title: Microtubule actin crosslinking factor 1 regulates the Balbiani body and animal-vegetal polarity of the zebrafish oocyte.
abstract: Although of fundamental importance in developmental biology, the genetic basis for the symmetry breaking events that polarize the vertebrate oocyte and egg are largely unknown. In vertebrates, the first morphological asymmetry in the oocyte is the Balbiani body, a highly conserved, transient structure found in vertebrates and invertebrates including Drosophila, Xenopus, human, and mouse. We report the identification of the zebrafish magellan (mgn) mutant, which exhibits a novel enlarged Balbiani body phenotype and a disruption of oocyte polarity. To determine the molecular identity of the mgn gene, we positionally cloned the gene, employing a novel DNA capture method to target region-specific genomic DNA of 600 kb for massively parallel sequencing. Using this technique, we were able to enrich for the genomic region linked to our mutation within one week and then identify the mutation in mgn using massively parallel sequencing. This is one of the first successful uses of genomic DNA enrichment combined with massively parallel sequencing to determine the molecular identity of a gene associated with a mutant phenotype. We anticipate that the combination of these technologies will have wide applicability for the efficient identification of mutant genes in all organisms. We identified the mutation in mgn as a deletion in the coding sequence of the zebrafish microtubule actin crosslinking factor 1 (macf1) gene. macf1 is a member of the highly conserved spectraplakin family of cytoskeletal linker proteins, which play diverse roles in polarized cells such as neurons, muscle cells, and epithelial cells. In mgn mutants, the oocyte nucleus is mislocalized; and the Balbiani body, localized mRNAs, and organelles are absent from the periphery of the oocyte, consistent with a function for macf1 in nuclear anchoring and cortical localization. These data provide the first evidence for a role for spectraplakins in polarization of the vertebrate oocyte and egg.
authors.affiliation: Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.
authors.forename: Tripti
authors.initials: T
authors.lastname: Gupta
authors.forename: Florence L
authors.initials: FL
authors.lastname: Marlow
authors.forename: Deborah
authors.initials: D
authors.lastname: Ferriola
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Mackiewicz
authors.forename: Johannes
authors.initials: J
authors.lastname: Dapprich
authors.forename: Dimitri
authors.initials: D
authors.lastname: Monos
authors.forename: Mary C
authors.initials: MC
authors.lastname: Mullins
mesh_terms: D000818:Animals; D019521:Body Patterning; D016764:Cell Polarity; D018507:Gene Expression Regulation, Developmental; D009154:Mutation; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism; D051179:Plakins / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D051179:Plakins; D029961:Zebrafish Proteins; C554444:macf1a protein, zebrafish
references.citation: Genetics. 1997 May;146(1):275-85
references.pmid: 9136017
references.citation: Dev Biol. 2008 Sep 1;321(1):40-50
references.pmid: 18582455
references.citation: J Embryol Exp Morphol. 1976 Dec;36(3):697-710
references.pmid: 188969
references.citation: Curr Top Dev Biol. 2004;59:1-36
references.pmid: 14975245
references.citation: Development. 1997 Aug;124(16):3157-65
references.pmid: 9272956
references.citation: J Cell Sci. 2006 Dec 15;119(Pt 24):5021-9
references.pmid: 17158909
references.citation: Cell. 1998 Jun 26;93(7):1147-58
references.pmid: 9657148
references.citation: Mamm Genome. 2009 Jul;20(7):424-36
references.pmid: 19629596
references.citation: Am J Anat. 1968 Jan;122(1):107-37
references.pmid: 5654499
references.citation: Dev Biol. 2004 Feb 1;266(1):43-61
references.pmid: 14729477
references.citation: Nucleic Acids Res. 2008 Sep;36(15):e94
references.pmid: 18611953
references.citation: J Cell Biol. 1998 Nov 30;143(5):1271-82
references.pmid: 9832555
references.citation: Nat Genet. 2007 Dec;39(12):1522-7
references.pmid: 17982454
references.citation: Nat Methods. 2007 Nov;4(11):903-5
references.pmid: 17934467
references.citation: Nat Genet. 1998 Apr;18(4):338-43
references.pmid: 9537415
references.citation: Curr Opin Cell Biol. 2006 Aug;18(4):358-64
references.pmid: 16806883
references.citation: Nat Methods. 2007 Nov;4(11):907-9
references.pmid: 17934469
references.citation: Cell. 2008 Oct 3;135(1):137-48
references.pmid: 18854161
references.citation: J Cell Sci. 2002 Nov 15;115(Pt 22):4215-25
references.pmid: 12376554
references.citation: Mol Biol Cell. 2005 Apr;16(4):1928-37
references.pmid: 15703217
references.citation: Development. 1993 Dec;119(4):1203-15
references.pmid: 8306883
references.citation: Mech Dev. 2007 Apr;124(4):279-89
references.pmid: 17293094
references.citation: Hum Mutat. 2010 Apr;31(4):494-9
references.pmid: 20151403
references.citation: J Cell Biol. 2005 Dec 5;171(5):799-810
references.pmid: 16330710
references.citation: Dev Cell. 2004 Jun;6(6):771-80
references.pmid: 15177026
references.citation: J Cell Sci. 2004 Sep 1;117(Pt 19):4537-49
references.pmid: 15316074
references.citation: Mech Dev. 2009 Jul;126(7):489-502
references.pmid: 19409984
references.citation: Dev Biol. 1984 Oct;105(2):458-69
references.pmid: 6541166
references.citation: Trends Cell Biol. 2002 Jan;12(1):37-45
references.pmid: 11854008
references.citation: Clin Chem. 2010 Jan;56(1):62-72
references.pmid: 19910506
references.citation: Mol Reprod Dev. 1996 Apr;43(4):536-47
references.pmid: 9052946
references.citation: Curr Opin Cell Biol. 2003 Feb;15(1):54-9
references.pmid: 12517704
references.citation: Proc Natl Acad Sci U S A. 2007 Jan 2;104(1):187-92
references.pmid: 17189423
references.citation: Cell. 1993 Jun 18;73(6):1137-53
references.pmid: 8513498
references.citation: Tissue Antigens. 2007 Feb;69(2):176-80
references.pmid: 17257321
references.citation: J Cell Biol. 1998 Nov 30;143(5):1259-70
references.pmid: 9832554
references.citation: Development. 2002 Mar;129(6):1509-20
references.pmid: 11880359
references.citation: Mol Cell Neurosci. 2010 May;44(1):1-14
references.pmid: 20170731
references.citation: J Neurosci. 2000 Feb 1;20(3):1096-108
references.pmid: 10648715
references.citation: Curr Biol. 2009 Mar 10;19(5):414-22
references.pmid: 19249209
references.citation: Int Rev Cytol. 1979;59:249-321
references.pmid: 385545
references.citation: Dev Biol. 1996 Oct 10;179(1):173-83
references.pmid: 8873762
references.citation: Am J Hum Genet. 2010 Mar 12;86(3):378-88
references.pmid: 20170899
references.citation: RNA. 2006 Apr;12(4):547-54
references.pmid: 16484376
references.citation: J Cell Biol. 2005 Feb 14;168(4):587-98
references.pmid: 15699215
references.citation: Genes Dev. 2006 Jul 15;20(14):1933-45
references.pmid: 16815997
references.citation: Nat Biotechnol. 2009 Feb;27(2):182-9
references.pmid: 19182786
references.citation: Development. 2003 Apr;130(8):1579-90
references.pmid: 12620983
references.citation: Am J Hum Genet. 2010 Feb 12;86(2):240-7
references.pmid: 20159111
references.citation: J Morphol. 1993 Nov;218(2):203-224
references.pmid: 29865471
references.citation: Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3219-24
references.pmid: 19218456
references.citation: Curr Biol. 2006 Nov 21;16(22):2259-64
references.pmid: 17113391
references.citation: Dev Growth Differ. 2008 Mar;50(3):189-201
references.pmid: 18312427
references.citation: Bioessays. 2005 Nov;27(11):1136-46
references.pmid: 16237665
references.citation: Traffic. 2006 May;7(5):499-507
references.pmid: 16643273
references.citation: Nat Rev Mol Cell Biol. 2008 Apr;9(4):309-22
references.pmid: 18322465
references.citation: J Cell Biol. 2000 Mar 20;148(6):1097-101
references.pmid: 10725321
references.citation: Cell Mol Life Sci. 2007 Feb;64(3):307-17
references.pmid: 17221167
references.citation: J Cell Biol. 1967 Aug;34(2):647-75
references.pmid: 4292010
references.citation: Traffic. 2006 Jun;7(6):639-46
references.pmid: 16683914
references.citation: J Cell Sci. 1998 Aug;111 ( Pt 16):2283-95
references.pmid: 9683624
references.citation: Development. 2003 Sep;130(17):3907-16
references.pmid: 12874114
references.citation: Development. 1995 Feb;121(2):287-97
references.pmid: 7539356
references.citation: Curr Biol. 2003 Jul 1;13(13):1086-95
references.pmid: 12842007
references.citation: Cell. 2003 Oct 31;115(3):343-54
references.pmid: 14636561
references.citation: Development. 2006 Sep;133(17):3371-7
references.pmid: 16887820
references.citation: Dev Cell. 2005 Nov;9(5):675-85
references.pmid: 16256742
references.citation: Hum Reprod. 2000 Jul;15 Suppl 2:129-47
references.pmid: 11041520
references.citation: Genetics. 2009 May;182(1):25-32
references.pmid: 19307605
references.citation: Curr Biol. 2004 Jan 20;14(2):99-110
references.pmid: 14738730
references.citation: PLoS Genet. 2006 Dec 15;2(12):e212
references.pmid: 17173485
references.citation: Trends Cell Biol. 2007 Oct;17(10):502-10
references.pmid: 17804238
references.citation: Traffic. 2007 Dec;8(12):1668-1675
references.pmid: 17944806
references.citation: Nat Rev Mol Cell Biol. 2004 Jul;5(7):542-53
references.pmid: 15232572