Nc. Schaad et al., SINGLE-CELL [CA2-CHARACTERIZATION OF PINEALOCYTES IMMOBILIZED WITH NOVEL ATTACHMENT PEPTIDE PREPARATION(]I ANALYSIS AND BIOCHEMICAL), Brain research, 614(1-2), 1993, pp. 251-256
Single-cell image analysis of rat pinealocytes has been difficult beca
use they do not attach readily to coated or uncoated surfaces and typi
cally adhere in clusters to fibroblast-like cells. In the present repo
rt, a new method for the rapid attachment of rat pinealocytes is descr
ibed. Cells were prepared using papain digestion and density centrifug
ation and then were placed on coverslips or slides coated with PepTite
-2000TM, a preparation containing the attachment peptide sequence Arg-
Gly-Asp. Cells immobilized with this preparation responded to norepine
phrine treatment with an increase in cyclic AMP and melatonin producti
on. Single-cell analysis of Fura-2-loaded cells revealed that norepine
phrine increased [Ca2+]i. This development makes it possible to conduc
t routine single-cell image analysis and other studies of freshly isol
ated rat pinealocytes.