ISSN:
1432-198X
Keywords:
Key words:Anoxia
;
Proximal convoluted tubules
;
Cytosolic free calcium
;
Kidney development
Source:
Springer Online Journal Archives 1860-2000
Topics:
Medicine
Notes:
Abstract. Calcium(Ca2+)-dependent processes mediate, in part, anoxic cell injury. These may account for the difference in sensitivity to anoxia between certain immature and mature renal cells. To address this question, we studied the effects of anoxia on cytosolic free Ca2+ concentration ([Ca2+]i), cell integrity, and transport functions in microdissected proximal convoluted tubules (PCT) of 〈3-week-old (newborn) and 〉12-week-old (adult) rabbits. Tubules were loaded with 10 μM fura-2 AM by incubation for 60 min at 37°C, and then superfused with isosmotic saline solution gassed with either 95%O2-5%CO2 (control group) or 95%N2-5%CO2 (anoxia group) for 30 min. [Ca2+]i was measured ratiometrically; cell damage was assessed by nuclear binding of propidium iodide (PI). Anoxia resulted in a fourfold increase in [Ca2+]i in adult tubules (from resting values of 245±10 to 975±100 nM, P 〈0.001), whereas in newborn tubules the rise was significantly less (from resting values of 137±5 to 165±5 nM, P 〈0.001 between anoxic groups). Transient exposure to 100 mM potassium chloride, which depolarizes the PCT cells, induced increases in [Ca2+]i from baseline, to 920±90 nM in tubules from adult and to 396±16 nM in those from newborn rabbits (P 〈0.001 between age groups). After exposure to ligands such as parathyroid hormone (PTH) and ATP, [Ca2+]i increased in both newborn and adult tubules, but to lower levels in newborn tubules. The response to PTH and ATP was transient in both age groups, [Ca2+]i returning to baseline levels after 2 min. Following anoxia, tubules from adult animals exhibited staining of all cell nuclei by 1 min exposure to PI, indicative of gross permeabilization of the cells. Nuclei of anoxic immatures tubules did not stain with PI. The sodium-dependent uptakes of a glucose analogue (14C-α-methyl-glucopyranoside) and phosphate (32Pi) were preserved in agarose-filled tubules of newborns after anoxia, whereas in those of adults recovery from anoxia was associated with drastic reduction in the uptake of these solutes. Overall, our results suggest that: (1) during anoxia, cell Ca2+ rises to critical levels in PCTs of adults compared with those of 〈3-week-old animals, (2) Ca2+ influx occurs via a pathway activated by exposure to high [K+]o, presumably voltage-sensitive Ca2+ channels or reversal of Na+-Ca2+ exchange, (3) these pathways are either less active or less abundant in proximal tubules of newborn compared with adult rabbits, and (4) secondary active transport activity and cellular integrity are well preserved after anoxia in PCT cells of newborn but not of adult rabbits.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s004670050171
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