R9AP overexpression alters phototransduction kinetics in iCre75 mice

Thomas R. Sundermeier, Frans Vinberg, Debarshi Mustafi, Xiaodong Bai, Vladimir J. Kefalov, Krzysztof Palczewski

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Purpose. Determine the impact of rod photoreceptor-specific expression of Cre recombinase on the kinetics of phototransduction in the mouse eye and identify changes in gene expression that underlie any observed phenotypic differences. Methods. Transretinal ERG and single-cell suction electrode recordings were used to measure the kinetics of phototransduction in a mouse line exhibiting rod photoreceptor-specific Cre recombinase expression, and the results were compared with those from control non-Creexpressing littermates. Gene expression changes were evaluated using RNA sequencing transcriptome analysis. The pattern of expression of Rgs9bp was determined by mapping sequencing reads to the mouse genome and performing 3'-rapid amplification of cDNA ends (3'-RACE). Results. Expression of the rod-specific iCre75 transgene was accompanied by accelerated phototransduction inactivation, likely due to overexpression of the Rgs9bp gene, which encodes the Rgs9 anchor protein (R9AP). R9AP upregulation stabilized the RGS9 GAP complex, altering phototransduction kinetics. 3'-Race identified an abundant, unexpected Rgs9bp-Prm1 fusion mRNA in Cre-expressing mouse retinas, which was determined to be derived from a second transgene present in the iCre75 line. Conclusions. Here we report the presence of a second, R9AP-expressing transgene in the iCre75 mouse line, leading to altered kinetics of phototransduction. These results highlight an important caveat that must be considered when utilizing this mouse line for rod photoreceptor-specific gene loss of function studies.

Original languageEnglish
Pages (from-to)1339-1347
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume55
Issue number3
DOIs
StatePublished - Feb 13 2014

Keywords

  • Cre recombinase
  • Mouse
  • Phototransduction
  • R9AP
  • Retina
  • Rod photoreceptors

Fingerprint

Dive into the research topics of 'R9AP overexpression alters phototransduction kinetics in iCre75 mice'. Together they form a unique fingerprint.

Cite this