Biomedical subjects

Rod J Sayer

Publications and source records attributed to Rod J Sayer.

3 recordsLinked to original sources

PubMed2004

Calbindin independence of calcium transport in developing teeth contradicts the calcium ferry dogma.

Cytosolic calcium-binding proteins termed calbindins are widely regarded as a key component of the machinery used to transport calcium safely across cells. Acting as mobile buffers, calbindins are thought to ferry calcium in bulk and simultaneously protect against its potentially cytotoxic effects. Here, we contradict this dogma by showing that teeth and bones were produced normally in null mutant mice lacking calbindin(28kDa). Structural analysis of dental enamel, the development of which depends critically on active calcium transport, showed that mineralization was unaffected in calbindin(28kDa)-null mutants. An unchanged rate of calcium transport was verified by measurements of (45)Ca incorporation into developing teeth in vivo. In enamel-forming cells, the absence of calbindin(28kDa) was not compensated by other cytosolic calcium-binding proteins as detectable by (45)Ca overlay, two-dimensional gel, and equilibrium binding analyses. Despite a 33% decrease in cytosolic buffer capacity, cytotoxicity was not evident in either the null mutant enamel or its formative cells. This is the first definitive evidence that calbindins are not required for active calcium transport, either as ferries or as facilitative buffers. Moreover, in challenging the broader notion of a cytosolic route for calcium, the findings support an alternative paradigm involving passage via calcium-tolerant organelles.

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PubMed2004

ERp29, a general endoplasmic reticulum marker, is highly expressed throughout the brain.

ERp29 is a recently discovered resident of the endoplasmic reticulum (ER) that is abundant in brain and most other mammalian tissues. Investigations of nonneural secretory tissues have implicated ERp29 in a major role producing export proteins, but a molecular activity remains wanting for this functional orphan. Intriguingly, ERp29 appears to be heavily utilized in the cerebellum, a brain region not conventionally regarded as neurosecretory. To elucidate this functional quandary, we used immunochemical approaches to characterize the regional, cellular, and subcellular distributions of ERp29 in rat brain. Immunohistochemistry revealed ubiquitous expression in neuronal and nonneuronal cells, with a distinctive variation in somatic ERp29 levels. Highly expressing cells were found in diverse locations, implying that ERp29 is not biased towards the cerebellum functionally. Using immunolocalization data mined from the literature, a proteomic profile was developed to assess the functional significance of ERp29's characteristic expression pattern. Surprisingly, ERp29 correlated poorly with classical markers of neurosecretion, but strongly with a variety of major membrane proteins. Together with immunogold localization of ERp29 to somatic ER, these observations led to a novel hypothesis that ERp29 is involved primarily in production of endomembrane proteins rather than proteins destined for export. This study establishes ERp29 as a general ER marker for brain cells and provides a stimulating clue about ERp29's enigmatic function. ERp29 appears to have broad significance for neural pathophysiology, given its ubiquitous distribution and prominence in brain over classical ER residents like BiP and protein disulfide isomerase.

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PubMed2002

Intracellular Ca2+ handling.

Intracellular Ca2+ is regulated within three major compartments: the cytosol, the endoplasmic reticulum and mitochondria. This Chapter reviews the mechanisms involved in handling of Ca2+ within these compartments with reference to potential strategies for neuroprotection. In the cytosol, Ca2+ buffering has a major influence on Ca2+ signals. Cytosolic Ca(2+)-binding proteins such as CB28 participate in Ca2+ buffering and may have a role in resistance to neurotoxicity. In the endoplasmic reticulum, a number of proteins are involved in Ca2+ uptake, lumenal buffering or release, and these may be of value as potential targets for therapeutic intervention. Mitochondria are receiving increasing attention for their role in Ca2+ storage and signaling, and as key players in the processes leading to cell death following Ca2+ overload. An improved understanding of how Ca2+ is controlled within these intracellular compartments, and how these compartments interact, will be important for neuroprotective strategies.

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