Genotypic and Phenotypic Characterization of P23H Line 1 Rat Model

نویسندگان

  • Elise Orhan
  • Deniz Dalkara
  • Marion Neuillé
  • Christophe Lechauve
  • Christelle Michiels
  • Serge Picaud
  • Thierry Léveillard
  • José-Alain Sahel
  • Muna I. Naash
  • Matthew M. Lavail
  • Christina Zeitz
  • Isabelle Audo
  • Knut Stieger
چکیده

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.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Genotypic and phenotypic characterization of enteroaggregative Escherichia coli (EAEC) isolates from diarrheic children: An unresolved diagnostic paradigm exists

Objective(s): The enteroaggregative Escherichia coli (EAEC) has been one of the most intriguing emerging bacterial pathogens in children that occur both in developing countries and the industrial world. Although various phenotypic and genotypic based protocols have been suggested for diagnosis of EAEC, they are not conclusive or practical to be used in most clinical la...

متن کامل

Phenotypic and Genotypic Characterization of Bifidobacterium Isolates from Healthy Adult Koreans

A total of twenty-two strict anaerobic and Gram-positive Bifidobacteria, identified as B. adolescentis, B.pseudocatenulatum, or B. longum, were isolated from healthy adult Koreans. We here investigated the cellmorphology, antimicrobial resistance patterns to novel antibiotics and genotypic differentiation ofBifidobacteria assessing repetitive DNA element PCR (rep-PCR) fingerpr...

متن کامل

Phenotypic characterizationand PCR-ribotyping of Pseudomonas fluorescensisolates, in tracking contamination routes in the production line of pasteurized milk

Contamination sites of Pseudomonas fluorescenswere traced in the production line for milk pasteurization in a large dairy plant in Shiraz, Iran. Samples of raw and pasteurized milk were collected at six sites along the line. All milk samples were incubated at 7°C until the aerobic plate count had reached 10 6 -10 7 cfu mL-1. Colonies were picked randomly and identified. No growth of gram negati...

متن کامل

P23H rhodopsin transgenic rat: correlation of retinal function with histopathology.

PURPOSE To correlate retinal functional changes with structural changes in P23H rhodopsin transgenic rats as a model of autosomal dominant retinitis pigmentosa. METHODS P23H heterozygote (lines 1 and 3) and Sprague-Dawley control rats were studied at 4 to 29 weeks by retinal histology, electroretinogram (ERG), and a-wave transduction modeling. RESULTS Both line 1 (faster degeneration) and l...

متن کامل

Hearing impairment in the P23H-1 retinal degeneration rat model

The transgenic P23H line 1 (P23H-1) rat expresses a variant of rhodopsin with a mutation that leads to loss of visual function. This rat strain is an experimental model usually employed to study photoreceptor degeneration. Although the mutated protein should not interfere with other sensory functions, observing severe loss of auditory reflexes in response to natural sounds led us to study audit...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 10  شماره 

صفحات  -

تاریخ انتشار 2015