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CC2D1A regulates human intellectual and social function as well as NF-κB signaling homeostasis.

Manzini MC, Xiong L, Shaheen R, Tambunan DE, Di Costanzo S, Mitisalis V, Tischfield DJ, Cinquino A, Ghaziuddin M, Christian M, Jiang Q, Laurent S, Nanjiani ZA, Rasheed S, Hill RS, Lizarraga SB, Gleason D, Sabbagh D, Salih MA, Alkuraya FS, Walsh CA - Cell Rep (2014)

Bottom Line: CC2D1A regulates multiple intracellular signaling pathways, and we found its strongest effect to be on the transcription factor nuclear factor κB (NF-κB).Cc2d1a gain and loss of function both increase activation of NF-κB, revealing a critical role of Cc2d1a in homeostatic control of intracellular signaling.Cc2d1a knockdown in neurons reduces dendritic complexity and increases NF-κB activity, and the effects of Cc2d1a depletion can be rescued by inhibiting NF-κB activity.

View Article: PubMed Central - PubMed

Affiliation: Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Boston, MA 02115, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815-6789, USA; Departments of Pediatrics and Neurology, Harvard Medical School, Boston, MA 02115, USA.

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Cc2d1a Knockdown Reduces Dendritic Complexity in Hippocampal Neurons(A) Expression levels of Cc2d1a increase during early neuronal development in the mouse brain. qPCR analysis was performed in different tissues (cortex and hippocampus) at the ages indicated. Mean and SEM were calculated from three independent samples per time point.(B) A simplified dendritic arbor was observed in cortical neurons at P11 following in utero electroporation of shRNA constructs at E15.5. Dendrites also appeared to be misoriented, terminating farther from the pial surface (red arrows). Scale bar, 50 μm.(C) Total dendritic length was reduced by approximately 40% (three electroporated animals).(D) Quantification of the disruption in dendrite orientation shown in (B). Although the nucleus was at the same distance from the pia, the apical dendritic arbor ended much farther from the surface.(E and F) Hippocampal neurons at 7 and 14 DIV following Cc2d1a knockdown at 1 DIV also showed a reduction in total dendritic length caused by a reduction in process number. Average process length was not affected (n > 3 independent cultures). Scale bars, 50 μm (7 DIV) and 100 μm (14 DIV).(G) The number of dendritic spines in hippocampal neurons (tested at 14 DIV) was reduced.(H) The type of spine (mature, spine with a neck and head; stubby, spine with no neck; filopodia, elongated filamentous protrusion) was not changed in the experimental group, suggesting that spine maturation was not affected (n = 6 for G and H).All graphs show mean ± SEM. *p < 0.5; **p < 0.01; ***p < 0.001.
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Figure 2: Cc2d1a Knockdown Reduces Dendritic Complexity in Hippocampal Neurons(A) Expression levels of Cc2d1a increase during early neuronal development in the mouse brain. qPCR analysis was performed in different tissues (cortex and hippocampus) at the ages indicated. Mean and SEM were calculated from three independent samples per time point.(B) A simplified dendritic arbor was observed in cortical neurons at P11 following in utero electroporation of shRNA constructs at E15.5. Dendrites also appeared to be misoriented, terminating farther from the pial surface (red arrows). Scale bar, 50 μm.(C) Total dendritic length was reduced by approximately 40% (three electroporated animals).(D) Quantification of the disruption in dendrite orientation shown in (B). Although the nucleus was at the same distance from the pia, the apical dendritic arbor ended much farther from the surface.(E and F) Hippocampal neurons at 7 and 14 DIV following Cc2d1a knockdown at 1 DIV also showed a reduction in total dendritic length caused by a reduction in process number. Average process length was not affected (n > 3 independent cultures). Scale bars, 50 μm (7 DIV) and 100 μm (14 DIV).(G) The number of dendritic spines in hippocampal neurons (tested at 14 DIV) was reduced.(H) The type of spine (mature, spine with a neck and head; stubby, spine with no neck; filopodia, elongated filamentous protrusion) was not changed in the experimental group, suggesting that spine maturation was not affected (n = 6 for G and H).All graphs show mean ± SEM. *p < 0.5; **p < 0.01; ***p < 0.001.

Mentions: Cc2d1a has been shown to be expressed throughout the brain (Basel-Vanagaite et al., 2006), and we asked whether expression is dynamic during development. Using quantitative PCR (qPCR), we determined that Cc2d1a mRNA expression in the developing murine cortex and hippocampus shows an increase during the postnatal period corresponding to early stages of postmitotic neuronal differentiation, between embryonic day 18.5 (E18.5) and postnatal day 1 (P1) in the cortex and hippocampus (Figure 2A), suggesting possible roles in neuronal morphogenesis or synapse formation.


CC2D1A regulates human intellectual and social function as well as NF-κB signaling homeostasis.

Manzini MC, Xiong L, Shaheen R, Tambunan DE, Di Costanzo S, Mitisalis V, Tischfield DJ, Cinquino A, Ghaziuddin M, Christian M, Jiang Q, Laurent S, Nanjiani ZA, Rasheed S, Hill RS, Lizarraga SB, Gleason D, Sabbagh D, Salih MA, Alkuraya FS, Walsh CA - Cell Rep (2014)

Cc2d1a Knockdown Reduces Dendritic Complexity in Hippocampal Neurons(A) Expression levels of Cc2d1a increase during early neuronal development in the mouse brain. qPCR analysis was performed in different tissues (cortex and hippocampus) at the ages indicated. Mean and SEM were calculated from three independent samples per time point.(B) A simplified dendritic arbor was observed in cortical neurons at P11 following in utero electroporation of shRNA constructs at E15.5. Dendrites also appeared to be misoriented, terminating farther from the pial surface (red arrows). Scale bar, 50 μm.(C) Total dendritic length was reduced by approximately 40% (three electroporated animals).(D) Quantification of the disruption in dendrite orientation shown in (B). Although the nucleus was at the same distance from the pia, the apical dendritic arbor ended much farther from the surface.(E and F) Hippocampal neurons at 7 and 14 DIV following Cc2d1a knockdown at 1 DIV also showed a reduction in total dendritic length caused by a reduction in process number. Average process length was not affected (n > 3 independent cultures). Scale bars, 50 μm (7 DIV) and 100 μm (14 DIV).(G) The number of dendritic spines in hippocampal neurons (tested at 14 DIV) was reduced.(H) The type of spine (mature, spine with a neck and head; stubby, spine with no neck; filopodia, elongated filamentous protrusion) was not changed in the experimental group, suggesting that spine maturation was not affected (n = 6 for G and H).All graphs show mean ± SEM. *p < 0.5; **p < 0.01; ***p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4334362&req=5

Figure 2: Cc2d1a Knockdown Reduces Dendritic Complexity in Hippocampal Neurons(A) Expression levels of Cc2d1a increase during early neuronal development in the mouse brain. qPCR analysis was performed in different tissues (cortex and hippocampus) at the ages indicated. Mean and SEM were calculated from three independent samples per time point.(B) A simplified dendritic arbor was observed in cortical neurons at P11 following in utero electroporation of shRNA constructs at E15.5. Dendrites also appeared to be misoriented, terminating farther from the pial surface (red arrows). Scale bar, 50 μm.(C) Total dendritic length was reduced by approximately 40% (three electroporated animals).(D) Quantification of the disruption in dendrite orientation shown in (B). Although the nucleus was at the same distance from the pia, the apical dendritic arbor ended much farther from the surface.(E and F) Hippocampal neurons at 7 and 14 DIV following Cc2d1a knockdown at 1 DIV also showed a reduction in total dendritic length caused by a reduction in process number. Average process length was not affected (n > 3 independent cultures). Scale bars, 50 μm (7 DIV) and 100 μm (14 DIV).(G) The number of dendritic spines in hippocampal neurons (tested at 14 DIV) was reduced.(H) The type of spine (mature, spine with a neck and head; stubby, spine with no neck; filopodia, elongated filamentous protrusion) was not changed in the experimental group, suggesting that spine maturation was not affected (n = 6 for G and H).All graphs show mean ± SEM. *p < 0.5; **p < 0.01; ***p < 0.001.
Mentions: Cc2d1a has been shown to be expressed throughout the brain (Basel-Vanagaite et al., 2006), and we asked whether expression is dynamic during development. Using quantitative PCR (qPCR), we determined that Cc2d1a mRNA expression in the developing murine cortex and hippocampus shows an increase during the postnatal period corresponding to early stages of postmitotic neuronal differentiation, between embryonic day 18.5 (E18.5) and postnatal day 1 (P1) in the cortex and hippocampus (Figure 2A), suggesting possible roles in neuronal morphogenesis or synapse formation.

Bottom Line: CC2D1A regulates multiple intracellular signaling pathways, and we found its strongest effect to be on the transcription factor nuclear factor κB (NF-κB).Cc2d1a gain and loss of function both increase activation of NF-κB, revealing a critical role of Cc2d1a in homeostatic control of intracellular signaling.Cc2d1a knockdown in neurons reduces dendritic complexity and increases NF-κB activity, and the effects of Cc2d1a depletion can be rescued by inhibiting NF-κB activity.

View Article: PubMed Central - PubMed

Affiliation: Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Boston, MA 02115, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815-6789, USA; Departments of Pediatrics and Neurology, Harvard Medical School, Boston, MA 02115, USA.

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Related in: MedlinePlus