Investigation of KATP Channel Subunits in the Peripheral and Central Nervous Systems

An image showing different cells viewed through a microscope.

Learn histology and imaging techniques.

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This project has reached full capacity for the current term. Please check back next semester for updates.

Project description

ATP-sensitive potassium channels (KATP) are downstream targets of opioid receptors in neurons in the peripheral and central nervous systems. KATP channels are composed of either Kir6.1 or Kir6.2 and SUR1, SUR2A, or SUR2B subunits in the nervous system. Studies of KATP channels highlight their importance in pain signaling and opioid tolerance. It remains unknown where the change in KATP channel expression occurs only in the peripheral and/or central nervous systems. This project will use RNA Scope technology to determine the expression of individual subunits in the dorsal root ganglia (DRG) and spinal cord of naïve and morphine tolerant mice. RNA Scope technology will be used to detect mRNA expression of the KATP channel subunits in neurons, astrocytes, and microglia. Two groups of mice, consisting of both males and females, will be utilized. Spinal cord and dorsal root ganglia will be collected from vehicle and morphine-treated (15 mg/kg, 2x daily, 5 days) mice and snap-frozen in liquid nitrogen and stored in a -80°C freezer. Tissues will be sectioned via cryostat and incubated with RNA Scope Fluorescent Multiplex assay reagents (ACD Bio). RNA Scope allows simultaneous detection of 3-4 mRNA targets within a single sample, each optimized for a different probe channel. Components within each multiplex probe are unique to individual mRNA targets. These results will then be presented at the end of semester and could possibly be used in future research projects. 

Project outcome

  • Develop skills in animal handling and tissue histology.
  • Collect samples and use confocal microscopy for imaging.
  • Understand the biochemistry and molecular biology rationale on how in situ hybridization works, including the preamplifiers, amplifiers and dyes are assigned to a specific probe channel.
  • Present data to laboratory members and other peers.
  • Gain experience discussing scientific studies and analyzing data.

Project details

Timing, eligibility and other details
Length of commitment Longer than a semester (about 6-9 months)
Start time Spring, Summer
In-person, remote, or hybrid? In-person
Level of collaboration Small group project (2-3 students)
Benefits Stipend
Who is eligible

Freshmen

Sophomores

Juniors

Project mentor

Amanda Klein

Associate Professor

Pharmacology and Toxicology

Phone: (716) 829-2453

Email: ahklein2@buffalo.edu

Start the project

  1. Email the project mentor using the contact information above to express your interest and get approval to work on the project. (Here are helpful tips on how to contact a project mentor.)
  2. After you receive approval from the mentor to start this project, click the button to start the digital badge. (Learn more about ELN's digital badge options.) 
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Preparation activities

Once you begin the digital badge series, you will have access to all the necessary activities and instructions. Your mentor has indicated they would like you to also complete the specific preparation activities below. After you’re approved to begin the project, your mentor will send the relevant materials. Please reference this when you get to Step 2 of the Preparation Phase. 

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Keywords

neuropharmacology, neuroscience, imaging, toxicology, pharmacology, jacobs school of medicine, nervous system