Protease inhibitor 15, a candidate gene for abdominal aortic internal elastic lamina ruptures in the rat

The inbred Brown Norway (BN) rat develops spontaneous ruptures of the internal elastic lamina (RIEL) of the abdominal aorta (AA) and iliac arteries. Prior studies with crosses of the BN/Orl RJ (susceptible) and LOU/M (resistant) showed the presence of a significant QTL on chromosome 5 and the production of congenic rats proved the involvement of this locus. In this study, we further dissected the above-mentioned QTL by creating a new panel of LOU.BN(chr5) congenic and sub-congenic lines and reduced the locus to 5.2 Mb. Then 1002 HS rats were studied, whose phenotyping revealed a low prevalence and high variability for RIEL. High resolution mapping in the HS panel detected the major locus on chromosome 5 (log P>35) and refined it to 1.4Mb. Subsequently, RNA-seq analysis on AA of BN, congenics and LOU revealed expression differences for only protease inhibitor15 (Pi15) gene and a putative long intergenic noncoding RNA (lincRNA) within the linkage region. The high abundance of lincRNA with respect to reduced Pi15 expression, in conjunction with exertion of longitudinal strain, may be related to RIEL, indicating the potential importance of proteases in biological processes related to defective aortic IEL structure. Similar mechanisms may be involved in aneurysm initiation in the human AA

Characteristics of the aortic elastic network and related phenotypes in seven inbred rat strains.

Extracellular matrix (ECM) molecules such as elastin and collagen provide mechanical support to the vessel wall and are essential for vascular function. Evidence that genetic factors influence aortic ECM composition and organization was concluded from our previous studies showing that the inbred Brown Norway (BN) rat differs significantly from the outbred Long-Evans (LE) and the inbred LOU rat with respect to both thoracic aortic elastin content and internal elastic lamina (IEL) rupture in the abdominal aorta and iliac arteries. Here, we measured aortic elastin and collagen contents as well as factors that may modulate these parameters [insulin growth factor (IGF)-I, transforming growth factor (TGF)-beta(1), and matrix metalloproteinase (MMP)-2] in seven inbred rat strains, including BN and LOU. We also investigated whether IEL ruptures occur in strains other than BN. We showed that LOU, LE, BN, and Fischer 344 (F344) rats were significantly different for aortic elastin content and elastin-to-collagen ratio, whereas LE, Lewis, WAG, and Wistar-Furth (WF) were similar for these parameters. BN and F344 had the lowest values. BN was the only strain to present numerous IEL ruptures, whereas F344, LE, and WF presented a few and the other strains presented none. In addition, IGF-I and TGF-beta(1) levels in the plasma and aorta differed significantly between strains, suggesting genetic control of their production. Because inbred rat strains provide interesting models for quantitative trait locus analysis, our results concerning elastin, collagen, IEL ruptures, and cytokines may provide a basis for the search for candidate genes involved in the control of these phenotypes

Localization of genetic loci controlling hydronephrosis in the Brown Norway rat and its association with hematuria

The aim of this study was to investigate the genetic basis of congenital hydronephrosis (HN), a poorly defined pathological entity, using a rat model. The Brown Norway (BN) strain presents spontaneously a high incidence of apparently asymptomatic HN, whereas the LOU strain does not. A backcross was established between these two strains [BNx(BNxLOU)] and a genome-wide scan performed with 193 microsatellite markers on 121 males and 118 females of this population, which had been phenotyped and scored for HN severity (defined as the degree of renal pelvic dilation), followed by linkage analysis using Mapmaker/QTL software. Bilateral HN score was significantly linked to a locus on chromosome 6 (Z-scores 4.4 and 4.8 for all rats and for females respectively). Suggestive loci were identified on chromosomes 2 (for only right-sided HN) and 4. This is the first study in rat to identify genetic loci for HN. 3 candidate genes present in these loci were sequenced and insertions detected in Id2 and Agtr1b genes in BN, which did not however lead to modified expression as measured by quantitative PCR. Production of a congenic line for part of the chromosome 6 locus confirmed its involvement in HN, but the phenotype was mild. Evidence of hematuria was observed in 9.6% of the backcross rats, mostly males and only in kidneys with HN, but not necessarily in the most severely affected. Hematuria also occurs in the BN colony used here, where it is due to papilloma-like lesions involving pelvic epithelial proliferation, but not in the LOU