Prostaglandin synthesis by isolated cells from the outer medulla and from the thick ascending loop of Henle of rabbit kidney

D. Schlondorff, R. Zanger, J. A. Satriano, Vaughn Wesley Folkert, J. Eveloff

Research output: Contribution to journalArticle

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Abstract

Prostaglandins (PGs) synthesized in the medulla, a major source of renal PGs, could have a direct effect on several tubular functions. In order to better characterize the site and pattern of PG synthesis in different medullary nephron segments, we examined PG synthesis by isolated medullary cells, a preparation enriched in medullary collecting tubule cells and isolated cells from the thick ascending limb of Henle's loop (TALH). Incubation of medullary cells and medullary collecting tubule cells in the presence of 5 μM [14C]arachidonic acid produced predominantly PGE2 with much less PGF2(α). In comparison, TALH cells synthesized only about 20% as much labeled PG composed of nearly equal amounts of PGE2 and PGF2(α). Specific radioimmunoassay after incubation in the absence of exogenous arachidonic acid revealed that medullary cells synthesized 83 ± 15 ng of PGE2/mg of protein x 15 min compared with 15 ± 5 ng of PGE2 for TALH cells. Furosemide (1 mM) inhibited PGE2 synthesis in medullary cells but had no effect on TALH cells. Bumetanide (1 mM) and amiloride (1 mM) had no effect. The calcium ionophore A23187 (2 μM) doubled PGE2 synthesis in both cell populations. The synthesis of PGF2(α) was comparable in all three preparations, but was only 5% of PGE2 production in medullary and medullary collecting tubule cells and about 20% of PGE2 production in TALH cells. Furosemide (1 mM) stimulated PGF2(α) synthesis in medullary cells and TALH cells (2.9 ± 0.7 to 6.8 ± 1.3 and 1.5 ± 0.7 to 5.5 ± 1.4 ng/mg of protein x 15 min, respectively). Hydrochlorothiazide was without effect. A23187 (2 μM) increased PGF2(α) synthesis 2- to 3-fold in both cell populations. These results indicate that cells from different nephron segments have different patterns of PG synthesis. They do not support a direct role for PGs in the action of loop diuretics, but do not rule out possible modulation of NaCl transport by PGs.

Original languageEnglish (US)
Pages (from-to)120-124
Number of pages5
JournalJournal of Pharmacology and Experimental Therapeutics
Volume223
Issue number1
StatePublished - 1982
Externally publishedYes

Fingerprint

Loop of Henle
Prostaglandins
Rabbits
Kidney
Dinoprostone
Dinoprost
Nephrons
Furosemide
Calcimycin
Arachidonic Acid
Sodium Potassium Chloride Symporter Inhibitors
Bumetanide
Hydrochlorothiazide
Calcium Ionophores
Amiloride

ASJC Scopus subject areas

  • Pharmacology

Cite this

Prostaglandin synthesis by isolated cells from the outer medulla and from the thick ascending loop of Henle of rabbit kidney. / Schlondorff, D.; Zanger, R.; Satriano, J. A.; Folkert, Vaughn Wesley; Eveloff, J.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 223, No. 1, 1982, p. 120-124.

Research output: Contribution to journalArticle

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title = "Prostaglandin synthesis by isolated cells from the outer medulla and from the thick ascending loop of Henle of rabbit kidney",
abstract = "Prostaglandins (PGs) synthesized in the medulla, a major source of renal PGs, could have a direct effect on several tubular functions. In order to better characterize the site and pattern of PG synthesis in different medullary nephron segments, we examined PG synthesis by isolated medullary cells, a preparation enriched in medullary collecting tubule cells and isolated cells from the thick ascending limb of Henle's loop (TALH). Incubation of medullary cells and medullary collecting tubule cells in the presence of 5 μM [14C]arachidonic acid produced predominantly PGE2 with much less PGF2(α). In comparison, TALH cells synthesized only about 20{\%} as much labeled PG composed of nearly equal amounts of PGE2 and PGF2(α). Specific radioimmunoassay after incubation in the absence of exogenous arachidonic acid revealed that medullary cells synthesized 83 ± 15 ng of PGE2/mg of protein x 15 min compared with 15 ± 5 ng of PGE2 for TALH cells. Furosemide (1 mM) inhibited PGE2 synthesis in medullary cells but had no effect on TALH cells. Bumetanide (1 mM) and amiloride (1 mM) had no effect. The calcium ionophore A23187 (2 μM) doubled PGE2 synthesis in both cell populations. The synthesis of PGF2(α) was comparable in all three preparations, but was only 5{\%} of PGE2 production in medullary and medullary collecting tubule cells and about 20{\%} of PGE2 production in TALH cells. Furosemide (1 mM) stimulated PGF2(α) synthesis in medullary cells and TALH cells (2.9 ± 0.7 to 6.8 ± 1.3 and 1.5 ± 0.7 to 5.5 ± 1.4 ng/mg of protein x 15 min, respectively). Hydrochlorothiazide was without effect. A23187 (2 μM) increased PGF2(α) synthesis 2- to 3-fold in both cell populations. These results indicate that cells from different nephron segments have different patterns of PG synthesis. They do not support a direct role for PGs in the action of loop diuretics, but do not rule out possible modulation of NaCl transport by PGs.",
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T1 - Prostaglandin synthesis by isolated cells from the outer medulla and from the thick ascending loop of Henle of rabbit kidney

AU - Schlondorff, D.

AU - Zanger, R.

AU - Satriano, J. A.

AU - Folkert, Vaughn Wesley

AU - Eveloff, J.

PY - 1982

Y1 - 1982

N2 - Prostaglandins (PGs) synthesized in the medulla, a major source of renal PGs, could have a direct effect on several tubular functions. In order to better characterize the site and pattern of PG synthesis in different medullary nephron segments, we examined PG synthesis by isolated medullary cells, a preparation enriched in medullary collecting tubule cells and isolated cells from the thick ascending limb of Henle's loop (TALH). Incubation of medullary cells and medullary collecting tubule cells in the presence of 5 μM [14C]arachidonic acid produced predominantly PGE2 with much less PGF2(α). In comparison, TALH cells synthesized only about 20% as much labeled PG composed of nearly equal amounts of PGE2 and PGF2(α). Specific radioimmunoassay after incubation in the absence of exogenous arachidonic acid revealed that medullary cells synthesized 83 ± 15 ng of PGE2/mg of protein x 15 min compared with 15 ± 5 ng of PGE2 for TALH cells. Furosemide (1 mM) inhibited PGE2 synthesis in medullary cells but had no effect on TALH cells. Bumetanide (1 mM) and amiloride (1 mM) had no effect. The calcium ionophore A23187 (2 μM) doubled PGE2 synthesis in both cell populations. The synthesis of PGF2(α) was comparable in all three preparations, but was only 5% of PGE2 production in medullary and medullary collecting tubule cells and about 20% of PGE2 production in TALH cells. Furosemide (1 mM) stimulated PGF2(α) synthesis in medullary cells and TALH cells (2.9 ± 0.7 to 6.8 ± 1.3 and 1.5 ± 0.7 to 5.5 ± 1.4 ng/mg of protein x 15 min, respectively). Hydrochlorothiazide was without effect. A23187 (2 μM) increased PGF2(α) synthesis 2- to 3-fold in both cell populations. These results indicate that cells from different nephron segments have different patterns of PG synthesis. They do not support a direct role for PGs in the action of loop diuretics, but do not rule out possible modulation of NaCl transport by PGs.

AB - Prostaglandins (PGs) synthesized in the medulla, a major source of renal PGs, could have a direct effect on several tubular functions. In order to better characterize the site and pattern of PG synthesis in different medullary nephron segments, we examined PG synthesis by isolated medullary cells, a preparation enriched in medullary collecting tubule cells and isolated cells from the thick ascending limb of Henle's loop (TALH). Incubation of medullary cells and medullary collecting tubule cells in the presence of 5 μM [14C]arachidonic acid produced predominantly PGE2 with much less PGF2(α). In comparison, TALH cells synthesized only about 20% as much labeled PG composed of nearly equal amounts of PGE2 and PGF2(α). Specific radioimmunoassay after incubation in the absence of exogenous arachidonic acid revealed that medullary cells synthesized 83 ± 15 ng of PGE2/mg of protein x 15 min compared with 15 ± 5 ng of PGE2 for TALH cells. Furosemide (1 mM) inhibited PGE2 synthesis in medullary cells but had no effect on TALH cells. Bumetanide (1 mM) and amiloride (1 mM) had no effect. The calcium ionophore A23187 (2 μM) doubled PGE2 synthesis in both cell populations. The synthesis of PGF2(α) was comparable in all three preparations, but was only 5% of PGE2 production in medullary and medullary collecting tubule cells and about 20% of PGE2 production in TALH cells. Furosemide (1 mM) stimulated PGF2(α) synthesis in medullary cells and TALH cells (2.9 ± 0.7 to 6.8 ± 1.3 and 1.5 ± 0.7 to 5.5 ± 1.4 ng/mg of protein x 15 min, respectively). Hydrochlorothiazide was without effect. A23187 (2 μM) increased PGF2(α) synthesis 2- to 3-fold in both cell populations. These results indicate that cells from different nephron segments have different patterns of PG synthesis. They do not support a direct role for PGs in the action of loop diuretics, but do not rule out possible modulation of NaCl transport by PGs.

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