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Castellani, F.

Publications and source records attributed to Castellani, F..

2 recordsLinked to original sources

Accurate spatiotemporal retinal responses require a color intensity balance fine-tuned to natural conditions

Color vision is vital for animal survival, essential for foraging and predator detection. In mice, as in other mammals, color vision originates in the retina, where photoreceptor signals are processed by neural circuits. However, retinal responses to stimuli involving multiple colors are still not well understood. One possible explanation of this knowledge gap is that previous studies have not thoroughly examined how neuronal activity adapts to a 30 seconds to a few minutes timescale when exposed to multiple color sources. To address this, we systematically varied the UV-to-green light balance with a custom-built stimulator targeting mice opsins spectra while recording retinal ganglion cell responses across the dorso-ventral axis of the retina using multielectrode arrays. Responses to full-field chirp and checkerboard stimulations with alternating UV and green light revealed that more than one order of magnitude of intensity difference favoring green M-opsin over UV S-opsin is needed for a balanced reliability in retinal ganglion cell responses in the ventral retina. An incorrect balance, with slightly increased UV light, silenced responses to green illumination. To determine if these values are consistent with natural conditions, we analyzed isomerisation rates in the mouse retina across different times of the day. We found that the M- to S-opsin activation ratio remains constant through the mesopic-photopic range, and that our empirically determined values in the ventral retina align well with these natural conditions. These lie far from a simple equalization of M- and S-opsin isomerisation rates, which we found only balances ganglion cell responses in the dorsal retina. In conclusion, a finely tuned color intensity balance matching natural light spectrum is essential for accurately measuring both fast temporal responses and detailed spatial receptive fields in the ventral retina.

neuroscience↗

Ultra-fast electroporation of giant unilamellar vesicles -- Experimental validation of a molecular model

Delivery of molecules to cells via electropermeabilization (electroporation) is a common procedure in laboratories and clinics. However, despite a long history of theoretical effort, electroporation protocols are still based on trial and error because the biomolecular structures and mechanisms underlying this phenomenon have not been established. Electroporation models, developed to explain observations of electrical breakdown of lipid membranes, describe the electric field-driven formation of pores in lipid bilayers. These transient pore models are consistent with molecular dynamics simulations, where field-stabilized lipid pores form within a few nanoseconds and collapse within tens of nanoseconds after the field is removed. Here we experimentally validate this nanoscale restructuring of bio-membranes by measuring the kinetics of transport of the impermeant fluorescent dye calcein into lipid vesicles exposed to ultrashort electric fields (6 ns and 2 ns), and by comparing these results to molecular simulations. Molecular transport after vesicle permeabilization induced by multiple pulses is additive for interpulse intervals as short as 50 ns, while the additive property of transport is no longer observed when the interval is reduced to 0 ns, consistent with the lifetimes of lipid electropores in molecular simulations. These results show that lipid vesicle responses to pulsed electric fields are significantly different from those of living cells where, for similar pulse properties, the uptake of fluorescent dye continues for several minutes. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=97 SRC="FIGDIR/small/890137v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@1397967org.highwire.dtl.DTLVardef@516b42org.highwire.dtl.DTLVardef@158c373org.highwire.dtl.DTLVardef@87c318_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗