Chromosome Spread Analyses of Meiotic Sex Chromosome Inactivation

Kris G Alavattam; Hironori Abe; Akihiko Sakashita; Satoshi H Namekawa

doi:10.1007/978-1-4939-8766-5_10

Chromosome Spread Analyses of Meiotic Sex Chromosome Inactivation

Methods Mol Biol. 2018:1861:113-129. doi: 10.1007/978-1-4939-8766-5_10.

Authors

Kris G Alavattam^{1

2

3}, Hironori Abe^{4

5

6}, Akihiko Sakashita^{4

5

6}, Satoshi H Namekawa^{7

8

9}

Affiliations

¹ Division of Reproductive Sciences, Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA. kristan.alavattam@cchmc.org.
² Division of Developmental Biology, Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA. kristan.alavattam@cchmc.org.
³ Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA. kristan.alavattam@cchmc.org.
⁴ Division of Reproductive Sciences, Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
⁵ Division of Developmental Biology, Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
⁶ Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
⁷ Division of Reproductive Sciences, Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA. satoshi.namekawa@cchmc.org.
⁸ Division of Developmental Biology, Perinatal Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA. satoshi.namekawa@cchmc.org.
⁹ Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA. satoshi.namekawa@cchmc.org.

Abstract

A distinct form of X chromosome inactivation takes place during male meiosis, when the male sex chromosomes undergo a phenomenon known as meiotic sex chromosome inactivation (MSCI). MSCI is directed by DNA damage response signaling independent of Xist RNA to silence the transcriptional activity of the sex chromosomes, an essential event in male germ cell development. Here, we present protocols for the preparation and analyses of chromosome spread slides of mouse meiotic spermatocytes, thereby enabling a quick, inexpensive, and powerful cytological method to complement gene expression studies.

Keywords: Cell staging; Cell suspension; Chromatin; Chromosomes; Cytology; DNA damage response; Immunofluorescence microscopy; Meiosis; Sex chromosomes; Slide protocol; Synapsis; Testis.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Epigenomics / methods
Fluorescent Antibody Technique / methods*
Male
Meiosis*
Mice
Microscopy, Fluorescence / methods
Sex Chromosomes*
Spermatocytes / cytology
Spermatocytes / metabolism*
X Chromosome Inactivation*

Grants and funding

R01 GM098605/GM/NIGMS NIH HHS/United States