Abstract
Nuc1 is a spontaneous rat mutant resulting from a mutation in the Cryba1 gene, coding for βA3/A1-crystallin. Our earlier studies with Nuc1 provided novel evidence that astrocytes, which express βA3/A1-crystallin, have a pivotal role in retinal remodeling. The role of astrocytes in the retina is only beginning to be explored. One of the limitations in the field is the lack of appropriate animal models to better investigate the function of astrocytes in retinal health and disease. We have now established transgenic mice that overexpress the Nuc1 mutant form of Cryba1, specifically in astrocytes. Astrocytes in wild type mice show normal compact stellate structure, producing a honeycomb-like network. In contrast, in transgenics over-expressing the mutant (Nuc1) Cryba1 in astrocytes, bundle-like structures with abnormal patterns and morphology were observed. In the nerve fiber layer of the transgenic mice, an additional layer of astrocytes adjacent to the vitreous is evident. This abnormal organization of astrocytes affects both the superficial and deep retinal vascular density and remodeling. Fluorescein angiography showed increased venous dilation and tortuosity of branches in the transgenic retina, as compared to wild type. Moreover, there appear to be fewer interactions between astrocytes and endothelial cells in the transgenic retina than in normal mouse retina. Further, astrocytes overexpressing the mutant βA3/A1-crystallin migrate into the vitreous, and ensheath the hyaloid artery, in a manner similar to that seen in the Nuc1 rat. Together, these data demonstrate that developmental abnormalities of astrocytes can affect the normal remodeling process of both fetal and retinal vessels of the eye and that βA3/A1-crystallin is essential for normal astrocyte function in the retina.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andley UP (2007) Crystallins in the eye: function and pathology. Prog Retin Eye Res 26:78–98
Bhat SP (2004) Transparency and non-refractive functions of crystallins-a proposal. Exp Eye Res 79:809–816
Brenner M (1994) Structure and transcriptional regulation of the GFAP gene. Brain Path 4:245–257
Cahoy JD, Emery B, Kaushal A, Foo LC, Zamanian JL, Christpherson KS, Xing Y, Lubischer JL, Krieg PA, Krupenko SA, Thompson WJ (2008) A transcriptome database for astrocytes, neurons, and oligodendrocytes: a new resource for understanding brain development and function. J Neurosci 28:264–278
Chan-Ling T, McLeod DS, Hughes S, Baxter L, Chu Y, Hasegawa T, Lutty GA (2004) Astrocyte-endothelial cell relationships during human retinal vascular development. Invest Ophthalmol Vis Sci 45:2020–2032
Delaney CL, Brenner M, Messing A (1996) Conditional ablation of cerebellar astrocytes in postnatal transgenic mice. J Neurosci 6:6908–6918
Edwards MM, McLeod DS, Grebe R, Heng C, Lefebvre O, Lutty GA (2011) Lama1 mutations lead to vitreoretinal blood vessel formation, persistence of the fetal vasculature, and epiretinal membrane formation in mice. BMC Dev Biol 11(1):60
Gehlbach P, Hose S, Lei B, Zhang C, Cano M, Arora M, Neal R, Barnstable C, Goldberg MF, Zigler JS Jr, Sinha D (2006) Developmental abnormalities in the Nuc1 rat retina: a spontaneous mutation that affects neuronal and vascular remodeling and retinal function. Neuroscience 137:447–461
Goldberg MF (1997) Persistant fetal vasculature (PFV): an integrated interpretation of signs and symptoms associated with persistent hyperplastic primary vitreous (PHPV) LIV Edward Jackson memorial lecture. Amer J Ophthalmol 124:587–626
Horwitz J (2003) Alpha-crystallin. Exp Eye Res 76(2):145–153
Hurskainen M, Eklund L, Hägg PO, Fruttiger M, Sormunen R, Ilves M, Pihlajaniemi T (2005) Abnormal maturation of the retinal vasculature in type XVIII collagen/endostatin deficient mice and changes in retinal glial cells due to lack of collagen types XV and XVIII. FASEB J 19:1564–1566
Jones SE, Jomary C, Grist J, Thomas MR, Neal MJ (1998) Expression of αB-crystallin in a mouse model of inherited retinal degeneration. NeuroReport 9:4161–4165
Kase S, He S, Sonoda S, Kitamurra M, Spee C, Wawrousek E, Ryan SJ, Kannan R, Hinton K (2010) αB-crystallin regulation of angiogenesis by modulation of VEGF. Blood 115:3398–3406
Krupenko SA, Oleinik NV (2002) 10-formyltetrahydrofolate dehydrogenase, one of the major folate enzymes, is down-regulated in tumor tissues and possesses suppressor effects on cancer cells. Cell Growth Differ 13(5):227–236
Ma B, Sen T, Asnaghi L, Valapala M, Yang F, Hose S, McLeod DS, Lu Y, Eberhart C, Zigler JS Jr, Sinha D (2011) βA3/A1-crystallin controls anoikis-mediated cell death in astrocytes by modulating PI3 K/AKT/mTOR and ERK survival pathways via the PDK/Bit1 signaling axis. Cell Death Dis 2:e217
McKay TL, Gedeon DJ, Vickerman MB, Hylton AG, Ribita D, Olar HH, Kaiser PK (2008) Parsons-Wingerter P (2008) Selective inhibition of angiogenesis in small blood vessels and decrease in vessel diameter throughout the vascular tree by triamcinolone acetonide. Invest Ophthalmol Vis Sci 49(3):1184–1190
Parthasarathy G, Ma B, Zhang C, Gongora C, Zigler JS Jr, Duncan MK, Sinha D (2011) Expression of βA3/A1-crystallin in the developing and adult rat eye. J Mol Histol 42:59–69
Piatigorsky J (2007) Gene sharing and evolution: the diversity of protein function. Harvard University Press, Cambridge, MA
Reneker LW, Overbeek PA (1996) Lens-specific expression of PDGF-A in Transgenic mice results in retinal astrocytic hamartomas. Invest Ophthalmol Vis Sci 37(12):2455–2466
Saint-Geniez M, D’Amore PA (2004) Development and pathology of the hyaloid, choroidal and retinal vasculature. Int J Dev Biol 48:1045–1058
Salvador-Silva M, Ricard CS, Agapora OA, Yang P, Hernandez MR (2001) Expression of small heat shock proteins and intermediate filaments in the human optic nerve head astrocytes exposed to elevated hydrostatic pressure in vitro. J Neurosci Res 66:59–73
Schnitzer J (1988) Astrocytes in the guinea pig, horse, and monkey retina: their occurrence coincides with the presence of blood vessels. Glia 1:74–89
Scott A, Powner MB, Gandhi P, Clarkin C, Gutmann DH, Johnson RS, Ferrara N, Fruttiger M (2010) Astrocyte-derived vascular endothelial growth factor stabilizes vessels in the developing retinal vasculature. PLoS One 5(7):e1 1863
Segovia J, Vergara P, Brenner M (1998) Astrocyte-specific expression of tyrosine hydroxylase after intracerebral gene transfer induces behavioral recovery in experimental parkinsonism. Gene Ther 5:1650–1655
Sinha D, Hose S, Zhang C, Neal R, Ghosh M, O’Brien TP, Sundin O, Goldberg MF, Robison WG Jr, Russell P, Lo WK, Zigler JS Jr (2005) A rat spontaneous mutation affects programmed cell death during the early development of the eye. Exp Eye Res 80:323–335
Sinha D, Klise A, Sergeev Y, Hose S, Bhutto IA, Hackler L Jr, Malpic-Llanos T, Samtani S, Grebe R, Goldberg MF, Hejtmancik JF, Nath A, Zack DJ, Fariss RN, McLeod DS, Sundin O, Broman KW, Lutty GA, Zigler JS Jr (2008) βA3/A1-crystallin in astroglial cells regulates retinal vascular remodeling during development. Mol Cell Neurosci 37:85–95
Stone J, Dreher Z (1987) Relationship between astrocytes, ganglion cells and vasculature of the retina. J Comp Neurol 255:35–49
Valapala M, Thamake SI, Vishwanatha JK (2011) A competitive hexapeptide inhibitor of annexin A2 prevents hypoxia-induced angiogenic events. J Cell Sci 124(Pt 9):1453–1464
Vandier D, Rixe O, Brenner M, Gouyette A, Besnard F (1998) Selective killing of glioma cell lines using an astrocyte-specific expression of the herpes simplex virus-thymidine kinase gene. Cancer Res 58:4577–4580
Werten PJL, Stege GJJ, de Jong WW (1999) The short 5′ untranslated region of the βA3/A1-crystallin mRNA is responsible for leaky ribosomal scanning. Mol Biol Reports 26:201–205
Wistow G (1995) Molecular biology and evolution of crystallins: gene recruitment and multifunctional proteins in the eye lens. Springer, New York
Yu AL, Fuchshofer R, Birha M, Priglinger SG, Eibl KH, Kampik A, Bloemendal H, Welge-Lussen U (2007) Hypoxia/reoxygenation and TGF-β increase αΒ-crystallin expression in human optic nerve head astrocytes. Exp Eye Res 84:694–706
Zhang C, Gehlbach P, Gongora C, Cano M, Fariss R, Hose S (2005) A potential role for β- and γ-crystallins in the vascular remodeling of the eye. Dev Dyn 234:36–47
Zhang C, Asnaghi L, Gongora C, Patek B, Hose S, Ma B, Fard MA, Brako L, Singh K, Goldberg MF, Handa JT, Lo WK, Eberhart CG, Zigler JS Jr, Sinha D (2011) A developmental defect in astrocytes inhibits programmed regression of the hyaloid vasculature in the mammalian eye. Eur J Cell Biol 90:440–448
Zigler JS Jr, Zhang C, Grebe R, Sehrawat G, Hackler L Jr, Adhya S, Hose S, McLeod DS, Bhutto I, Barbour W, Parthasarathy G, Zack DJ, Sergeev Y, Lutty GA, Handa JT, Sinha D (2011) Mutation in the βA3/A1-crystallin gene impairs phagosome degradation in the retinal pigmented epithelium of the rat. J Cell Sci 124:523–531
Acknowledgments
DS is thankful to the National Eye Institute for support of his IPA. This work was also supported by National Institutes of Health: EY018636 (DS), EY019037 (DS), EY019037-S (DS), Wilmer Pooled Professors Fund (DS), Helena Rubinstein Foundation (DS), EY01765 (Wilmer Imaging Core), Intramural Research Program, National Eye Institute (EFW and JSZ), and Research to Prevent Blindness (an unrestricted grant to The Wilmer Eye Institute). We also wish to thank the Norman Raab Foundation and the Wilmer Pooled Professors Fund for purchasing the Micron III imaging system. We thank Steven Lee and Carl Haugen for their technical help with the transgenic mouse production and to the Staff Members at Spring Valley Laboratories, Woodbine, MD, for taking care of the experimental animals. We thank Drs. Morton F. Goldberg and Bhaja K. Padhi for critical reading and discussion regarding this manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sinha, D., Valapala, M., Bhutto, I. et al. βA3/A1-crystallin is required for proper astrocyte template formation and vascular remodeling in the retina. Transgenic Res 21, 1033–1042 (2012). https://doi.org/10.1007/s11248-012-9608-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-012-9608-0