A Closer Look at Protein Transport During Synaptic Development

Posted May 26th, 2010 at 2:52 pm.

Maja Hadziomerovic and Allison Petonic

Mentor: Dr. Karen Greif

Professor Karen Greif’s lab has examined the normal patterns of development of synaptic molecules in the rat Central Nervous System (CNS) and in sympathetic ganglia. Much is known about the possible functions of synaptic vesicle proteins. The assembly and transport of these proteins to developing terminals will be examined. The purpose of this specific research project is to determine whether the proteins syntaxin and synaptotagmin are co-localized in developing synapses in the pineal gland of neonatal rats.

A previous study in this lab has shown that synaptotagmin-yellow fluorescent protein (syt-YFP) is transported by axons prior to synapse formation in the pineal gland (Narayan and Greif, 2000, Soc Neurosci Abstr 2000; 26:301). Other evidence suggests that synaptic proteins are grouped together in axons before they reach their targets. Superior cervical ganglions (SCGs) of rats will be injected at postnatal day 1 and postnatal day 7 with the adenovirus syt-YFP construct. Animals will be kept alive for 48 hours before the SCGs and pineals are removed, frozen, and sectioned. Proteins will be labeled with fluorescently-labeled secondary antibodies via the techniques of immunocytochemistry. Confocal microscopy will be used to analyze and compare the patterns of syt-YFP expression with that of other proteins. Data analysis will focus on distribution and relative intensity of labeling.

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