CDKL5's role in microtubule-based transport and cognitive function

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 29, 2024

Abstract Cyclin-dependent kinase like 5 (CDKL5) is a serine-threonine highly enriched in mammalian neurons. CDKL5 located on the X-chromosome and its loss-of-function leads to severe neurodevelopmental disorder called deficiency (CDD). phosphorylates microtubule-associated protein MAP1S regulates binding microtubules. How phosphorylation affects microtubule function not well understood. To address this question, we generated phosphomutant mice, which sites S786 S812 are mutated Alanine (MAP1S S786/812A or SA). Using co-sedimentation assay, showed that dynein microtubules severely reduced knockout (KO) SA brains. Time-lapse imaging primary neurons impaired motility both Cdkl5 KO SA. Dynein-driven cargo transport was affected mutant neuron dendrites, including delivery of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. We next studied tubulin tyrosination found mutants had when compared WT Since dynein-dynactin has higher affinity for tyrosinated microtubules, hypothesized mice could be mechanistic cause motility. In support this, show upon expression tyrosine TTL, rescued defects Hippocampal derived from revealed significant reduction spine density synapses, altered morphology. Finally, behavioral phenotyping increased anxiety, motor performance, social memory deficits, mirroring some extent clinical manifestations present CDD patients. Our results reveal an important contributor dynein-mediated synapse formation. Graphical abstract

Язык: Английский

Quantitative Comparison of Monomeric StayGold Variants Using Protein Nanocages in Living Cells

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 17, 2024

Abstract To standardize comparison of fluorescent proteins and independently determine which monomeric StayGold variant is best for live microscopy, we analyzed protein tagged I3-01 peptides that self-assemble into stable sixty subunit dodecahedrons inside cells. We find mStayGold 3-fold brighter more photostable compared with EGFP superior to other variants in mammalian cytoplasm. In addition, analysis intracellular nanocage diffusion confirms the nature mStayGold.

Язык: Английский