High potassium promotes differentiation of retinal neurons but does not favor rod differentiation.
Neural retinal cells of newborn rats were cultured under dissociated culture conditions. Differentiation of several types of retinal cells was confirmed by immunohistochemical detection of type-specific neural phenotypes. We used Thy-1.1 antigen as a ganglion cell marker, HPC-1 or GABA as an amacrine cell marker and rhodopsin as a rod cell marker. With a high concentration of potassium (38 mM), expression of the respective neural phenotypes were differentially affected. High K+ increased the number of Thy-1.1 positive cells 6 to 8 fold, and drastically promoted their neurite extension. The same culture conditions, however, reduced considerably the number of rhodopsin positive cells, possibly due to the unique membrane properties of photoreceptors. A high K+ concentration also promoted differentiation of HPC-1 positive and GABA positive cells, but to a lesser extent than the Thy-1.1 positive cells. Several possibilities were examined to understand the effect of a high K+ concentration on retinal neural cells. The total cell number in cultures with a high K+ concentration was approximately half of that in control cultures at day 3 and slightly smaller at day 11, suggesting that high K+ did not have a positive general effect on the proliferation or survival of retinal cells. Naturally occurring neuronal death (apoptosis) is a well-known phenomenon during retinal development. A histochemical method for detecting DNA fragmentation, a step preceding apoptosis, showed that high K+ had no preventive effect. BrdU (bromodeoxyuridine) immunohistochemistry showed that high K+ did not seem to enhance proliferation of neural precursor cells. These results indicate that a high K+ concentration promotes the expression of neuronal phenotypes but is not a favorable condition for rod differentiation. Since a high K+ concentration is considered to induce depolarization of nerve cells, the present results suggest an anterograde influence from surrounding neuronal cells, through chronic depolarization by elevated K+, is essential for the differentiation and maturation of retinal cells.
Araki
M
M
Laboratory of Neurobiology, Osaka, Japan.
Tonè
S
S
Akagawa
K
K
Kimura
H
H
D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000737:chemistry; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004249:DNA Damage; D007124:Immunoenzyme Techniques; D008858:Microscopy, Phase-Contrast; D009474:Neurons / Q000737:chemistry / Q000187:drug effects*; D011188:Potassium / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D017948:Retinal Rod Photoreceptor Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D005680:gamma-Aminobutyric Acid / Q000032:analysis
D015415:Biomarkers; D005680:gamma-Aminobutyric Acid; D011188:Potassium