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The wmN1 Enhancer Region of the Mouse Myelin Proteolipid Protein Gene (mPlp1) is Indispensable for Expression of an mPlp1-lacZ Transgene in Both the CNS and PNS

  • Pankaj Patyal
  • Neriman T. Kockara
  • Patricia A. WightEmail author
Original Paper
  • 15 Downloads

Abstract

The myelin proteolipid protein gene (PLP1) encodes the most abundant protein in CNS myelin. Expression of the gene must be strictly regulated, as evidenced by human X-linked leukodystrophies resulting from variations in PLP1 copy number, including elevated dosages as well as deletions. Recently, we showed that the wmN1 region in human PLP1 (hPLP1) intron 1 is required to promote high levels of an hPLP1-lacZ transgene in mice, using a Cre-lox approach. The current study tests whether loss of the wmN1 region from a related transgene containing mouse Plp1 (mPlp1) DNA produces similar results. In addition, we investigated the effects of loss of another region (ASE) in mPlp1 intron 1. Previous studies have shown that the ASE is required to promote high levels of mPlp1-lacZ expression by transfection analysis, but had no effect when removed from the native gene in mouse. Whether this is due to compensation by another regulatory element in mPlp1 that was not included in the mPlp1-lacZ constructs, or to differences in methodology, is unclear. Two transgenic mouse lines were generated that harbor mPLP(+)Z/FL. The parental transgene utilizes mPlp1 sequences (proximal 2.3 kb of 5ʹ-flanking DNA to the first 37 bp of exon 2) to drive expression of a lacZ reporter cassette. Here we demonstrate that mPLP(+)Z/FL is expressed in oligodendrocytes, oligodendrocyte precursor cells, olfactory ensheathing cells and neurons in brain, and Schwann cells in sciatic nerve. Loss of the wmN1 region from the parental transgene abolished expression, whereas removal of the ASE had no effect.

Keywords

Myelin proteolipid protein gene (Plp1Gene regulation Enhancer Cre/loxP and Flp/Frt recombination lacZ Transgenic mouse 

Notes

Acknowledgements

The authors are grateful to Joseph J. Goellner and Dr. Charles A. O’Brien at the UAMS Transgenic Mouse Core Facility for generation of the mPLP(+)Z/FL founder mice. The core is supported by a COBRE grant from the National Institutes of Health (P20GM125503) and Tobacco Settlement Funds.

Funding

This work was supported by National Institutes of Health Grants R01NS106179 (PAW) and UL1TR000039, and a gift from the Rampy MS Research Foundation.

Compliance with Ethical Standards

Conflict of interest

The authors declare that there are no conflicts of interest.

Supplementary material

11064_2019_2919_MOESM1_ESM.pptx (1.1 mb)
Supplementary material 1 (PPTX 1101 kb)
11064_2019_2919_MOESM2_ESM.pptx (527 kb)
Supplementary material 2 (PPTX 527 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Physiology and BiophysicsUniversity of Arkansas for Medical SciencesLittle RockUSA

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