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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.

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Related in: MedlinePlus

CBP protein expression was decreased in the cerebellum from ethanol-exposed developing rats.(A) Image of one of the western blots showing differential CBP expression in cerebellum isolated from control pups (exposed to air) analyzed at P2 to P12. (B) Representative CBP western blot analysis of cerebellum isolated from ethanol-exposed pups analyzed at P2 to P12. (C) Quantitative analysis showing that CBP protein levels (normalized to β-actin) are lower in the ethanol group compared to the control group between P2 and P10, but not at P12. Three independently exposed liters were analyzed and each was counted as n = 1. Multiple western blots were analyzed for each exposure to minimize the variation between western blots. (D) Representative CBP immunohisological images of control and ethanol exposed brains at P8. CBP, red;’ Dapi, blue Scale bar = 50 µm (E) Quantitative analysis of CBP immunohistological signal intensity demonstrates that CBP expression is reduced in ethanol exposed cerebellum compared to controls. N = 3 individual animal for P2 and P4/condition. N = 4 individual animal for P6 –P12/condition. *, P<0.05, t-test.
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pone-0019351-g004: CBP protein expression was decreased in the cerebellum from ethanol-exposed developing rats.(A) Image of one of the western blots showing differential CBP expression in cerebellum isolated from control pups (exposed to air) analyzed at P2 to P12. (B) Representative CBP western blot analysis of cerebellum isolated from ethanol-exposed pups analyzed at P2 to P12. (C) Quantitative analysis showing that CBP protein levels (normalized to β-actin) are lower in the ethanol group compared to the control group between P2 and P10, but not at P12. Three independently exposed liters were analyzed and each was counted as n = 1. Multiple western blots were analyzed for each exposure to minimize the variation between western blots. (D) Representative CBP immunohisological images of control and ethanol exposed brains at P8. CBP, red;’ Dapi, blue Scale bar = 50 µm (E) Quantitative analysis of CBP immunohistological signal intensity demonstrates that CBP expression is reduced in ethanol exposed cerebellum compared to controls. N = 3 individual animal for P2 and P4/condition. N = 4 individual animal for P6 –P12/condition. *, P<0.05, t-test.

Mentions: We next tested the effect of ethanol exposure on cerebellar CBP expression. Exposure to ethanol vapor resulted in serum ethanol concentrations in the pups that were similar across postnatal days (overall average = 54±4 mM; n = 48; Fig. 3A; for comparison, 80 mg/dl = 17.4 mM). Furthermore, pup weight gain was similar in the air and ethanol groups (Fig. 3B). There was only one case of neonatal mortality and it occurred in a litter exposed to air. In cerebellar homogenates from air-exposed animals, CBP expression increased approximately 4-fold between postnatal day 2 and postnatal day 4, and then remained relatively stable until postnatal day 12, the last day of our analysis (Fig. 4A, C). In cerebellar homogenates from ethanol-exposed animals, CBP levels were significantly lower compared to control pups between postnatal days 2 and 10 (Fig. 4B, C). Although both control and ethanol groups had similar CBP expression levels at postnatal day 12 (Fig. 4A–C), the developmental stage-dependent upregulation of CBP was much delayed in ethanol-exposed animals compared to control animals. Immunohistochemical experiments also detected a decrease in CBP levels in IGC and PCL at postnatal day 8 (Fig. 4D, E). Therefore, 3rd trimester equivalent ethanol exposure led to reduced CBP expression in the developing cerebellum.


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 protein expression was decreased in the cerebellum from ethanol-exposed developing rats.(A) Image of one of the western blots showing differential CBP expression in cerebellum isolated from control pups (exposed to air) analyzed at P2 to P12. (B) Representative CBP western blot analysis of cerebellum isolated from ethanol-exposed pups analyzed at P2 to P12. (C) Quantitative analysis showing that CBP protein levels (normalized to β-actin) are lower in the ethanol group compared to the control group between P2 and P10, but not at P12. Three independently exposed liters were analyzed and each was counted as n = 1. Multiple western blots were analyzed for each exposure to minimize the variation between western blots. (D) Representative CBP immunohisological images of control and ethanol exposed brains at P8. CBP, red;’ Dapi, blue Scale bar = 50 µm (E) Quantitative analysis of CBP immunohistological signal intensity demonstrates that CBP expression is reduced in ethanol exposed cerebellum compared to controls. N = 3 individual animal for P2 and P4/condition. N = 4 individual animal for P6 –P12/condition. *, P<0.05, t-test.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3104983&req=5

pone-0019351-g004: CBP protein expression was decreased in the cerebellum from ethanol-exposed developing rats.(A) Image of one of the western blots showing differential CBP expression in cerebellum isolated from control pups (exposed to air) analyzed at P2 to P12. (B) Representative CBP western blot analysis of cerebellum isolated from ethanol-exposed pups analyzed at P2 to P12. (C) Quantitative analysis showing that CBP protein levels (normalized to β-actin) are lower in the ethanol group compared to the control group between P2 and P10, but not at P12. Three independently exposed liters were analyzed and each was counted as n = 1. Multiple western blots were analyzed for each exposure to minimize the variation between western blots. (D) Representative CBP immunohisological images of control and ethanol exposed brains at P8. CBP, red;’ Dapi, blue Scale bar = 50 µm (E) Quantitative analysis of CBP immunohistological signal intensity demonstrates that CBP expression is reduced in ethanol exposed cerebellum compared to controls. N = 3 individual animal for P2 and P4/condition. N = 4 individual animal for P6 –P12/condition. *, P<0.05, t-test.
Mentions: We next tested the effect of ethanol exposure on cerebellar CBP expression. Exposure to ethanol vapor resulted in serum ethanol concentrations in the pups that were similar across postnatal days (overall average = 54±4 mM; n = 48; Fig. 3A; for comparison, 80 mg/dl = 17.4 mM). Furthermore, pup weight gain was similar in the air and ethanol groups (Fig. 3B). There was only one case of neonatal mortality and it occurred in a litter exposed to air. In cerebellar homogenates from air-exposed animals, CBP expression increased approximately 4-fold between postnatal day 2 and postnatal day 4, and then remained relatively stable until postnatal day 12, the last day of our analysis (Fig. 4A, C). In cerebellar homogenates from ethanol-exposed animals, CBP levels were significantly lower compared to control pups between postnatal days 2 and 10 (Fig. 4B, C). Although both control and ethanol groups had similar CBP expression levels at postnatal day 12 (Fig. 4A–C), the developmental stage-dependent upregulation of CBP was much delayed in ethanol-exposed animals compared to control animals. Immunohistochemical experiments also detected a decrease in CBP levels in IGC and PCL at postnatal day 8 (Fig. 4D, E). Therefore, 3rd trimester equivalent ethanol exposure led to reduced CBP expression in the developing cerebellum.

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