Beware of your Cre-Ation: lacZ expression impairs neuronal integrity and hippocampus-dependent memory

Hippocampus. 2016 Oct;26(10):1250-64. doi: 10.1002/hipo.22601. Epub 2016 May 24.

Abstract

Expression of the lacZ-sequence is a widely used reporter-tool to assess the transgenic and/or transfection efficacy of a target gene in mice. Once activated, lacZ is permanently expressed. However, protein accumulation is one of the hallmarks of neurodegenerative diseases. Furthermore, the protein product of the bacterial lacZ gene is ß-galactosidase, an analog to the mammalian senescence-associated ß-galactosidase, a molecular marker for aging. Therefore we studied the behavioral, structural and molecular consequences of lacZ expression in distinct neuronal sub-populations. lacZ expression in cortical glutamatergic neurons resulted in severe impairments in hippocampus-dependent memory accompanied by marked structural alterations throughout the CNS. In contrast, GFP expression or the expression of the ChR2/YFP fusion product in the same cell populations did not result in either cognitive or structural deficits. GABAergic lacZ expression caused significantly decreased hyper-arousal and mild cognitive deficits. Attenuated structural and behavioral consequences of lacZ expression could also be induced in adulthood, and lacZ transfection in neuronal cell cultures significantly decreased their viability. Our findings provide a strong caveat against the use of lacZ reporter mice for phenotyping studies and point to a particular sensitivity of the hippocampus formation to detrimental consequences of lacZ expression. © 2016 Wiley Periodicals, Inc.

Keywords: cognition; hippocampus; memory impairment; neurodegeneration; neurotoxicity; transgenic; volume loss.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Cell Survival / physiology
  • Cerebral Cortex / diagnostic imaging
  • Cerebral Cortex / metabolism
  • Cerebral Cortex / pathology
  • Gene Expression
  • Glutamic Acid / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hippocampus / diagnostic imaging
  • Hippocampus / metabolism*
  • Hippocampus / pathology
  • Integrases / genetics
  • Integrases / metabolism
  • Lac Operon*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Memory / physiology*
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Neurons / metabolism*
  • Neurons / pathology
  • Recombinant Fusion Proteins / metabolism
  • beta-Galactosidase / metabolism*
  • gamma-Aminobutyric Acid / metabolism

Substances

  • Bacterial Proteins
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria
  • Green Fluorescent Proteins
  • Glutamic Acid
  • gamma-Aminobutyric Acid
  • Cre recombinase
  • Integrases
  • beta-Galactosidase