GenieEffect’s first two work packages (WP1 and WP2) reflect the two main objectives of the research project. WP1 was dedicated to experimental work and WP2 was dedicated to the modeling of the PFC circuit.
In W1, the objective was to manipulate inhibitory signaling in the mouse prefrontal cortex (PFC) by silencing parvalbumin-positive (PV) inhibitory neurons using genetic techniques. The impact of this manipulation was assessed in a two-choice assay testing flexible behavior. However, the extensive training required for the complex techniques and the need to standardize multiple experimental steps limited the sample size of the mice used, resulting in inconclusive results. Nonetheless, the thorough testing and standardization of the surgical procedures and the behavioral paradigm conducted during this study will significantly benefit future research endeavors.
At the same time, I had the opportunity to standardize an alternative to the typical water deprivation during the days when no behavioral experiments were taking place. On these days water deprivation was replaced with giving citric acid water (first introduced for mice experiments by Urai et al, 2021) to the animal ad libitum. This alteration, while not impending the behavioral training process, ameliorated the animal welfare by helping it maintain a healthier weight, and reduced the labor intensity required from the experimenter on a daily basis. The standardization of this method according to our specific experimental requirements primarily benefited the host lab, while also serving as a valuable example for other research facilities looking to adopt this approach.
During my secondment at Prof. Pitsouli's lab at the University of Cyprus, I received comprehensive training in staining procedures, advanced microscopy techniques, and utilization of relevant software tools. This included proficiency in immunohistochemical staining of tissues with mitosis, nuclear, and tracheal markers, operating the Leica SP8 confocal microscope, and image processing using FIJI (ImageJ) software. Upon my return, I successfully transferred this knowledge to my host lab.
Regarding the WP2, I utilized Dendrify 2.1.4 an open-source Python package developed by a colleague within our lab, to develop a neocortex pyramidal neuron model, that features seven dendritic compartments encompassing dendritic properties. This model can be used to develop an artificial network of this region in order to simulate physiological processes and predict behavioral outcomes.
