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Steyger, P. S.

Publications and source records attributed to Steyger, P. S..

Calbindin and parvalbumin are early markers of non-mitotically regenerating hair cells in the bullfrog vestibular otolith organs

Earlier studies have demonstrated hair cell regeneration in the absence of cell proliferation, and suggested that supporting cells could phenotypically convert into hair cells following hair cell loss. Because calcium-binding proteins are involved in gene up-regulation, cell growth, and cell differentiation, we wished to determine if these proteins were up-regulated in scar formations and regenerating hair cells following gentamicin treatment. Calbindin and parvalbumin immunolabeling was examined in control or gentamicin-treated (GT) bullfrog saccular and utricular explants cultured for 3 days in amphibian culture medium or amphibian culture medium supplemented with aphidicolin, a blocker of nuclear DNA replication in eukaryotic cells. In control cultures, calbindin and parvalbumin immunolabeled the hair bundles and, less intensely, the cell bodies of mature hair cells. In GT or mitotically-blocked GT (MBGT) cultures, calbindin and parvalbumin immunolabeling was also seen in the hair bundles, cuticular plates, and cell bodies of hair cells with immature hair bundles. Thus, these antigens were useful markers for both normal and regenerating hair cells. Supporting cell immunolabeling was not seen in control cultures nor in the majority of supporting cells in GT cultures. In MBGT cultures, calbindin and parvalbumin immunolabeling was up-regulated in the cytosol of single supporting cells participating in scar formations and in supporting cells with hair cell-like characteristics. These data provide further evidence that non-mitotic hair cell regeneration in cultures can be accomplished by the conversion of supporting cells into hair cells.

Non-NASA Center↗

Intracellular distributions and putative functions of calcium-binding proteins in the bullfrog vestibular otolith organs

Hair cells in the bullfrog vestibular otolith organs were immunolabeled by monoclonal and polyclonal antisera against calbindin (CaB), calmodulin (CaM), calretinin (CaR), and parvalbumin (PA). S-100, previously shown to immunolabel striolar hair cells in fish vestibular organs, only weakly immunolabeled hair cells in the bullfrog vestibular otolith organs. Immunolabeling was not detected in supporting cells. With the exception of CaR, myelinated axons and unmyelinated nerve terminals were immunolabeled by all of the above antisera. Immunolabeling was seen in all saccular hair cells, although hair cells at the macular margins were immunolabeled more intensely for CaB, CaM, and PA than more centrally located hair cells. As the macula margins are known to be a growth zone, this labeling pattern suggests that marginal hair cells up-regulate their calcium-binding proteins during hair cell development. In the utriculus, immunolabeling for CaM and PA was generally restricted to striolar hair cells. CaR immunolabeling was restricted to the stereociliary array. Immunolabeling for other calcium-binding proteins was generally seen in both the cell body and hair bundles of hair cells, although this labeling was often localized to the stereociliary array and the apical portion of the cell body. CaM and PA immunolabeling in the stereociliary array in saccular and utricular striolar cells suggests a functional role for these proteins in mechanoelectric transduction and adaptation.

NASA Discipline Number 40-10↗

Visualization of newt aragonitic otoconial matrices using transmission electron microscopy

Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. These protein matrices are thought to originate from the supporting or hair cells in the macula during development. Previous studies of mammalian calcitic, barrel-shaped otoconia revealed an organized protein matrix consisting of a thin peripheral layer, a well-defined organic core and a flocculent matrix inbetween. No studies have reported the microscopic organization of the aragonitic otoconial matrix, despite its protein characterization. Pote et al. (1993b) used densitometric methods and inferred that prismatic (aragonitic) otoconia have a peripheral protein distribution, compared to that described for the barrel-shaped, calcitic otoconia of birds, mammals, and the amphibian utricle. By using tannic acid as a negative stain, we observed three kinds of organic matrices in preparations of fixed, decalcified saccular otoconia from the adult newt: (1) fusiform shapes with a homogenous electron-dense matrix; (2) singular and multiple strands of matrix; and (3) more significantly, prismatic shapes outlined by a peripheral organic matrix. These prismatic shapes remain following removal of the gelatinous matrix, revealing an internal array of organic matter. We conclude that prismatic otoconia have a largely peripheral otoconial matrix, as inferred by densitometry.

NASA Discipline Neuroscience↗

Atomic force microscope observations of otoconia in the newt

Calcitic and aragonitic otoconia from the Japanese red-bellied newt, Cynops pyrrhogaster, were examined using an atomic force microscope. The surface structure of both otoconial polymorphs consisted of arrays of elements approximately 50 nm in diameter. Elements were generally round and were separated by shallow depressions of no more than 20 nm. The elements are suggested to be single crystals of calcium carbonate. The relationship of these observations to theories of otoconial genesis is discussed.

NASA Discipline Developmental Biology↗

The morphogenic features of otoconia during larval development of Cynops pyrrhogaster, the Japanese red-bellied newt

Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. Mammalian otoconia are barrel-shaped with triplanar facets at each end. Reptilian otoconia are commonly prismatic or fusiform in shape. Amphibians have all three otoconial morphologies, barrel-shaped otoconia within the utricle, with prismatic and fusiform otoconia in the saccule. Scanning electron microscopy revealed a sequential appearance of all three otoconial morphologies during larval development of the newt, Cynops pyrrhogaster. The first otoconia appear within a single, developing otolith, and some resemble adult barrel-shaped otoconia. As the larvae hatch, around stages 39-42, the single otolith divides into two anatomically separate regions, the utricle and saccule, and both contain otoconia similar to those seen in the single otolith. Throughout development, these otoconia may have variable morphologies, with serrated surfaces, or circumferential striations with either separated facets or adjacent facets in the triplanar end-regions. Small fusiform otoconia occur later, at stage 51, and only in the saccule. Prismatic otoconia appear later still, at stage 55, and again only in the saccule. Thus, although prismatic otoconia are the most numerous in adult newts, it is the last vestibular otoconial morphology to be expressed.

NASA Discipline Neuroscience↗