The primary goal of this competitive grant renewal remains to identify genes and regulatory non-coding sequences that generate species-specific differences in development and organization of the cerebral cortex, particularly association areas, such as the prefrontal cortex. The prefrontal cortex is considered the crucible of human cognitive capacities with well-documented neuroanatomical connectivity and unique cellular features that are thought to be undermined in neuropsychiatric disorders and hijacked by drugs of abuse. We start with the assumption that basic principles of cortical development in all mammals are remarkably similar. It is, however, reasonable to expect important quantitative (e.g. the number of neurons, tempo and sequence of cellular events) and qualitative changes (e.g. the introduction of new neuronal subtypes, elaboration of synaptic connections and addition of functionally specialized cortical areas) since primates split from the rodent lineage about 100 million years ago. Thus, our strategy has been to study in parallel developmental events in the rodent (mouse), non-human primate (macaque) and human embryonic telencephalon by using the most advanced molecular and cell biological methods available, including comparative high-resolution mRNA-sequencing, in utero and ex utero gene manipulation (Loss or Gain of function) and heterologous transplantation of neural stem cells. We will complete and further augment our ongoing high-resolution mRNA-sequencing, confirm results by qRT-PCR, and gain further insight through bioinformatics network analysis in three species, which was initiated in the first cycle of this grant (Aim #1). Then, based on our discovery of several primate-specific cortical neuronal subtypes, we now plan to identify their genetic determinants by performing lineage analysis and use of ex-utero electroporation to overexpress and/or knockdown selected genes to examine their interactions and identify downstream transcription factors (Aim #2). Finally, we will proceed to the next stage of this research by identifying regulatory changes driving human and nonhuman primate-specific gene expression and binding sites on downstream targets followed by comparison of ChIP-seq data to differential gene expression (Aim #3). Although the proposed research is extremely time-consuming, logistically difficult and costly, it is realistic based on our published record and our progress, which shows that we succeeded to establish the unique facilities, master and modify essential methodology and already have obtained a substantial amount of feasibility data. We argue that abnormal genesis and initial formation of the evolutionarily novel, human- specific traits of the cerebral cortex may be particularly vulnerable to genetic mutations and environmental influences each of which alone, or in combination, can give rise to elusive neuropsychiatric disorders and neuronal response to prenatal exposure to drugs of therapy and abuse.

Public Health Relevance

The proposed research is a continuation of the longitudinal studies of developmental molecular and cellular events that generate differences between cortical organization of humans, non-human primates and rodents. The expected set of unique data on the genetic origin and nature of species-specific differences are not onl of theoretical significance, but also of considerable clinical importance, as many of the newly evolved traits may be involved in neuropsychiatric disorders as well as in response to drugs used in therapy and abuse in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
2R01DA023999-06
Application #
8631539
Study Section
Developmental Brain Disorders Study Section (DBD)
Program Officer
Wu, Da-Yu
Project Start
2007-12-01
Project End
2019-02-28
Budget Start
2014-03-15
Budget End
2015-02-28
Support Year
6
Fiscal Year
2014
Total Cost
$729,725
Indirect Cost
$274,477
Name
Yale University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Rash, Brian G; Ackman, James B; Rakic, Pasko (2016) Bidirectional radial Ca(2+) activity regulates neurogenesis and migration during early cortical column formation. Sci Adv 2:e1501733
Son, Alexander I; Hashimoto-Torii, Kazue; Rakic, Pasko et al. (2016) EphA4 has distinct functionality from EphA7 in the corticothalamic system during mouse brain development. J Comp Neurol 524:2080-92
Duque, Alvaro; Krsnik, Zeljka; Kostović, Ivica et al. (2016) Secondary expansion of the transient subplate zone in the developing cerebrum of human and nonhuman primates. Proc Natl Acad Sci U S A 113:9892-7
Benoit, Jamie; Ayoub, Albert; Rakic, Pasko (2016) Epigenetic stability in the adult mouse cortex under conditions of pharmacologically induced histone acetylation. Brain Struct Funct 221:3963-3978
Morozov, Yury M; Sun, Yu-Yo; Kuan, Chia-Yi et al. (2016) Alteration of SLP2-like immunolabeling in mitochondria signifies early cellular damage in developing and adult mouse brain. Eur J Neurosci 43:245-57
Reilly, Steven K; Yin, Jun; Ayoub, Albert E et al. (2015) Evolutionary genomics. Evolutionary changes in promoter and enhancer activity during human corticogenesis. Science 347:1155-9
Ayoub, Albert E; Rakic, Pasko (2015) Neuronal misplacement in schizophrenia. Biol Psychiatry 77:925-6
Brennand, K; Savas, J N; Kim, Y et al. (2015) Phenotypic differences in hiPSC NPCs derived from patients with schizophrenia. Mol Psychiatry 20:361-8
Benoit, Jamie; Ayoub, Albert E; Rakic, Pasko (2015) Transcriptomics of critical period of visual cortical plasticity in mice. Proc Natl Acad Sci U S A 112:8094-9
Koch, Marco; Varela, Luis; Kim, Jae Geun et al. (2015) Hypothalamic POMC neurons promote cannabinoid-induced feeding. Nature 519:45-50

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