Laser Capture Microdissection of Nematode Feeding Cells

Obligate plant-parasitic nematodes, such as cyst nematodes (Heterodera and Globodera spp.) and root-knot nematodes (Meloidogyne spp.), form specialized feeding cells in host plant roots. These feeding cells provide the sole source of nutrition for the growth and reproduction of the nematode to complete its life cycle. Feeding cell formation involves complex physiological and morphological changes to normal root cells and is accompanied by dramatic changes in plant gene expression. The distinct features of feeding cells suggest that their formation entails a unique gene expression profile, a better understanding of which will assist in building models to explain signaling pathways that modulate transcriptional changes in response to nematodes. Ultimately, this knowledge can be used to design strategies to develop resistance against nematodes in crop plants. Feeding cells comprise a small fraction of the total root cell population, and identification of plant gene expression changes specific to these cells is difficult. Until recently, the specific isolation of nematode feeding cells could be accomplished only by manual dissection or microaspiration. These approaches are limited in that only mature feeding cells can be isolated. These limitations in tissue accessibility for macromolecule isolation at different stages of feeding cell development can be overcome through the use of laser microdissection (LM), a technique that enables the specific isolation of feeding cells from early to late stages for RNA isolation, amplification, and downstream analysis.