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ethnic differences in ovarian reserve markers
RESULTS: A significant correlation between CMA3 positive cells and DNA damage was observed when damage was greater than 15% in the OAT and T groups. A negative correlation was found between CMA3 positive cells and sperm morphology. CONCLUSION: Decreased protamination positively correlates with DNA damage that exceeds 15% in OAT and T patients. Sperm DNA damage increases with deteriorating semen parameters. Assessment of sperm DNA damage, especially in oligoasthenoteratozoospermic and asthenoteratozoospermic patients, may be beneficial in assisted reproduction.
O-211 Wednesday, October 27, 2010 04:45 PM QUANTITATIVE SHOTGUN PROTEOMIC ANALYSIS OF SEMINAL PLASMA FROM MEN WITH SPINAL CORD INJURY-INDUCED ANEJACULATION. B. F. da Silva, C. R. Ferreira, M. N. Eberlin, J. S. Garcia, G. H. M. F. Souza, R. P. Bertolla. Division of Urology, Human Reproduction Section, Sao Paulo Federal University, Sao Paulo, SP, Brazil; ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, Campinas, SP, Brazil; Mass Spectrometry Applications Research and Development Laboratory, Waters Corporation, Alphaville, SP, Brazil; Department of Exact Sciences, Alfenas Federal University, Alfenas, MG, Brazil. OBJECTIVE: Most spinal cord injured (SCI) patients present a semen profile mainly characterized by normal sperm count and abnormal sperm motility. A possible correlation between this problem and abnormal semen constituents seems to exist. To further explore this information and to potentially identify proteins involved in SCI infertility, a quantitative shotgun proteomics strategy was employed to observe the seminal plasma proteome in these men. DESIGN: Prospective study. MATERIALS AND METHODS: Semen was obtained from men with SCI. Electroejaculation (EEJ) was performed in 6 patients, penile vibratory stimulation (PVS) in 6 other. 10 patients without SCI presenting normal semen were included as controls. Samples were pooled by group and submitted to nanoUPLC tandem nanoESI-MSE quantitative shotgun proteomics. Statistically identified proteins were analyzed for interactome networks using Cytoscape and classified according to Gene Ontology. RESULTS: A total of 881 proteins were identified, 325 differentially expressed between the three groups – 98 exclusively in the EEJ group, 135 in the PVS group, and 40 in controls. Moreover, 31 were overexpressed in men with EEJ, 15 in PVS, and 6 in controls. Cytoscape analysis demonstrated 8 interactome clusters, of which one was present only in men with PVS, responsible for response to hypoxia. A main cluster responsible for reproduction and ion binding was observed in all groups, but with 6 different proteins in men with EEJ and 4 in men with PVS, demonstrating putative pathways for altered semen quality. CONCLUSION: Men presenting with anejaculation due to SCI present alteration in the expression of a number of proteins when compared to controls with normal semen. The use of a quantitative shotgun proteomics approach has increased sensitivity and the dynamic range for protein identification. The information from this study may allow us to (i) better understand the pathways leading to normal and/or altered semen quality and (b) determine biomarkers for therapeutic intervention. Supported by: CNPq.
MATERIALS AND METHODS: 20 pubescent male Wistar Hannover rats were fed deionized water containing 0 or 100 mg/L Cd ad libitum for 1 or 8 weeks. Whole testis RNA was template for cRNA synthesis. Biotinylated cRNAs were hybridized to Affymetrix Rat 230.2 microarrays, and datamined by GeneSpring GX 7.3.1. Five microarrays per time/dose combination condition (one per rat) were statistically analyzed. RESULTS: Actin-related genes studied fell into two groups: [1] expression independent of Cd but changed with time, and [2] expression down-regulated by Cd at 1 week or 8 weeks. At 1 week, Cd-regulated genes largely controlled shape/morphogenesis, and kinases/phosphatases. At 8 weeks, Cd-modulated genes controlled actin filaments (interacting proteins; polymerization/depolymerization), cell projection formation and cell motility/migration. CONCLUSION: Cd differentially affected adolescent vs. adult testes. Our data are consistent with Cd-effected actin gene expression disruption contributing to apoptosis and decreased spermatogenesis in rat testes. Studies of gene expression in human testes biopsies should determine relevance to varicocele-associated infertility. Supported by: PHS Grant ES10496 (SHB); FIMR Faculty Research Award (SHB).
SOCIETY FOR REPRODUCTIVE ENDOCRINOLOGY AND INFERTILITY O-213 Wednesday, October 27, 2010 03:45 PM RACIAL/ETHNIC DIFFERENCES IN OVARIAN RESERVE MARKERS. M. P. Rosen, E. B. Johnstone, C. Addauan-Andersen, B. Sternfeld, C. McCulloch, M. I. Cedars. Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA; Division of Research, Kaiser Permanente, Oakland, CA; Epidemiology & Biostatistics, University of California San Francisco, San Francisco, CA. OBJECTIVE: Historical evidence suggests that African Americans and Hispanics have earlier ages of menopause than Caucasians, and Asians, later. We utilize a large, population-based cohort to evaluate racial/ethnic differences in markers of ovarian age. DESIGN: Cross-sectional. MATERIALS AND METHODS: Women aged 25-45 with ovulatory menses at 22-35 day intervals, enrolled in the population-based Ovarian Aging study, underwent transvaginal ultrasound for antral follicle count (AFC) and blood testing for antimullerian hormone (AMH) between cycle day 2 and 4. Generalized linear models were used to assess for ethnic differences in AMH and AFC, adjusted for age. RESULTS: This study includes 694 subjects: Caucasians (n¼244), African American (n¼175), Chinese (n¼131), and Latinas(n¼144). In Table 1 are the means and SDs for AMH and antral follicle count (AFC), stratified by age and ethnicity. AMH and AFC decline with age. Based on the regression analysis, AMH and AFC both differ by ethnicity: Caucasian women have higher levels of AMH compared to Latina women (p ¼ 0.006) while AFC is significantly higher in African-Americans than Chinese (p ¼ 0.03).
Mean SDs for AMH and AFC by Age
Age
Caucasian
Latina
AA
Chinese
42 19 22 10 36 24 18 9 26 22 13 8 10 7 85
40 18 23 9 25 17 17 9 17 12 12 7 11 8 94
35 8 23 11 28 6 18 10 28 1 14 9 15 6 96
37 27 18 8 36 28 17 8 19 14 11 6 11 8 84
O-212 Wednesday, October 27, 2010 05:00 PM CADMIUM ALTERS TESTICULAR EXPRESSION OF GENES REGULATING THE STRUCTURE AND FUNCTION OF THE ACTIN CYTOSKELETON. S. H. Benoff, J. L. Marmar, G. M. Centola, I. R. Hurley. Research, Feinstein Institute for Medical Research, Manhasset, NY; Surgery, Cooper University Hospital, Camden, NJ; Clinical Laboratory, New England Cryogenic Center, Newton, MA. OBJECTIVE: Actin dynamics affects cell cycle progression, cell junction function, spermiogenesis and spermiation. Actin immunoreactivity is low in testis of infertile men with varicocele. Varicocele actin loss may lead to sloughing of germ cells, increased apoptosis, and abnormal sperm head shape. Testes cadmium (Cd) is elevated in infertile men with varicoceles. Athough Cd can directly disassemble and degrade actin, it may also reduce actin by altering gene expression. In this study, we asked if Cd changed actinrelated gene expression in mammalian testes. DESIGN: We modeled chronic Cd exposures in Cd-sensitive rats, previously reporting time and dose dependent increases in testicular Cd and germ cell apoptosis, and decreased epididymal sperm counts. Now we examined expression of 105 cytoskeleton regulating genes.
S62
Abstracts
25-30 31-35 36-40 41-45
AMH AFC AMH AFC AMH AFC AMH AFC
CONCLUSION: This is the first study to estimate normative values for AMH and AFC in the general population and to compare these markers across ethnicities. Latina women show lower AMH than Caucasians despite similar AFC, while Chinese women have lower AFCs than African Americans, a finding not anticipated by the reported later age at menopause. These findings suggest ethnic differences in ovarian biology that are not explained by standard known confounders or available epidemiological evidence. Supported by: NICHD/NIA R01HD044876 & NIH/NCRR UL1 RR024131.
Vol. 94., No. 4, Supplement, September 2010
O-211 Wednesday, October 27, 2010 04:45 PM QUANTITATIVE SHOTGUN PROTEOMIC ANALYSIS OF SEMINAL PLASMA FROM MEN WITH SPINAL CORD INJURY-INDUCED ANEJACULATION. B. F. da Silva, C. R. Ferreira, M. N. Eberlin, J. S. Garcia, G. H. M. F. Souza, R. P. Bertolla. Division of Urology, Human Reproduction Section, Sao Paulo Federal University, Sao Paulo, SP, Brazil; ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, Campinas, SP, Brazil; Mass Spectrometry Applications Research and Development Laboratory, Waters Corporation, Alphaville, SP, Brazil; Department of Exact Sciences, Alfenas Federal University, Alfenas, MG, Brazil. OBJECTIVE: Most spinal cord injured (SCI) patients present a semen profile mainly characterized by normal sperm count and abnormal sperm motility. A possible correlation between this problem and abnormal semen constituents seems to exist. To further explore this information and to potentially identify proteins involved in SCI infertility, a quantitative shotgun proteomics strategy was employed to observe the seminal plasma proteome in these men. DESIGN: Prospective study. MATERIALS AND METHODS: Semen was obtained from men with SCI. Electroejaculation (EEJ) was performed in 6 patients, penile vibratory stimulation (PVS) in 6 other. 10 patients without SCI presenting normal semen were included as controls. Samples were pooled by group and submitted to nanoUPLC tandem nanoESI-MSE quantitative shotgun proteomics. Statistically identified proteins were analyzed for interactome networks using Cytoscape and classified according to Gene Ontology. RESULTS: A total of 881 proteins were identified, 325 differentially expressed between the three groups – 98 exclusively in the EEJ group, 135 in the PVS group, and 40 in controls. Moreover, 31 were overexpressed in men with EEJ, 15 in PVS, and 6 in controls. Cytoscape analysis demonstrated 8 interactome clusters, of which one was present only in men with PVS, responsible for response to hypoxia. A main cluster responsible for reproduction and ion binding was observed in all groups, but with 6 different proteins in men with EEJ and 4 in men with PVS, demonstrating putative pathways for altered semen quality. CONCLUSION: Men presenting with anejaculation due to SCI present alteration in the expression of a number of proteins when compared to controls with normal semen. The use of a quantitative shotgun proteomics approach has increased sensitivity and the dynamic range for protein identification. The information from this study may allow us to (i) better understand the pathways leading to normal and/or altered semen quality and (b) determine biomarkers for therapeutic intervention. Supported by: CNPq.
MATERIALS AND METHODS: 20 pubescent male Wistar Hannover rats were fed deionized water containing 0 or 100 mg/L Cd ad libitum for 1 or 8 weeks. Whole testis RNA was template for cRNA synthesis. Biotinylated cRNAs were hybridized to Affymetrix Rat 230.2 microarrays, and datamined by GeneSpring GX 7.3.1. Five microarrays per time/dose combination condition (one per rat) were statistically analyzed. RESULTS: Actin-related genes studied fell into two groups: [1] expression independent of Cd but changed with time, and [2] expression down-regulated by Cd at 1 week or 8 weeks. At 1 week, Cd-regulated genes largely controlled shape/morphogenesis, and kinases/phosphatases. At 8 weeks, Cd-modulated genes controlled actin filaments (interacting proteins; polymerization/depolymerization), cell projection formation and cell motility/migration. CONCLUSION: Cd differentially affected adolescent vs. adult testes. Our data are consistent with Cd-effected actin gene expression disruption contributing to apoptosis and decreased spermatogenesis in rat testes. Studies of gene expression in human testes biopsies should determine relevance to varicocele-associated infertility. Supported by: PHS Grant ES10496 (SHB); FIMR Faculty Research Award (SHB).
SOCIETY FOR REPRODUCTIVE ENDOCRINOLOGY AND INFERTILITY O-213 Wednesday, October 27, 2010 03:45 PM RACIAL/ETHNIC DIFFERENCES IN OVARIAN RESERVE MARKERS. M. P. Rosen, E. B. Johnstone, C. Addauan-Andersen, B. Sternfeld, C. McCulloch, M. I. Cedars. Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA; Division of Research, Kaiser Permanente, Oakland, CA; Epidemiology & Biostatistics, University of California San Francisco, San Francisco, CA. OBJECTIVE: Historical evidence suggests that African Americans and Hispanics have earlier ages of menopause than Caucasians, and Asians, later. We utilize a large, population-based cohort to evaluate racial/ethnic differences in markers of ovarian age. DESIGN: Cross-sectional. MATERIALS AND METHODS: Women aged 25-45 with ovulatory menses at 22-35 day intervals, enrolled in the population-based Ovarian Aging study, underwent transvaginal ultrasound for antral follicle count (AFC) and blood testing for antimullerian hormone (AMH) between cycle day 2 and 4. Generalized linear models were used to assess for ethnic differences in AMH and AFC, adjusted for age. RESULTS: This study includes 694 subjects: Caucasians (n¼244), African American (n¼175), Chinese (n¼131), and Latinas(n¼144). In Table 1 are the means and SDs for AMH and antral follicle count (AFC), stratified by age and ethnicity. AMH and AFC decline with age. Based on the regression analysis, AMH and AFC both differ by ethnicity: Caucasian women have higher levels of AMH compared to Latina women (p ¼ 0.006) while AFC is significantly higher in African-Americans than Chinese (p ¼ 0.03).
Mean SDs for AMH and AFC by Age
Age
Caucasian
Latina
AA
Chinese
42 19 22 10 36 24 18 9 26 22 13 8 10 7 85
40 18 23 9 25 17 17 9 17 12 12 7 11 8 94
35 8 23 11 28 6 18 10 28 1 14 9 15 6 96
37 27 18 8 36 28 17 8 19 14 11 6 11 8 84
O-212 Wednesday, October 27, 2010 05:00 PM CADMIUM ALTERS TESTICULAR EXPRESSION OF GENES REGULATING THE STRUCTURE AND FUNCTION OF THE ACTIN CYTOSKELETON. S. H. Benoff, J. L. Marmar, G. M. Centola, I. R. Hurley. Research, Feinstein Institute for Medical Research, Manhasset, NY; Surgery, Cooper University Hospital, Camden, NJ; Clinical Laboratory, New England Cryogenic Center, Newton, MA. OBJECTIVE: Actin dynamics affects cell cycle progression, cell junction function, spermiogenesis and spermiation. Actin immunoreactivity is low in testis of infertile men with varicocele. Varicocele actin loss may lead to sloughing of germ cells, increased apoptosis, and abnormal sperm head shape. Testes cadmium (Cd) is elevated in infertile men with varicoceles. Athough Cd can directly disassemble and degrade actin, it may also reduce actin by altering gene expression. In this study, we asked if Cd changed actinrelated gene expression in mammalian testes. DESIGN: We modeled chronic Cd exposures in Cd-sensitive rats, previously reporting time and dose dependent increases in testicular Cd and germ cell apoptosis, and decreased epididymal sperm counts. Now we examined expression of 105 cytoskeleton regulating genes.
S62
Abstracts
25-30 31-35 36-40 41-45
AMH AFC AMH AFC AMH AFC AMH AFC
CONCLUSION: This is the first study to estimate normative values for AMH and AFC in the general population and to compare these markers across ethnicities. Latina women show lower AMH than Caucasians despite similar AFC, while Chinese women have lower AFCs than African Americans, a finding not anticipated by the reported later age at menopause. These findings suggest ethnic differences in ovarian biology that are not explained by standard known confounders or available epidemiological evidence. Supported by: NICHD/NIA R01HD044876 & NIH/NCRR UL1 RR024131.
Vol. 94., No. 4, Supplement, September 2010