Limits...
A key role for poly(ADP-ribose) polymerase 3 in ectodermal specification and neural crest development.

Rouleau M, Saxena V, Rodrigue A, Paquet ER, Gagnon A, Hendzel MJ, Masson JY, Ekker M, Poirier GG - PLoS ONE (2011)

Bottom Line: We have used several in vitro and in vivo approaches to examine the possible functions of PARP3 as a transcriptional regulator, a function suggested from its previously reported association with several Polycomb group (PcG) proteins.It delays pigmentation and severely impedes the development of the median fin fold and tail bud.Our findings demonstrate that Parp3 is crucial in the early stages of zebrafish development, possibly by exerting its transcriptional regulatory functions as early as during the specification of the neural plate border.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Laboratory, CHUQ Research Center, Centre Hospitalier de l'Université Laval, Québec, Canada.

ABSTRACT

Background: The PARP family member poly(ADP-ribose) polymerase 3 (PARP3) is structurally related to the well characterized PARP1 that orchestrates cellular responses to DNA strand breaks and cell death by the synthesis of poly(ADP-ribose). In contrast to PARP1 and PARP2, the functions of PARP3 are undefined. Here, we reveal critical functions for PARP3 during vertebrate development.

Principal findings: We have used several in vitro and in vivo approaches to examine the possible functions of PARP3 as a transcriptional regulator, a function suggested from its previously reported association with several Polycomb group (PcG) proteins. We demonstrate that PARP3 gene occupancy in the human neuroblastoma cell line SK-N-SH occurs preferentially with developmental genes regulating cell fate specification, tissue patterning, craniofacial development and neurogenesis. Addressing the significance of this association during zebrafish development, we show that morpholino oligonucleotide-directed inhibition of parp3 expression in zebrafish impairs the expression of the neural crest cell specifier sox9a and of dlx3b/dlx4b, the formation of cranial sensory placodes, inner ears and pectoral fins. It delays pigmentation and severely impedes the development of the median fin fold and tail bud.

Conclusion: Our findings demonstrate that Parp3 is crucial in the early stages of zebrafish development, possibly by exerting its transcriptional regulatory functions as early as during the specification of the neural plate border.

Show MeSH

Related in: MedlinePlus

Impaired expression of sox9a, dlx3b and dlx4b in parp3 morphants.Zebrafish embryos were untreated (WT) or were injected with 4 ng parp3 MO1. Gene expression was detected by in situ hybridization. A–F. The expression of sox9a is drastically reduced in the otic placodes (small arrows) at 10 hpf and in the otic vesicles (arrowheads) at 16 hpf. Expression of sox9a in somite cells (small arrows in B and E) appears diffuse in parp3 morphants. Expression of sox9a is almost completely abolished in the head region at 24 hpf (C, F). Expressions of dlx3b (G–L) and dlx4b (M–R) are minimally affected by parp3 MO in ectodermal cells at 10 hpf (G, J, M, P) but are significantly reduced in the otic vesicles (arrowheads), olfactory placodes (large arrows) and branchial arches (white arrows) of parp3 morphants at 16 hpf (H, K, N, Q) and 24 hpf (I, L, O, R). The expression of dlx3b and dlx4b is abolished in the median fin fold of 24 hpf parp3 morphant embryos (small arrows in I). Dorsal views of embryos with anterior to the bottom in A, D, G, J, M, P and lateral views with anterior to the left, dorsal to the top, in B, C, E, F, H, I, K, L, N, O, Q and R. Scale bars represent 10 µm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3022025&req=5

pone-0015834-g006: Impaired expression of sox9a, dlx3b and dlx4b in parp3 morphants.Zebrafish embryos were untreated (WT) or were injected with 4 ng parp3 MO1. Gene expression was detected by in situ hybridization. A–F. The expression of sox9a is drastically reduced in the otic placodes (small arrows) at 10 hpf and in the otic vesicles (arrowheads) at 16 hpf. Expression of sox9a in somite cells (small arrows in B and E) appears diffuse in parp3 morphants. Expression of sox9a is almost completely abolished in the head region at 24 hpf (C, F). Expressions of dlx3b (G–L) and dlx4b (M–R) are minimally affected by parp3 MO in ectodermal cells at 10 hpf (G, J, M, P) but are significantly reduced in the otic vesicles (arrowheads), olfactory placodes (large arrows) and branchial arches (white arrows) of parp3 morphants at 16 hpf (H, K, N, Q) and 24 hpf (I, L, O, R). The expression of dlx3b and dlx4b is abolished in the median fin fold of 24 hpf parp3 morphant embryos (small arrows in I). Dorsal views of embryos with anterior to the bottom in A, D, G, J, M, P and lateral views with anterior to the left, dorsal to the top, in B, C, E, F, H, I, K, L, N, O, Q and R. Scale bars represent 10 µm.

Mentions: Having determined that parp3 morphants have defective neural crest development, we next characterized the expression of sox9a, dlx3b and dlx4b. By ISH, we find that the expression of the neural crest specifier sox9a is indeed reduced in parp3 morphants at 10, 16 and 24 hpf (Fig. 6A–F). The high level of expression of sox9a in the otic placode (Fig. 6A) and later in the otic vesicle (Fig. 6B) of wild type embryos is drastically reduced in parp3 morphants (Fig. 6D, E). At 16 hpf, the expression of sox9a in the morphant paraxial cells is more diffuse suggesting that the somitogenesis is possibly disturbed (Fig. 6B, E). At 24 hpf, sox9a is normally expressed in the trunk and in three major areas of the head: the forebrain, the midbrain-hindbrain boundary and the pharyngeal arches (Fig. 6C). Expression in these three major sites is lost in the morphants and there is barely any expression in the trunk (Fig. 6F).


A key role for poly(ADP-ribose) polymerase 3 in ectodermal specification and neural crest development.

Rouleau M, Saxena V, Rodrigue A, Paquet ER, Gagnon A, Hendzel MJ, Masson JY, Ekker M, Poirier GG - PLoS ONE (2011)

Impaired expression of sox9a, dlx3b and dlx4b in parp3 morphants.Zebrafish embryos were untreated (WT) or were injected with 4 ng parp3 MO1. Gene expression was detected by in situ hybridization. A–F. The expression of sox9a is drastically reduced in the otic placodes (small arrows) at 10 hpf and in the otic vesicles (arrowheads) at 16 hpf. Expression of sox9a in somite cells (small arrows in B and E) appears diffuse in parp3 morphants. Expression of sox9a is almost completely abolished in the head region at 24 hpf (C, F). Expressions of dlx3b (G–L) and dlx4b (M–R) are minimally affected by parp3 MO in ectodermal cells at 10 hpf (G, J, M, P) but are significantly reduced in the otic vesicles (arrowheads), olfactory placodes (large arrows) and branchial arches (white arrows) of parp3 morphants at 16 hpf (H, K, N, Q) and 24 hpf (I, L, O, R). The expression of dlx3b and dlx4b is abolished in the median fin fold of 24 hpf parp3 morphant embryos (small arrows in I). Dorsal views of embryos with anterior to the bottom in A, D, G, J, M, P and lateral views with anterior to the left, dorsal to the top, in B, C, E, F, H, I, K, L, N, O, Q and R. Scale bars represent 10 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3022025&req=5

pone-0015834-g006: Impaired expression of sox9a, dlx3b and dlx4b in parp3 morphants.Zebrafish embryos were untreated (WT) or were injected with 4 ng parp3 MO1. Gene expression was detected by in situ hybridization. A–F. The expression of sox9a is drastically reduced in the otic placodes (small arrows) at 10 hpf and in the otic vesicles (arrowheads) at 16 hpf. Expression of sox9a in somite cells (small arrows in B and E) appears diffuse in parp3 morphants. Expression of sox9a is almost completely abolished in the head region at 24 hpf (C, F). Expressions of dlx3b (G–L) and dlx4b (M–R) are minimally affected by parp3 MO in ectodermal cells at 10 hpf (G, J, M, P) but are significantly reduced in the otic vesicles (arrowheads), olfactory placodes (large arrows) and branchial arches (white arrows) of parp3 morphants at 16 hpf (H, K, N, Q) and 24 hpf (I, L, O, R). The expression of dlx3b and dlx4b is abolished in the median fin fold of 24 hpf parp3 morphant embryos (small arrows in I). Dorsal views of embryos with anterior to the bottom in A, D, G, J, M, P and lateral views with anterior to the left, dorsal to the top, in B, C, E, F, H, I, K, L, N, O, Q and R. Scale bars represent 10 µm.
Mentions: Having determined that parp3 morphants have defective neural crest development, we next characterized the expression of sox9a, dlx3b and dlx4b. By ISH, we find that the expression of the neural crest specifier sox9a is indeed reduced in parp3 morphants at 10, 16 and 24 hpf (Fig. 6A–F). The high level of expression of sox9a in the otic placode (Fig. 6A) and later in the otic vesicle (Fig. 6B) of wild type embryos is drastically reduced in parp3 morphants (Fig. 6D, E). At 16 hpf, the expression of sox9a in the morphant paraxial cells is more diffuse suggesting that the somitogenesis is possibly disturbed (Fig. 6B, E). At 24 hpf, sox9a is normally expressed in the trunk and in three major areas of the head: the forebrain, the midbrain-hindbrain boundary and the pharyngeal arches (Fig. 6C). Expression in these three major sites is lost in the morphants and there is barely any expression in the trunk (Fig. 6F).

Bottom Line: We have used several in vitro and in vivo approaches to examine the possible functions of PARP3 as a transcriptional regulator, a function suggested from its previously reported association with several Polycomb group (PcG) proteins.It delays pigmentation and severely impedes the development of the median fin fold and tail bud.Our findings demonstrate that Parp3 is crucial in the early stages of zebrafish development, possibly by exerting its transcriptional regulatory functions as early as during the specification of the neural plate border.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Laboratory, CHUQ Research Center, Centre Hospitalier de l'Université Laval, Québec, Canada.

ABSTRACT

Background: The PARP family member poly(ADP-ribose) polymerase 3 (PARP3) is structurally related to the well characterized PARP1 that orchestrates cellular responses to DNA strand breaks and cell death by the synthesis of poly(ADP-ribose). In contrast to PARP1 and PARP2, the functions of PARP3 are undefined. Here, we reveal critical functions for PARP3 during vertebrate development.

Principal findings: We have used several in vitro and in vivo approaches to examine the possible functions of PARP3 as a transcriptional regulator, a function suggested from its previously reported association with several Polycomb group (PcG) proteins. We demonstrate that PARP3 gene occupancy in the human neuroblastoma cell line SK-N-SH occurs preferentially with developmental genes regulating cell fate specification, tissue patterning, craniofacial development and neurogenesis. Addressing the significance of this association during zebrafish development, we show that morpholino oligonucleotide-directed inhibition of parp3 expression in zebrafish impairs the expression of the neural crest cell specifier sox9a and of dlx3b/dlx4b, the formation of cranial sensory placodes, inner ears and pectoral fins. It delays pigmentation and severely impedes the development of the median fin fold and tail bud.

Conclusion: Our findings demonstrate that Parp3 is crucial in the early stages of zebrafish development, possibly by exerting its transcriptional regulatory functions as early as during the specification of the neural plate border.

Show MeSH
Related in: MedlinePlus