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DOES THE MALE AGEING INFLUENCE CLINICAL OUTCOMES ON ICSI CYCLES?
620
THE JOURNAL OF UROLOGY®
by electroporation. Our results suggest that this approach could have great relevance for the study of sperm function and male infertility. Source of Funding: None
1805 TROPHININ-BINDING PEPTIDE, GWRQ, PROMOTES HUMAN SPERM MOTILITY Shingo Hatakeyama*, Noritaka Kamimura, Takuya Koie, Kazuyuki Mori, Yasuhiro Hashimoto, Ikuya Iwabuchi, Takahiro Yoneyama, Atsushi Imai, Akiko Okamoto, Hayato Yamamoto, Chikara Ohyama, Michiko N Fukuda. La Jolla, CA, and Hirosaki, Japan. INTRODUCTION AND OBJECTIVE: Trophinin mediates a homophilic cell adhesion by trophinin-trophinin binding between trophectoderm and endometrial epithelium. (Genes Dev 1995) Previously we found that multivalent peptide, GWRQ-MAPS, binds to trophinin on cell surface, mimicking trophinin-mediated cell adhesion and subsequent signal transduction for trophectoderm activation.(PNAS 2007) Here we report that trophinin and trophinin-associated bystin and tastin proteins are found in the tail of human sperm, and that GWRQ-MAPS promoted motility of sperm. METHODS: Sperm samples were collected from 23 healthy males. Sperm mobility was compared by counting the numbers in WKH SUHVHQFH RI *:540$36 RU FRQWURO SHSWLGH /RFDOL]DWLRQ RI trophinin, bystin, tastin and GWRQ-peptide binding were determined by immunocytochemistry using anti-trophinin, anti-bystin, anti-tastin antibody and GWRQ-peptide displaying phage. ATP levels and intercellular calcium concentration were measured by luminometer. Swim down assay was assumed by transwell chamber with 8 µm pores from 3 healthy males. Cells moved from upper to bottom chamber were counted. 5(68/76 ,PPXQRF\WRFKHPLVWU\VXJJHVWHGFRORFDOL]DWLRQ of trophinin, bystin and tastin and the binding of GWRQ-peptide displaying phage to sperm tail was detected by immunostaining. (Fig. 1) In the presence of GWRQ-MAPS, the motility of human sperm was VLJQL¿FDQWO\LQFUHDVHG S )LJ DQGFHOOVUHGXFHG$73 S DQGH[KLELWHGVLJQL¿FDQWXSWDNHRIFDOFLXP S 6LQFHWDVWLQELQGV to Tctex-1 or microtubule-associated motor dynein ATPase (Biochem J 2002), trophinin bound with GWRQ may result in an activation of microtubule-associated ATPase. Since GWRQ-MAPS treated human sperm traveled through 8µm pores at higher frequency than those treated with control peptide, GWRQ-MAPS also promotes directed motility for a short distance. CONCLUSIONS: These results suggest that GWRQ-peptide enhances the sperm mobility. In normal human sperm, it is possible that trophinin plays a role in suppressing ATPase from premature over activation.
Vol. 179, No. 4, Supplement, Wednesday, May 21, 2008
1806 DOES THE MALE AGEING INFLUENCE CLINICAL OUTCOMES ON ICSI CYCLES? Renata C Ferreira, Tatiana C S Bonetti, Daniela Braga, Priscila Queiroz, Fabio F Pasqualotto, Assumpto Iaconelli, Edson Borges*. Sao Paulo, Brazil. INTRODUCTION AND OBJECTIVE: Lately an increased number of older couples requesting assisted reproductive treatment (ART) have been observed. It is clearly established the reduced fertility potential of older women, and alterations in semen quality associated with male ageing. Advanced male age in ART cycles provides a model to study biological paternal effects on embryo development and clinical outcomes, however the results are still controversy. The aim of this study was to evaluate the effect of male ageing on clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles, according to semen parameters. METHODS: Retrospective analysis included 760 ICSI cycles ZLWKIUHVKVSHUPDWR]RDDQGPRUHWKDQ0,,RRF\WHUHFRYHUHG:RPHQ received pituitary down regulation and controlled ovarian stimulation with recombinant-FSH. Follicle aspiration was done 35-36 hours after hCG trigger and MII oocytes were injected. Semen samples were collected E\ PDVWXUEDWLRQ DQG DQDO\]HG DFFRUGLQJ WR :+2 7KH F\FOHV ZHUH divided depending on sperm concentration: <20x106sperm/ml (n=220) DQG[6sperm/ml (n=540). In order to evaluate the effect of male ageing on clinical outcomes, adjusted to variations in women age, multiple linear regression analysis was performed. RESULTS: When sperm concentration were <20x106sperm/ml, there was a negative correlation between male age and implantation rate U S DQGPXOWLYDULDWHUHJUHVVLRQDGMXVWHGWRPDWHUQDO DJHVKRZHGWKHLQÀXHQFHRIH[FOXVLYHO\PDOHDJHLQJRQLPSODQWDWLRQ S :KHQWKHVSHUPFRQFHQWUDWLRQZHUH[6sperm/ml, there ZDVQRFRUUHODWLRQEHWZHHQPDOHDJHDQGLPSODQWDWLRQUDWH U S EXWRQWKHRWKHUKDQGWKHPDWHUQDODJHQHJDWLYHO\LQÀXHQFHG implantation rates (p<0.0001). CONCLUSIONS: The male ageing is associated with a VLJQL¿FDQW GHFOLQH LQ LPSODQWDWLRQ UDWHV LQ ,&6, F\FOHV ZKHQ VSHUP concentration was lower than 20x106sperm/ml, independently on maternal age. On the other hand, when sperm concentration was normal, HYHQWKRXJKLWZDVQRWREVHUYHGLQÀXHQFHRIPDOHDJHWKHPDWHUQDO age had an inverse correlation with implantation. The semen quality associated with advanced paternal age has an important impact in ART RXWFRPH7KHVH¿QGLQJVVXJJHVWWKDWWKHPDOHDJHLQJPD\QHJDWLYHO\ LQÀXHQFH $57 RXWFRPHV ZKHQ WKH VSHUPDWR]RD FRQFHQWUDWLRQ LV reduced. Source of Funding: None
1807
Source of Funding: DOD Prostate Cancer Research IDEA grant W81XWH-04-1-0917.
EFFECT OF ALLOPURINOL ON GERM CELL APOPTOSIS FOLLOWING TESTICULAR ISCHEMIA-REPERFUSION INJURY IN A RAT Gil Meyer*, Igor Sukhotnik, Jorge G Mogilner, Boaz Moskovitz, Ofer Nativ. Haifa, Israel. INTRODUCTION AND OBJECTIVE: Recent evidence suggests that apoptosis is involved in germ cell loss following testicular ischemia-reperfusion (IR) injury. Allopurinol (Allo) is as a free radical scavenger which prevents tissue damage caused by reperfusion and R[\JHQDWLRQDIWHULVFKHPLD KRZHYHULWVHIIHFWRQDSRSWRVLVLQWKLVW\SH of injury has not been studied. Aim: To examine the effect of allopurinol on germ cell apoptosis following testicular IR in a rat. METHODS: Forty rats were divided randomly into four experimental groups of 10 rats each: Group A (Sham)- Sham operated DQLPDOV *URXS % 6KDP$OOR 6KDP RSHUDWHG UDWV WUHDWHG ZLWK allopurinol given PO (by gavage) at a dose of 200 mg/kg, once daily, LPPHGLDWHO\EHIRUHDQGKRXUVIROORZLQJRSHUDWLRQ *URXS& ,5 UDWV underwent 90 minutes of unilateral testicular ischemia and 48 hours of UHSHUIXVLRQ *URXS' ,5$OOR UDWVXQGHUZHQW,5DQGZHUHWUHDWHGZLWK allopurinol similar to Group B. The ipsilateral and contralateral testes were harvested 48 hours following operation. Johnsen’s criteria and the QXPEHURIJHUPLQDOFHOOOD\HUVZHUHXVHGWRFDWHJRUL]HVSHUPDWRJHQHVLV TUNEL assay was used to determine germ cell apoptosis. Statistical
THE JOURNAL OF UROLOGY®
by electroporation. Our results suggest that this approach could have great relevance for the study of sperm function and male infertility. Source of Funding: None
1805 TROPHININ-BINDING PEPTIDE, GWRQ, PROMOTES HUMAN SPERM MOTILITY Shingo Hatakeyama*, Noritaka Kamimura, Takuya Koie, Kazuyuki Mori, Yasuhiro Hashimoto, Ikuya Iwabuchi, Takahiro Yoneyama, Atsushi Imai, Akiko Okamoto, Hayato Yamamoto, Chikara Ohyama, Michiko N Fukuda. La Jolla, CA, and Hirosaki, Japan. INTRODUCTION AND OBJECTIVE: Trophinin mediates a homophilic cell adhesion by trophinin-trophinin binding between trophectoderm and endometrial epithelium. (Genes Dev 1995) Previously we found that multivalent peptide, GWRQ-MAPS, binds to trophinin on cell surface, mimicking trophinin-mediated cell adhesion and subsequent signal transduction for trophectoderm activation.(PNAS 2007) Here we report that trophinin and trophinin-associated bystin and tastin proteins are found in the tail of human sperm, and that GWRQ-MAPS promoted motility of sperm. METHODS: Sperm samples were collected from 23 healthy males. Sperm mobility was compared by counting the numbers in WKH SUHVHQFH RI *:540$36 RU FRQWURO SHSWLGH /RFDOL]DWLRQ RI trophinin, bystin, tastin and GWRQ-peptide binding were determined by immunocytochemistry using anti-trophinin, anti-bystin, anti-tastin antibody and GWRQ-peptide displaying phage. ATP levels and intercellular calcium concentration were measured by luminometer. Swim down assay was assumed by transwell chamber with 8 µm pores from 3 healthy males. Cells moved from upper to bottom chamber were counted. 5(68/76 ,PPXQRF\WRFKHPLVWU\VXJJHVWHGFRORFDOL]DWLRQ of trophinin, bystin and tastin and the binding of GWRQ-peptide displaying phage to sperm tail was detected by immunostaining. (Fig. 1) In the presence of GWRQ-MAPS, the motility of human sperm was VLJQL¿FDQWO\LQFUHDVHG S )LJ DQGFHOOVUHGXFHG$73 S DQGH[KLELWHGVLJQL¿FDQWXSWDNHRIFDOFLXP S 6LQFHWDVWLQELQGV to Tctex-1 or microtubule-associated motor dynein ATPase (Biochem J 2002), trophinin bound with GWRQ may result in an activation of microtubule-associated ATPase. Since GWRQ-MAPS treated human sperm traveled through 8µm pores at higher frequency than those treated with control peptide, GWRQ-MAPS also promotes directed motility for a short distance. CONCLUSIONS: These results suggest that GWRQ-peptide enhances the sperm mobility. In normal human sperm, it is possible that trophinin plays a role in suppressing ATPase from premature over activation.
Vol. 179, No. 4, Supplement, Wednesday, May 21, 2008
1806 DOES THE MALE AGEING INFLUENCE CLINICAL OUTCOMES ON ICSI CYCLES? Renata C Ferreira, Tatiana C S Bonetti, Daniela Braga, Priscila Queiroz, Fabio F Pasqualotto, Assumpto Iaconelli, Edson Borges*. Sao Paulo, Brazil. INTRODUCTION AND OBJECTIVE: Lately an increased number of older couples requesting assisted reproductive treatment (ART) have been observed. It is clearly established the reduced fertility potential of older women, and alterations in semen quality associated with male ageing. Advanced male age in ART cycles provides a model to study biological paternal effects on embryo development and clinical outcomes, however the results are still controversy. The aim of this study was to evaluate the effect of male ageing on clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles, according to semen parameters. METHODS: Retrospective analysis included 760 ICSI cycles ZLWKIUHVKVSHUPDWR]RDDQGPRUHWKDQ0,,RRF\WHUHFRYHUHG:RPHQ received pituitary down regulation and controlled ovarian stimulation with recombinant-FSH. Follicle aspiration was done 35-36 hours after hCG trigger and MII oocytes were injected. Semen samples were collected E\ PDVWXUEDWLRQ DQG DQDO\]HG DFFRUGLQJ WR :+2 7KH F\FOHV ZHUH divided depending on sperm concentration: <20x106sperm/ml (n=220) DQG[6sperm/ml (n=540). In order to evaluate the effect of male ageing on clinical outcomes, adjusted to variations in women age, multiple linear regression analysis was performed. RESULTS: When sperm concentration were <20x106sperm/ml, there was a negative correlation between male age and implantation rate U S DQGPXOWLYDULDWHUHJUHVVLRQDGMXVWHGWRPDWHUQDO DJHVKRZHGWKHLQÀXHQFHRIH[FOXVLYHO\PDOHDJHLQJRQLPSODQWDWLRQ S :KHQWKHVSHUPFRQFHQWUDWLRQZHUH[6sperm/ml, there ZDVQRFRUUHODWLRQEHWZHHQPDOHDJHDQGLPSODQWDWLRQUDWH U S EXWRQWKHRWKHUKDQGWKHPDWHUQDODJHQHJDWLYHO\LQÀXHQFHG implantation rates (p<0.0001). CONCLUSIONS: The male ageing is associated with a VLJQL¿FDQW GHFOLQH LQ LPSODQWDWLRQ UDWHV LQ ,&6, F\FOHV ZKHQ VSHUP concentration was lower than 20x106sperm/ml, independently on maternal age. On the other hand, when sperm concentration was normal, HYHQWKRXJKLWZDVQRWREVHUYHGLQÀXHQFHRIPDOHDJHWKHPDWHUQDO age had an inverse correlation with implantation. The semen quality associated with advanced paternal age has an important impact in ART RXWFRPH7KHVH¿QGLQJVVXJJHVWWKDWWKHPDOHDJHLQJPD\QHJDWLYHO\ LQÀXHQFH $57 RXWFRPHV ZKHQ WKH VSHUPDWR]RD FRQFHQWUDWLRQ LV reduced. Source of Funding: None
1807
Source of Funding: DOD Prostate Cancer Research IDEA grant W81XWH-04-1-0917.
EFFECT OF ALLOPURINOL ON GERM CELL APOPTOSIS FOLLOWING TESTICULAR ISCHEMIA-REPERFUSION INJURY IN A RAT Gil Meyer*, Igor Sukhotnik, Jorge G Mogilner, Boaz Moskovitz, Ofer Nativ. Haifa, Israel. INTRODUCTION AND OBJECTIVE: Recent evidence suggests that apoptosis is involved in germ cell loss following testicular ischemia-reperfusion (IR) injury. Allopurinol (Allo) is as a free radical scavenger which prevents tissue damage caused by reperfusion and R[\JHQDWLRQDIWHULVFKHPLD KRZHYHULWVHIIHFWRQDSRSWRVLVLQWKLVW\SH of injury has not been studied. Aim: To examine the effect of allopurinol on germ cell apoptosis following testicular IR in a rat. METHODS: Forty rats were divided randomly into four experimental groups of 10 rats each: Group A (Sham)- Sham operated DQLPDOV *URXS % 6KDP$OOR 6KDP RSHUDWHG UDWV WUHDWHG ZLWK allopurinol given PO (by gavage) at a dose of 200 mg/kg, once daily, LPPHGLDWHO\EHIRUHDQGKRXUVIROORZLQJRSHUDWLRQ *URXS& ,5 UDWV underwent 90 minutes of unilateral testicular ischemia and 48 hours of UHSHUIXVLRQ *URXS' ,5$OOR UDWVXQGHUZHQW,5DQGZHUHWUHDWHGZLWK allopurinol similar to Group B. The ipsilateral and contralateral testes were harvested 48 hours following operation. Johnsen’s criteria and the QXPEHURIJHUPLQDOFHOOOD\HUVZHUHXVHGWRFDWHJRUL]HVSHUPDWRJHQHVLV TUNEL assay was used to determine germ cell apoptosis. Statistical