SINGLE-CELL [CA2-CHARACTERIZATION OF PINEALOCYTES IMMOBILIZED WITH NOVEL ATTACHMENT PEPTIDE PREPARATION(]I ANALYSIS AND BIOCHEMICAL)

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.