Dec 09, 2024

Public workspaceRodent ear punch assay

DOI
dx.doi.org/10.17504/protocols.io.bp2l6dx65vqe/v1
  • Ashley Seifert1
  • 1University of Kentucky
Ashley Seifert
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DOI: dx.doi.org/10.17504/protocols.io.bp2l6dx65vqe/v1
Protocol Citation: Ashley Seifert 2024. Rodent ear punch assay. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6dx65vqe/v1
Manuscript citation:
Gawriluk, T., Simkin, J., Thompson, K. et al. Comparative analysis of ear-hole closure identifies epimorphic regeneration as a discrete trait in mammals. Nat Commun 7, 11164 (2016). https://doi.org/10.1038/ncomms11164
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 06, 2024
Last Modified: December 09, 2024
Protocol Integer ID: 114524
Keywords: ASAPCRN
Abstract
This protocol describes the procedure used by the Seifert Lab to assay tissue regeneration following a 4mm ear punch in rodents, including Acomys spiny mice.

Note that any protocol involving animals should be reviewed and approved by your Institutional Animal Care and Use Committee (IACUC) before use.
Protocol materials
ReagentIsoflurane USPHenry Schein (Butler Schein)Catalog #017579
Reagent4 mm biopsy punchSklarCatalog #96-1146
Reagent8 mm biopsy punchSklarCatalog #96-1152
Anesthetize animal with Concentration3 % (v/v) vaporized ReagentIsoflurane USPHenry Schein (Butler Schein)Catalog #017579 at Pressure1 psi oxygen flow rate

Create a Thikness4 mm through-and-through hole in the center of the ear pinna with a Reagent4 mm biopsy punchSklarCatalog #96-1146

At desired time after injury, collect Thikness8 mm tissue surrounding the hole using an Reagent8 mm biopsy punchSklarCatalog #96-1152 for analysis
Note
If you are interested in assessing the timeline of wound closure, you can anesthetize the animal at desired intervals (e.g., every 5 days) and use calipers to measure the diameter of the ear hole in two dimensions: along the proximal-distal axis (DPD) and along the anterior-posterior axis (DAP). The area of the hole can then be calculated as an ellipse (to account for unevenness in closure between the two axes) by: