Fig. 4: DNRAbs induced neuronal loss and dendritic pruning in CA1 pyramidal neurons persist in mice-lacking GluN2B, but not in those lacking GluN2A. | Nature Communications

Fig. 4: DNRAbs induced neuronal loss and dendritic pruning in CA1 pyramidal neurons persist in mice-lacking GluN2B, but not in those lacking GluN2A.

From: Lupus autoantibodies act as positive allosteric modulators at GluN2A-containing NMDA receptors and impair spatial memory

Fig. 4

a Schematic of NMDAR composition expected in hippocampal pyramidal neurons in the various genetic backgrounds (Supplementary Fig. 2). b Micrographs of hippocampal CA1 neurons from DNRAb+ or DNRAb− N2B cKO or N2A KO mice. Scale bar, 20 µm. c Quantification of CA1 neurons (mean ± SEM, n = 24 sections per group). Each dot represents a CA1 pyramidal neuron field counted using a standard unbiased stereological protocol (eight runs of systematic random sampling in three animals per group) (***p < 0.001, two-sided Mann–Whitney U test) (left to right, p = 0.0000156, 0.765). d, e Dendritic complexity of Golgi-stained neurons. Left, tracings of CA1 pyramidal neurons from N2B cKO (d) or N2A KO (e) mice. Right, Sholl analysis for N2B cKO (d) or N2A KO (e) mice (***p < 0.001, two-sided Kolmogorov–Smirnov test for data converted to cumulative distribution function) (left to right, p = 2.754 × 10−16, 0.61). Values at concentric rings are shown as mean ± SEM. For each group, 10–20 neurons per animal, 4 animals per group. f, g Representative images of microglia from N2B cKO (a) or N2A KO (b) mice. Microglia are labeled with antibody to Iba1 (red, Alexa 594) and to CD68 (green, Alexa-488). Scale (white bar): 5 µm. Right panels, quantification of CD68 + score (mean ± SEM) (***p < 0.001, Kruskal–Wallis ANOVA test) (left to right, p = 0.00000000222, 0.0793) (see Supplementary Methods). Three animals per group; number of quantified microglia: N2B cKO, DNRAb− (n = 42), DNRAb+ (n = 87), N2A KO, DNRAb− (n = 57), DNRAb+ (n = 58). h, i Microglia process complexity and length. Left panels, Sholl analysis of process length of microglia from N2B cKO (h) or N2A KO (i) mice. Right panels, cumulative probability distribution curves of microglia process length from N2B cKO (h) or N2A KO (i) (***p < 0.001, two-sided Kolmogorov–Smirnov test) (top to bottom, p = 7.16 × 10−8, 0.999). Three animals per group; number of quantified microglia: N2B cKO, DNRAb− (n = 33), DNRAb+ (n = 34); N2A KO, DNRAb− (n = 39), DNRAb+ (n = 40).

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