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. 2002 Aug 2;2(1):8.
doi: 10.1186/1472-6874-2-8.

Investigation of KIT gene mutations in women with 46,XX spontaneous premature ovarian failure

Kyoko Shibanuma  1 , Zhi-Bin Tong, Vien H Vanderhoof, Konstantina Vanevski, Lawrence M Nelson
Affiliations

Investigation of KIT gene mutations in women with 46,XX spontaneous premature ovarian failure

Kyoko Shibanuma et al. BMC Womens Health. .

Abstract

BACKGROUND: Spontaneous premature ovarian failure presents most commonly with secondary amenorrhea. Young women with the disorder are infertile and experience the symptoms and sequelae of estrogen deficiency. The mechanisms that give rise to spontaneous premature ovarian failure are largely unknown, but many reports suggest a genetic mechanism in some cases. The small family size associated with infertility makes genetic linkage analysis studies extremely difficult. Another approach that has proven successful has been to examine candidate genes based on known genetic phenotypes in other species. Studies in mice have demonstrated that c-kit, a transmembrane tyrosine kinase receptor, plays a critical role in gametogenesis. Here we test the hypothesis that human KIT mutations might be a cause of spontaneous premature ovarian failure. METHODS AND RESULTS: We examined 42 women with spontaneous premature ovarian failure and found partial X monosomy in two of them. In the remaining 40 women with known 46,XX spontaneous premature ovarian failure we evaluated the entire coding region of the KIT gene. We did this using polymerase chain reaction based single-stranded conformational polymorphism analysis and DNA sequencing. We did not identify a single mutation that would alter the amino acid sequence of the c-KIT protein in any of 40 patients (upper 95% confidence limit is 7.2%). We found one silent mutation at codon 798 and two intronic polymorphisms. CONCLUSION: Mutations in the coding regions of the KIT gene appear not to be a common cause of 46,XX spontaneous premature ovarian failure in North American women.

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Figures

Figure 1
Figure 1
Human KIT organization and PCR amplification of its exons. A. Schematic representation of exon-intron map of human KIT. It is composed of 21 exons and 20 introns. Vertical bars and horizontal solid lines represent the exons and introns, respectively. The // indicates that these introns are not in scale. Arabic numbers above each of the vertical bars indicate the exon number. Modified from Giebel et al.[27]. B. Analysis of PCR products of the human KIT gene. Human genomic DNA spanning each of the human KIT exons were amplified using specific primer pairs. PCR products for each of the exons were separated by 1.5% agarose-electrophoresis and stained with ethidium-bromide. The Arabic numbers above each of the PCR products represent the exon number. DNA size markers (φ X174 RF DNA/HaeIII) are shown at the left of the panel (M).
Figure 2
Figure 2
SSCP analysis of KIT PCR products The PCR products were separated on 20% TBE-acrylamide gel electrophoresis and stained with ethidium bromide. SSCP analysis of samples from three normal control women and three patients with spontaneous premature ovarian failure are shown for exon 10,16, and 17. DNA mobility of PCR products from exons 10 and 16 was altered in one patient (Lane 4). One patient showed an alternation of DNA mobility in the PCR product from exon 17 (Lane 5). Also shown is the DNA mobility of samples from normal women (Lanes 1–3) and from a representative patient with no changes in DNA mobility (Lane 6).
Figure 3
Figure 3
DNA sequence analysis of a KIT intronic region variant downstream of exon 16 Electropherogram displaying the sequence of the KIT variant of the intronic sequence downstream of exon 16 compared to the wild-type sequence. Arrows indicate a deletion of 5T nucleotides in the variant as compared to a normal series of 22 consecutive T nucleotides in the wild-type.
Figure 4
Figure 4
Restriction fragment length polymorphism analysis of a KIT variant in exon 17 using Ssp I Undigested DNA and wild-type DNA digested with Ssp I gives a single band of 390 bp. Homozygosity for the variant yields two fragments of 207 and 183 bp. A heterozygote carrier would have demonstrated all three of the fragment lengths. Therefore, this patient is homozygous for this variant.
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References

    1. Bondy CA, Nelson LM, Kalantaridou SN. The genetic origins of ovarian failure. J Womens Health. 1998;7:1225–1229. - PubMed
    1. Kalantaridou SN, Davis SR, Nelson LM. Premature ovarian failure. Endocrinology and Metabolism Clinics of North America. 1998;27:989–1005. - PubMed
    1. van Kasteren YM, Hundscheid RD, Smits AP, Cremers FP, van Zonneveld P, Braat DD. Familial idiopathic premature ovarian failure: an overrated and underestimated genetic disease? Hum Reprod. 1999;14:2455–2459. doi: 10.1093/humrep/14.10.2455. - DOI - PubMed
    1. Davis CJ, Davison RM, Payne NN, Rodeck CH, Conway GS. Female sex preponderance for idiopathic familial premature ovarian failure suggests an X chromosome defect: opinion. Hum Reprod. 2000;15:2418–2422. doi: 10.1093/humrep/15.11.2418. - DOI - PubMed
    1. Wylie C. Germ cells. Cell. 1999;96:165–174. - PubMed

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