Morphological and immunocytochemical heterogeneity of cultured pinealocytes from one‐week‐and two‐month‐old rats: Planimetric and densitometric investigations

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Abstract
In vitro preparations of rat pinealocytes are widely used for biochemical analyses of signal transduction processes. This paper deals with morphological and immunocytochemical features of such preparations. Special attention was paid to the problems of whether pinealocytes represent a heterogeneous cell population and how such heterogeneity may develop during ontogeny. The investigations were performed with cells which were obtained from the pineal organ of one‐week‐and two‐month‐old rats, attached to synthetic peptide‐coated coverslips or tissue culture chamber slides, and maintained under in vitro conditions overnight. The attached cells were then fixed with paraformaldehyde. These preparations yielded monolayers of spherical cells of different sizes; most cells were isolated, but some of them were aggregated and formed small clusters. On the average, the cells from the one‐week‐old animals were smaller than the cells from the two‐month‐old animals. Immunocytochemical demonstration of S‐antigen, a pinealocyte‐specific marker, showed that the majority of the cells from two‐month‐old animals were intensely or moderately labelled. Pinealocytes from one‐week‐old animals were less S‐antigen immunoreactive. Only very few cells (less than 1% displayed glial fibrillary acidic protein (GFAP)‐immunoreactivity. Planimetric investigations of the cell size and semiquantitative densitometric investigations of the intensity of the S‐antigen immunoreaction revealed that (i) pinealocytes kept in vitro form a heterogeneous cell population, and that (ii) this heterogeneity increases during postnatal development from one‐week‐old to two‐month‐old animals. Two groups of pinealocytes can be distinguished based on their developmental fate: pinealocytes of one group grow dramatically, but show only a moderate increase in S‐antigen immunoreactivity, and pinealocytes of the other group retain their size, but display a distinct increment in S‐antigen immunoreacti vitv.

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