Differential expression of AP-2a and AP-2b in the developing chick retina: repression of R-FABP promoter activity by AP-2

Bisgrove, Dwayne A.; Monckton, Elizabeth A.; Godbout, Roseline

doi:10.1139/bcb-77-4-384

Cited by 15 publications

(24 citation statements)

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“…AP‐2α knockout mice exhibited perinatal death with severe skeletal defects in craniofacial and limb bud development (shortened limb length, loss of radius, and loss or transformation of the first digit of the forelimb) in addition to other developmental defects (neural tube closure, body wall formation, and eye formation). This finding was consistent with the detection of AP‐2α and γ in the developing limb bud during early embryogenesis (21, 26) . Subsequently, chimeric mice studies confirmed all the major defects found in AP‐2α knockout mice.…”

Section: Introductionsupporting

confidence: 88%

Negative Regulation of Chondrocyte Differentiation by Transcription Factor AP-2α

Huang

Sandell

2004

Journal of Bone and Mineral Research

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This study investigated the role of transcription factor AP-2␣ in chondrocyte differentiation in vitro. AP-2␣ mRNA declined during differentiation, and overexpression of AP-2␣ inhibited differentiation. The results demonstrated that AP-2␣ plays a negative role in chondrocyte differentiation.Introduction: Transcription factor AP-2␣ has been detected in growth plate and articular chondrocytes and has been shown to regulate cartilage matrix gene expression in vitro. However, the precise functional role of AP-2␣ in chondrocyte differentiation is not known. In this study, we assessed the expression and the function of AP-2␣ in chondrocyte differentiation of ATDC5 cells. Materials and Methods: Chondrocyte differentiation of ATDC5 cells was induced with insulin or transforming growth factor ␤ (TGF-␤). Proteoglycan production was assessed by alcian blue staining, and expression levels of chondrocyte marker genes and AP-2 gene family were determined by quantitative real time reverse transcriptasepolymerase chain reaction (RT-PCR). Overexpression of AP-2␣ in ATDC5 cells was accomplished by retroviral infection. Infected cells were selected for G418 resistance and pooled for further analysis. Results and Conclusions:Quantitative real time RT-PCR analysis showed that among the four members of the AP-2 gene family, AP-2␣ mRNA was the most abundant. AP-2␣ mRNA levels progressively declined during the differentiation induced by either insulin or TGF-␤ treatment. Retroviral expression of AP-2␣ in ATDC5 cells prevented the formation of cartilage nodules, suppressed the proteoglycan production, and inhibited the expression of type II collagen, aggrecan, and type X collagen. Expression profile analysis of key transcription factors involved in chondrogenesis showed that overexpression of AP-2␣ maintained the expression of Sox9 but suppressed the expression of Sox5 and Sox6. Taken together, we provide, for the first time, molecular and cellular evidence suggesting that AP-2␣ is a negative regulator of chondrocyte differentiation.

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Section: Introductionsupporting

confidence: 88%

Negative Regulation of Chondrocyte Differentiation by Transcription Factor AP-2α

Huang

Sandell

2004

Journal of Bone and Mineral Research

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“…These markers included calretinin, expressed in horizontal and ganglion cells and some amacrine cells [43,44]; general neural marker MAP2 present in all retinal neurons except bipolar cells [45]; ganglion cell markers RA4 [46] and Islet-1 [47], which also labels bipolar cells at late developmental stages, amacrine markers AP2α [48] and GAD67 [49]; bipolar cell markers ath3 and chx10 [50]; horizontal cell marker LIM [51]; photoreceptor markers red opsin, IRBP [52,53], and visinin [54], whose immunoreactivity is also detected weakly in developing amacrine cells in the chick retina [55,56]. …”

Section: Resultsmentioning

confidence: 99%

Reprogramming Retinal Pigment Epithelium to Differentiate Toward Retinal Neurons with Sox2

Yan

et al. 2009

Stem Cells

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Guiding non-neural, retinal pigment epithelium (RPE) to produce retinal neurons may offer a source of developing neurons for cell-replacement. Sox2 plays important roles in maintaining neural progenitor/stem cell properties and in converting fibroblasts into pluripotent stem cells. This study tests the possibility of using Sox2 to reprogram RPE to differentiate toward retinal neurons in vivo and in vitro. Expression of Sox2 in the chick retina was detected in progenitor cells, in cells at a discrete location in the layers of amacrine and ganglion cells, and in Mu †ller glia. Overexpression of Sox2 in the developing eye resulted in hypopigmentation of the RPE. In the affected regions, expression of retinal ganglion cell markers became apparent in the RPE layer. In RPE cell culture, Sox2 promoted the expression of retinal ganglion and amacrine markers, and suppressed the expression of genes associated with RPE properties. Mechanistic investigation using the developing retina revealed a coexpression of Sox2 and basic fibroblast growth factor (bFGF), a growth factor commonly used in stem cell culture and capable of inducing RPE-to-retina transdifferentiation (or reprogramming) during early development. Similar patterns of changes in Sox2 expression and in bFGF expression were observed in atrophic retina and in injured retina. In RPE cell culture, Sox2 and bFGF mutually enhanced one another's expression. Upregulation of bFGF expression by Sox2 also occurred in the retina. These results suggest that Sox2 can initiate a reprogramming of RPE cells to differentiate toward retinal neurons and may engage bFGF during the process.

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“…The antibody specifically recognized a 50‐kDa protein (corresponding to AP2α) on a Western blot of nuclear extract from Hela cells engineered to express AP2α. Immunohistochemical analyses of the chick retina carried out in different laboratories (including our own) show that the antibody specifically labels amacrine and horizontal cells, all other cell types being immunonegative (Bisgrove and Godbout,1999; Li C‐M et al,2001; Mao et al,2009). A mouse monoclonal antibody against AP2α (clone 3B5) also specifically labels amacrine and horizontal cells in the chick retina (Bisgrove and Godbout,1999; Li C‐M et al,2001; Fischer et al,2007; Mao et al,2009; Ma et al,2009b).…”

Section: Methodsmentioning

confidence: 94%

“…Immunohistochemical analyses of the chick retina carried out in different laboratories (including our own) show that the antibody specifically labels amacrine and horizontal cells, all other cell types being immunonegative (Bisgrove and Godbout,1999; Li C‐M et al,2001; Mao et al,2009). A mouse monoclonal antibody against AP2α (clone 3B5) also specifically labels amacrine and horizontal cells in the chick retina (Bisgrove and Godbout,1999; Li C‐M et al,2001; Fischer et al,2007; Mao et al,2009; Ma et al,2009b). Monoclonal antibody against Brn3A specifically recognized Brn3A, with no reactivity to Brn3B or Brn3C by Western blots, and showed no reactivity to Brn3A knockout mice (data sheet from Chemicon/Millipore, Temecula, CA).…”

Section: Methodsmentioning

confidence: 99%

Neurogenin1 effectively reprograms cultured chick retinal pigment epithelial cells to differentiate toward photoreceptors

Yan

Liang

et al. 2009

J of Comparative Neurology

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Photoreceptors are highly specialized sensory neurons in the retina, and their degeneration results in blindness. Replacement with developing photoreceptor cells promises to be an effective therapy, but it requires a supply of new photoreceptors, because the neural retina in human eyes lacks regeneration capability. We report efficient generation of differentiating, photoreceptor-like neurons from chick retinal pigment epithelial (RPE) cells propagated in culture through reprogramming with neurogenin1 (ngn1). In reprogrammed culture, a large number of the cells (85.0 ± 5.9%) began to differentiate towards photoreceptors. Reprogrammed cells expressed transcription factors that set in motion photoreceptor differentiation, including Crx, Nr2E3, NeuroD, and RXRγ, and phototransduction pathway components, including transducin, cGMP-gated channel, and red opsin of cone photoreceptors (equivalent to rhodopsin of rod photoreceptors). They developed inner segments rich in mitochondria. Furthermore, they responded to light by decreasing their cellular free calcium (Ca2+) levels and responded to 9-cis-retinal by increasing their Ca2+ levels after photobleaching, hallmarks of photoreceptor physiology. The high efficiency and the advanced photoreceptor differentiation indicate ngn1 as a gene of choice to reprogram RPE progeny cells to differentiate into photoreceptor neurons in future cell replacement studies.

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Differential expression of AP-2a and AP-2b in the developing chick retina: repression of R-FABP promoter activity by AP-2

Cited by 15 publications

References 0 publications

Negative Regulation of Chondrocyte Differentiation by Transcription Factor AP-2α

Negative Regulation of Chondrocyte Differentiation by Transcription Factor AP-2α

Reprogramming Retinal Pigment Epithelium to Differentiate Toward Retinal Neurons with Sox2

Neurogenin1 effectively reprograms cultured chick retinal pigment epithelial cells to differentiate toward photoreceptors

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