#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Microtubule Actin Crosslinking Factor 1 Regulates the Balbiani Body and Animal-Vegetal Polarity of the Zebrafish Oocyte


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.


Vyšlo v časopise: Microtubule Actin Crosslinking Factor 1 Regulates the Balbiani Body and Animal-Vegetal Polarity of the Zebrafish Oocyte. PLoS Genet 6(8): e32767. doi:10.1371/journal.pgen.1001073
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1001073

Souhrn

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.


Zdroje

1. GurayaSS

1979 Recent advances in the morphology, cytochemistry, and function of Balbiani's vitelline body in animal oocytes. Int Rev Cytol 59 249 321

2. KlocM

BilinskiS

EtkinLD

2004 The Balbiani body and germ cell determinants: 150 years later. Curr Top Dev Biol 59 1 36

3. CoxRT

SpradlingAC

2003 A Balbiani body and the fusome mediate mitochondrial inheritance during Drosophila oogenesis. Development 130 1579 1590

4. BillettFS

AdamE

1976 The structure of the mitochondrial cloud of Xenopus laevis oocytes. J Embryol Exp Morphol 36 697 710

5. HeasmanJ

QuarmbyJ

WylieCC

1984 The mitochondrial cloud of Xenopus oocytes: the source of germinal granule material. Dev Biol 105 458 469

6. PeplingME

WilhelmJE

O'HaraAL

GephardtGW

SpradlingAC

2007 Mouse oocytes within germ cell cysts and primordial follicles contain a Balbiani body. Proc Natl Acad Sci U S A 104 187 192

7. HertigAT

AdamsEC

1967 Studies on the human oocyte and its follicle. I. Ultrastructural and histochemical observations on the primordial follicle stage. J Cell Biol 34 647 675

8. HertigAT

1968 The primary human oocyte: some observations on the fine structure of Balbiani's vitelline body and the origin of the annulate lamellae. Am J Anat 122 107 137

9. MottaPM

NottolaSA

MakabeS

HeynR

2000 Mitochondrial morphology in human fetal and adult female germ cells. Hum Reprod 15 Suppl 2 129 147

10. WilhelmJE

BuszczakM

SaylesS

2005 Efficient protein trafficking requires Trailer Hitch, a component of a ribonucleoprotein complex localized to the ER in Drosophila. Dev Cell 9 675 685

11. YangWH

YuJH

GulickT

BlochKD

BlochDB

2006 RNA-associated protein 55 (RAP55) localizes to mRNA processing bodies and stress granules. RNA 12 547 554

12. KlocM

EtkinLD

1995 Two distinct pathways for the localization of RNAs at the vegetal cortex in Xenopus oocytes. Development 121 287 297

13. ZhouY

KingML

1996 RNA transport to the vegetal cortex of Xenopus oocytes. Dev Biol 179 173 183

14. KosakaK

KawakamiK

SakamotoH

InoueK

2007 Spatiotemporal localization of germ plasm RNAs during zebrafish oogenesis. Mech Dev 124 279 289

15. MarlowFL

MullinsMC

2008 Bucky ball functions in Balbiani body assembly and animal-vegetal polarity in the oocyte and follicle cell layer in zebrafish. Dev Biol 321 40 50

16. ZhangYZ

OuyangYC

HouY

SchattenH

ChenDY

2008 Mitochondrial behavior during oogenesis in zebrafish: a confocal microscopy analysis. Dev Growth Differ 50 189 201

17. BontemsF

SteinA

MarlowF

LyauteyJ

GuptaT

2009 Bucky ball organizes germ plasm assembly in zebrafish. Curr Biol 19 414 422

18. KlocM

BilinskiS

DoughertyMT

BreyEM

EtkinLD

2004 Formation, architecture and polarity of female germline cyst in Xenopus. Dev Biol 266 43 61

19. DoschR

WagnerDS

MintzerKA

RunkeG

WiemeltAP

2004 Maternal control of vertebrate development before the midblastula transition: mutants from the zebrafish I. Dev Cell 6 771 780

20. SelmanK

Wallace

RA

Sarka

A

Xiaoping

Q

1993 Stages of Development in the Zebrafish, Brachydanio rerio. Journal of Morphology 218 203 224

21. RoperK

GregorySL

BrownNH

2002 The ‘spectraplakins’: cytoskeletal giants with characteristics of both spectrin and plakin families. J Cell Sci 115 4215 4225

22. DapprichJ

FerriolaD

MagiraEE

KunkelM

MonosD

2008 SNP-specific extraction of haplotype-resolved targeted genomic regions. Nucleic Acids Res 36 e94

23. LeungCL

GreenKJ

LiemRK

2002 Plakins: a family of versatile cytolinker proteins. Trends Cell Biol 12 37 45

24. JeffersonJJ

LeungCL

LiemRK

2004 Plakins: goliaths that link cell junctions and the cytoskeleton. Nat Rev Mol Cell Biol 5 542 553

25. VactorDV

SinkH

FambroughD

TsooR

GoodmanCS

1993 Genes that control neuromuscular specificity in Drosophila. Cell 73 1137 1153

26. LeeS

HarrisKL

WhitingtonPM

KolodziejPA

2000 short stop is allelic to kakapo, and encodes rod-like cytoskeletal-associated proteins required for axon extension. J Neurosci 20 1096 1108

27. ProutM

DamaniaZ

SoongJ

FristromD

FristromJW

1997 Autosomal mutations affecting adhesion between wing surfaces in Drosophila melanogaster. Genetics 146 275 285

28. GregorySL

BrownNH

1998 kakapo, a gene required for adhesion between and within cell layers in Drosophila, encodes a large cytoskeletal linker protein related to plectin and dystrophin. J Cell Biol 143 1271 1282

29. StrumpfD

VolkT

1998 Kakapo, a novel cytoskeletal-associated protein is essential for the restricted localization of the neuregulin-like factor, Vein, at the muscle-tendon junction site. J Cell Biol 143 1259 1270

30. RoperK

BrownNH

2004 A spectraplakin is enriched on the fusome and organizes microtubules during oocyte specification in Drosophila. Curr Biol 14 99 110

31. LeeS

KolodziejPA

2002 The plakin Short Stop and the RhoA GTPase are required for E-cadherin-dependent apical surface remodeling during tracheal tube fusion. Development 129 1509 1520

32. BottenbergW

Sanchez-SorianoN

Alves-SilvaJ

HahnI

MendeM

2009 Context-specific requirements of functional domains of the Spectraplakin Short stop in vivo. Mech Dev 126 489 502

33. KodamaA

KarakesisoglouI

WongE

VaeziA

FuchsE

2003 ACF7: an essential integrator of microtubule dynamics. Cell 115 343 354

34. ChenHJ

LinCM

LinCS

Perez-OlleR

LeungCL

2006 The role of microtubule actin cross-linking factor 1 (MACF1) in the Wnt signaling pathway. Genes Dev 20 1933 1945

35. DrabekK

van HamM

StepanovaT

DraegesteinK

van HorssenR

2006 Role of CLASP2 in microtubule stabilization and the regulation of persistent motility. Curr Biol 16 2259 2264

36. CoxRT

SpradlingAC

2006 Milton controls the early acquisition of mitochondria by Drosophila oocytes. Development 133 3371 3377

37. ReinschS

GonczyP

1998 Mechanisms of nuclear positioning. J Cell Sci 111 Pt 16 2283 2295

38. MorrisNR

2000 Nuclear migration. From fungi to the mammalian brain. J Cell Biol 148 1097 1101

39. MorrisNR

2003 Nuclear positioning: the means is at the ends. Curr Opin Cell Biol 15 54 59

40. StarrDA

FischerJA

2005 KASH 'n Karry: the KASH domain family of cargo-specific cytoskeletal adaptor proteins. Bioessays 27 1136 1146

41. WilhelmsenK

KetemaM

TruongH

SonnenbergA

2006 KASH-domain proteins in nuclear migration, anchorage and other processes. J Cell Sci 119 5021 5029

42. WilhelmsenK

LitjensSH

KuikmanI

TshimbalangaN

JanssenH

2005 Nesprin-3, a novel outer nuclear membrane protein, associates with the cytoskeletal linker protein plectin. J Cell Biol 171 799 810

43. JaegerV-M

2009 Analyse der MACF1 Isoform-3 Funktionen, einem Spektraplakin Protein mit dualen Rollen im Zytoplasma und dem Nukleus. PHD thesis Universität zu Köln

44. VedrenneC

HauriHP

2006 Morphogenesis of the endoplasmic reticulum: beyond active membrane expansion. Traffic 7 639 646

45. BoldoghIR

PonLA

2007 Mitochondria on the move. Trends Cell Biol 17 502 510

46. FrederickRL

ShawJM

2007 Moving mitochondria: establishing distribution of an essential organelle. Traffic 8 1668 1675

47. SantamaN

ErCP

OngLL

YuH

2004 Distribution and functions of kinectin isoforms. J Cell Sci 117 4537 4549

48. TanakaY

KanaiY

OkadaY

NonakaS

TakedaS

1998 Targeted disruption of mouse conventional kinesin heavy chain, kif5B, results in abnormal perinuclear clustering of mitochondria. Cell 93 1147 1158

49. BorgeseN

FrancoliniM

SnappE

2006 Endoplasmic reticulum architecture: structures in flux. Curr Opin Cell Biol 18 358 364

50. VedrenneC

KlopfensteinDR

HauriHP

2005 Phosphorylation controls CLIMP-63-mediated anchoring of the endoplasmic reticulum to microtubules. Mol Biol Cell 16 1928 1937

51. LansbergenG

AkhmanovaA

2006 Microtubule plus end: a hub of cellular activities. Traffic 7 499 507

52. AkhmanovaA

SteinmetzMO

2008 Tracking the ends: a dynamic protein network controls the fate of microtubule tips. Nat Rev Mol Cell Biol 9 309 322

53. SubramanianA

ProkopA

YamamotoM

SugimuraK

UemuraT

2003 Shortstop recruits EB1/APC1 and promotes microtubule assembly at the muscle-tendon junction. Curr Biol 13 1086 1095

54. MorrisonEE

2007 Action and interactions at microtubule ends. Cell Mol Life Sci 64 307 317

55. SlepKC

RogersSL

ElliottSL

OhkuraH

KolodziejPA

2005 Structural determinants for EB1-mediated recruitment of APC and spectraplakins to the microtubule plus end. J Cell Biol 168 587 598

56. BlumenstielJP

NollAC

GriffithsJA

PereraAG

WaltonKN

2009 Identification of EMS-induced mutations in Drosophila melanogaster by whole-genome sequencing. Genetics 182 25 32

57. ZhangZ

AlpertD

FrancisR

ChatterjeeB

YuQ

2009 Massively parallel sequencing identifies the gene Megf8 with ENU-induced mutation causing heterotaxy. Proc Natl Acad Sci U S A 106 3219 3224

58. AlbertTJ

MollaMN

MuznyDM

NazarethL

WheelerD

2007 Direct selection of human genomic loci by microarray hybridization. Nat Methods 4 903 905

59. HodgesE

XuanZ

BalijaV

KramerM

MollaMN

2007 Genome-wide in situ exon capture for selective resequencing. Nat Genet 39 1522 1527

60. OkouDT

SteinbergKM

MiddleC

CutlerDJ

AlbertTJ

2007 Microarray-based genomic selection for high-throughput resequencing. Nat Methods 4 907 909

61. D'AscenzoM

MeachamC

KitzmanJ

MiddleC

KnightJ

2009 Mutation discovery in the mouse using genetically guided array capture and resequencing. Mamm Genome 20 424 36

62. ChouLS

LiuCS

BoeseB

ZhangX

MaoR

2010 DNA sequence capture and enrichment by microarray followed by next-generation sequencing for targeted resequencing: neurofibromatosis type 1 gene as a model. Clin Chem 56 62 72

63. HoischenA

GilissenC

ArtsP

WieskampN

van der VlietW

2010 Massively parallel sequencing of ataxia genes after array-based enrichment. Hum Mutat 31 494 499

64. NikopoulosK

GilissenC

HoischenA

van NouhuysCE

BoonstraFN

2010 Next-generation sequencing of a 40 Mb linkage interval reveals TSPAN12 mutations in patients with familial exudative vitreoretinopathy. Am J Hum Genet 86 240 247

65. RehmanAU

MorellRJ

BelyantsevaIA

KhanSY

BogerET

2010 Targeted capture and next-generation sequencing identifies C9orf75, encoding Taperin, as the mutated gene in nonsyndromic deafness DFNB79. Am J Hum Genet 86 378 388

66. GnirkeA

MelnikovA

MaguireJ

RogovP

LeProustEM

2009 Solution hybrid selection with ultra-long oligonucleotides for massively parallel targeted sequencing. Nat Biotechnol 27 182 189

67. WuX

KodamaA

FuchsE

2008 ACF7 regulates cytoskeletal-focal adhesion dynamics and migration and has ATPase activity. Cell 135 137 148

68. GoryunovD

HeCZ

LinCS

LeungCL

LiemRK

2010 Nervous-tissue-specific elimination of microtubule-actin crosslinking factor 1a results in multiple developmental defects in the mouse brain. Mol Cell Neurosci 44 1 14

69. ThisseC

ThisseB

SchillingTF

PostlethwaitJH

1993 Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos. Development 119 1203 1215

70. YoonC

KawakamiK

HopkinsN

1997 Zebrafish vasa homologue RNA is localized to the cleavage planes of 2- and 4-cell-stage embryos and is expressed in the primordial germ cells. Development 124 3157 3165

71. KnapikEW

GoodmanA

EkkerM

ChevretteM

DelgadoJ

1998 A microsatellite genetic linkage map for zebrafish (Danio rerio). Nat Genet 18 338 343

72. GabrielA

DapprichJ

KunkelM

GreshamD

PrattSC

2006 Global mapping of transposon location. PLoS Genet 2 e212 doi:10.1371/journal.pgen.0020212

73. NagyM

EntzP

OtrembaP

SchoenemannC

MurphyN

2007 Haplotype-specific extraction: a universal method to resolve ambiguous genotypes and detect new alleles - demonstrated on HLA-B. Tissue Antigens 69 176 180

74. DekensMP

PelegriFJ

MaischeinHM

Nusslein-VolhardC

2003 The maternal-effect gene futile cycle is essential for pronuclear congression and mitotic spindle assembly in the zebrafish zygote. Development 130 3907 3916

75. BeckerKA

HartNH

1996 The cortical actin cytoskeleton of unactivated zebrafish eggs: spatial organization and distribution of filamentous actin, nonfilamentous actin, and myosin-II. Mol Reprod Dev 43 536 547

76. HayatM

1986 Basic Techniques for Transmission Electron Microscopy. San Diego Academic Press

77. HayatM

2000 Principles and Techniques of Electron Microscopy: Biological Applications. Cambridge Cambridge University Press

Štítky
Genetika Reprodukčná medicína

Článok vyšiel v časopise

PLOS Genetics


2010 Číslo 8
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#