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K Rarey

Publications and source records attributed to K Rarey.

5 recordsLinked to original sources

Na,K-ATPase alpha- and beta-isoforms in the developing cochlea of the mouse.

Immunohistochemistry was used to investigate the presence of Na,K-ATPase alpha- and beta-subunits isoforms (alpha 1, alpha 2, alpha 3, beta 1 and beta 2) in the cochlea of the mouse at different ages between embryological day (E) 19 and postnatal day (P) + 30. alpha 1 was mainly found in the stria vascularis and in the spiral ligament; it increased steadily from p+4. These data correlates well with the morphological and electrophysiological maturation of the cochlea. alpha 3 predominated in the spiral ganglia and the cochlear nerve. This finding is well in accordance with reports that alpha 3 seems to be associated with the nervous system. The beta-subunit was found mainly in those tissues where staining of the alpha-subunit also was seen. Both subunits were localized in tissue regions where fluid regulation is expected to play an important role. For some isoforms, the expression pattern of Na,K-ATPase during development in the mouse is different from that in the rat. The expression of Na,K-ATPase and that of glucocorticoid receptors during development in the inner ear of the mouse show a similar pattern, which may indicate that glucocorticoid receptors could be involved in regulating the expression of Na,K-ATPase.

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Appearance of glucocorticoid receptors in the inner ear of the mouse during development.

CBA mice were sacrificed at different ages of developments at embryonic day 13 (E13), E14, E16, E19, E20 and postnatal day 1 (P1), P2, P3, P4, P6, P8, P10, P14, P16, P18, P20 and P30. The temporal bones were quickly removed and deep frozen in order to prepare cryosections for immunohistochemical staining with polyclonal antibodies against glucocorticoid receptors. The avidin-biotin, ABC-method was used to visualize binding. Both the vestibular and the cochlear regions of the inner ear were analysed. A faint staining of the crista ampullaris, the utricle and the cochlear duct was seen at E19; and staining became clearly visible at P1. A decrease in labelling was found at day 2-5 post partum whereafter an increased staining was again noticed until postnatal day 14 when an adult pattern was observed. The appearance of glucocorticoid receptors in the inner ear during development does not follow a linear curve. Further, the labelling pattern may indicate an impact of glucocorticoid receptors on the embryologic maturation itself as well as a functional role in the adult ear.

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Primary culture of capillary endothelial cells from the spiral ligament and stria vascularis of bovine inner ear. Retention of several endothelial cell properties in vitro.

Methods for isolation and culture of microvascular endothelial cells of the inner ear were devised to provide an in-vitro system for studying endothelial functions in this tissue. Capillaries from the stria vascularis and spiral ligament were treated enzymatically to free them from surrounding tissue. Contamination by extraneous tissue was minimized by banding capillary segments in Percoll gradients and culture in plasma-derived serum on a fibronectin-coated substrate. Although only small amounts of inner ear tissue were available, tritiated thymidine autoradiography demonstrated that considerable growth in culture was possible. Addition of heparin and endothelial cell growth supplement to the medium enhanced proliferation. The endothelial origin of the cultured cells was confirmed by immunofluorescent demonstration of the presence of Factor VIII-related antigen and angiotensin-converting enzyme. In addition, tight junctions between cells were observed in both thin sections and platinum replicas obtained by freeze-fracture techniques. Endothelial cells from neither the stria vascularis nor the spiral ligament allowed passage of horseradish peroxidase across the monolayer during a 5-min period. However, endothelial cells from the stria vascularis exhibited a greater amount of pinocytotic activity than those of the spiral ligament, a difference that is also observed in vivo. Methods for expanding a small population of endothelial cells with retention of specialized properties into one of sufficient size for morphologic and biochemical studies have been demonstrated for the inner ear.

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