Obstetrics Gynecology & Reprod Scisyale University
47 College Street, Ste 203
new Haven, Ct 065208047
Grant 5R01HD013587-14 from National Institute Of Child Health And Human Development IRG: BCE
Project start date: 1980-09-30
Project end date: 1996-03-31
5R01HD013587-14 (1994): $312646