Gap junctional intercellular communication between cultured ependymal cells, revealed by lucifer yellow CH transfer and freeze‐fracture

Abstract

In order to analyze intercellular communication between ependymal cells in mammalian brain, we have studied gap junctional communication of ependymal and glial cells in long term primary cultures derived from fetal mouse or rat hypothalamus and choroid plexus obtained in serum supplemented media with two complementary methods: 1) dye transfer of Lucifer Yellow CH after intracellular microinjection of the different cellular types, and 2) freeze-fracture of the same cultured ependymal cells. In our culture conditions, we have shown that the GJIC capacity to transfer dye was very different according to cellular types microinjected with Lucifer Yellow CH in the following respects: 1) in ependymal cells, GJIC was always important: ciliated ependymal cells, which are numerous in hypothalamic ependymal cultures (10-120 coupled cells), choroidal ependymocytes in plexus cultures (15-250 coupled cells), and non-choroidal ependymocytes in diencephalic roof cultures (10-30 coupled cells), and 2) in astroglial cells found in these primary cultures, no GJIC was observed in spite of the presence of welldifferentiated gap junctions revealed by freeze-fracture replicas. All these results show a strong GJIC in ependymal cells and indicate the very good functional state of these cells in vitro.

Gap junction proteins and cell-cell communication in the three functional zones of the adrenal gland

“…Most of this evidence comes from in vitro studies of primary cell cultures or cell lines (Murray & Fletcher 1984, Bouille et al 1991, Baldwin & Calabrese 1994, Munari-Silem et al 1995, Veenstra 1996, Willecke & Haubrich 1996, Tomai et al 1999. Because cell functions in cultures of immortalized cell lines are, by definition, significantly altered as are the three-dimensional relationships of primary cells in culture compared with the intact gland, assessment of in vivo conditions is often compromised.…”

Culture of Choroid Plexus Epithelial Cells and In Vitro Model of Blood–CSF Barrier

“…The establishment of choroidal epithelial cell cultures from various sources has been well documented in the literature. These include cells from mouse (19, 20), rat (20–22), rabbit (12), sheep (23), and cow (24), each having its own particular set of advantages and disadvantages. …”

Primary culture of choroidal epithelial cells: Characterization of an in vitro model of blood-CSF barrier

“…The establishment of choroidal epithelial cell culture has been described in the literature for the cells from various sources, including mouse (2,6,19), rat (2,6,19,24), rabbit (15), sheep (10), cow (3,25), and human (8). Some of these reports have used transferrin (24), Na + , K + -ATPase (19,25), G proteins (19), ciliary rootlet proteins (6), and the absence of antihemophilic factor (AHF) antigen (3) as the cellular markers to characterize the origin of the cultures.…”