A rat H1t-GFP transgene recapitulates endogenous H1t expression pattern in mouse

Genesis. 2020 May;58(5):e23355. doi: 10.1002/dvg.23355. Epub 2020 Jan 28.

Abstract

H1 histones bind to linker DNA. H1t (H1f6), a testis-specific linker histone variant, is present in pachytene spermatocytes and spermatids. The expression of H1t histone coincides with the acquisition of metaphase I competence in pachytene spermatocytes. Here we report the generation of H1t-GFP transgenic mice. The H1t-GFP (H1 histone testis-green fluorescence protein) fusion protein expression recapitulates the endogenous H1t expression pattern. This protein appears first in mid pachytene spermatocytes in stage V seminiferous tubules, persists in round spermatids and elongating spermatids, but is absent in elongated spermatids. The strong green fluorescence signal, due to the high abundance of H1t-GFP, is maintained in spermatocytes after induction towards metaphase I through treatment with okadaic acid. Therefore, H1t-GFP can be used as a visual marker for monitoring the progression of meiosis in vitro and in vivo, as well as fluorescence-activated cell sorting (FACS) sorting of germ cells.

Publication types

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

MeSH terms

  • Animals
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Histones / genetics*
  • Histones / metabolism
  • Male
  • Mice
  • Mice, Transgenic
  • Okadaic Acid / pharmacology
  • Rats
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Spermatids / metabolism*
  • Testis / drug effects
  • Testis / metabolism
  • Transgenes*

Substances

  • Histones
  • Recombinant Proteins
  • Green Fluorescent Proteins
  • Okadaic Acid