fig1

Figure 1. CRISPR/Cas9-mediated SHANK3 deletion results in neuronal loss in the monkey brain. (A) Western blot analysis of SHANK3 protein in the PFC and striatum of SHANK3-mutant monkeys. Ctl1 and Ctl2 are age-matched controls at gestational day 135 and full-term gestation, respectively; (B) Altered expressions of postsynaptic receptors and scaffold proteins in SHANK3M1 brain. Normal expressions of GluN1, GluA2, Homer1b/c, and pan-Homer but decreased expressions of GluN2B, mGluR5, and PSD95 were observed in the PFC of SHANK3M1; (C) Western blot analysis of NeuN, GFAP, and DCX in whole-cell lysates from PFC. SHANK3M1 showed decreased levels of NeuN and DCX but increased levels of GFAP in the PFC. Western blot analysis was repeated at least three times independently; (D) NeuN and GFAP staining (brown) in layers I-VI of PFC. NeuN staining showed a strong signal in the nucleus and a weak signal in the cytosol. Nuclei were stained with hematoxylin (blue). There were fewer NeuN+ neurons and more GFAP+ astrocytes in the PFC of SHANK3M1. Scale bar, 100 μm^[51]. DCX: Doublecortin; GFAP: glial fibrillary acidic protein; NeuN: neuronal nuclei; PFC: prefrontal cortex; PSD: postsynaptic density; SHANK3: SH3 and multiple ankyrin repeat domains 3.