Bulk isolation of renal PCT and PST. II. Differential responses to anoxia or hypoxia.
Innate biochemical responses of rabbit renal proximal convoluted (PCT) and straight (PST) segments following in vitro exposure to anoxia or hypoxia were investigated to delineate the mechanisms responsible for segment-selective injury in vivo. After bulk isolation, suspensions (1 mg/ml) enriched in either PCT or PST were preincubated in Dulbecco's modified Eagle's-Ham's F-12 medium for 1 h before being exposed to either 40 min of anoxia (N2) or 120 min of hypoxia (1% O2) and 1 h of recovery under air-CO2 conditions. After recovery from anoxia, the percent of control values for each viability indicator in PCT and PST, respectively, were as follows: O2 consumption (QO2), 30/50; ATP content, 22/49; K+ content, 60/70; and percent lactate dehydrogenase (LDH) release, 66/45. Likewise, following recovery from hypoxia, the percent of control values for PCT and PST, respectively, were as follows: QO2, 50/90; ATP, 16/57; K+, 52/79; LDH, 45/17. These differential responses indicate that PCT segments were innately more susceptible to anoxic and hypoxic injury than PST segments. Because ATP content was significantly higher in PST segments immediately after anoxia and hypoxia, we investigated glucose-dependent responses during anoxia by exposing these segments to 30 min of anoxia in nutrient buffer with or without glucose. Results from these experiments demonstrate that the PST protection from anoxia was glucose dependent because removal of glucose from the nutrient buffers during anoxia abolishes the differential responses between PCT and PST. The in vitro PCT sensitivity observed here contrasts with the PST sensitivity observed following in vivo ischemia, suggesting that hemodynamic factors present in vivo may ultimately determine the overall susceptibility of PST segments in situ.
Ruegg
C E
CE
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710.
Mandel
L J
LJ
D000227:Adenine Nucleotides / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D005260:Female; D005947:Glucose / Q000378:metabolism / Q000502:physiology; D000860:Hypoxia / Q000378:metabolism / Q000503:physiopathology*; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D010101:Oxygen Consumption / Q000502:physiology; D011817:Rabbits
D000227:Adenine Nucleotides; D005947:Glucose