Limits...
Alcohol exposure decreases CREB binding protein expression and histone acetylation in the developing cerebellum.

Guo W, Crossey EL, Zhang L, Zucca S, George OL, Valenzuela CF, Zhao X - PLoS ONE (2011)

Bottom Line: However, the underlying molecular mechanism is unclear.CBP is a histone acetyltransferase, a component of the epigenetic mechanism controlling neuronal gene expression.This effect of ethanol may be responsible for the motor coordination deficits that characterize fetal alcohol spectrum disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosciences, University of New Mexico School of Medicine, Albuquerque, New Mexico, United States of America.

ABSTRACT

Background: Fetal alcohol exposure affects 1 in 100 children making it the leading cause of mental retardation in the US. It has long been known that alcohol affects cerebellum development and function. However, the underlying molecular mechanism is unclear.

Methodology/principal findings: We demonstrate that CREB binding protein (CBP) is widely expressed in granule and Purkinje neurons of the developing cerebellar cortex of naïve rats. We also show that exposure to ethanol during the 3(rd) trimester-equivalent of human pregnancy reduces CBP levels. CBP is a histone acetyltransferase, a component of the epigenetic mechanism controlling neuronal gene expression. We further demonstrate that the acetylation of both histone H3 and H4 is reduced in the cerebellum of ethanol-treated rats.

Conclusions/significance: These findings indicate that ethanol exposure decreases the expression and function of CBP in the developing cerebellum. This effect of ethanol may be responsible for the motor coordination deficits that characterize fetal alcohol spectrum disorders.

Show MeSH

Related in: MedlinePlus

CBP expression is low in immature cells of the cerebellum at postnatal day 8.(A) CBP is nearly undetectable in Nestin+ ECL cells. Note that Nestin is expressed in the cells of the ML and PCL (see text for discussion). CBP, red; Nestin, green; Dapi, blue. EGL, external granular layer; ML, Molecular Layer; PCL, Purkinje cell layer; IGL, internal granular layer. Scale bar = 20 µm. (B) CBP is expressed at relatively low level in doublecortin (DCX+) young neurons in the ML. CBP, red; DCX, green; Dapi, blue. Scale bar = 20 µm. Boxes in A and B show the regions where magnification photos (C–E) are from. (C) High magnification confocal images showing that CBP expression is undetectable in Nestin+ EGL cells and very low in Nestin+ IGL cells. Note that in IGL, CBP is expressed at high level by a nearby Nestin-negative cell. Scale bar = 5 µm (D) High magnification confocal images showing that CBP is expressed by DCX+ cells in the IGL. Scale bar = 5 µm (E) High magnification confocal images showing that CBP is in both nucleus and cytosol of a Purkinje cell in the PCL. Scale bar = 10 µm. (F) A Dapi-stained cerebellar section showing the region for immunohistological analysis for Figures 1 and 2. Scale bar = 100 µm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3104983&req=5

pone-0019351-g001: CBP expression is low in immature cells of the cerebellum at postnatal day 8.(A) CBP is nearly undetectable in Nestin+ ECL cells. Note that Nestin is expressed in the cells of the ML and PCL (see text for discussion). CBP, red; Nestin, green; Dapi, blue. EGL, external granular layer; ML, Molecular Layer; PCL, Purkinje cell layer; IGL, internal granular layer. Scale bar = 20 µm. (B) CBP is expressed at relatively low level in doublecortin (DCX+) young neurons in the ML. CBP, red; DCX, green; Dapi, blue. Scale bar = 20 µm. Boxes in A and B show the regions where magnification photos (C–E) are from. (C) High magnification confocal images showing that CBP expression is undetectable in Nestin+ EGL cells and very low in Nestin+ IGL cells. Note that in IGL, CBP is expressed at high level by a nearby Nestin-negative cell. Scale bar = 5 µm (D) High magnification confocal images showing that CBP is expressed by DCX+ cells in the IGL. Scale bar = 5 µm (E) High magnification confocal images showing that CBP is in both nucleus and cytosol of a Purkinje cell in the PCL. Scale bar = 10 µm. (F) A Dapi-stained cerebellar section showing the region for immunohistological analysis for Figures 1 and 2. Scale bar = 100 µm.

Mentions: To determine whether CBP is involved in cerebellar development, we first investigated which neuronal populations express CBP in the developing cerebellum from postnatal day 8 (P8) rats, by using immunohistochemical studies. At this age, the cerebellar cortex consists of four layers: external granule layer (EGL), molecular layer (ML), Purkinje cell layer (PCL), and the internal granule layer (IGL) [42], [43]. The EGL is mainly composed of immature proliferating granule cells. The ML contains Purkinje cell dendrites and migrating molecular layer interneurons. The Purkinje layer contains the cell bodies of these neurons, which are arranged in multiple layers at this developmental stage. The IGL consists of more mature granule cells, Golgi cells and mossy fibers. At P8, immature granule cells in the EGL are migrating (passing through ML and PCL) to become more mature granule neurons at IGL. As shown in Figure 1, CBP expression was nearly undetectable in the nestin+ EGL cells (Fig. 1A, 1C, and 1F), and low in doublecortin-positive (DCX+) ML neurons (Fig. 1B, 1D, and 1F), but high in PCL and IGL neurons (Fig. 1A, 1B, and 1F). We found that the young Purkinje cells also express nestin (Fig. 1), a finding that is consistent with the literature[44] and the Allan Brain Atlas (http://www.brain-map.org/). Interestingly, while CBP is located in granule neuron nuclei at IGL, it is both in the nucleus and cysotol of Purkinje cells in PCL (Fig. 1E). Using a mature neuronal marker, NeuN, we found that CBP was predominantly expressed in relatively more mature, non-migrating neurons, located in the IGL and PCL (Fig. 2A and D). A subpopulation of GFAP astroctyes also expressed CBP (Fig. 2B and E). In addition, CBP expression was detected in MBP+ myelin fibers (Fig. 2C and F), suggesting its expression in oligodendrocytes. In summary, CBP is expressed at high levels in neurons and also glial cells of the developing cerebellar cortex.


Alcohol exposure decreases CREB binding protein expression and histone acetylation in the developing cerebellum.

Guo W, Crossey EL, Zhang L, Zucca S, George OL, Valenzuela CF, Zhao X - PLoS ONE (2011)

CBP expression is low in immature cells of the cerebellum at postnatal day 8.(A) CBP is nearly undetectable in Nestin+ ECL cells. Note that Nestin is expressed in the cells of the ML and PCL (see text for discussion). CBP, red; Nestin, green; Dapi, blue. EGL, external granular layer; ML, Molecular Layer; PCL, Purkinje cell layer; IGL, internal granular layer. Scale bar = 20 µm. (B) CBP is expressed at relatively low level in doublecortin (DCX+) young neurons in the ML. CBP, red; DCX, green; Dapi, blue. Scale bar = 20 µm. Boxes in A and B show the regions where magnification photos (C–E) are from. (C) High magnification confocal images showing that CBP expression is undetectable in Nestin+ EGL cells and very low in Nestin+ IGL cells. Note that in IGL, CBP is expressed at high level by a nearby Nestin-negative cell. Scale bar = 5 µm (D) High magnification confocal images showing that CBP is expressed by DCX+ cells in the IGL. Scale bar = 5 µm (E) High magnification confocal images showing that CBP is in both nucleus and cytosol of a Purkinje cell in the PCL. Scale bar = 10 µm. (F) A Dapi-stained cerebellar section showing the region for immunohistological analysis for Figures 1 and 2. Scale bar = 100 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3104983&req=5

pone-0019351-g001: CBP expression is low in immature cells of the cerebellum at postnatal day 8.(A) CBP is nearly undetectable in Nestin+ ECL cells. Note that Nestin is expressed in the cells of the ML and PCL (see text for discussion). CBP, red; Nestin, green; Dapi, blue. EGL, external granular layer; ML, Molecular Layer; PCL, Purkinje cell layer; IGL, internal granular layer. Scale bar = 20 µm. (B) CBP is expressed at relatively low level in doublecortin (DCX+) young neurons in the ML. CBP, red; DCX, green; Dapi, blue. Scale bar = 20 µm. Boxes in A and B show the regions where magnification photos (C–E) are from. (C) High magnification confocal images showing that CBP expression is undetectable in Nestin+ EGL cells and very low in Nestin+ IGL cells. Note that in IGL, CBP is expressed at high level by a nearby Nestin-negative cell. Scale bar = 5 µm (D) High magnification confocal images showing that CBP is expressed by DCX+ cells in the IGL. Scale bar = 5 µm (E) High magnification confocal images showing that CBP is in both nucleus and cytosol of a Purkinje cell in the PCL. Scale bar = 10 µm. (F) A Dapi-stained cerebellar section showing the region for immunohistological analysis for Figures 1 and 2. Scale bar = 100 µm.
Mentions: To determine whether CBP is involved in cerebellar development, we first investigated which neuronal populations express CBP in the developing cerebellum from postnatal day 8 (P8) rats, by using immunohistochemical studies. At this age, the cerebellar cortex consists of four layers: external granule layer (EGL), molecular layer (ML), Purkinje cell layer (PCL), and the internal granule layer (IGL) [42], [43]. The EGL is mainly composed of immature proliferating granule cells. The ML contains Purkinje cell dendrites and migrating molecular layer interneurons. The Purkinje layer contains the cell bodies of these neurons, which are arranged in multiple layers at this developmental stage. The IGL consists of more mature granule cells, Golgi cells and mossy fibers. At P8, immature granule cells in the EGL are migrating (passing through ML and PCL) to become more mature granule neurons at IGL. As shown in Figure 1, CBP expression was nearly undetectable in the nestin+ EGL cells (Fig. 1A, 1C, and 1F), and low in doublecortin-positive (DCX+) ML neurons (Fig. 1B, 1D, and 1F), but high in PCL and IGL neurons (Fig. 1A, 1B, and 1F). We found that the young Purkinje cells also express nestin (Fig. 1), a finding that is consistent with the literature[44] and the Allan Brain Atlas (http://www.brain-map.org/). Interestingly, while CBP is located in granule neuron nuclei at IGL, it is both in the nucleus and cysotol of Purkinje cells in PCL (Fig. 1E). Using a mature neuronal marker, NeuN, we found that CBP was predominantly expressed in relatively more mature, non-migrating neurons, located in the IGL and PCL (Fig. 2A and D). A subpopulation of GFAP astroctyes also expressed CBP (Fig. 2B and E). In addition, CBP expression was detected in MBP+ myelin fibers (Fig. 2C and F), suggesting its expression in oligodendrocytes. In summary, CBP is expressed at high levels in neurons and also glial cells of the developing cerebellar cortex.

Bottom Line: However, the underlying molecular mechanism is unclear.CBP is a histone acetyltransferase, a component of the epigenetic mechanism controlling neuronal gene expression.This effect of ethanol may be responsible for the motor coordination deficits that characterize fetal alcohol spectrum disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosciences, University of New Mexico School of Medicine, Albuquerque, New Mexico, United States of America.

ABSTRACT

Background: Fetal alcohol exposure affects 1 in 100 children making it the leading cause of mental retardation in the US. It has long been known that alcohol affects cerebellum development and function. However, the underlying molecular mechanism is unclear.

Methodology/principal findings: We demonstrate that CREB binding protein (CBP) is widely expressed in granule and Purkinje neurons of the developing cerebellar cortex of naïve rats. We also show that exposure to ethanol during the 3(rd) trimester-equivalent of human pregnancy reduces CBP levels. CBP is a histone acetyltransferase, a component of the epigenetic mechanism controlling neuronal gene expression. We further demonstrate that the acetylation of both histone H3 and H4 is reduced in the cerebellum of ethanol-treated rats.

Conclusions/significance: These findings indicate that ethanol exposure decreases the expression and function of CBP in the developing cerebellum. This effect of ethanol may be responsible for the motor coordination deficits that characterize fetal alcohol spectrum disorders.

Show MeSH
Related in: MedlinePlus