A Model to Study NMDA Receptors in Early Nervous System Development

• Published in: The Journal of neuroscience Vol. 40; no. 18; pp. 3631 - 3645
• Main Authors: Zoodsma, Josiah D, Chan, Kelvin, Bhandiwad, Ashwin A, Golann, David R, Liu, Guangmei, Syed, Shoaib A, Napoli, Amalia J, Burgess, Harold A, Sirotkin, Howard I, Wollmuth, Lonnie P
• Format: Journal Article
• Language: English
• Published: United States Society for Neuroscience 29.04.2020
• Subjects: Acute effects; Calcium ions; Calcium permeability; CRISPR; Danio rerio; Defects; Dizocilpine; Embryos; Fertilization; Glutamic acid receptors (ionotropic); Hyperactivity; Ion channels; Larvae; Light effects; Mutants; N-Methyl-D-aspartic acid receptors; Nervous system; Prey; Receptors; Stimuli; Synaptic transmission; Vertebrates; Zebrafish; visual acuity; prey-capture; zebrafish; habituation; CRISPR-Cas9; calcium permeability
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/JNEUROSCI.3025-19.2020

-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, and , which encode the obligatory GluN1 subunits. While receptors containing or show high Ca permeability, like their mammalian counterpart, is expressed earlier and more broadly in development than Both and zebrafish are viable. Unlike in rodents, where the knockout is embryonic lethal, double-mutant fish ( ), 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 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. 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.