A Study on Hippocampal Neurons

Jeannina María Villalobos, Truman College

Mahesh Gurung is the faculty sponsor of this poster.

Description

Mitochondrial movement and distribution during primary axonal and dendritic elongation will be investigated over time in E18 harvested rat embryonic hippocampus neurons grown in primary culture. Filming under high resolution microscopy and phase contrast will be used to follow mitochondrial trafficking in the soma and growing axons and dendrites over a period of three weeks. Freshly harvested rat embryonic hippocampus will be obtained from “Brain Bits” with a total of 10^6 viable cells attainable per specimen pair (right and left sides). Micro-tracker Red staining will be employed periodically to verify mitochondrial identification and its intensity will be followed over time by imaging every 10 seconds to assess mitochondrial membrane potentials. Cells will also be stained with DAPI and Alexa488-Phalloidin to confirm nucleus location and actin production and orientation. As how time allows, we will determine the optimal timing for maximum differentiation of mitochondrial activity, since over-staining may render all of these in a relatively similar state of saturation. During this same period, we will attempt to correlate staining intensity with mitochondrial motility to assess the length of time they remain active following exposure to the stain. This will give us an idea of the window of opportunity we have to assess location and motility of positively identified mitochondria.

 
Apr 19th, 11:00 AM

A Study on Hippocampal Neurons

Mitochondrial movement and distribution during primary axonal and dendritic elongation will be investigated over time in E18 harvested rat embryonic hippocampus neurons grown in primary culture. Filming under high resolution microscopy and phase contrast will be used to follow mitochondrial trafficking in the soma and growing axons and dendrites over a period of three weeks. Freshly harvested rat embryonic hippocampus will be obtained from “Brain Bits” with a total of 10^6 viable cells attainable per specimen pair (right and left sides). Micro-tracker Red staining will be employed periodically to verify mitochondrial identification and its intensity will be followed over time by imaging every 10 seconds to assess mitochondrial membrane potentials. Cells will also be stained with DAPI and Alexa488-Phalloidin to confirm nucleus location and actin production and orientation. As how time allows, we will determine the optimal timing for maximum differentiation of mitochondrial activity, since over-staining may render all of these in a relatively similar state of saturation. During this same period, we will attempt to correlate staining intensity with mitochondrial motility to assess the length of time they remain active following exposure to the stain. This will give us an idea of the window of opportunity we have to assess location and motility of positively identified mitochondria.