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A C‐terminal motif containing a PKC phosphorylation site regulates γ‐Protocadherin‐mediated dendrite arborization in the cerebral cortex in vivo
Journal article   Open access   Peer reviewed

A C‐terminal motif containing a PKC phosphorylation site regulates γ‐Protocadherin‐mediated dendrite arborization in the cerebral cortex in vivo

Camille M. Hanes, Kar Men Mah, David M. Steffen, Cathy M. McLeod, Charles G. Marcucci, Leah C. Fuller, Robert W. Burgess, Andrew M. Garrett and Joshua A. Weiner
Developmental neurobiology (Hoboken, N.J.), Vol.84(3), pp.217-235
07/15/2024
DOI: 10.1002/dneu.22950
PMCID: PMC11251855
PMID: 38837880
Appears in  UI Libraries Support Open Access
url
https://doi.org/10.1002/dneu.22950View
Published (Version of record) Open Access

Abstract

The Pcdhg gene cluster encodes 22 γ‐Protocadherin (γ‐Pcdh) cell adhesion molecules that critically regulate multiple aspects of neural development, including neuronal survival, dendritic and axonal arborization, and synapse formation and maturation. Each γ‐Pcdh isoform has unique protein domains—a homophilically interacting extracellular domain and a juxtamembrane cytoplasmic domain—as well as a C‐terminal cytoplasmic domain shared by all isoforms. The extent to which isoform‐specific versus shared domains regulate distinct γ‐Pcdh functions remains incompletely understood. Our previous in vitro studies identified protein kinase C (PKC) phosphorylation of a serine residue within a shared C‐terminal motif as a mechanism through which γ‐Pcdh promotion of dendrite arborization via myristoylated alanine‐rich C‐kinase substrate (MARCKS) is abrogated. Here, we used CRISPR/Cas9 genome editing to generate two new mouse lines expressing only non‐phosphorylatable γ‐Pcdhs, due either to a serine‐to‐alanine mutation ( Pcdhg S/A ) or to a 15‐amino acid C‐terminal deletion resulting from insertion of an early stop codon ( Pcdhg CTD ). Both lines are viable and fertile, and the density and maturation of dendritic spines remain unchanged in both Pcdhg S/A and Pcdhg CTD cortex. Dendrite arborization of cortical pyramidal neurons, however, is significantly increased in both lines, as are levels of active MARCKS. Intriguingly, despite having significantly reduced levels of γ‐Pcdh proteins, the Pcdhg CTD mutation yields the strongest phenotype, with even heterozygous mutants exhibiting increased arborization. The present study confirms that phosphorylation of a shared C‐terminal motif is a key γ‐Pcdh negative regulation point and contributes to a converging understanding of γ‐Pcdh family function in which distinct roles are played by both individual isoforms and discrete protein domains.
 cell adhesion molecules cell signaling CRISPR dendritic arborization dendritic spines UIOWA OA Agreement

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