Tissue printed cells from teleost electrosensory and cerebellar structures

Area of Science:

• Neuroscience
• Cell Biology
• Fish Biology

Background:

• The electrosensory lateral line lobe (ELL) and cerebellum are crucial for sensory processing in electric fish.
• Studying these complex neural circuits requires methods for isolating and culturing intact cells.

Purpose of the Study:

• To develop and validate a modified tissue printing technique for isolating and culturing cells from specific brain regions of adult weakly electric fish.
• To assess the preservation of cellular morphology and the potential for studying neuronal and glial interactions in vitro.

Main Methods:

• A modified tissue printing technique was employed to isolate cells from the ELL, corpus cerebelli (CCb), and eminentia granularis pars posterior (EGp) of Apteronotus leptorhynchus.
• Cells were cultured as a monolayer on glass coverslips without proteolytic enzymes, preserving cellular structure and allowing for organotypic distribution.
• Immunocytochemistry, including for gamma-aminobutyric acid (GABA), was used to identify cell types and map neural networks.

Main Results:

• The technique successfully isolated cells with excellent preservation of soma-dendritic structure, enabling identification of electrosensory and glial cell classes.
• Tissue-printed cells maintained an organotypic distribution for up to 27 days in culture.
• A previously unrecognized network of GABAergic axonal processes was identified in the CCb and EGp.

Conclusions:

• The modified tissue printing technique provides a robust method for isolating and culturing intact electrosensory cells from the adult fish central nervous system.
• This approach facilitates electrophysiological analyses of membrane properties and synaptic interactions between identified cells.
• The method allows for detailed investigation of neural circuits and cell types in the ELL and cerebellum.