title: Probing potassium channel function in vivo by intracellular delivery of antibodies in a rat model of retinal neurodegeneration.
abstract: Inward rectifying potassium (Kir) channels participate in regulating potassium concentration (K(+)) in the central nervous system (CNS), including in the retina. We explored the contribution of Kir channels to retinal function by delivering Kir antibodies (Kir-Abs) into the rat eye in vivo to interrupt channel activity. Kir-Abs were coupled to a peptide carrier to reach intracellular epitopes. Functional effects were evaluated by recording the scotopic threshold response (STR) and photopic negative response (PhNR) of the electroretinogram (ERG) noninvasively with an electrode on the cornea to determine activity of the rod and cone pathways, respectively. Intravitreal delivery of Kir2.1-Ab coupled to the peptide carrier diminished these ERG responses equivalent to dimming the stimulus 10- to 100-fold. Immunohistochemistry (IHC) showed Kir2.1 immunostaining of retinal bipolar cells (BCs) matching the labeling pattern obtained with conventional IHC of applying Kir2.1-Ab to fixed retinal sections postmortem. Whole-cell voltage-clamp BC recordings in rat acute retinal slices showed suppression of barium-sensitive Kir2.1 currents upon inclusion of Kir2.1-Ab in the patch pipette. The in vivo functional and structural results implicate a contribution of Kir2.1 channel activity in these electronegative ERG potentials. Studies with Kir4.1-Ab administered in vivo also suppressed the ERG components and showed immunostaining of Müller cells. The strategy of administering Kir antibodies in vivo, coupled to a peptide carrier to facilitate intracellular delivery, identifies roles for Kir2.1 and Kir4.1 in ERG components arising in the proximal retina and suggests this approach could be of further value in research.
authors.affiliation: Section for Translational Research in Retinal and Macular Degeneration, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Dorit
authors.initials: D
authors.lastname: Raz-Prag
authors.forename: William N
authors.initials: WN
authors.lastname: Grimes
authors.forename: Robert N
authors.initials: RN
authors.lastname: Fariss
authors.forename: Camasamudram
authors.initials: C
authors.lastname: Vijayasarathy
authors.forename: Maria M
authors.initials: MM
authors.lastname: Campos
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Bush
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Diamond
authors.forename: Paul A
authors.initials: PA
authors.lastname: Sieving
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology; D001464:Barium / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology; D002490:Central Nervous System / Q000378:metabolism / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism / Q000502:physiology; D004596:Electroretinography; D009410:Nerve Degeneration / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism / Q000502:physiology; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology; D015221:Potassium Channels / Q000378:metabolism* / Q000494:pharmacology / Q000502:physiology; D024661:Potassium Channels, Inwardly Rectifying; D051381:Rats; D011922:Rats, Mutant Strains; D012160:Retina / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D012172:Retinaldehyde / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology
chemical_list: D000906:Antibodies; C489461:Kir2.1 channel; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D001464:Barium; D012172:Retinaldehyde; D011188:Potassium