PMID- 32202531
OWN - NLM
STAT- MEDLINE
DCOM- 20200922
LR  - 20201114
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 157
DP  - 2020 Mar 3
TI  - Preparation of Meiotic Chromosome Spreads from Zebrafish Spermatocytes.
LID - 10.3791/60671 [doi]
AB  - Meiosis is the key cellular process required to create haploid gametes for sexual 
      reproduction. Model organisms have been instrumental in understanding the 
      chromosome events that take place during meiotic prophase, including the pairing, 
      synapsis, and recombination events that ensure proper chromosome segregation. 
      While the mouse has been an important model for understanding the molecular 
      mechanisms underlying these processes, not all meiotic events in this system are 
      analogous to human meiosis. We recently demonstrated the exciting potential of 
      the zebrafish as a model of human spermatogenesis. Here we describe, in detail, 
      our methods to visualize meiotic chromosomes and associated proteins in 
      chromosome spread preparations. These preparations have the advantage of allowing 
      high resolution analysis of chromosome structures. First, we describe the 
      procedure for dissecting testes from adult zebrafish, followed by cell 
      dissociation, lysis, and spreading of the chromosomes. Next, we describe the 
      procedure for detecting the localization of meiotic chromosome proteins, by 
      immunofluorescence detection, and nucleic acid sequences, by fluorescence in situ 
      hybridization (FISH). These techniques comprise a useful set of tools for the 
      cytological analysis of meiotic chromatin architecture in the zebrafish system. 
      Researchers in the zebrafish community should be able to quickly master these 
      techniques and incorporate them into their standard analyses of reproductive 
      function.
FAU - Blokhina, Yana P
AU  - Blokhina YP
AD  - Department of Molecular and Cellular Biology, University of California, Davis; 
      Integrative Genetics and Genomics Graduate Group, University of California, 
      Davis.
FAU - Olaya, Ivan
AU  - Olaya I
AD  - Department of Molecular and Cellular Biology, University of California, Davis; 
      Integrative Genetics and Genomics Graduate Group, University of California, 
      Davis.
FAU - Burgess, Sean M
AU  - Burgess SM
AD  - Department of Molecular and Cellular Biology, University of California, Davis; 
      smburgess@ucdavis.edu.
LA  - eng
GR  - R01 GM075119/GM/NIGMS NIH HHS/United States
GR  - R25 GM116690/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Video-Audio Media
DEP - 20200303
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Chromatin)
SB  - IM
MH  - Animals
MH  - Chromatin/metabolism
MH  - Chromosome Pairing
MH  - Chromosome Segregation
MH  - Chromosomes/*ultrastructure
MH  - Fluorescent Antibody Technique
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - *Meiosis
MH  - Spermatocytes/*physiology
MH  - Testis/pathology
MH  - Zebrafish/*genetics
PMC - PMC7375709
MID - NIHMS1582156
EDAT- 2020/03/24 06:00
MHDA- 2020/09/23 06:00
PMCR- 2020/07/22
CRDT- 2020/03/24 06:00
PHST- 2020/03/24 06:00 [entrez]
PHST- 2020/03/24 06:00 [pubmed]
PHST- 2020/09/23 06:00 [medline]
PHST- 2020/07/22 00:00 [pmc-release]
AID - 10.3791/60671 [doi]
PST - epublish
SO  - J Vis Exp. 2020 Mar 3;(157):10.3791/60671. doi: 10.3791/60671.