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Global metabolic and gene expression changes in adult tissues of mice conceived by in vitro fertilization
with PCOS compared to white women from NHANES was 1.52 (95% CI 1.03-2.23), while the same comparison for black women was 2.42 (95% CI 1.52-3.82). After controlling for age and BMI, black women with PCOS had an increased prevalence of low HDL and high glucose. CONCLUSION: This is the first study to comprehensively demonstrate increased risk of both Met Syn and general CVD in black women with PCOS compared to white women with PCOS. This racial disparity exists in the PCOS population compared to NHANES controls. Our findings highlight the importance of screening and need for longitudinal follow-up for cardiometabolic risk in young black women with PCOS. Supported by: FOCUS Medical Student Fellowship in Women’s Health Supported by the Edna G. Kynett Memorial Foundation. O-329 Wednesday, October 16, 2013 12:15 PM GLOBAL METABOLIC AND GENE EXPRESSION CHANGES IN ADULT TISSUES OF MICE CONCEIVED BY IN VITRO FERTILIZATION. S. K. Feuer, X. Liu, W. Lin, R. Simbulan, A. Donjacour, P. F. Rinaudo. OB/GYN & Reproductive Sciences, University of California, San Francisco, San Francisco, CA. OBJECTIVE: Mice conceived by in vitro fertilization (IVF) display evidence of altered growth, fat metabolism, and glucose homeostasis. This study investigates the mechanisms underlying these metabolic alterations observed in adult IVF mice. DESIGN: Experimental laboratory study in C57Bl/6J mice. MATERIALS AND METHODS: In vitro-fertilized embryos were cultured in KSOM with amino acids under 5% O2 and 5% CO2 until the blastocyst stage before transfer into pseudopregnant recipient dams, using in vivo-conceived blastocysts flushed and transferred into surrogates as a control. At 29wks of age, microarray analysis was conducted on liver, gonadal fat, skeletal muscle, and pancreatic islets from IVF and control-conceived female animals. Unbiased metabolic profiling of serum, liver, and gonadal fat was concurrently performed using a triple platform non-targeted mass spectrometry analysis (Metabolon, Inc) to identify metabolites differentially present between the in vivo and IVF conception conditions. RESULTS: Global metabolomic analysis revealed changes in glutathione and redox homeostasis, glycolytic and TCA cycle derivatives, the pentose phosphate pathway, branched chain amino acid oxidation, bile acid metabolism, ketogenesis, and purine nucleotide precursors like AMP, all of which may influence the metabolic alterations observed in IVF animals compared to in vivo controls. Microarray analyses uncovered differential gene expression signatures affecting lipid metabolism, insulin signaling, and mitochondrial function in IVF animals. Genes differentially expressed across multiple tissues were functionally enriched in transcription and chromatin remodeling, mRNA processing, and metabolism. CONCLUSION: Conception by IVF leads to long-term metabolic stress in a mouse model, manifesting as impaired redox and glucose homeostasis reflected in both gene expression and metabolite profiles. Supported by: R01 HD 062803 – 01A1 to PFR, NIH #T32DK007418-31 to SKF.
O-330 Wednesday, October 16, 2013 12:30 PM TELOMERE ORGANIZATION WITHIN THE SPERM NUCLEUS: AN ESSENTIAL PREREQUISITE FOR NORMAL FERTILIZATION AND EMBRYOGENESIS? N. M. Millan,a E. Jordan,a D. Ioannou,a H. G. Tempest.a aHuman and Molecular Genetics, Florida International University, Miami, FL; bIVF Florida Reproductive Associates, Margate, FL. OBJECTIVE: Evaluate the organization of telomeres within human spermatozoa. DESIGN: Cross-sectional study in a laboratory setting. MATERIALS AND METHODS: Nine normozoospermic males were recruited. Telomere organization for chromosomes 18, 19, 21, 22 and X within spermatozoa was performed utilizing fluorescence in-situ hybridization (FISH). Telomere organization was assessed via two means: radial (interior-peripheral) and polar (head-tail). A total of 9,000 sperm nuclei were examined (100 cells/chromosome/subject/analysis). The Chi-squared goodness-of-fit test was performed to determine nonrandom organization of telomeres. RESULTS: All telomeres tested exhibited nonrandom organization for both radial and polar analysis (p¼<0.05), with the exception of the telomeres for chromosome 18 (displayed a random organization in one subject). A distinct hierarchy of telomere position was identified both radially and longi-
FERTILITY & STERILITYÒ
tudinally. Our findings indicate a preferential radial nuclear organization of telomeres with respect to the nuclear interior to the periphery (22, X, 19, 21 and 18 respectively) and longitudinally from the sperm head to the tail (19, X, 22, 21 & 18 respectively). CONCLUSION: Paternal chromosomes are hypothesized to be withdrawn from the nucleus in a sequential order allowing gradual exposure of chromosomes to the ooplasm. This gradual decondensation and remodeling of specific regions could contribute to the differential gene activation patterns within the early embryo. This study provides evidence of nonrandom organization of human telomeres within the sperm nuclei. This hierarchical organization of chromatin suggests that those telomeric regions occupying the apical head region and the nuclear periphery are more likely to be exposed and activated first. We hypothesize that alterations within this organization may be associated with infertility and abnormal embryogenesis. Supported by: NM NIH/NIGMS R25 GM061347; HT & EJ DoD US Army Medical Research & Material Command W81XWH-10-1-0732.
ENDOMETRIOSIS II O-331 Wednesday, October 16, 2013 04:00 PM LOCALIZATION OF NERVE FIBERS IN NATURAL AND INDUCED ENDOMETRIOSIS IN RHESUS MACAQUES (MACACA MULATTA). O. D. Slayden,a J. Kawi,a C. Hergert,a L. D. Martin,b a a L. A. Holden. Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, Beaverton, OR; bDivision of Comparative Medicine, Oregon National Primate Research Center, Beaverton, OR. OBJECTIVE: To determine if changes in endometrial nerve fiber (NF) density and matrix metalloproteinase-7 (MMP-7) could be detected in a nonhuman primate model of induced endometriosis. DESIGN: In vivo treatments in a National Primate Research Center setting. MATERIALS AND METHODS: Endometriosis was induced in diseasefree rhesus macaques (n¼5) by seeding of menstrual tissue 72 hours following progesterone withdrawal, concordant with day one of menstruation. Menstrual tissue was collected by needle aspiration of the endometrial space and immediately transferred into the intraperitoneal cavity. Menstrual seeding was repeated for three menstrual cycles. Lesions and reproductive tract tissues were collected in the late proliferative phase of the sixth cycle, and assayed by immunohistochemistry for NF and MMP-7. Endometriosisfree animals (n¼4) were assayed as healthy controls and animals with advanced, naturally occurring, endometriosis (n¼4) acted as natural disease controls. RESULTS: Inoculations with menstrual debris created endometriosis in all 5 animals. Abundant nerve fibers were identified in the basalis zone of 3/5 animals with induced endometriosis. Nerve fibers were also abundant in 3/ 4 of the animals with naturally occurring advanced endometriosis. Nerve fibers were absent in the functionalis zone of control animals. MMP-7 was increased in the glands of the basalis zone of animals with endometriosis. MMP-7 was absent in control endometria. Nerve fibers and MMP-7 expression was observed in the ectopic lesions. CONCLUSION: Aberrant expression of nerve fibers is reported in the endometrium of patients with endometriosis who present with pelvic pain. Creating endometriosis in disease free macaques also results in aberrant nerve fiber development in the endometrium in most, but not all, animals. Coexpression of MMP-7 with abundant nerves in the endometrium and endometriotic lesions may be associated with endometriosis-related pelvic pain. Supported by: NIH P51 OD 011092.
O-332 Wednesday, October 16, 2013 04:15 PM CAN BRAIN-DERIVED NEUROTROPHIC FACTOR BE A CLINICAL MARKER FOR ENDOMETRIOSIS? J. M. Wessels,a N. A. Leyland,a S. Agarwal,b A. Murji,a W. G. Foster.a aObstetrics and Gynecology, McMaster University, Hamilton, ON, Canada; bReproductive Medicine, University of California, San Diego, La Jolla, CA. OBJECTIVE: To measure circulating brain-derived neurotrophic factor (BDNF) in women with endometriosis and disease-free controls and examine the relation to disease severity, pain, and menstrual cycle stage.
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O-330 Wednesday, October 16, 2013 12:30 PM TELOMERE ORGANIZATION WITHIN THE SPERM NUCLEUS: AN ESSENTIAL PREREQUISITE FOR NORMAL FERTILIZATION AND EMBRYOGENESIS? N. M. Millan,a E. Jordan,a D. Ioannou,a H. G. Tempest.a aHuman and Molecular Genetics, Florida International University, Miami, FL; bIVF Florida Reproductive Associates, Margate, FL. OBJECTIVE: Evaluate the organization of telomeres within human spermatozoa. DESIGN: Cross-sectional study in a laboratory setting. MATERIALS AND METHODS: Nine normozoospermic males were recruited. Telomere organization for chromosomes 18, 19, 21, 22 and X within spermatozoa was performed utilizing fluorescence in-situ hybridization (FISH). Telomere organization was assessed via two means: radial (interior-peripheral) and polar (head-tail). A total of 9,000 sperm nuclei were examined (100 cells/chromosome/subject/analysis). The Chi-squared goodness-of-fit test was performed to determine nonrandom organization of telomeres. RESULTS: All telomeres tested exhibited nonrandom organization for both radial and polar analysis (p¼<0.05), with the exception of the telomeres for chromosome 18 (displayed a random organization in one subject). A distinct hierarchy of telomere position was identified both radially and longi-
FERTILITY & STERILITYÒ
tudinally. Our findings indicate a preferential radial nuclear organization of telomeres with respect to the nuclear interior to the periphery (22, X, 19, 21 and 18 respectively) and longitudinally from the sperm head to the tail (19, X, 22, 21 & 18 respectively). CONCLUSION: Paternal chromosomes are hypothesized to be withdrawn from the nucleus in a sequential order allowing gradual exposure of chromosomes to the ooplasm. This gradual decondensation and remodeling of specific regions could contribute to the differential gene activation patterns within the early embryo. This study provides evidence of nonrandom organization of human telomeres within the sperm nuclei. This hierarchical organization of chromatin suggests that those telomeric regions occupying the apical head region and the nuclear periphery are more likely to be exposed and activated first. We hypothesize that alterations within this organization may be associated with infertility and abnormal embryogenesis. Supported by: NM NIH/NIGMS R25 GM061347; HT & EJ DoD US Army Medical Research & Material Command W81XWH-10-1-0732.
ENDOMETRIOSIS II O-331 Wednesday, October 16, 2013 04:00 PM LOCALIZATION OF NERVE FIBERS IN NATURAL AND INDUCED ENDOMETRIOSIS IN RHESUS MACAQUES (MACACA MULATTA). O. D. Slayden,a J. Kawi,a C. Hergert,a L. D. Martin,b a a L. A. Holden. Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, Beaverton, OR; bDivision of Comparative Medicine, Oregon National Primate Research Center, Beaverton, OR. OBJECTIVE: To determine if changes in endometrial nerve fiber (NF) density and matrix metalloproteinase-7 (MMP-7) could be detected in a nonhuman primate model of induced endometriosis. DESIGN: In vivo treatments in a National Primate Research Center setting. MATERIALS AND METHODS: Endometriosis was induced in diseasefree rhesus macaques (n¼5) by seeding of menstrual tissue 72 hours following progesterone withdrawal, concordant with day one of menstruation. Menstrual tissue was collected by needle aspiration of the endometrial space and immediately transferred into the intraperitoneal cavity. Menstrual seeding was repeated for three menstrual cycles. Lesions and reproductive tract tissues were collected in the late proliferative phase of the sixth cycle, and assayed by immunohistochemistry for NF and MMP-7. Endometriosisfree animals (n¼4) were assayed as healthy controls and animals with advanced, naturally occurring, endometriosis (n¼4) acted as natural disease controls. RESULTS: Inoculations with menstrual debris created endometriosis in all 5 animals. Abundant nerve fibers were identified in the basalis zone of 3/5 animals with induced endometriosis. Nerve fibers were also abundant in 3/ 4 of the animals with naturally occurring advanced endometriosis. Nerve fibers were absent in the functionalis zone of control animals. MMP-7 was increased in the glands of the basalis zone of animals with endometriosis. MMP-7 was absent in control endometria. Nerve fibers and MMP-7 expression was observed in the ectopic lesions. CONCLUSION: Aberrant expression of nerve fibers is reported in the endometrium of patients with endometriosis who present with pelvic pain. Creating endometriosis in disease free macaques also results in aberrant nerve fiber development in the endometrium in most, but not all, animals. Coexpression of MMP-7 with abundant nerves in the endometrium and endometriotic lesions may be associated with endometriosis-related pelvic pain. Supported by: NIH P51 OD 011092.
O-332 Wednesday, October 16, 2013 04:15 PM CAN BRAIN-DERIVED NEUROTROPHIC FACTOR BE A CLINICAL MARKER FOR ENDOMETRIOSIS? J. M. Wessels,a N. A. Leyland,a S. Agarwal,b A. Murji,a W. G. Foster.a aObstetrics and Gynecology, McMaster University, Hamilton, ON, Canada; bReproductive Medicine, University of California, San Diego, La Jolla, CA. OBJECTIVE: To measure circulating brain-derived neurotrophic factor (BDNF) in women with endometriosis and disease-free controls and examine the relation to disease severity, pain, and menstrual cycle stage.
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