N-methyl-D-aspartate Receptors In Development Research Articles

A Model to Study NMDA Receptors in Early Nervous System Development.

N-methyl-D-aspartate receptors (NMDARs) are glutamate-gated ion channels that play critical roles in neuronal development and nervous system function. Here, we developed a model to study NMDARs in early development in zebrafish, by generating CRISPR-mediated lesions in the NMDAR genes, grin1a and grin1b, which encode the obligatory GluN1 subunits. While receptors containing grin1a or grin1b show high Ca2+ permeability, like their mammalian counterpart, grin1a is expressed earlier and more broadly in development than grin1b Both grin1a-/- and grin1b-/- zebrafish are viable. Unlike in rodents, where the grin1 knockout is embryonic lethal, grin1 double-mutant fish (grin1a-/-; grin1b-/-), which lack all NMDAR-mediated synaptic transmission, survive until ∼10 d dpf (days post fertilization), providing a unique opportunity to explore NMDAR function during development and in generating behaviors. Many behavioral defects in the grin1 double-mutant larvae, including abnormal evoked responses to light and acoustic stimuli, prey-capture deficits, and a failure to habituate to acoustic stimuli, are replicated by short-term treatment with the NMDAR antagonist MK-801, suggesting that they arise from acute effects of compromised NMDAR-mediated transmission. Other defects, however, such as periods of hyperactivity and alterations in place preference, are not phenocopied by MK-801, suggesting a developmental origin. Together, we have developed a unique model to study NMDARs in the developing vertebrate nervous system.SIGNIFICANCE STATEMENT Rapid communication between cells in the nervous system depends on ion channels that are directly activated by chemical neurotransmitters. One such ligand-gated ion channel, the NMDAR, impacts nearly all forms of nervous system function. It has been challenging, however, to study the prolonged absence of NMDARs in vertebrates, and hence their role in nervous system development, due to experimental limitations. Here, we demonstrate that zebrafish lacking all NMDAR transmission are viable through early development and are capable of a wide range of stereotypic behaviors. As such, this zebrafish model provides a unique opportunity to study the role of NMDAR in the development of the early vertebrate nervous system.

NMDA Receptors and Epileptogenesis in Human Cortical Dysplasia: A Metaanalysis

Cortical dysplasia (CD) is a cerebral disorder caused by disruption of neuronal migration and disorganization of cortex development. The mechanisms of epileptogenicity in CD have been investigated yet still remain unknown. One of the possible reasons is the changes of expression of Nmethyl-D-aspartate (NMDA) receptors. Emerging evidences have shown that NR2A/B in the region of focal cortical dysplasia (FCD) cortex may play an important role in the risk of epilepsy and NR1 remains unchanged during epilepsy pathogenesis but recent published studies showed inconclusive results. This meta-analysis aimed to derive a more precise estimation of the associations between NMDA receptors and CD related epilepsy risk. A literature search of PubMed, Embase, Web of Science and China BioMedicine (CBM) databases was conducted on articles published before July 1st, 2013. Crude odds ratio (OR) with 95% confidence intervals (CI) were calculated. Ten studies were included with a total of 170 subjects. 104 of them were diagnosed with FCD and epilepsy while 66 resected specimen were non-CD cerebral tissue. The meta-analysis results showed that the expression of NR2B is increased in FCD cortex, which is indicated might be an important role in FCD related epilepsy pathogenesis. On the other hand, the expression of NR1 showed no significant difference between FCD group and controls, therefore the NR1 might not be a relevant factor. The function of NMDA receptors in development of FCD is not clear yet, a further explanation is still needed. We assume that NMDA receptors might interact with other substances in different signaling pathways to initiate and promote the epileptogenic process. Meanwhile we suggest more research to focus on the causal relationship between NMDA receptors and epileptogenisis.

NMDA induces persistent inward and outward currents that cause rhythmic bursting in adult rodent motoneurons

N-methyl-d-aspartate (NMDA) receptors are of critical importance for locomotion in the developing neonatal spinal cord in rats and mice. However, due to profound changes in the expression of NMDA receptors in development between the neonatal stages and adulthood, it is unclear whether NMDA receptors are still an important component of locomotion in the adult rodent spinal cord. To shed light on this issue, we have taken advantage of recently developed preparations allowing the intracellular recording of adult motoneurons that control the tail in the sacrocaudal spinal cord of adult mice and rats. We show that in the adult sacrocaudal spinal cord, NMDA induces rhythmic activity recorded on the ventral roots, often coordinated from left to right, as in swimming motions with the tail (fictive locomotion). The adult motoneurons themselves are intrinsically sensitive to NMDA application. That is, when motoneurons are synaptically isolated with TTX, NMDA still causes spontaneous bursts of rhythmic activity, depending on the membrane potential. We show that these bursts in motoneurons depend on an NMDA-mediated persistent inward current and are terminated by the progressive activation of a persistent outward current. These results indicate that motoneurons, along with the central pattern generator, can actively participate in the production of swimminglike locomotor activity in adult rodents.

Role of Altered Structure and Function of NMDA Receptors in Development of Alcohol Dependence

Long-term alcohol exposure gives rise to development of physical dependence on alcohol in consequence of changes in certain neurotransmitter functions. Accumulating evidence suggests that the glutamatergic neurotransmitter system, especially the N-methyl-D-aspartate (NMDA) type of glutamate receptors is a particularly important site of ethanol's action, since ethanol is a potent inhibitor of the NMDA receptors (NMDARs) and prolonged ethanol exposition leads to a compensatory "upregulation" of NMDAR mediated functions supposedly contributing to the occurrence of ethanol tolerance, dependence as well as the acute and delayed signs of ethanol withdrawal.Recently, expression of different types of NMDAR subunits was found altered after long-term ethanol exposure. Especially, the expression of the NR2B and certain splice variant forms of the NR1 subunits were increased in primary neuronal cultures treated intermittently with ethanol. Since NMDA ion channels with such an altered subunit composition have increased permeability for calcium ions, increased agonist sensitivity, and relatively slow closing kinetics, the abovementioned alterations may underlie the enhanced NMDAR activation observed after long-term ethanol exposure. In accordance with these changes, the inhibitory potential of NR2B subunit-selective NMDAR antagonists is also increased, demonstrating excellent potency against alcohol withdrawal-induced in vitro cytotoxicity. Although in vivo data are few with these compounds, according to the effectiveness of the classic NMDAR antagonists in attenuation, not only the physical symptoms, but also some affective and motivational components of alcohol withdrawal, novel NR2B subunit selective NMDAR antagonists may offer a preferable alternative in the pharmacotherapy of alcohol dependence.

Role of NMDA receptors in development of respiratory control

The N-methyl-D-aspartate (NMDA) receptor has many functions throughout the central nervous system (CNS) including its role within the centers controlling respiration. Although NMDA receptors are important for normal breathing, they are specifically active under conditions of stress, such as hypoxia. Consistent with its role in other neurological functions, the NMDA receptor is also important to the prenatal development of normal neurological pathways for the control of ventilation. The importance of NMDA receptors to both normal breathing and stress responses is demonstrated by recent observations of antenatal effects of disturbances to the NMDA receptor which disrupts normal breathing as well as causing reduced ventilatory responses during stress in newborns. These characteristics fit with the known NMDA influences on neuronal development and plasticity. The methods used to evaluate these functions have mainly included pharmacological agents for activation (agonists) or depression (antagonists) of NMDA receptors. NMDA receptor expression has also been measured histologically, and more recently knockout animal models have been used to provide additional functional information.

NMDA receptors in cortical development are essential for the generation of coordinated increases in [Ca2+](i) in "neuronal domains".

Spontaneous correlated activity regulates the precision of developing neural circuits. A synchronized elevation of intracellular calcium ion concentration, [Ca(2+)](i), occurred in 5-50 adjacent neurons--known as a "neuronal domain"--in developing neocortex. This coordinated response of neuronal cells is mediated by the diffusion of inositol trisphosphate (IP(3)) via gap-junction channels. In this study, we utilized the N-methyl-D-aspartate (NMDA)-type glutamate receptor epsilon 2 (GluR epsilon 2/NR2B)(-/-) mouse, which does not possess any functional NMDA receptors in the developing neocortex, and showed that NMDA receptors are essential for the generation of "neuronal domains". First, the frequency of spontaneously occurring neuronal domains in brain slices from GluR epsilon 2(-/-) mice was significantly reduced compared to that seen in brain slices from wild-type mice. Secondly, IP(3) injection into a single neuron in a cortical slice from a GluR epsilon 2(-/-) brain resulted in very few neuronal domains being observed, but an injection similarly made into a neuron in a wild-type slice promptly resulted in neuronal domains. Even in the GluR epsilon 2(-/-) brain, the elevation of intracellular [Ca2+](i) was observed frequently in single neurons and microinjection of IP(3) produced an elevation of [Ca2+](i) in the injected cells. These results suggest that the diffusion of IP(3) into the surrounding neurons via gap junctions is almost completely absent in the GluR epsilon 2(-/-) brain. Our results may reflect the critical role of NMDA receptors in the formation of cortical circuitry, probably via the regulation of gap-junction channels between immature cortical neurons.