Biology subjects

Tomanic, T.

Publications and source records attributed to Tomanic, T..

2 recordsLinked to original sources

bioRxiv2025

Fluorescence recovery after photobleaching reveals different 1 behaviour2 of tropomyosin isoforms Tpm3.1 and Tpm4.2 in dendritic spines

Actin is the predominant cytoskeletal structure in both the pre- and the post-synaptic compartment of excitatory synapses in the brain, which are formed between the distal part of the axon and the distal site of dendritic spines. Tropomyosin (Tpm) is regarded a master regulator of actin dynamics in mammalian cells. Tpm isoforms, found in neurons are encoded by the Tpm1, Tpm3 and Tpm4 genes, and have a distinct temporal and spatial distribution of expression. Tpm3 and Tpm4 gene products have been found to segregate to the postsynaptic region of central nervous system synapses. Functional differences between Tpm3.1 and Tpm4.2 in neurons have been reported in previous studies. However, these were lacking a detailed analysis of the molecular mobility and dynamics of these two Tpm isoforms in the dendritic compartment. Here, we investigated the kinetic properties of Tpm3.1 and Tpm4.2 via a Fluorescent Recovery After Photobleaching (FRAP) approach and have discovered that Tpm3.1 and Tpm4.2 have distinct kinetic features in dendritic spines. Moreover, we investigated the dynamics of actin in the presence of either Tpm3.1 or Tpm4.2 isoform overexpression, using F-tractin as a reporter of filamentous actin. We have shown that the kinetics of actin turnover is significantly different in response to Tpm3.1 overexpression when compared the actin turnover in response to Tpm4.2 overexpression. Our study further elucidates the roles of Tpm3.1 and Tpm4.2 and provides important conclusions for future studies that are focused on discerning the molecular pathways of Tpm3.1 and Tpm4.2 segregation into different neuronal compartments.

cell biology↗

bioRxiv2025

Knock-out of Tpm4.2/actin filaments alters neuronal signaling, neurite outgrowth and behavioral phenotypes in mice.

Tropomyosins (Tpm) are master regulators of actin dynamics through forming co-polymers with filamentous actin. Despite the well-understood function of muscle Tpms in the contractile apparatus of muscle cells, much less is known about the diverse physiological function of cytoplasmic Tpms in eukaryotic cells. Here, we investigated the role of the Tpm4.2 isoform in neuronal processes including signaling, neurite outgrowth and receptor recycling using primary neurons from Tpm4.2 knock-out mice. Live imaging of calcium and electro-physiology data demonstrated increased frequency, yet reduced strength of single neuron spikes. Calcium imaging further showed increase in neuronal networks. In vitro assays of Tpm4.2 knock-out neurons displayed impaired recycling of the AMPA neurotransmitter receptor subunit GluA1. Morphometric analysis of neurite growth showed increased dendritic complexity and altered dendritic spine morphology in Tpm4.2 knock-out primary neurons. Behavioral analysis of Tpm4.2 knock-out mice displayed heightened anxiety in Open field test whilst Elevated Plus maze displayed heightened anxiety only in females. A sex-dependent phenotype was also seen in the Social Preference test with impaired social memory and socialization in female Tpm4.2 knock-out mice, whilst male and female knock-out mice had impaired recognition memory in a Novel Object Recognition test. Our study depicts the multi-faceted role of the Tpm4.2 isoform and its co-polymer F-actin population in neurons, with potential implications for better understanding diseases of the nervous system which involve actin cytoskeleton dys-function.

neuroscience↗