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Cell Selective Conditional Null Mutations of Serine Racemase Demonstrate a Predominate Localization in Cortical Glutamatergic Neurons

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Abstract

d-Serine, which is synthesized by the enzyme serine racemase (SR), is a co-agonist at the N-methyl-d-aspartate receptor (NMDAR). Crucial to an understanding of the signaling functions of d-serine is defining the sites responsible for its synthesis and release. In order to quantify the contributions of astrocytes and neurons to SR and d-serine localization, we used recombinant DNA techniques to effect cell type selective suppression of SR expression in astrocytes (aSRCKO) and in forebrain glutamatergic neurons (nSRCKO). The majority of SR is expressed in neurons: SR expression was reduced by ~65% in nSRCKO cerebral cortex and hippocampus, but only ~15% in aSRCKO as quantified by western blots. In contrast, nSRCKO is associated with only modest decreases in d-serine levels as quantified by HPLC, whereas d-serine levels were unaffected in aSRCKO mice. Liver expression of SR was increased by 35% in the nSRCKO, suggesting a role for peripheral SR in the maintenance of brain d-serine. Electrophysiologic studies of long-term potentiation (LTP) at the Schaffer collateral–CA1 pyramidal neuron synapse revealed no alterations in the aSRCKO mice versus wild-type. LTP induced by a single tetanic stimulus was reduced by nearly 70% in the nSRCKO mice. Furthermore, the mini-excitatory post-synaptic currents mediated by NMDA receptors but not by AMPA receptors were significantly reduced in nSRCKO mice. Our findings indicate that in forebrain, where d-serine appears to be the endogenous co-agonist at NMDA receptors, SR is predominantly expressed in glutamatergic neurons, and co-release of glutamate and d-serine is required for optimal activation of post-synaptic NMDA receptors.

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Abbreviations

NMDA:

N-methyl-d-aspartate

SR:

Serine racemase

aSRCKO:

Astrocyte-specific SR conditional knockout

nSRCKO:

Neuron-specific SR conditional knockout

GFAP:

Glial fibrillary acidic protein

TAM:

Tamoxifen

CaMKIIα:

α-Subunit of Ca2+/calmodulin-dependent kinase II

WT:

Wild-type

HPLC:

High-performance liquid chromatography

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Acknowledgments

This work was funded by NIH R01MH051290 and P50MH0G0450. We thank Jiamin Feng and Alexander S. Roseman for animal colony maintenance and genotyping. Cre transgenic mouse strains were generously provided by Yu Yamaguchi and Ken D. McCarthy.

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Authors and Affiliations

  1. McLean Hospital, MRC 114, 115 Mill St., Belmont, MA, 02478, USA

    Michael A. Benneyworth

  2. McLean Hospital, MRC 206, 115 Mill St., Belmont, MA, 02478, USA

    Yan Li

  3. McLean Hospital, MRC 117, 115 Mill St., Belmont, MA, 02478, USA

    Alo C. Basu

  4. McLean Hospital, MRC 209, 115 Mill St., Belmont, MA, 02478, USA

    Vadim Y. Bolshakov

  5. McLean Hospital, MRC 122, 115 Mill St., Belmont, MA, 02478, USA

    Joseph T. Coyle

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  1. Michael A. Benneyworth
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  2. Yan Li
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  3. Alo C. Basu
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  4. Vadim Y. Bolshakov
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  5. Joseph T. Coyle
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Correspondence to Joseph T. Coyle.

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Benneyworth, M.A., Li, Y., Basu, A.C. et al. Cell Selective Conditional Null Mutations of Serine Racemase Demonstrate a Predominate Localization in Cortical Glutamatergic Neurons. Cell Mol Neurobiol 32, 613–624 (2012). https://doi.org/10.1007/s10571-012-9808-4

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  • DOI: https://doi.org/10.1007/s10571-012-9808-4

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