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Suture of ovary after laparoscopic excision of ovarian cysts preserves more ovarian function than bipolar electro-coagulation
genes, with higher expression of Cox2 in women >35 years of age (P<0.05). The level of expression of StAR, Cox2 and IL-1b was lower in follicular cells derived from women with endometriosis as compared to indication related to male factor and tubal factor with an inverse correlation with level of mir-17, mir-542 and mir-211, respectively (P<0.05). There was no correlation between the expression of these miRNAs and their target genes with Day-3 and 10 FSH or Day-3 E2 serum levels. CONCLUSIONS: This is the first demonstration of the expression of miRNAs and their target genes in human ovarian cells. Since these and other miRNAs regulate the expression of a number of genes functionally related to cell growth, differentiation, apoptosis and inflammation, altered expression of mir-17, mir-211 and mir-542 in ovarian cells-derived from women experiencing infertility imply a key regulatory function for these miRNAs in cellular activities that leads to ovarian dysfunction (supported in part by NIH grant HD37432). Supported by: Supported in part by NIH grant HD37432.
P-175 MICROARRAY ANALYSIS OF GENE EXPRESSION IN THE PRIMATE OVULATORY FOLLICLE: SELECTED GENE TRANSCRIPTS INVOLVED IN CUMULUS–OOCYTE EXPANSION. F. Xu, J. D. Hennebold, J. Mueller, B. Lindenthal, M. A. Sackett, R. L. Stouffer. Division of Reproductive Sciences, Oregon National Primate Research Center, Beaverton, OR; Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR; TRG Gynecology and Andrology, Bayer Schering Pharma AG, Berlin, Germany. OBJECTIVE: To identify, using a genome-wide approach, genes whose expression is regulated by LH in the primate ovulatory follicle. Particular attention was given to those genes implicated in cumulus-oocyte expansion. DESIGN: Longitudinal study of gene expression at specific timepoints during controlled ovulation (COv) of the naturally selected, dominant follicle in rhesus monkeys during the menstrual cycle. MATERIALS AND METHODS: The preovulatory follicle (n ¼ 4–8/time point) was collected before (0 hr) and 12, 24, and 36 hr after injection of an ovulatory bolus of hCG. Since ovulated follicles were observed at 36 hr, samples at this timepoint were divided into unruptured and ruptured follicles (n ¼ 4/group). Total RNA was prepared from individual follicles. An aliquot used for DNA microarray analysis (Affymetrix GeneChipÒ Macaque Genome Array) and the remainder stored for validation of expression patterns for selected genes by real-time PCR. The latter data were analyzed by oneway ANOVA followed by S-N-K tests. RESULTS: Between timepoint comparisons revealed profound changes (P<0.05) in mRNA levels. Those for an enzyme catalyzing hyaluronan synthesis (HAS2) and for a component (tumor necrosis factor-alpha induced protein, TNFAIP6) linking hyaluronan in the cumulus, were low at 0 hr, increased markedly (5–10 fold) at 12 hr, then declined to baseline by 36 hr before rupture. Also, mRNA levels for the epidermal growth factor family members, amphiregulin and epiregulin (not betacellulin) also increased up to 100 fold by 12 hr, and decreased markedly before rupture. In contrast, mRNA levels for the rate-limiting enzyme in prostaglandin synthesis (PTGS2) increased modestly at 12 hr, followed by a second rise (>10fold) at 36 hrs in the ruptured follicle. The four receptors for PGE (PTGER1–4) were expressed, but mRNA levels did not change markedly (2–3 fold increase, PTGER2 and 3, P<0.05) following hCG treatment. Notably, mRNA levels for several of these genes (e.g., HAS2, TNFAIP6, amphiregulin, epiregulin) also increased 5–10 fold at 36 hr in ruptured compared to unruptured follicles. CONCLUSIONS: Several genes encoding factors involved in oocyte maturation and cumulus-oocyte expansion in rodents are also expressed in a pattern that is consistent with similar activities in the primate ovulatory follicle. Their subsequent expression in the ruptured follicle suggests an additional role during development of the functional corpus luteum. Supported by: Bayer Schering Pharma, Female AMPPA; U54 CDRC HD55744; M.J. Murdock Charitable Trust; RR00163.
M.-Y. Chang, C.-S. Shiau, T.-T. Hsieh. Obstetrics and Gynecology, Chang Gung Memorial Hospital, Taipei, Taiwan. OBJECTIVE: To compare the functional ovarian damage associated with the use of bipolar coagulation vs. ovarian suture after laparoscopic excision of unilateral ovarian cysts. DESIGN: A retrospective study was performed from June 2000 to May 2005 in Taipei-CGMH. MATERIALS AND METHODS: 69 consecutive women who underwent laparoscopic stripping of the unilateral benign ovarian cyst. 40 patients had bipolar electrocoagulation (group A) while 29 had suturing of the ovary (group B). Plasma follicle-stimulating hormone (FSH), transvaginal ultrasound monitoring of antral follicle count (AFC) and basal ovarian volume in day 3 (D3) of the cycle were determined before surgery and re-evaluated at 12-month follow-up. RESULTS: Significant difference of AFC between the operated and contralateral ovaries in group A (6.70 0.45 vs. 7.95 0.48, P¼0.02) in 12month follow-up. There was a trend of higher variation percentage of AFC in the suturing group. Significant difference of postsurgical ovarian volume was observed in group A (5.27 0.59 vs. 8.43 1.04, P<0.004). No significant reduction of D3 FSH was shown in each group. TABLE. Ovarian function of operated and contralateral ovaries in both groups in 12-month follow up
Operated ovary Contralateral ovary Group A (n ¼ 40) AF count Variation percentage of AF count (%) Ovarian volume (cm3) Group B (n ¼ 29) AF count Variation percentage of AF count (%) Ovarian volume (cm3)
P
6.70 0.45 46.12 7.58
7.95 0.65 10.60 8.23
0.02 <0.001
5.27 0.59
8.43 1.04
<0.004
7.20 0.67 65.23 9.40
9.14 0.46 11.21 3.84
0.09 <0.001
7.19 0.93
8.09 1.09
0.54
Values are mean SEM.
Figure
CONCLUSIONS: Bipolar electrocoagulation of the ovarian parenchyma adversely affects ovarian function and suturing would be the proper way for homeostasis and preserving residual ovarian reserve. Supported by: None.
P-176
P-177
SUTURE OF OVARY AFTER LAPAROSCOPIC EXCISION OF OVARIAN CYSTS PRESERVES MORE OVARIAN FUNCTION THAN BIPOLAR ELECTRO-COAGULATION. C.-L. Hsieh,
THE EXPRESSION OF microRNA (miRNA) mir-23a AND 23b AND THEIR TARGET GENE, CYP19A1 (AROMATASE) IN FOLLICULAR CELLS OBTAINED FROM WOMEN UNDERGOING ART.
S166
Abstracts
Vol. 88, Suppl 1, September 2007
P-175 MICROARRAY ANALYSIS OF GENE EXPRESSION IN THE PRIMATE OVULATORY FOLLICLE: SELECTED GENE TRANSCRIPTS INVOLVED IN CUMULUS–OOCYTE EXPANSION. F. Xu, J. D. Hennebold, J. Mueller, B. Lindenthal, M. A. Sackett, R. L. Stouffer. Division of Reproductive Sciences, Oregon National Primate Research Center, Beaverton, OR; Department of Obstetrics and Gynecology, Oregon Health & Science University, Portland, OR; TRG Gynecology and Andrology, Bayer Schering Pharma AG, Berlin, Germany. OBJECTIVE: To identify, using a genome-wide approach, genes whose expression is regulated by LH in the primate ovulatory follicle. Particular attention was given to those genes implicated in cumulus-oocyte expansion. DESIGN: Longitudinal study of gene expression at specific timepoints during controlled ovulation (COv) of the naturally selected, dominant follicle in rhesus monkeys during the menstrual cycle. MATERIALS AND METHODS: The preovulatory follicle (n ¼ 4–8/time point) was collected before (0 hr) and 12, 24, and 36 hr after injection of an ovulatory bolus of hCG. Since ovulated follicles were observed at 36 hr, samples at this timepoint were divided into unruptured and ruptured follicles (n ¼ 4/group). Total RNA was prepared from individual follicles. An aliquot used for DNA microarray analysis (Affymetrix GeneChipÒ Macaque Genome Array) and the remainder stored for validation of expression patterns for selected genes by real-time PCR. The latter data were analyzed by oneway ANOVA followed by S-N-K tests. RESULTS: Between timepoint comparisons revealed profound changes (P<0.05) in mRNA levels. Those for an enzyme catalyzing hyaluronan synthesis (HAS2) and for a component (tumor necrosis factor-alpha induced protein, TNFAIP6) linking hyaluronan in the cumulus, were low at 0 hr, increased markedly (5–10 fold) at 12 hr, then declined to baseline by 36 hr before rupture. Also, mRNA levels for the epidermal growth factor family members, amphiregulin and epiregulin (not betacellulin) also increased up to 100 fold by 12 hr, and decreased markedly before rupture. In contrast, mRNA levels for the rate-limiting enzyme in prostaglandin synthesis (PTGS2) increased modestly at 12 hr, followed by a second rise (>10fold) at 36 hrs in the ruptured follicle. The four receptors for PGE (PTGER1–4) were expressed, but mRNA levels did not change markedly (2–3 fold increase, PTGER2 and 3, P<0.05) following hCG treatment. Notably, mRNA levels for several of these genes (e.g., HAS2, TNFAIP6, amphiregulin, epiregulin) also increased 5–10 fold at 36 hr in ruptured compared to unruptured follicles. CONCLUSIONS: Several genes encoding factors involved in oocyte maturation and cumulus-oocyte expansion in rodents are also expressed in a pattern that is consistent with similar activities in the primate ovulatory follicle. Their subsequent expression in the ruptured follicle suggests an additional role during development of the functional corpus luteum. Supported by: Bayer Schering Pharma, Female AMPPA; U54 CDRC HD55744; M.J. Murdock Charitable Trust; RR00163.
M.-Y. Chang, C.-S. Shiau, T.-T. Hsieh. Obstetrics and Gynecology, Chang Gung Memorial Hospital, Taipei, Taiwan. OBJECTIVE: To compare the functional ovarian damage associated with the use of bipolar coagulation vs. ovarian suture after laparoscopic excision of unilateral ovarian cysts. DESIGN: A retrospective study was performed from June 2000 to May 2005 in Taipei-CGMH. MATERIALS AND METHODS: 69 consecutive women who underwent laparoscopic stripping of the unilateral benign ovarian cyst. 40 patients had bipolar electrocoagulation (group A) while 29 had suturing of the ovary (group B). Plasma follicle-stimulating hormone (FSH), transvaginal ultrasound monitoring of antral follicle count (AFC) and basal ovarian volume in day 3 (D3) of the cycle were determined before surgery and re-evaluated at 12-month follow-up. RESULTS: Significant difference of AFC between the operated and contralateral ovaries in group A (6.70 0.45 vs. 7.95 0.48, P¼0.02) in 12month follow-up. There was a trend of higher variation percentage of AFC in the suturing group. Significant difference of postsurgical ovarian volume was observed in group A (5.27 0.59 vs. 8.43 1.04, P<0.004). No significant reduction of D3 FSH was shown in each group. TABLE. Ovarian function of operated and contralateral ovaries in both groups in 12-month follow up
Operated ovary Contralateral ovary Group A (n ¼ 40) AF count Variation percentage of AF count (%) Ovarian volume (cm3) Group B (n ¼ 29) AF count Variation percentage of AF count (%) Ovarian volume (cm3)
P
6.70 0.45 46.12 7.58
7.95 0.65 10.60 8.23
0.02 <0.001
5.27 0.59
8.43 1.04
<0.004
7.20 0.67 65.23 9.40
9.14 0.46 11.21 3.84
0.09 <0.001
7.19 0.93
8.09 1.09
0.54
Values are mean SEM.
Figure
CONCLUSIONS: Bipolar electrocoagulation of the ovarian parenchyma adversely affects ovarian function and suturing would be the proper way for homeostasis and preserving residual ovarian reserve. Supported by: None.
P-176
P-177
SUTURE OF OVARY AFTER LAPAROSCOPIC EXCISION OF OVARIAN CYSTS PRESERVES MORE OVARIAN FUNCTION THAN BIPOLAR ELECTRO-COAGULATION. C.-L. Hsieh,
THE EXPRESSION OF microRNA (miRNA) mir-23a AND 23b AND THEIR TARGET GENE, CYP19A1 (AROMATASE) IN FOLLICULAR CELLS OBTAINED FROM WOMEN UNDERGOING ART.
S166
Abstracts
Vol. 88, Suppl 1, September 2007