Study of the role of Protein 4.1/Coracle in the cortical cytoskeleton of Drosophila melanogaster.

BRUNO G; GAZAL NG; RAMOS FO; ZAMPONI N; GOROSTIZA A; UNSAIN N

San Luis

Congreso; XXXVIII Congreso Reunión Anual - Sociedad Argentina de Investigación en Neurociencia. San Luis, Argentina; 2023

Sociedad Argentina de Investigación en Neurociencia

Actin, spectrin, and associated proteins form a periodic membrane-associated skeleton (MPS) that is ubiquitously present in axons and dendrites of all neuronal types. The protein 4.1 stabilizes the spectrin-actin interaction and their connection to the plasma membrane. The protein Coracle (D4.1) in Drosophila melanogaster is the sole ortholog of mammalian protein 4.1. Due to limited knowledge about Coracle in Dm neurons and the fact that protein 4.1 has not been described as part of the neuronal MPS, we will first determine the structure, function, and evolution of Coracle by integrating information from its primary sequence, secondary structure, and protein-protein interactions. To assess whether different Coracle mutants affect the turnover dynamics of spectrin in the MPS, we will evaluate the β-spectrin-GFP fly using FRAP and FCS techniques. Furthermore, super-resolution microscopy techniques ExM and STED will be employed to determine the localization and potential function of D4.1 in the MPS of axons and dendrites in different neuronal populations. Additionally, we will study whether the loss or gain of Coracle function in these specific neuronal populations of larvae and adults modifies the development and/or maintenance of the MPS. We believe that this work, besides establishing the groundwork for understanding Coracle in neurons, will enable subsequent studies associated with other Coracle functions related to MPS dynamics.