Chronic lithium treatment antagonizes glutamate-induced decrease of phosphorylated CREB in neurons via reducing protein phosphatase 1 and increasing MEK activities

doi:10.1016/S0306-4522(02)00573-0

Neuroscience

Volume 116, Issue 2, 31 January 2003, Pages 425-435

https://doi.org/10.1016/S0306-4522(02)00573-0 Get rights and content

Abstract

The cyclic AMP response element binding protein (CREB) has major roles in mediating adaptive responses at glutamatergic synapses and in the neuroprotective effects of neurotrophins. CREB has been implicated as a potential mediator of antidepressant actions. In vitro, chronic lithium treatment has been shown to promote neuronal cell survival. In the present study, we have used cultures of cerebellar granule neurons to analyze the effects of acute and chronic lithium treatment on the response to toxic concentrations of glutamate. Such concentrations of glutamate decrease the phosphorylation of CREB at serine¹³³ in an N-methyl-d-aspartate (NMDA) receptor-dependent manner. Chronic, but not acute, lithium treatment suppresses glutamate-induced decreases in phosphorylated CREB, and transfection studies indicate that chronic lithium, in the presence of a glutamate stimulus, markedly increases CRE-driven gene expression. Experiments with selected pharmacological reagents indicate that the glutamate-induced decreases in phosphorylated CREB are regulated primarily by protein phosphatase 1. Chronic lithium treatment not only decreases protein phosphatase 1 activity under these circumstances, but also augments glutamate-induced increases in MEK activity. PD 98059, a MEK inhibitor, prevents chronic lithium treatment from increasing phosphorylated CREB levels in glutamate-treated neurons. We conclude from these results that chronic lithium treatment is permissive for maintaining higher phosphorylated CREB levels in the presence of glutamate in part by decreasing protein phosphatase 1 activity and in part by increasing MEK activity. Higher levels of phosphorylated CREB and CRE-responsive genes such as bcl-2 may be responsible for lithium’s reported effects on neuronal survival.

Section snippets

Cell culture and treatments

Primary cultures of cerebellar granule neurons were prepared from 8-day-old Sprague–Dawley rat pups. Briefly, cerebella were chopped into segments of 400 μm, and the cells dissociated by trypsinization, treated with DNase and triturated before being plated onto poly-l-lysine-coated 60-mm culture dishes or 24-well plates. The plating media consisted of basal Eagle’s medium containing 10% fetal calf serum, 2-mM glutamine, 50 μg/ml gentamicin, and 25 mM KCl. Cytosine β-d-arabinoside (10 μM) was

Results

Exposure of rat cerebellar granule cultures after 8 days in vitro (DIV) to different concentrations of glutamate and for varying periods of time caused a time- and dose-dependent change in pCREB at Ser¹³³. At 5 μM glutamate, pCREB levels were significantly increased at all time points tested but there was little or no effect on total CREB levels (Fig. 1A). Treatment with 50 μM glutamate caused a transient but insignificant increase in pCREB after 2 min of exposure. After 15, 30 and 60 min of

Discussion

Our results demonstrate that acute glutamate and chronic lithium treatment can modulate pCREB levels in cultures of cerebellar granule cells prepared from 8-day-old rats. pCREB levels can be increased by relatively low concentrations of glutamate in a time-dependent, neuroprotective manner. Conversely, pCREB levels are decreased by higher concentrations of glutamate that are neurotoxic and act through NMDA receptors. Acute lithium treatment has no apparent effect on the glutamate-induced loss

Acknowledgements

We thank Jessica Madert for her excellent technical assistance and Peter Leeds for his assistance in many aspects of manuscript preparation.

References (49)

S. Ahn et al.
A late phase of cerebellar long-term depression requires activation of CaMKIV and CREB
Neuron
(1999)
G. Alvarez et al.
Lithium protects cultured neurons against beta-amyloid-induced neurodegeneration
FEBS Lett
(1999)
H. Bito et al.
CREB phosphorylation and dephosphorylationa Ca(2+)- and stimulus duration-dependent switch for hippocampal gene expression
Cell
(1996)
R.W. Chen et al.
Long term lithium treatment suppresses p53 and Bax expression but increases Bcl-2 expressiona prominent role in neuroprotection against excitotoxicity
J Biol Chem
(1999)
S.R. D’Mello et al.
Lithium induces apoptosis in immature cerebellar granule cells but promotes survival of mature neurons
Exp Cell Res
(1994)
S. Finkbeiner
CREB couples neurotrophin signals to survival messages
Neuron
(2000)
D.D. Ginty et al.
Nerve growth factor activates a Ras-dependent protein kinase that stimulates c-fos transcription via phosphorylation of CREB
Cell
(1994)
G.A. Gonzalez et al.
Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133
Cell
(1989)
M. Hagiwara et al.
Transcriptional attenuation following cAMP induction requires PP-1-mediated dephosphorylation of CREB
Cell
(1992)
R. Hashimoto et al.
Lithium induces brain-derived neurotrophic factor and activates TrkB in rodent cortical neurons: an essential step for neuroprotection against glutamate excitotoxicity
Neuropharmacology
(2002)

M. Hong et al.

Lithium reduces tau phosphorylation by inhibition of glycogen synthase kinase-3

J Biol Chem

(1997)

H.K. Manji et al.

Lithium at 50have the neuroprotective effects of this unique cation been overlooked?

Biol Psychiatry

(1999)

G.J. Moore et al.

Lithium-induced increase in human brain grey matter

Lancet

(2000)

J.R. Munoz-Montano et al.

Lithium inhibits Alzheimer’s disease-like tau protein phosphorylation in neurons

FEBS Lett

(1997)

P.B. Shieh et al.

Identification of a signaling pathway involved in calcium regulation of BDNF expression

Neuron

(1998)

X. Tao et al.

Ca2+ influx regulates BDNF transcription by a CREB family transcription factor-dependent mechanism

Neuron

(1998)

J.F. Wang et al.

Cyclic AMP responsive element binding protein phosphorylation and DNA binding is decreased by chronic lithium but not valproate treatment of SH-SY5Y neuroblastoma cells

Neuroscience

(1999)

H. Wei et al.

beta-amyloid peptide-induced death of PC 12 cells and cerebellar granule cell neurons is inhibited by long-term lithium treatment

Eur J Pharmacol

(2000)

P.-X. Yuan et al.

The mood stabilizer valproic acid activates mitogen-activated protein kinases and promotes neurite growth

J Biol Chem

(2001)

A.S. Alberts et al.

Expression of a peptide inhibitor of protein phosphatase 1 increases phosphorylation and activity of CREB in NIH 3T3 fibroblasts

Mol Cell Biol

(1994)

A. Bonni et al.

Cell survival promoted by the Ras-MAPK signaling pathway by transcription-dependent and -independent mechanisms

Science

(1999)

E. Chalecka-Franaszek et al.

Lithium activates the serine/threonine kinase Akt-1 and suppresses glutamate-induced inhibition of Akt-1 activity in neurons

Proc Natl Acad Sci USA

(1999)

G. Chen et al.

The mood-stabilizing agents lithium and valproate robustly increase the levels of the neuroprotective protein bcl-2 in the CNS

J Neurochem

(1999)

P. Cohen

The structure and regulation of protein phosphatases

Annu Rev Biochem

(1989)

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Citation Excerpt :
Post mortem studies reported altered CREB levels in MDD subjects and different CREB levels between treated and untreated patients (Dowlatshahi et al., 1998, 1999; Laifenfeld et al., 2005; Uslaner et al., 2009). Furthermore, CREB1 seems to be involved in antidepressant (Golan et al., 2011; Hisaoka et al., 2008; Qi et al., 2008), mood stabilizer (Boer et al., 2007, 2008; Casu et al., 2007; Hammonds and Shim, 2009; Kopnisky et al., 2003; Liang et al., 2008; Mai et al., 2002; Mamdani et al., 2008) and antipsychotic mechanisms of action in vivo and in vitro (Lee et al., 2010; Yang et al., 2004). Interestingly, previous research indicates that the MAPK/CREB signal system may be involved in the molecular mechanism of depression (Drago et al., 2011; Qi et al., 2006).
Treatment resistant depression (TRD) is a significant clinical and public health problem. Among others, neuroplasticity and inflammatory pathways seem to play a crucial role in the pathomechanisms of antidepressant efficacy.
The primary aim of this study was to investigate whether a set of single nucleotide polymorphisms (SNPs) within two genes implicated in neuroplasticity and inflammatory processes (the mitogen activated protein kinase 1, MAPK1 (rs3810608, rs6928, rs13515 and rs8136867), and the cyclic AMP responsive element binding protein 1, CREB1 (rs889895, rs6740584, rs2551922 and rs2254137)) was associated with antidepressant treatment resistance (according to two different definitions), in 285 Major Depressive Disorder (MDD) patients. As secondary aims, we investigated the genetic modulation of the same SNPs on response, remission and other clinical features both in MDD patients and in a larger sample including 82 Bipolar Disorder (BD) patients as well. All patients were screened in the context of a European multicenter project.
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Published by Elsevier Ltd.

Original contributionChronic lithium treatment antagonizes glutamate-induced decrease of phosphorylated CREB in neurons via reducing protein phosphatase 1 and increasing MEK activities

Abstract

Section snippets

Cell culture and treatments

Results

Discussion

Acknowledgements

Neuron

FEBS Lett

Cell

J Biol Chem

Exp Cell Res

Neuron

Cell

Cell

Cell

Neuropharmacology

J Biol Chem

Biol Psychiatry

Lancet

FEBS Lett

Neuron

Neuron

Neuroscience

Eur J Pharmacol

J Biol Chem

Expression of a peptide inhibitor of protein phosphatase 1 increases phosphorylation and activity of CREB in NIH 3T3 fibroblasts

Mol Cell Biol

Cell survival promoted by the Ras-MAPK signaling pathway by transcription-dependent and -independent mechanisms

Science

Lithium activates the serine/threonine kinase Akt-1 and suppresses glutamate-induced inhibition of Akt-1 activity in neurons

Proc Natl Acad Sci USA

The mood-stabilizing agents lithium and valproate robustly increase the levels of the neuroprotective protein bcl-2 in the CNS

J Neurochem

The structure and regulation of protein phosphatases

Annu Rev Biochem

Original contribution
Chronic lithium treatment antagonizes glutamate-induced decrease of phosphorylated CREB in neurons via reducing protein phosphatase 1 and increasing MEK activities