NO inhibits Na<sup>+</sup>-K<sup>+</sup>-2Cl<sup>−</sup>cotransport via a cytochrome<i>P</i>-450-dependent pathway in renal epithelial cells (MMDD1)
Bibliographic record
Abstract
Nitric oxide (NO) exerts direct effects on nephron transport. We determined the effect of NO on Na(+)-K(+)-2Cl(-) cotransport in a cell line (MMDD1) with properties of macula densa. Na(+)-K(+)-2Cl(-) cotransport was measured as bumetanide-sensitive (86)Rb(+) uptake in the presence of ouabain. MMDD1 cells expressed mRNA for the neuronal isoform of nitric oxide synthase, as well as NKCC1 and NKCC2(B) isoforms of the Na(+)-K(+)-2Cl(-) cotransporter. Preincubation of cells with the NO donors sodium nitroprusside (SNP) or S-nitroso-N-acetylpenicillamine (SNAP) caused concentration-dependent inhibition of Na(+)-K(+)-2Cl(-) cotransport. Both apical and basolateral Na(+)-K(+)-2Cl(-) cotransport was inhibited by NO donors. SNP or SNAP had no significant effect on cellular levels of cGMP, cAMP, cytosolic calcium, or phosphorylation of ERK1 and ERK2. In contrast, the inhibitors of cytochrome P-450, 1-aminobenzotriazole (ABT; 10(-3) M) or ketoconazole (1.5 x 10(-5) M), completely reversed the inhibitory effect of SNAP on apical or basolateral Na(+)-K(+)-2Cl(-) cotransport [apical: control 1.18 +/- 0.15 vs. SNAP (10(-4) M) 0.41 +/- 0.05 pmol x mg(-1) x 5 min(-1); P < 0.001; SNAP (10(-4) M) + ABT 1.32 +/- 0.10 pmol x mg(-1) x 5 min(-1); P = not significant vs. control; n = 5]. The cytochrome P-450 epoxyeicosatrienoic acid (EET) metabolite 14,15-EET (5 x 10(-7) M) inhibited both apical and basolateral cotransport, whereas 8,9-EET and 11,12-EET had no significant effect. Although 20-hydroxyeicosatetraenoic acid inhibited apical cotransport, the inhibitor of omega-hydroxylase activity HET0016 did not reverse SNAP-mediated inhibition of apical cotransport. These data indicate that NO inhibits apical and basolateral Na(+)-K(+)-2Cl(-) cotransport in MMDD1 cells. The results suggest that the inhibitory pathway is independent of cGMP and might involve stimulation of a cytochrome P-450-dependent pathway.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.002 | 0.001 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.001 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".