WORKSHOP BOOK Fluorescent in Situ Hybridisation - FISH - on Human Sperm

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<strong>FISH</strong> Results Interpretation Sperm <strong>FISH</strong> def<strong>in</strong>es the proportion of aneuploidy present <strong>in</strong> the ejaculates of males with other (genetic/cl<strong>in</strong>ical) problems The goal: to identify “errors” and the “at-risk” males Quantitative vs Qualitative analyses “...<strong>FISH</strong> analysis of spermatozoa should be used as a tool of genetic screen<strong>in</strong>g <strong>in</strong> <strong>in</strong>fertile patients. Significant differences <strong>in</strong> the rates of chromosome abnormalities with respect to controls should be taken <strong>in</strong>to consideration regardless of the numerical value. When abnormal results are obta<strong>in</strong>ed, <strong>in</strong>dividuals should be identified as ‘‘at risk’’ and the couple should be advised about the available techniques of preimplantation and prenatal genetic diagnosis” Sarrate et al, 2010 35 36
Figure 5. Model of chromosome organization <strong>in</strong> human spermatozoa. Compact CTs (filled contours) have overall hairp<strong>in</strong> conformations (chromosome paths <strong>in</strong>dicated by dashed l<strong>in</strong>es) with the p and q telomere/subtelomere doma<strong>in</strong>s (orange circles) form<strong>in</strong>g dimers at nuclear periphery. Gene-rich CHRs – rosy, gene-poor – 37 <strong>in</strong>digo. CTs are connected via centromeres/peri-centromeres (green circles and l<strong>in</strong>es) <strong>in</strong>to arrays and have a fixed l<strong>in</strong>ear order which determ<strong>in</strong>es the longitud<strong>in</strong>al position<strong>in</strong>g of chromosomes. Mudrak OS, B. Nazarov I, Jones EL, Zalensky AO (2012) Position<strong>in</strong>g of Chromosomes <strong>in</strong> Human Spermatozoa Is Determ<strong>in</strong>ed by Ordered Centromere Arrangement. PLoS ONE 7(12): e52944. doi:10.1371/journal.pone.0052944 http://www.plosone.org/article/<strong>in</strong>fo:doi/10.1371/journal.pone.0052944 38
Page 1 and 2: 1 WORKSHOP <strong
Page 3 and 4: 3 Content Workshop Book Embryolab A
Page 5 and 6: 5 Hands-on Workshop 1 Fluor
Page 7 and 8: 7 Programme 8:30 - 8:45 Pick- up fr
Page 9 and 10: Aneuploidy in male
Page 11 and 12: Spermatogenesis Hands-on Workshops,
Page 13 and 14: Paternally derived aneuploidies A 2
Page 15 and 16: Reciprocal Translocation The most f
Page 17 and 18: Interchromosomal Effect (ICE) Aneup
Page 19 and 20: Aneuploidy in male
Page 21 and 22: Sperm morphology Data regard<strong
Page 23 and 24: Infertile males (46,XY) Cryptorhidi
Page 25 and 26: DNA Fragmentation Index (DFI) Sever
Page 27 and 28: Occupational/Life style hazards Stu
Page 29 and 30: 41 Mechanisms again</strong
Page 31 and 32: Take home message At present, PGD i
Page 33 and 34: In summary • Multicolour
Page 35 and 36: FISH on Spermatozo
Page 37 and 38: FISH: What can it
Page 39 and 40: A DNA probe finds
Page 41 and 42: Hybridisation targ
Page 43 and 44: Sperm-FISH protoco
Page 45 and 46: 1. Preparation of sample Nuclear De
Page 47 and 48: 2. Co-Denaturation (target DNA & pr
Page 49 and 50: 5. Examine slides
Page 51: FISH Advantages )
Page 55 and 56: 55 FISH technique:
Page 57 and 58: Tip for spreading
Page 59 and 60: Post Hybridization Wash The aim of
Page 61 and 62: Sperm FISH Externa
Page 63 and 64: Issue Date: 05/12/2012 Version: 1 R
Page 71 and 72: Our generous Sponsors 71
Page 73 and 74: 73 Certificate of attendance A Cert

WORKSHOP BOOK Fluorescent in Situ Hybridisation - FISH - on Human Sperm

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