Microtubule actin crosslinking factor 1 regulates the balbiani body and animal-vegetal polarity of the zebrafish oocyte

Tripti Gupta, Florence L. Marlow, Deborah Ferriola, Katarzyna Mackiewicz, Johannes Dapprich, Dimitri Monos, Mary C. Mullins

Research output: Contribution to journalArticle

68 Citations (Scopus)

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.

Original languageEnglish (US)
JournalPLoS Genetics
Volume6
Issue number8
DOIs
StatePublished - Aug 2010
Externally publishedYes

Fingerprint

Zebrafish
Danio rerio
crosslinking
microtubules
Oocytes
actin
oocytes
vertebrate
High-Throughput Nucleotide Sequencing
Vertebrates
gene
animal
mutation
genomics
vertebrates
mutants
animals
DNA
Genes
phenotype

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Ecology, Evolution, Behavior and Systematics
  • Cancer Research
  • Genetics(clinical)

Cite this

Microtubule actin crosslinking factor 1 regulates the balbiani body and animal-vegetal polarity of the zebrafish oocyte. / Gupta, Tripti; Marlow, Florence L.; Ferriola, Deborah; Mackiewicz, Katarzyna; Dapprich, Johannes; Monos, Dimitri; Mullins, Mary C.

In: PLoS Genetics, Vol. 6, No. 8, 08.2010.

Research output: Contribution to journalArticle

Gupta, Tripti ; Marlow, Florence L. ; Ferriola, Deborah ; Mackiewicz, Katarzyna ; Dapprich, Johannes ; Monos, Dimitri ; Mullins, Mary C. / Microtubule actin crosslinking factor 1 regulates the balbiani body and animal-vegetal polarity of the zebrafish oocyte. In: PLoS Genetics. 2010 ; Vol. 6, No. 8.
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