Influence of the cell wall on intracellular delivery to algal cells by electroporation and sonication.
To assess the cell wall's role as a barrier to intracellular delivery, wild-type Chlamydomonas reinhardtii algal cells and mutant cells lacking a cell wall were exposed to electroporation or sonication. Flow cytometry determined intracellular uptake of calcein and bovine serum albumin (BSA) and loss of cell viability as functions of electroporation transmembrane potential and acoustic energy. Electroporation of wild-type cells increased calcein uptake with increasing transmembrane potential, but delivered much less BSA. Electroporation of wall-deficient cells had similar effects on calcein uptake, but increased BSA uptake as much as 7.5-fold relative to wild-type cells, which indicated that the cell wall was a significant barrier to BSA delivery during electroporation. Sonication of wild-type cells caused calcein and BSA uptake at similar levels. This suggests that the cell wall barrier to BSA delivery can be overcome by sonication. Increased electroporation transmembrane potential or acoustic energy also caused increased loss of cell viability, where wall-deficient cells were especially susceptible to lysis. Overall, we believe this is the first study to compare directly the effects of electroporation and sonication in any cell type. Specifically, these findings suggest that electroporation primarily transports molecules across the plasma membrane because its mechanism is specific to lipid bilayer disruption, whereas sonication transports molecules across both the plasma membrane and cell wall, because it nonspecifically disrupts cell-surface barriers.
Azencott
Harold R
HR
School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA.
Peter
Gary F
GF
Prausnitz
Mark R
MR
D000162:Acoustics; D000818:Animals; D002463:Cell Membrane Permeability / Q000502:physiology; D002473:Cell Wall / Q000378:metabolism* / Q000502:physiology; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D018274:Electroporation / Q000379:methods*; D005452:Fluoresceins / Q000493:pharmacokinetics; D008564:Membrane Potentials / Q000502:physiology; D009154:Mutation; D012710:Serum Albumin, Bovine / Q000493:pharmacokinetics; D013010:Sonication
D005452:Fluoresceins; D012710:Serum Albumin, Bovine; C007740:fluorexon