PMID- 29286418
OWN - NLM
STAT- MEDLINE
DCOM- 20180619
LR  - 20181207
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 130
DP  - 2017 Dec 6
TI  - Using Fluorescence In Situ Hybridization (FISH) to Monitor the State of Arm 
      Cohesion in Prometaphase and Metaphase I Drosophila Oocytes.
LID - 10.3791/56802 [doi]
LID - 56802
AB  - In humans, chromosome segregation errors in oocytes are responsible for the 
      majority of miscarriages and birth defects. Moreover, as women age, their risk of 
      conceiving an aneuploid fetus increases dramatically and this phenomenon is known 
      as the maternal age effect. One requirement for accurate chromosome segregation 
      during the meiotic divisions is maintenance of sister chromatid cohesion during 
      the extended prophase period that oocytes experience. Cytological evidence in 
      both humans and model organisms suggests that meiotic cohesion deteriorates 
      during the aging process. In addition, segregation errors in human oocytes are 
      most prevalent during meiosis I, consistent with premature loss of arm cohesion. 
      The use of model organisms is critical for unraveling the mechanisms that 
      underlie age-dependent loss of cohesion. Drosophila melanogaster offers several 
      advantages for studying the regulation of meiotic cohesion in oocytes. However, 
      until recently, only genetic tests were available to assay for loss of arm 
      cohesion in oocytes of different genotypes or under different experimental 
      conditions. Here, a detailed protocol is provided for using fluorescence in situ 
      hybridization (FISH) to directly visualize defects in arm cohesion in 
      prometaphase I and metaphase I arrested Drosophila oocytes. By generating a FISH 
      probe that hybridizes to the distal arm of the X chromosome and collecting 
      confocal Z stacks, a researcher can visualize the number of individual FISH 
      signals in three dimensions and determine whether sister chromatid arms are 
      separated. The procedure outlined makes it possible to quantify arm cohesion 
      defects in hundreds of Drosophila oocytes. As such, this method provides an 
      important tool for investigating the mechanisms that contribute to cohesion 
      maintenance as well as the factors that lead to its demise during the aging 
      process.
FAU - Perkins, Adrienne T
AU  - Perkins AT
AD  - Department of Biological Sciences, Dartmouth College.
FAU - Bickel, Sharon E
AU  - Bickel SE
AD  - Department of Biological Sciences, Dartmouth College; 
      Sharon.E.Bickel@Dartmouth.edu.
LA  - eng
GR  - R01 GM059354/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Video-Audio Media
DEP - 20171206
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Drosophila/cytology/*physiology
MH  - Female
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Metaphase/physiology
MH  - Oocytes/*cytology
MH  - Prometaphase/physiology
PMC - PMC5755544
EDAT- 2017/12/30 06:00
MHDA- 2018/06/21 06:00
PMCR- 2018/12/06
CRDT- 2017/12/30 06:00
PHST- 2017/12/30 06:00 [entrez]
PHST- 2017/12/30 06:00 [pubmed]
PHST- 2018/06/21 06:00 [medline]
PHST- 2018/12/06 00:00 [pmc-release]
AID - 56802 [pii]
AID - 10.3791/56802 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Dec 6;(130):56802. doi: 10.3791/56802.