Heavy metals such as lead (Pb), mercury (Hg), and methyl mercury (MeHg) impair cell functions. For bone it is known that Pb changes bone formation rates, which depend on intracellular free calcium concentration ([Ca2+](i)). Since heavy metals compete with Ca2+ at multiple sites and increased [Ca2+](i) reduces gap junctional coupling between bone cells, we analyzed the effects of extracellular (e) and intracellular (i) application of Pb, Hg, and MeHg on these channels. Using primary cultures of osteoblast- like cells, relative changes of [Ca2+](i) were studied in Fura-2/AM loaded cells. Parallel intracellular recordings of neighboring cells were obtained using a conventional and a patch electrode. Pb((e)) (5 μmol/liter; n = 3) and Hg((e)) (5 μmol/liter; n = 3) as well as Pb((i)) (25 μmol/liter; n = 7) did not change the coupling (ΔMP2/ΔMP1). In contrast, MeHg((e)) (1-10 μmol/liter; n = 6) and Hg((i)) (≤5 μmol/liter; n = 8) reduced the coupling to 79.5 ± 19.3% and 62.4 ± 15.3%, respectively, within 15-20 minutes. The reduction of coupling followed individual time courses, and in no case was a steady state of decoupling reached within 20 minutes. Extracellular application of Pb((e)) (5 μmol/liter, n = 74) for 20 minutes, linearly elevated the Fura emission ratio reflecting transmembrane Pb permeation rather than [Ca2+](i) in- crease. Hg((e)) (n = 48) slightly increased [Ca2+](i) from 100 to ≤200 nmol/liter, whereas MeHg((e)) (5 μmol/liter, n = 52) released Ca2+ from internal stores, thus increasing [Ca2+](i) up to 2 μmol/liter. In conclusion, Pb((e)), Pb((i)) and Hg((e)) do not affect gap junctional coupling per se. Since MeHg((e)) and Hg((i)) deplete calcium stores, the decrease of the electric coupling is attributable to increased [Ca2+](i), which affects gap junction channels.
- Free intracellular Ca
- Gap junctions
- Methyl mercury
ASJC Scopus subject areas
- Endocrinology, Diabetes and Metabolism
- Orthopedics and Sports Medicine