title: Genotypic and phenotypic characterization of P23H line 1 rat model.
abstract: Rod-cone dystrophy, also known as retinitis pigmentosa (RP), is the most common inherited degenerative photoreceptor disease, for which no therapy is currently available. The P23H rat is one of the most commonly used autosomal dominant RP models. It has been created by incorporation of a mutated mouse rhodopsin (Rho) transgene in the wild-type (WT) Sprague Dawley rat. Detailed genetic characterization of this transgenic animal has however never been fully reported. Here we filled this knowledge gap on P23H Line 1 rat (P23H-1) and provide additional phenotypic information applying non-invasive and state-of-the-art in vivo techniques that are relevant for preclinical therapeutic evaluations. Transgene sequence was analyzed by Sanger sequencing. Using quantitative PCR, transgene copy number was calculated and its expression measured in retinal tissue. Full field electroretinography (ERG) and spectral domain optical coherence tomography (SD-OCT) were performed at 1-, 2-, 3- and 6-months of age. Sanger sequencing revealed that P23H-1 rat carries the mutated mouse genomic Rho sequence from the promoter to the 3' UTR. Transgene copy numbers were estimated at 9 and 18 copies in the hemizygous and homozygous rats respectively. In 1-month-old hemizygous P23H-1 rats, transgene expression represented 43% of all Rho expressed alleles. ERG showed a progressive rod-cone dysfunction peaking at 6 months-of-age. SD-OCT confirmed a progressive thinning of the photoreceptor cell layer leading to the disappearance of the outer retina by 6 months with additional morphological changes in the inner retinal cell layers in hemizygous P23H-1 rats. These results provide precise genotypic information of the P23H-1 rat with additional phenotypic characterization that will serve basis for therapeutic interventions, especially for those aiming at gene editing.
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Elise
authors.initials: E
authors.lastname: Orhan
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Deniz
authors.initials: D
authors.lastname: Dalkara
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Neuillé
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Lechauve
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Michiels
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Picaud
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Léveillard
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France; Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, DHU ViewMaintain, INSERM-DHOS CIC 1423, Paris, France; Institute of Ophthalmology, University College of London, London, United Kingdom; Fondation Ophtalmologique Adolphe de Rothschild, Paris, France; Academie des Sciences, Institut de France, Paris, France.
authors.forename: José-Alain
authors.initials: JA
authors.lastname: Sahel
authors.affiliation: Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
authors.forename: Muna I
authors.initials: MI
authors.lastname: Naash
authors.affiliation: Department of Ophthalmology, University of California San Francisco, San Francisco, California, United States of America; Department of Anatomy, University of California San Francisco, San Francisco, California United States of America.
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Lavail
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France.
authors.forename: Christina
authors.initials: C
authors.lastname: Zeitz
authors.affiliation: INSERM, U968, Paris, France; CNRS, UMR_7210, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, France; Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, DHU ViewMaintain, INSERM-DHOS CIC 1423, Paris, France; Institute of Ophthalmology, University College of London, London, United Kingdom.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Audo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055253:Color Vision; D004195:Disease Models, Animal; D004596:Electroretinography; D018628:Gene Dosage; D057898:Hemizygote; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D012160:Retina / Q000473:pathology; D012174:Retinitis Pigmentosa / Q000235:genetics*; D012243:Rhodopsin / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA; D041623:Tomography, Optical Coherence; D019076:Transgenes
chemical_list: D012243:Rhodopsin