1. THE POLYMORPHISM OF FSH RECEPTOR GENE IN INDONESIAN WOMEN UNDERGOING ASSISTED REPRODUCTION DWI ANITA SURYANDARI

2. ROLEOFFSH ROLEOFFSH

3. Structure of FSH reseptor (FSHR) molecule member of the family of G-protein coupled receptors The FSHR molecule consist of 3 domains a. Extracellular domain b. Transmembrane domain c. Intracellular domain a bcbc

4. Single gene of 2p21, 54 kbp, consisting 10 exons and 9 introns Thre307Ala Asn680Ser FSH Receptor gene

5. Objective: 1). To assess the incidence of different FSHR genotypes in Indonesian women undergoing assisted reproduction (anovulatory patients) and those with normo ovulatory controls 2). To correlate the FSHR genotypes with baseline serum FSH levels. and their response to exogenous FSH prior to super ovulation induction in anovulatory patients.

6. Design : Cross sectional study Samples : 130 normoovulatory controls and 187 normogonadotropic anovulatory women who participated in assisted reproduction in Bunda Hospital, Jakarta. Main outcome Measure(s): Prevalence of FSHR polymorphisms, Baseline serum FSH levels and dosage of FSH administered.

7. DNA ISOLATION DNA AMPLIFICATION PCR (Sampel DNA; PCR buffer 1X; 1,5 mM MgCl 2 ; 0,2 mM dNTP; 0,4 pmol/µl primer L8211 dan H8310; 0,25 unit enzim DNA polimerase Taq; dan ddH 2 O steril ) total volume 25 µl 95 o C 1’ 58 o C 1’ 72 o C 5’ Denaturasi Ekstensi 30 siklus 95 o C 5’ 4 o C  72 o C 1’30’’ Annealing Denaturasi awal Ekstensi akhir ELECTROPHORESIS agarosa /acrylamide Loading:PCR produk (6  l produk PCR + 2  l Loading Buffer) - DNA Marker  X174RF/HaeIII Electrophoresis 80 V, 1,5 H Visualisasi of DNA gel by Gel-Doc 1000 [Bio Rad]

8. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Determination of FSHR gene polymorphisms in 307 position by SSCP using 20% acrylamide gel. Ala/Thre Ala/Ala Thre/Thre

9. Asn/AsnSer/Ser Ser/Asn PCR-RFLP of FSHR gene in 680 position by 2% gel agarose

10. The distribution of FSHR genotypes in anovulatory patients (n= 187) and normo-ovulatory controls (n=130) was significantly different for both genotypes (p< 0,05; Chi- square test). The distribution of FSHR gene Polymorphisms in normoovulatory controls (n= 130) and in anovulatory patients (n = 187)

11. The baseline serum FSH level in ser/ser genotypes was significantly different from Ser/Asn or Asn/Asn genotypes (p< 0,01; Anova). The baseline serum FSH levels in normoovulatory controls

12. The baseline serum FSH levels and the dosage of FSH stimulation was not significantly different among women with 307 FSHR genotypes (p> 0,05; Kruskal-Wallis test). The baseline serum FSH levels and the dose of FSH stimulation in anovulatory patients with 307 FSHR genotypes

13. The baseline serum FSH levels and the dosage of FSH stimulation in women with Ser/Ser genotype was significantly higher than those in Ser/Asn and Asn/Asn genotypes in 680 position (p< 0,01; Kruskal-Wallis test). The baseline serum FSH levels and the dosage of FSH stimulation in anovulatory patients with 680 FSHR genotypes

14. The ovarian response to FSH stimulation is dependent to the FSH receptor genotypes Each FSHR isoform may show different expression on the cell surface, determining the variation modes of interaction with the hormone. Each receptor genotypes may show different turnover or down regulation rate. Each FSHR isoform may have different response to FSH stimulation. Gene expression test ?

15. CONCLUTIONS 1.There is no linkage of polymorphic nucleotide 307 with 680 of FSHR gene detectable in Indonesian women. 2. The FSHR genotypes of 680 position determine ovarium response to FSH stimulation.

17. * Polymorphism of FSHR gene not linkage between position 307 and 680, making 9 isoform. Polymorphism of FSHR gene in Indonesian, South of Asia, Arabian, Surinames, and Holland women