Protocol

Surgical Manipulations on Pupal Wings from the African Butterfly Bicyclus anynana: Grafts

Institute of Biology, Leiden University, 2300 RA Leiden, The Netherlands

¹Corresponding author (p.m.brakefield{at}biology.leidenuniv.nl)

This article is also available in Emerging Model Organisms: A Laboratory Manual, Vol. 1. CSHL Press, Cold Spring Harbor, NY, USA, 2009.

INTRODUCTION

This protocol describes a procedure for transplanting tissue of the eyespot organizer in early pupae of the African butterfly Bicyclus anynana. In very young pupae, before apolysis, the pupal wing case is attached to the sheet of cells that will form the dorsal surface of the adult forewing. Therefore, if a piece of pupal cuticle is removed and transplanted to a novel site on the developing wing (where a corresponding patch has been removed), the underlying cells of the dorsal forewing are transferred together with it. The pattern determination effects of the transplanted wing tissue on the surrounding cells can be examined by comparing the experimental and control wings of the adult butterfly. Square grafts can be turned 180° (or 90°) before implantation to distinguish donor and host tissue, because the orientation of the scale cells is determined before grafting.