Supplemental data

Files in this Data Supplement:

• supplemental material - Supplemental Figure 1. Juvenile Ddo�/� mice display elevated D-aspartate levels A, HPLC amino acid analysis of 4-week brain D-aspartate Ddo+/+ vs. Ddo�/�; p = 0.0004. B, 4-week adrenal D-aspartate Ddo+/+ vs. Ddo�/�; p = 0.005, and C, 4-week testicular D-aspartate Ddo+/+ vs. Ddo�/�; p = 0.0001 and L-aspartate Ddo+/+ vs. Ddo�/�; p = 0.015. (*, p < 0.05; **, p < 0.01; ***, p < 0.001; comparison using two-tailed t-test; data are expressed as mean � SEM).
• supplemental material - Supplemental Figure 2. D-Aspartate immunohistochemistry in Ddo�/� and wild-type brain. D-Aspartate immunohistochemistry in wild-type mice resembles rat findings with light immunoreactivity in cerebral cortical neurons (A, arrow), neurons in the hilum of the dentate gyrus (B, arrow), hippocampal CA3/CA2 neurons (C), hypothalamic paraventricular nucleus (D), hypothalamic supraoptic nucleus (E), and pineal gland (F). D-Aspartate is not observed in CA1 neurons in wild-type or Ddo�/� hippocampus (C). Additionally, central nervous system cells normally containing D-aspartate are more strongly stained in the Ddo�/� mice as seen in cerebral cortical neurons (A; arrowhead), neurons in the hilum of the dentate gyrus (B, arrowhead), hippocampal CA3/CA2 neurons (C), and pineal gland (F). Each experiment was performed two independent times at 4 weeks and at 4 months with similar results. (Scale bars = 100 μm; 3V = 3rd ventricle; O.N. = optic nerve; DG = dentate gyrus; CA = cornu ammonis).
• supplemental material - Supplemental Figure 3. D-Aspartate immunohistochemistry in peripheral organs of Ddo�/� and wild-type mice. Wild-type kidney is normally devoid of D-aspartate; however, D-aspartate appears in both Ddo�/� mouse kidney cortex and medulla (A & B). Ddo�/� renal cortical staining is restricted to Bowman�s capsule (A, arrow). Ddo�/� renal medullary staining is found in the thin limbs of the Loops of Henle (B, arrow) as evidenced by immunopositive cells having a thin epithelia, nuclei bulging into the cytoplasm, and round shape on transverse section (data not shown). Unlike rat, mouse D-aspartate is not observed in the 4-week wild-type adrenal medulla (C) but is present in epinephrine-producing chromaffin cells of the Ddo�/� adrenal medulla (C, asterisk). Similar to the rat, wild-type D-aspartate in mice localizes to the innermost rim of the seminiferous tubules suggestive of elongate spermatids (D, asterisk). Ddo�/� testes display this pattern (D, double asterisk) and show new immunoreactivity in Leydig cells (D, arrow). Each experiment was done two independent times at 4 weeks and 4 months yielding similar results. (Scale bars = 100 �m).