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Neural crest specification and migration independently require NSD3-related lysine methyltransferase activity.

Jacques-Fricke BT, Gammill LS - Mol. Biol. Cell (2014)

Bottom Line: Here we show that the lysine methyltransferase NSD3 is abundantly and specifically expressed in premigratory and migratory neural crest cells.Nevertheless, only Sox10 histone H3 lysine 36 dimethylation requires NSD3, revealing unexpected complexity in NSD3-dependent neural crest gene regulation.These results identify NSD3 as the first protein methyltransferase essential for neural crest gene expression during specification and show that NSD3-related methyltransferase activity independently regulates migration.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN 55455.

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NSD3 or a related methyltransferase regulates neural crest migration independent of specification. (A) Whereas electroporation at stage 4+ knocks down NSD3 or interferes with NSD3-related activity throughout early neural crest development, electroporation at stages 7–8 limits NSD3 loss of function to migratory stages. (B, C) Embryos were unilaterally electroporated at one to four somites with 6–8 μg/μl GFP bicistronic expression plasmid pMES (B) or pMES driving NSD3Δ1707 (C). After incubation to migration stages, neural crest cells were visualized by Sox10 in situ hybridization. Bracket, targeted cells; white arrow, neural crest cells that have not migrated. (B′, C′) GFP construct targeting. Dorsal view, anterior up. White arrowhead, targeted side; black arrowhead, untargeted side; s, somite. (D) Embryos were categorized by the distance MO-targeted neural crest cells migrated compared with the untargeted control side. All embryos electroporated with NSD3Δ1707 had impaired neural crest migration.
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Figure 7: NSD3 or a related methyltransferase regulates neural crest migration independent of specification. (A) Whereas electroporation at stage 4+ knocks down NSD3 or interferes with NSD3-related activity throughout early neural crest development, electroporation at stages 7–8 limits NSD3 loss of function to migratory stages. (B, C) Embryos were unilaterally electroporated at one to four somites with 6–8 μg/μl GFP bicistronic expression plasmid pMES (B) or pMES driving NSD3Δ1707 (C). After incubation to migration stages, neural crest cells were visualized by Sox10 in situ hybridization. Bracket, targeted cells; white arrow, neural crest cells that have not migrated. (B′, C′) GFP construct targeting. Dorsal view, anterior up. White arrowhead, targeted side; black arrowhead, untargeted side; s, somite. (D) Embryos were categorized by the distance MO-targeted neural crest cells migrated compared with the untargeted control side. All embryos electroporated with NSD3Δ1707 had impaired neural crest migration.

Mentions: We electroporated NSD3Δ1707 or pMES into one- to four-somite cranial neural folds (HH stages 7–8; Figure 7A). After incubating electroporated embryos to migratory stages, we visualized migratory neural crest cells using Sox10 in situ hybridization. Delays in neural crest migration were apparent in 100% of NSD3Δ1707-electroporated embryos (Figure 7, C and D). Of these, 75% exhibited moderate to severe defects, compared with mild disruptions in only 33.3% of pMES electroporated embryos (Figure 7, B and D, p = 3.8 × 10−4). NSD3Δ1707-expressing neural crest cells emigrated from the neural tube but appeared to halt en route (Figure 7C), suggesting that dominant-negative activity commenced in actively migrating cells. This effect was sometimes apparent throughout the population, so that the extent of migration was reduced compared with the untargeted side of the embryo (Figure 7C, white arrowhead), as seen after gastrula-stage MO electroporation (Figure 6, B, E, and F). Alternatively, groups of cells trailed the rest of the normally migrating population or did not migrate away from the midline (Figure 7C, white arrow). These data indicate that methylation by NSD3 or a related methyltransferase directly regulates neural crest migration.


Neural crest specification and migration independently require NSD3-related lysine methyltransferase activity.

Jacques-Fricke BT, Gammill LS - Mol. Biol. Cell (2014)

NSD3 or a related methyltransferase regulates neural crest migration independent of specification. (A) Whereas electroporation at stage 4+ knocks down NSD3 or interferes with NSD3-related activity throughout early neural crest development, electroporation at stages 7–8 limits NSD3 loss of function to migratory stages. (B, C) Embryos were unilaterally electroporated at one to four somites with 6–8 μg/μl GFP bicistronic expression plasmid pMES (B) or pMES driving NSD3Δ1707 (C). After incubation to migration stages, neural crest cells were visualized by Sox10 in situ hybridization. Bracket, targeted cells; white arrow, neural crest cells that have not migrated. (B′, C′) GFP construct targeting. Dorsal view, anterior up. White arrowhead, targeted side; black arrowhead, untargeted side; s, somite. (D) Embryos were categorized by the distance MO-targeted neural crest cells migrated compared with the untargeted control side. All embryos electroporated with NSD3Δ1707 had impaired neural crest migration.
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Related In: Results  -  Collection

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Figure 7: NSD3 or a related methyltransferase regulates neural crest migration independent of specification. (A) Whereas electroporation at stage 4+ knocks down NSD3 or interferes with NSD3-related activity throughout early neural crest development, electroporation at stages 7–8 limits NSD3 loss of function to migratory stages. (B, C) Embryos were unilaterally electroporated at one to four somites with 6–8 μg/μl GFP bicistronic expression plasmid pMES (B) or pMES driving NSD3Δ1707 (C). After incubation to migration stages, neural crest cells were visualized by Sox10 in situ hybridization. Bracket, targeted cells; white arrow, neural crest cells that have not migrated. (B′, C′) GFP construct targeting. Dorsal view, anterior up. White arrowhead, targeted side; black arrowhead, untargeted side; s, somite. (D) Embryos were categorized by the distance MO-targeted neural crest cells migrated compared with the untargeted control side. All embryos electroporated with NSD3Δ1707 had impaired neural crest migration.
Mentions: We electroporated NSD3Δ1707 or pMES into one- to four-somite cranial neural folds (HH stages 7–8; Figure 7A). After incubating electroporated embryos to migratory stages, we visualized migratory neural crest cells using Sox10 in situ hybridization. Delays in neural crest migration were apparent in 100% of NSD3Δ1707-electroporated embryos (Figure 7, C and D). Of these, 75% exhibited moderate to severe defects, compared with mild disruptions in only 33.3% of pMES electroporated embryos (Figure 7, B and D, p = 3.8 × 10−4). NSD3Δ1707-expressing neural crest cells emigrated from the neural tube but appeared to halt en route (Figure 7C), suggesting that dominant-negative activity commenced in actively migrating cells. This effect was sometimes apparent throughout the population, so that the extent of migration was reduced compared with the untargeted side of the embryo (Figure 7C, white arrowhead), as seen after gastrula-stage MO electroporation (Figure 6, B, E, and F). Alternatively, groups of cells trailed the rest of the normally migrating population or did not migrate away from the midline (Figure 7C, white arrow). These data indicate that methylation by NSD3 or a related methyltransferase directly regulates neural crest migration.

Bottom Line: Here we show that the lysine methyltransferase NSD3 is abundantly and specifically expressed in premigratory and migratory neural crest cells.Nevertheless, only Sox10 histone H3 lysine 36 dimethylation requires NSD3, revealing unexpected complexity in NSD3-dependent neural crest gene regulation.These results identify NSD3 as the first protein methyltransferase essential for neural crest gene expression during specification and show that NSD3-related methyltransferase activity independently regulates migration.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN 55455.

Show MeSH
Related in: MedlinePlus