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We use the Li-cor Odyssesy scanner to lable multiple proteins in brian tissue. When this is used to image pan and phospho proteins molecular signaling in brain regions can be measures. When necessary we use confocal microscopes in the Bioimaging center at the Delaware Biotechnology Institute (DBI) to lable multiple proteins in neurons. This technique can be used to measure neural activity in specific groups of neurons as well as verify expression of reporter genes. For see all the imaging capabilities at the Bioimaging center at DBI please click on Bioimaging Center
Double labeling of septal cholinergic neurons that project to the ventral hippocampus
Double labeling of septal cholinergic neurons that project to the ventral hippocampus
In this proof of concept experiment we injected cholera toxin b-488 (ctb-488) into the ventral hippocampus and then used immunohistochemistry to label cholinergic neurons in the medial septum that project to the ventral hippocampus. We also labeled c-Fos in septal neurons, which allowed us to visualize active neurons in the medial septum that project to the ventral hippocampus. Confocal microscopes were used to images neurons.
Labeling AMPARs and NMDARs in multiple brain regions
Labeling AMPARs and NMDARs in multiple brain regions
In this study we used the Li-cor Odyssey scanner to image AMPARs and NMDARs across multiple brain regions in the same tissue. This allowed us to construct AMPAR/NMDAR ratios after fear conditioning across multiple brain regions. Green shows GluR2 subunits, red NR2A subunits and the yellow images are the two channels merged. This research was published in the article below.
Shultz, B*., Farkash, A. ·,, Collins, B. ·,, Mohammadmirzaei, N.*, and Knox., D. (2022). Fear learning-induced changes in AMPAR and NMDAR expression in the fear circuit. Learning & Memory, 29(3): 83-92.
Using double labeling westerns to measure Akt signaling
Using double labeling westerns to measure Akt signaling
In this study we used the Li-cor scanner to perform westerns probing panAkt and pAkt from homogenates obtained from multiple brain regions (e.g. PFC, amygdala) in order to determine how single prolonged stress affected Akt signaling in these brain regions. Molecular marker is to the left, bands in green are panAkt and red are pAkt and beta-actin (lower band). Beta-actin was used as a reference protein. This research was published in the article below.
Knox, D., Della Valle, R*. Mohammadmirzaei, N.*, Shultz, B.*, Biddle, M., Farkash, A. ·, Chamness, M., and Moulton, E ·. (2021). PI3K-Akt signaling in the basolateral amygdala facilitates traumatic stress enhancements in fear memory. International Journal Of Neuropsychopharmacology. 24(3):229-238.
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