Advanced optical and computational methods combined with IEGs expression, allows to analyze the activity in the entire brain with single-neuron resolution, and extract salient and robust knowledge from large datasets, to link interconnected functional networks to their corresponding anatomical brain circuits. In the work presented in this dissertation, I applied whole-brain c-Fos imaging using light sheet fluorescence microscopy to assess how the mouse brain represents and distinguish familiar and unfamiliar olfactory stimuli from conspecifics of the opposite sex. Brains from exposed mice were collected and underwent whole-brain c-Fos staining, iDISCO tissue clearing, and light-sheet recording. To reconstruct the entire 3D brain volume, the post-processing of the different tiles was done using the BigStitcher software and ultimately analyzed by ClearMap. By applying this analytical pipeline we identified a subset of brain regions increasing their c-Fos expression profile in response to the olfactory stimulation. Specifically, increased c-Fos+ expression is evident in regions involved in processing sexual odorants, such as the Main Olfactory Bulb, the Accessory Olfactory Bulb, the Medial Amygdalar Nucleus, and the Ventral Premammillary Nucleus. Moreover, certain brain regions exhibit increased c-Fos expression in response to unfamiliar stimuli only, with the Supramammillary Nucleus being notable among them. Finally, correlation matrices and graph theoretical analysis were applied to better characterize the functional network underlying odor representation, highlighting substantial differences between the three experimental groups in their internal functional connections. In conclusion, the work presented in this dissertation contributed to identify specific brain regions involved in discrimination of odors from familiar and unfamiliar conspecifics, paving the way for future investigations aimed to causally link selected hub areas with mouse behavior.
Advanced optical and computational methods combined with IEGs expression, allows to analyze the activity in the entire brain with single-neuron resolution, and extract salient and robust knowledge from large datasets, to link interconnected functional networks to their corresponding anatomical brain circuits. In the work presented in this dissertation, I applied whole-brain c-Fos imaging using light sheet fluorescence microscopy to assess how the mouse brain represents and distinguish familiar and unfamiliar olfactory stimuli from conspecifics of the opposite sex. Brains from exposed mice were collected and underwent whole-brain c-Fos staining, iDISCO tissue clearing, and light-sheet recording. To reconstruct the entire 3D brain volume, the post-processing of the different tiles was done using the BigStitcher software and ultimately analyzed by ClearMap. By applying this analytical pipeline we identified a subset of brain regions increasing their c-Fos expression profile in response to the olfactory stimulation. Specifically, increased c-Fos+ expression is evident in regions involved in processing sexual odorants, such as the Main Olfactory Bulb, the Accessory Olfactory Bulb, the Medial Amygdalar Nucleus, and the Ventral Premammillary Nucleus. Moreover, certain brain regions exhibit increased c-Fos expression in response to unfamiliar stimuli only, with the Supramammillary Nucleus being notable among them. Finally, correlation matrices and graph theoretical analysis were applied to better characterize the functional network underlying odor representation, highlighting substantial differences between the three experimental groups in their internal functional connections. In conclusion, the work presented in this dissertation contributed to identify specific brain regions involved in discrimination of odors from familiar and unfamiliar conspecifics, paving the way for future investigations aimed to causally link selected hub areas with mouse behavior.
Mouse whole-brain analysis of sexually relevant stimuli perception through light-sheet microscopy
MANCIN, NICOLE
2022/2023
Abstract
Advanced optical and computational methods combined with IEGs expression, allows to analyze the activity in the entire brain with single-neuron resolution, and extract salient and robust knowledge from large datasets, to link interconnected functional networks to their corresponding anatomical brain circuits. In the work presented in this dissertation, I applied whole-brain c-Fos imaging using light sheet fluorescence microscopy to assess how the mouse brain represents and distinguish familiar and unfamiliar olfactory stimuli from conspecifics of the opposite sex. Brains from exposed mice were collected and underwent whole-brain c-Fos staining, iDISCO tissue clearing, and light-sheet recording. To reconstruct the entire 3D brain volume, the post-processing of the different tiles was done using the BigStitcher software and ultimately analyzed by ClearMap. By applying this analytical pipeline we identified a subset of brain regions increasing their c-Fos expression profile in response to the olfactory stimulation. Specifically, increased c-Fos+ expression is evident in regions involved in processing sexual odorants, such as the Main Olfactory Bulb, the Accessory Olfactory Bulb, the Medial Amygdalar Nucleus, and the Ventral Premammillary Nucleus. Moreover, certain brain regions exhibit increased c-Fos expression in response to unfamiliar stimuli only, with the Supramammillary Nucleus being notable among them. Finally, correlation matrices and graph theoretical analysis were applied to better characterize the functional network underlying odor representation, highlighting substantial differences between the three experimental groups in their internal functional connections. In conclusion, the work presented in this dissertation contributed to identify specific brain regions involved in discrimination of odors from familiar and unfamiliar conspecifics, paving the way for future investigations aimed to causally link selected hub areas with mouse behavior.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.14240/8239