PMID- 15589873
OWN - NLM
STAT- MEDLINE
DCOM- 20050125
LR  - 20061115
IS  - 0015-0282 (Print)
IS  - 0015-0282 (Linking)
VI  - 82
IP  - 6
DP  - 2004 Dec
TI  - Simultaneous analysis of cytoskeletal patterns and chromosome positioning in 
      human fertilization failures.
PG  - 1654-9
AB  - OBJECTIVE: To sequentially and reliably apply both tubulin immunocytochemistry 
      (ICC) and fluorescence in situ hybridization (FISH) to human fertilization 
      failures, thus providing a tool for a multiple analysis of arrest. DESIGN: 
      Analysis of human fertilization failures at several stages of arrest. SETTING: 
      Academic and clinical institutions. PATIENT(S): Consenting patients undergoing 
      assisted reproduction technologies. INTERVENTION(S): Failed fertilizations 
      displaying signs of activation without pronuclear development, or with the 
      absence of polar body emission or cleavage 48 hours after insemination or 
      microinjection were analyzed. Fertilization failures were fixed and processed for 
      ICC. After data was collected the same samples were then subjected to FISH 
      analysis using probes for chromosomes X, Y, and 18. MAIN OUTCOME MEASURE(S): 
      Simultaneous ICC and FISH data on the same sample. RESULT(S): Sequential 
      application of straightforward standard ICC and FISH techniques was not possible, 
      as the morphologic features had been altered, microtubular patterns were not 
      preserved, and many samples were rendered opaque. Only chromatin at the cell 
      surface or outside the oocyte/zygote, such as metaphase II spindles or polar body 
      nuclei, could be routinely probed for FISH after ICC. However, an increase in 
      detergent-induced sample permeabilization as well as the removal of several steps 
      usually performed for FISH made it possible to directly compare microtubular 
      patterns and chromosome position, regardless of chromatin status. CONCLUSION(S): 
      Analysis of specific proteins by immunocytochemistry and of chromosome 
      status/positioning by FISH can be carried out sequentially in human fertilization 
      failures, irrespective of the stage of arrest.
FAU - Ramalho-Santos, Joao
AU  - Ramalho-Santos J
AD  - Center for Neuroscience and Cell Biology, Department of Zoology, University of 
      Coimbra, Coimbra, Portugal. jramalho@ci.uc.pt
FAU - Amaral, Alexandra
AU  - Amaral A
FAU - Brito, Raquel
AU  - Brito R
FAU - Freitas, Mariana
AU  - Freitas M
FAU - Almeida Santos, Teresa
AU  - Almeida Santos T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Fertil Steril
JT  - Fertility and sterility
JID - 0372772
RN  - 0 (Chromatin)
SB  - IM
MH  - Blastocyst/cytology/ultrastructure
MH  - Cell Membrane/ultrastructure
MH  - Chromatin/ultrastructure
MH  - Chromosomes, Human/*ultrastructure
MH  - Cytoskeleton/*ultrastructure
MH  - Female
MH  - *Fertilization
MH  - *Fertilization in Vitro
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - Metaphase
MH  - Microtubules/ultrastructure
MH  - Oocytes/ultrastructure
MH  - *Sperm Injections, Intracytoplasmic
MH  - Treatment Failure
EDAT- 2004/12/14 09:00
MHDA- 2005/01/26 09:00
CRDT- 2004/12/14 09:00
PHST- 2004/01/07 00:00 [received]
PHST- 2004/05/04 00:00 [revised]
PHST- 2004/05/04 00:00 [accepted]
PHST- 2004/12/14 09:00 [pubmed]
PHST- 2005/01/26 09:00 [medline]
PHST- 2004/12/14 09:00 [entrez]
AID - S0015-0282(04)02411-2 [pii]
AID - 10.1016/j.fertnstert.2004.05.086 [doi]
PST - ppublish
SO  - Fertil Steril. 2004 Dec;82(6):1654-9. doi: 10.1016/j.fertnstert.2004.05.086.