Enhanced asynchronous Ca(2+) oscillations associated with impaired glutamate transport in cortical astrocytes expressing Fmr1 gene premutation expansion.

Abstract

BACKGROUND:

FMR1 CGG expansion repeats in the premutation range have not been linked to astrocyte pathophysiology.

RESULTS:

Premutation cortical astrocytes display decreased Glu transporter expression/activity and enhanced asynchronous Ca(2+) oscillations.

CONCLUSION:

Glu transport and Ca(2+) signaling defects in premutation astrocytes could contribute to FXTAS neuropathology.

SIGNIFICANCE:

Premutation astrocytes may have an etiological role in FXTAS neuropathology. Premutation CGG repeat expansions (55-200 CGG repeats; preCGG) within the fragile X mental retardation 1 (FMR1) gene can cause fragile X-associated tremor/ataxia syndrome. Defects in early neuronal migration and morphology, electrophysiological activity, and mitochondria trafficking have been described in a premutation mouse model, but whether preCGG mutations also affect astrocyte function remains unknown. PreCGG cortical astrocytes (∼170 CGG repeats) displayed 3-fold higher Fmr1 mRNA and 30% lower FMR1 protein (FMRP) when compared with WT. PreCGG astrocytes showed modest reductions in expression of glutamate (Glu) transporters GLT-1 and GLAST and attenuated Glu uptake (p < 0.01). Consistent with astrocyte cultures in vitro, aged preCGG mice cerebral cortex also displayed reduced GLAST and GLT-1 expression. Approximately 65% of the WT and preCGG cortical astrocytes displayed spontaneous asynchronous Ca(2+) oscillations. PreCGG astrocytes exhibited nearly 50% higher frequency of asynchronous Ca(2+) oscillations (p < 0.01) than WT, a difference mimicked by chronic exposure of WT astrocytes to l-trans-pyrrolidine-2,4-dicarboxylic acid (l-trans-PDC) or by partial suppression of GLAST using siRNA interference. Acute challenge with Glu augmented the frequency of Ca(2+) oscillations in both genotypes. Additionally, 10 μm Glu elicited a sustained intracellular Ca(2+) rise in a higher portion of preCGG astrocytes when compared with WT. Pharmacological studies showed that mGluR5, but not NMDA receptor, contributed to Glu hypersensitivity in preCGG astrocytes. These functional defects in preCGG astrocytes, especially in Glu signaling, may contribute to fragile X-associated tremor/ataxia syndrome neuropathology.

• Authors: Zhengyu Cao, Susan Hulsizer, Yanjun Cui, Dalyir L Pretto, Kyung Ho Kim, Paul J Hagerman, Flora Tassone, Isaac N Pessah
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 288 Issue 19 Pg. 13831-41 (May 10 2013) ISSN: 1083-351X [Electronic] United States
• PMID: 23553633 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Dicarboxylic Acids
• Excitatory Amino Acid Agonists
• Excitatory Amino Acid Transporter 1
• Excitatory Amino Acid Transporter 2
• Fmr1 protein, mouse
• Gluk1 kainate receptor
• Neurotransmitter Uptake Inhibitors
• Pyrrolidines
• Receptors, Glutamate
• Receptors, Kainic Acid
• Receptors, N-Methyl-D-Aspartate
• Slc1a3 protein, mouse
• Fragile X Mental Retardation Protein
• Glutamic Acid
• N-Methylaspartate
• pyrrolidine-2,4-dicarboxylic acid

Topics

• Animals
• Astrocytes (drug effects, metabolism)
• Ataxia (genetics)
• Biological Transport
• Calcium Signaling
• Cells, Cultured
• Cerebral Cortex (pathology)
• Dicarboxylic Acids (pharmacology)
• Excitatory Amino Acid Agonists (pharmacology)
• Excitatory Amino Acid Transporter 1 (metabolism)
• Excitatory Amino Acid Transporter 2 (metabolism)
• Female
• Fragile X Mental Retardation Protein (genetics, metabolism)
• Fragile X Syndrome (genetics)
• Glutamic Acid (metabolism, physiology)
• Kinetics
• Male
• Mice
• Mice, Inbred C57BL
• N-Methylaspartate (pharmacology)
• Neurotransmitter Uptake Inhibitors (pharmacology)
• Pyrrolidines (pharmacology)
• Receptors, Glutamate (metabolism)
• Receptors, Kainic Acid (metabolism)
• Receptors, N-Methyl-D-Aspartate (metabolism)
• Tremor (genetics)
• Trinucleotide Repeat Expansion