The common modification in alphaA-crystallin in the lens, N101D, is associated with increased opacity in a mouse model.
To elucidate the morphological and cellular changes due to introduction of a charge during development and the possible mechanism that underlies cataract development in humans as a consequence of an additional charge, we generated a transgenic mouse model mimicking deamidation of Asn at position 101. The mouse model expresses a human αA-crystallin gene in which Asn-101 was replaced with Asp, which is referred to as αAN101D-transgene and is considered to be "deamidated" in this study. Mice expressing αAN101D-transgene are referred to here CRYAA(N101D) mice. All of the lines showed the expression of αAN101D-transgene. Compared with the lenses of mice expressing wild-type (WT) αA-transgene (referred to as CRYAA(WT) mice), the lenses of CRYAA(N101D) mice showed (a) altered αA-crystallin membrane protein (aquaporin-0 (AQP0), a specific lens membrane protein) interaction, (b) extracellular spaces between outer cortical fiber cells, (c) attenuated denucleation during confocal microscopic examination, (d) disrupted normal fiber cell organization and structure during scanning electron microscopic examination, (e) distorted posterior suture lines by bright field microscopy, and (f) development of a mild anterior lens opacity in the superior cortical region during the optical coherence tomography scan analysis. Relative to lenses with WT αA-crystallin, the lenses containing the deamidated αA-crystallin also showed an aggregation of αA-crystallin and a higher level of water-insoluble proteins, suggesting that the morphological and cellular changes in these lenses are due to the N101D mutation. This study provides evidence for the first time that expression of deamidated αA-crystallin caused disruption of fiber cell structural integrity, protein aggregation, insolubilization, and mild cortical lens opacity.
Department of Vision Sciences, University of Alabama at Birmingham, Birmingham, Alabama 35294-4390, USA.
Ratna
R
Gupta
Chinwe O
CO
Asomugha
Om P
OP
Srivastava
D019943:Amino Acid Substitution; D000818:Animals; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism*; D002386:Cataract / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D003459:Crystallins / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008822:Mice, Transgenic; D020125:Mutation, Missense
D020346:Aquaporins; C511246:CRYAA protein, human; D003459:Crystallins; D005136:Eye Proteins; C030328:aquaporin 0
Exp Eye Res. 2004 Mar;78(3):673-87
15106947
Nature. 1970 Aug 15;227(5259):680-5
5432063
J Cell Sci. 2003 Mar 15;116(Pt 6):1073-85
12584250
Mol Vis. 2008 Jul 10;14:1282-7
18618007
FASEB J. 2006 May;20(7):846-57
16675842
Exp Eye Res. 1994 May;58(5):595-603
7925697
Int J Dev Biol. 2004;48(8-9):845-56
15558476
Exp Eye Res. 2003 Mar;76(3):385-91
12573667
Exp Eye Res. 1998 Aug;67(2):163-77
9733583
Invest Ophthalmol Vis Sci. 1991 Jun;32(7):2119-29
2055702
Exp Eye Res. 1998 Jul;67(1):21-30
9702175
Eur J Cell Biol. 1990 Oct;53(1):59-74
2076709
Invest Ophthalmol Vis Sci. 2010 Jun;51(6):3107-14
20053973
BMC Ophthalmol. 2003 Jan 24;3:3
12546709
Curr Eye Res. 1992 Sep;11(9):929-33
1424733
Biochem Biophys Res Commun. 2009 Dec 18;390(3):1034-9
19857466
Nature. 1983 Mar 31-Apr 6;302(5907):415-7
6835373
Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10449-53
1438232
Invest Ophthalmol Vis Sci. 2001 Mar;42(3):735-42
11222535
Int Rev Cytol. 2000;192:171-87
10553279
Invest Ophthalmol Vis Sci. 2006 May;47(5):2036-44
16639013
Exp Eye Res. 1994 Nov;59(5):505-20
9492753
J Proteome Res. 2006 Oct;5(10):2554-66
17022627
BMC Ophthalmol. 2009 Jul 20;9:4
19619312
Exp Eye Res. 2003 Feb;76(2):175-82
12565805
Exp Eye Res. 1996 Mar;62(3):237-51
8690033
Dev Biol. 2002 May 1;245(1):13-27
11969252
Curr Eye Res. 1998 Mar;17(3):247-50
9543632
Mol Vis. 2006 Aug 10;12:879-84
16917488
J Proteome Res. 2007 Oct;6(10):3935-43
17824632
BMC Ophthalmol. 2007 Dec 20;7:19
18096063
Invest Ophthalmol Vis Sci. 2005 Jan;46(1):223-34
15623777
Biochemistry. 2006 Mar 14;45(10):3146-53
16519509
Exp Eye Res. 2000 Aug;71(2):195-207
10930324
Biochemistry. 2002 Jan 15;41(2):483-90
11781086
Biochemistry. 2002 Nov 26;41(47):14076-84
12437365
Eye (Lond). 1989;3 ( Pt 5):597-605
2630337
Exp Eye Res. 2003 Nov;77(5):567-74
14550398
Mol Vis. 2000 Sep 05;6:164-8
10976112
Dev Biol. 2008 Sep 15;321(2):420-33
18652817
Proc Natl Acad Sci U S A. 1982 May;79(10):3208-12
6954471
Mol Vis. 2010 Mar 23;16:476-94
20352024
Invest Ophthalmol Vis Sci. 2004 Oct;45(10):3608-19
15452068
Invest Ophthalmol Vis Sci. 2004 Jan;45(1):206-14
14691175
Dev Biol. 2005 Apr 1;280(1):1-14
15766743
Mol Vis. 2000 May 01;6:63-71
10811949
Int J Biochem Cell Biol. 2008;40(3):317-23
18093866
Biochemistry. 2000 Dec 26;39(51):15791-8
11123904
Invest Ophthalmol Vis Sci. 2001 Mar;42(3):601-5
11222516
Br J Ophthalmol. 2009 Jan;93(1):61-5
18838411
Biochemistry. 2008 Sep 23;47(38):10069-83
18754677
Protein Sci. 2008 Sep;17(9):1565-75
18567786
J Cell Sci. 2004 Feb 29;117(Pt 6):871-80
14762116
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
388439
Exp Eye Res. 2007 Mar;84(3):513-28
17217947
Exp Eye Res. 1998 Jul;67(1):31-43
9702176
BMC Ophthalmol. 2003 Jan 06;3:1
12515578
Eye (Lond). 1999 Jun;13 ( Pt 3b):417-24
10627819
Prog Biophys Mol Biol. 2004 Nov;86(3):407-85
15302206
News Physiol Sci. 2001 Jun;16:118-23
11443230
J Biol Chem. 2004 Oct 22;279(43):44258-69
15284238