The<i>Rd8</i>Mutation of the<i>Crb1</i>Gene Is Present in Vendor Lines of C57BL/6N Mice and Embryonic Stem Cells, and Confounds Ocular Induced Mutant Phenotypes

Mattapallil, Mary J.; Wawrousek, Eric F.; Chan, Chi‐Chao; Zhao, Hui; Roychoudhury, Jayeeta; Ferguson, T.; Caspi, Rachel R

doi:10.1167/iovs.12-9662

Cited by 596 publications

(545 citation statements)

References 10 publications

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“…As reported by Mattapallil et al, 17 the rd8 mutation in the Crb1 gene is found in the C57BL/6N substrain but not in the C57BL/6J substrain. This mutation results in ocular spots and retinal lesions such as retinal folding, dysplasia of the nuclear layers, retinal degeneration, and vacuolation of the retinal pigment epithelium.…”

Section: Rd8 Mutationsupporting

confidence: 66%

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Severe ocular phenotypes in Rbp4-deficient mice in the C57BL/6 genetic background

Shen

Shi

Suzuki

et al. 2016

Laboratory Investigation

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Retinol-binding protein 4 (RBP4) is a specific carrier for retinol in the blood. In hepatocytes, newly synthesized RBP4 associates with retinol and transthyretin and is secreted into the blood. The ternary transthyretin-RBP4-retinol complex transports retinol in the circulation and delivers it to target tissues. Rbp4-deficient mice in a mixed genetic background (129xC57BL/6J) have decreased sensitivity to light in the b-wave amplitude on electroretinogram. Sensitivity progressively improves and approaches that of wild-type mice at 24 weeks of age. In the present study, we produced Rbp4-deficient mice in the C57BL/6 genetic background. These mice displayed more severe phenotypes. They had decreased a-and b-wave amplitudes on electroretinograms. In accordance with these abnormalities, we found structural changes in these mice, such as loss of the peripheral choroid and photoreceptor layer in the peripheral retinas. In the central retinas, the distance between the inner limiting membrane and the outer plexiform layer was much shorter with fewer ganglion cells and fewer synapses in the inner plexiform layer. Furthermore, ocular developmental defects of retinal depigmentation, optic disc abnormality, and persistent hyaloid artery were also observed. All these abnormalities had not recovered even at 40 weeks of age. Our Rbp4-deficient mice accumulated retinol in the liver but it was undetectable in the serum, indicating an inverse relation between serum and liver retinol levels. Our results suggest that RBP4 is critical for the mobilization of retinol from hepatic storage pools, and that such mobilization is necessary for ocular development and visual function.

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Section: Rd8 Mutationsupporting

confidence: 66%

“…Mattapallil et al 17 showed that a series of commercially available ES cells of C57BL/6N origin and C57BL/6N mouse lines used to generate ES cells contained the rd8 mutation in the Crb1 gene. This mutation results in a form of retinal dysplasia and degeneration.…”

Section: Southern Blot Analyses For Isolation Of Targeted Es Cell Clonesmentioning

confidence: 99%

Severe ocular phenotypes in Rbp4-deficient mice in the C57BL/6 genetic background

Shen

Shi

Suzuki

et al. 2016

Laboratory Investigation

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“…Chimeric mice were mated to 129/SvEv mice, and the colony was maintained in C57/129 mixed background. The C57BL/6N strain carries the Crb1 rd8 mutation (43), and the rd8 mutation was eliminated by breeding. Lztfl1 genotype was determined by PCR, using the following primers: F-common: TAA-CAT-GCC-ACT-TGG-ACA-TCA-TGG, R-WT: ATT-CCA-TGA-AAG-CTG-GTG-TTG-TGA, and R-Mut: CCA-CAA-CGG-GTT-CTT-CTG-TTA-GTC.…”

Section: Methodsmentioning

confidence: 99%

“…Lztfl1 genotype was determined by PCR, using the following primers: F-common: TAA-CAT-GCC-ACT-TGG-ACA-TCA-TGG, R-WT: ATT-CCA-TGA-AAG-CTG-GTG-TTG-TGA, and R-Mut: CCA-CAA-CGG-GTT-CTT-CTG-TTA-GTC. Bbs1 M390R knock-in mice and rd8 genotyping were previously described (27,43).…”

Section: Methodsmentioning

confidence: 99%

Accumulation of non-outer segment proteins in the outer segment underlies photoreceptor degeneration in Bardet–Biedl syndrome

Datta

Allamargot

Hudson

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

132

128

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Compartmentalization and polarized protein trafficking are essential for many cellular functions. The photoreceptor outer segment (OS) is a sensory compartment specialized for phototransduction, and it shares many features with primary cilia. As expected, mutations disrupting protein trafficking to cilia often disrupt protein trafficking to the OS and cause photoreceptor degeneration. Bardet-Biedl syndrome (BBS) is one of the ciliopathies associated with defective ciliary trafficking and photoreceptor degeneration. However, precise roles of BBS proteins in photoreceptor cells and the underlying mechanisms of photoreceptor degeneration in BBS are not well understood. Here, we show that accumulation of non-OS proteins in the OS underlies photoreceptor degeneration in BBS. Using a newly developed BBS mouse model [Leucine zipper transcription factor-like 1 (Lztfl1)/Bbs17 mutant], isolated OSs, and quantitative proteomics, we determined 138 proteins that are enriched more than threefold in BBS mutant OS. In contrast, only eight proteins showed a more than threefold reduction. We found striking accumulation of Stx3 and Stxbp1/Munc18-1 and loss of polarized localization of Prom1 within the Lztfl1 and Bbs1 mutant OS. Ultrastructural analysis revealed that large vesicles are formed in the BBS OS, disrupting the lamellar structure of the OS. Our findings suggest that accumulation (and consequent sequestration) of non-OS proteins in the OS is likely the primary cause of photoreceptor degeneration in BBS. Our data also suggest that a major function of BBS proteins in photoreceptors is to transport proteins from the OS to the cell body or to prevent entry of non-OS proteins into the OS.photoreceptor degeneration | trafficking | primary cilia | outer segment | retinitis pigmentosa

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“…In addition, they were free of the rd8 mutation by PCR analysis. 55 129/SvImJ mice (strain # 002448) were purchased from Jackson Laboratories. Mice were housed in a barrier facility operated under standard 12 h light/12 h dark cycle (referred to as normal L/D) with the luminance ranging from 60 to 100 lux during the daily/light cycle.…”

Section: Methodsmentioning

confidence: 99%

Autophagy supports survival and phototransduction protein levels in rod photoreceptors

et al. 2015

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Damage and loss of the postmitotic photoreceptors is a leading cause of blindness in many diseases of the eye. Although the mechanisms of photoreceptor death have been extensively studied, few studies have addressed mechanisms that help sustain these non-replicating neurons for the life of an organism. Autophagy is an intracellular pathway where cytoplasmic constituents are delivered to the lysosomal pathway for degradation. It is not only a major pathway activated in response to cellular stress, but is also important for cytoplasmic turnover and to supply the structural and energy needs of cells. We examined the importance of autophagy in photoreceptors by deleting the essential autophagy gene Atg5 specifically in rods. Loss of autophagy led to progressive degeneration of rod photoreceptors beginning at 8 weeks of age such that by 44 weeks few rods remained. Cone photoreceptor numbers were only slightly diminished following rod degeneration but their function was significantly decreased. Rod cell death was apoptotic but was not dependent on daily light exposure or accelerated by intense light. Although the light-regulated translocation of the phototransduction proteins arrestin and transducin were unaffected in rods lacking autophagy, Atg5-deficient rods accumulated transducin-α as they degenerated suggesting autophagy might regulate the level of this protein.This was confirmed when the light-induced decrease in transducin was abolished in Atg5-deficient rods and the inhibition of autophagy in retinal explants cultures prevented its degradation. These results demonstrate that basal autophagy is essential to the long-term health of rod photoreceptors and a critical process for maintaining optimal levels of the phototransduction protein transducin-α. As the lack of autophagy is associated with retinal degeneration and altered phototransduction protein degradation in the absence of harmful gene products, this process may be a viable therapeutic target where rod cell loss is the primary pathologic event. Autophagy is an intracellular pathway where cytoplasmic constituents are delivered to the lysosomes for degradation. Defective autophagy can contribute to the age-dependent accumulation of damaged proteins and organelles leading to altered cellular homeostasis and loss of function. [1][2][3][4][5] The metabolic roles of autophagy can be classified into two types, basal and induced. In nutrient-rich conditions, autophagy is suppressed but still occurs at low levels (basal autophagy); however, when cells are subjected to stress (starvation, injury, hypoxia), autophagy is activated immediately (induced autophagy). 6 Induced autophagy maintains the amino acid pool inside cells to adapt to starvation while constitutive autophagy has been shown to function as a cell-repair mechanism that is important for long-lived postmitotic cells. 7-11 Defects in autophagy have been associated with neurodegenerative diseases, 12-15 diabetes, 16,17 lysosomal storage disease 18 and the loss of vision. 19 In addition to macroautophagy, m...

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TheRd8Mutation of theCrb1Gene Is Present in Vendor Lines of C57BL/6N Mice and Embryonic Stem Cells, and Confounds Ocular Induced Mutant Phenotypes

Cited by 596 publications

References 10 publications

Severe ocular phenotypes in Rbp4-deficient mice in the C57BL/6 genetic background

Severe ocular phenotypes in Rbp4-deficient mice in the C57BL/6 genetic background

Accumulation of non-outer segment proteins in the outer segment underlies photoreceptor degeneration in Bardet–Biedl syndrome

Autophagy supports survival and phototransduction protein levels in rod photoreceptors

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