Generation and characterization of a human iPSC line and gene-corrected isogenic line derived from a patient with a CELF2 gene mutation.

The identification of neurodevelopmental defects in a patient harboring a heterozygous de novo missense variant (NM_006561.4, c.1517G > A, p.Arg506His) within the CELF2 gene. Here, we describe the establishment of a patient-derived induced pluripotent stem cell (iPSC) line, alongside an isogenic gene-corrected iPSC line, achieved through CRISPR/Cas9 genome editing. These lines exhibit the expression of pluripotency markers, demonstrate differentiation potential into all three germ layers, and maintain a normal karyotype. These iPSC lines serve as valuable tools for investigating the consequences of CELF2 related neurodevelopmental disorders.

Immunohistochemical analysis of the developing mouse cortex.

The cerebral cortex is the most complex structure in the mammalian brain, whose development requires coordinated proliferation of neural stem/precursor cells (NPCs) and their differentiation into neurons and glia. Perturbations in NPC homeostasis can lead to abnormal cortical development which is frequently seen in neurodevelopmental disorders. In this chapter, we describe the preparation of cortical tissues from mice and step-by-step protocol for immunohistochemistry to study cortical development. With this technique, we employ commonly used molecular markers and thymidine analog methods to analyze NPC populations. We also discuss assay conditions that can be optimized according to the specific needs to improve experimental outcomes.

Ubiquitination and deubiquitination of 4E-T regulate neural progenitor cell maintenance and neurogenesis by controlling P-body formation.

During embryogenesis, neural stem/progenitor cells (NPCs) proliferate and differentiate to form brain tissues. Here, we show that in the developing murine cerebral cortex, the balance between the NPC maintenance and differentiation is coordinated by ubiquitin signals that control the formation of processing bodies (P-bodies), cytoplasmic membraneless organelles critical for cell state regulation. We find that the deubiquitinase Otud4 and the E3 ligase Trim56 counter-regulate the ubiquitination status of a core P-body protein 4E-T to orchestrate the assembly of P-bodies in NPCs. Aberrant induction of 4E-T ubiquitination promotes P-body assembly in NPCs and causes a delay in their cell cycle progression and differentiation. In contrast, loss of 4E-T ubiquitination abrogates P-bodies and results in premature neurogenesis. Thus, our results reveal a critical role of ubiquitin-dependent regulation of P-body formation in NPC maintenance and neurogenesis during brain development.

Parent-child cultural orientations and child adjustment in Chinese American immigrant families.

Direct and indirect/mediated relations of (a) children's and parents' cultural orientations and (b) parent-child gaps in cultural orientations to children's psychological adjustment were examined in a socioeconomically diverse sample of 258 Chinese American children (age = 6-9 years) from immigrant families. Parents reported on children's and their own Chinese and American orientations in language proficiency, media use, and social relationships. Parents and teachers rated children's externalizing and internalizing problems and social competence. Using structural equation modeling, we found evidence for both the effects of children's and parents' cultural orientations and the effects of parent-child gaps. Specifically, children's American orientations across domains were associated with their better adjustment (especially social competence). These associations were partly mediated by authoritative parenting. Parents' English and Chinese media use were both associated with higher authoritative parenting, which in turn was associated with children's better adjustment. Furthermore, greater gaps in parent-child Chinese proficiency were associated with children's poorer adjustment, and these relations were partly mediated by authoritative parenting. Together, the findings underscore the complex relations between immigrant families' dual orientations to the host and heritage cultures and children's psychological adjustment.