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F P Diecke

Publications and source records attributed to F P Diecke.

At least 19 recordsLinked to original sources
PubMed2001

Corneal endothelial NKCC: molecular identification, location, and contribution to fluid transport.

Although Na(+)-K(+)-2Cl(-) cotransport has been demonstrated in cultured bovine corneal endothelial cells, its presence and role in the native tissue have been disputed. Using RT-PCR we have now identified a partial clone of the cotransporter protein in freshly dissected as well as in cultured corneal endothelial and epithelial cells. The deduced amino acid sequence of this protein segment is 99% identical to that of the bovine isoform (bNKCC1). [(3)H]bumetanide binding shows that the cotransporter sites are located in the basolateral membrane region at a density of 1.6 pmol/mg of protein, close to that in lung epithelium. Immunocytochemistry confirms the basolateral location of the cotransporter. We calculate the turnover rate of the cotransporter to be 83 s(-1). Transendothelial fluid transport, determined from deepithelialized rabbit corneal thickness measurements, is partially inhibited (30%) by bumetanide in a dose-dependent manner. Our results demonstrate that Na(+)-K(+)-2Cl(-) cotransporters are present in the basolateral domain of freshly dissected bovine corneal endothelial cells and contribute to fluid transport across corneal endothelial preparations.

Amino Acid Sequence↗
PubMed2001

Immunocytochemical localization of aquaporin-1 in bovine corneal endothelial cells and keratocytes.

For immunocytochemistry, cultured bovine corneal endothelial cells (CBCEC) and bovine corneal cryosections were utilized. Preparations were fixed, permeabilized, and incubated with primary rabbit anti-rat aquaporin 1 (AQP1) antibody followed by rhodamine-conjugated secondary antibody, and were counter-stained with Sytox nuclear acid stain. Confocal microscopy of CBCEC in the x, y, and z planes showed rhodamine fluorescence, indicating the presence of AQP1 antibody localized to the apical and basolateral domains of the plasma membrane, but not to the membranes of intracellular compartments or other subcellular locations. Preabsorption with control antigenic peptide yielded no positive staining. Similar results were obtained using freshly dissected bovine corneas; in addition, these images showed AQP1 distributed to the plasma membranes of keratocytes. No AQP1 staining was seen in corneal epithelium, and no staining was observed in CBCEC layers exposed to AQP3, AQP4, and AQP5 antibodies.

Animals↗
PubMed1999

Transport of fluid by lens epithelium.

We report for the first time that cultured lens epithelial cell layers and rabbit lenses in vitro transport fluid. Layers of the alphaTN4 mouse cell line and bovine cell cultures were grown to confluence on permeable membrane inserts. Fluid movement across cultured layers and excised rabbit lenses was determined by volume clamp (37 degrees C). Cultured layers transported fluid from their basal to their apical sides against a pressure head of 3 cmH2O. Rates were (in microliter. h-1. cm-2) 3.3 +/- 0.3 for alphaTN4 cells (n = 27) and 4.7 +/- 1.0 for bovine layers (n = 6). Quinidine, a blocker of K+ channels, and p-chloromercuribenzenesulfonate and HgCl2, inhibitors of aquaporins, inhibited fluid transport. Rabbit lenses transported fluid from their anterior to their posterior sides against a 2.5-cmH2O pressure head at 10.3 +/- 0.62 microliter. h-1. lens-1 (n = 5) and along the same pressure head at 12.5 +/- 1.1 microliter. h-1. lens-1 (n = 6). We calculate that this flow could wash the lens extracellular space by convection about once every 2 h and therefore might contribute to lens homeostasis and transparency.

4-Chloromercuribenzenesulfonate↗
PubMed1998

Sodium, potassium, two chloride cotransport in corneal endothelium: characterization and possible role in volume regulation and fluid transport.

PURPOSE: To search for membrane transporter proteins that could contribute to volume regulation and fluid transport by corneal endothelium. As an initial step, the authors have focused on Na+-K+-2Cl- cotransporters. METHODS: Bovine corneal endothelial cells were cultured to confluence. 86Rubidium was used as a tracer for K+ uptake determinations; uptake values were normalized per milligram of cell protein. RESULTS: Three components of K+ uptake were characterized: ouabain (1 mM) sensitive, bumetanide (0.1 mM) sensitive, and ouabain-bumetanide insensitive. Both the ouabain-sensitive and bumetanide-sensitive components increased in the presence of 26.2 mM HCO3-; 0.5 mM 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid abolished this increase. The bumetanide-sensitive component was completely inhibited in the absence of Na+ or Cl-. This component was increased 33% by a 33% hypertonic solution and was decreased 38% by a 33% hypotonic solution. The protein kinase C activator phorbol 12-myristate 13-acetate decreased the activity of the cotransporter, whereas forskolin, in the presence of isobutylmethylxanthine, decreased it. Calyculin A (100 nM), an inhibitor of phosphatases 1 and 2a, produced a large (97%) activation of this component. CONCLUSIONS: These results provided for the first time conclusive evidence for the presence of a Na+-K+-2Cl- cotransporter in corneal endothelium and of its possible involvement in volume-regulatory processes in these cells. Given the uptake values reported here, such cotransporter could contribute significantly to electrolyte transport and hence to fluid transport across this preparation.

1-Methyl-3-isobutylxanthine↗
PubMed1997

A mechanism for regulatory volume decrease in cultured lens epithelial cells.

PURPOSE: To identify mechanisms contributing to regulatory volume decrease in lens epithelial cells. METHODS: Cells of the lens epithelial cell line alpha TN4 were cultured in four-well culture dishes in Dulbecco's Modified Eagle Medium containing 10% fetal bovine serum. After confluence cell water space was determined by measuring the equilibrium distribution of 3-O-methylglucose. Potassium influx and efflux in isotonic and hypotonic solutions were measured using 86rubidium (86Rb) as tracer. Total cell potassium and sodium content were determined with atomic absorption spectroscopy. Protein content per well was assayed with a modified Lowry assay and flux data and ion concentrations were normalized per mg of protein. RESULTS: Lens epithelial cells responded to hypotonic solutions with rapid swelling followed by regulatory volume decrease (RVD). During swelling and subsequent volume decrease the unidirectional Rb efflux was increased proportionaly to the osmotic challenge. Rubidium efflux was highly sensitive to changes in extracellular osmolarity and responded with a measurable activation to changes of 12.5 mOsm. No changes in 86Rb influx were observed with small changes (< 20%) in osmolarity and only relatively small changes occurred with larger changes in osmolarity. The resulting net loss of 86Rb and potassium (K+) was demonstrated by measuring the change of intracellular [K+] in hypotonic solutions using atomic absorption spectroscopy. The K(+)-channel blockers quinine-HCl and BaCl2 and the Cl(-)-channel blockers diphenyl-2-carboxylate (DPC) and 5-nitro-2-(3-phenyl propylamino) benzoic acid (NPPB) did not significantly affect the 86Rb efflux induced by hypotonic solutions. However, [(dihydroindenyl)oxy]alkanoic acid (DIOA), reported to be a specific inhibitor of the K-Cl cotransporter, inhibited the activation of 86Rb efflux. 86Rb efflux could be activated in isosmotic solutions by the addition of 1 mM N-ethylmaleimide (NEM). This activation of Rb efflux could be prevented by the addition of 1 mM dithiothreitol and could be 90% blocked by DIOA. The activation of rubidium efflux by NEM led to a significant decrease of the intracellular water content. The volume regulatory changes in NEM and in hypotonic solutions could be inhibited in DIOA. CONCLUSIONS: The observations are consistent with the presence in lens epithelial cells of a K-Cl cotransporter serving as a mechanism for regulatory volume decrease.

3-O-Methylglucose↗
PubMed1997

Comparison of the effects of Zopolrestat and Sorbinil on lens myo-inositol influx.

The effects of two structurally dissimilar aldose reductase inhibitors, Zopolrestat and Sorbinil, were investigated on the sodium-dependent, myo-inositol (MI) cotransporter in rat lenses maintained in either normal (5.5 mmol/l) or high sugar medium (35.5 mmol/l glucose or 30 mmol/l galactose). MI influx was compared to the lens polyol content. The effects of Sorbinil (10, 20 and 40 mumol/l) were determined on normal lens MI influx. At all concentrations, Sorbinil had no effect on normal MI influx; therefore, there was no direct effect on the MI transporter. Acute exposure (4-hour incubation) in either high D- or L-glucose media significantly inhibited lens MI influx, which was attributed to competitive inhibition by either D- or L-glucose with MI cotransporter. Due to the short incubation period and rapid metabolism of D-glucose to fructose, there was a low level of polyol (sorbitol) in these lenses. Thus, concomitant administration of Sorbinil (10, 20 and 40 mumol/l) had no significant effect on MI influx in this short-term experiment. Sorbinil had no effect in the presence of L-glucose because L-glucose was not metabolized; thus the polyol content remained normal. To investigate the effects of large accumulations of polyol, lenses were preincubated for 8, 12 and 16 h in 30 mmol/l galactose medium. Large amounts of polyol (galactitol) rapidly accumulated because galactitol was not metabolized. Galactose served as substrate for aldose reductase, and lens polyol (galactitol) content increased markedly. Inhibition of MI influx directly correlated with the increased lens polyol content. Lens polyol accumulation resulted in noncompetitive inhibition of MI influx. Coadministration of 40 mumol/l Sorbinil inhibited 80% of polyol formation and protected 80% of MI influx. Furthermore, in the presence of Sorbinil, lens galactose increased rapidly and equilibrated with galactose in the medium further indicating that Sorbinil inhibited aldose reductase. The effects of 40 mumol/l Sorbinil were compared to 40 mumol/l Zopolrestat. Zopolrestat was as effective as Sorbinil; both aldose reductase inhibitors maintained MI influx at approximately 80% of control values after 12- and 16-hour incubations in high galactose medium. In conclusion, Sorbinil did not exert a direct effect on the sodium-dependent, MI cotransport system or prevent the direct competitive inhibition of either D- or L-glucose. Sorbinil and Zopolrestat inhibited lens polyol formation, thereby eliminating noncompetitive inhibition of MI influx.

Aldehyde Reductase↗
PubMed1997

Effect of pyruvate on lens myo-inositol transport and polyol formation in diabetic cataract.

In diabetic cataract, sorbitol pathway flux perturbs intracellular metabolism by two putative mechanisms. The osmolyte hypothesis implicates the aldose reductase enzyme, increased rate of reduction of glucose of sorbitol and reciprocal osmoregulatory depletion of organic osmolytes (myo-inositol). Redox hypothesis favors alterations in the ratios (NADP+/NADPH and/or NADH/NAD+ as the primary cause of glucose-induced aldose reductase related defects. Increase in NADH/NAD+ promotes increased oxidation of sorbitol to fructose by polyol dehydrogenase; potential normalization of this ratio by coadministration of pyruvate (which reoxidizes NADH to NAD+ via lactate dehydrogenases reaction) was investigated. Effects of exogenous pyruvate on lens polyol formation and sodium-dependent myo-inositol (MI) cotransporter using two in vitro models of sugar cataract were determined. Rat lenses were incubated for 16 h in either normal (5.5 mM) or high sugar medium, 35.5 mM glucose or 30 mM galactose. Then lens MI influx was compared to polyol, MI and fructose content. Pyruvate did not affect MI influx or sorbitol content in lenses incubated in control medium. In 35.5 mM glucose, coadministration of pyruvate maintained lens MI influx at 76% of control values vs. 43% for lenses without pyruvate. Furthermore, pyruvate treatment diminished lens sorbitol content by 50% and increased lens sugar content (myo-inositol, fructose, lactate) and media lactate levels. Lenses incubated in high galactose medium formed galactitol with a corresponding decreased MI content. Coadministration of pyruvate had no effect on either lens sugar content (galactitol, myo-inositol, fructose) or MI influx, consistent with the fact that galactitol was not metabolized to fructose. In conclusion, pyruvate did not exert a direct effect on the MI co-transporter or prevent galactitol inhibition of MI influx. Coadministration of pyruvate with high glucose altered lens metabolism and promoted reduction of pyruvate to lactate, increased fructose, decreased sorbitol, enhanced MI influx, maintained lens MI content, implicating both osmotic and redox systems.

Animals↗
PubMed1996

Force development with inosine triphosphate and uridine triphosphate in chemically skinned vascular smooth muscle.

The contraction of vascular smooth muscle is thought to be regulated by reversible phosphorylation of the 20,000 dalton light chains of myosin, catalyzed by myosin light chain kinase that is dependent on calcium and calmodulin. With phosphorylation, there is a coincident increase in the actin-activated myosin NTPase activity, cross bridge interaction and contractile activity. However, this myosin phosphorylation mechanism may not be the sole factor controlling actin-myosin interaction in vascular smooth muscle. Other mechanisms may function in addition to this myosin-linked regulation. A calcium-insensitive regulation of contraction was observed in helical strips of chemically skinned (Triton X-100) arterial smooth muscle. Millimolar concentrations of inosine triphosphate and uridine triphosphate supported concentration dependent force development in the absence of calcium. Force development was a function of the MgNTP concentration. At high free calcium concentrations, an additional component of force was observed. ITP and UTP, in contrast to ATP, are less effective substrates for the myosin light chain kinase, and their effect on actin-myosin interaction is thus less than that of ATP. They are, however, utilized by the myosin NTPase after treatment by ATP-gamma-S. The efficacy of the substrate for the activated NTPase is greater for UTP than ITP than for ATP.

Acid Anhydride Hydrolases↗
PubMed1996

Zopolrestat prevention of proteinuria, albuminuria and cataractogenesis in diabetes mellitus.

The aldose reductase inhibitor, Zopolrestat, reduced proteinuria and albuminuria in streptozocin-induced diabetic rats compared with both untreated diabetic and age-matched controls. Daily administration of Zopolrestat (100 mg/kg) for 4 months decreased 24 h total protein excretion to 15.07 +/- 2.17 mg from 49.97 +/- 7.94 mg/day in untreated diabetic rats. Zopolrestat protected against excretion of any array of urinary proteins with molecular weights between 30 and 100 kD. These effects were sustained throughout the 5th and 6th months of treatment. At the end of 6 months, Zopolrestat-treated diabetic rats excreted 22.77 +/- 4.39 mg/day compared to untreated diabetic rats (67.05 +/- 14.03 mg/day), a 6-fold increase in urinary protein excretion compared to age-matched nondiabetic controls (11.65 +/- 1.71 mg/day). Zopolrestat treatment for 6 months produced therapeutic effects in the lens: transparency and myo-inositol content were maintained and lens sorbitol diminished, despite elevated lens glucose. In contrast, untreated diabetic rats had opaque lenses which exhibited a 40-fold increase in sorbitol and myo-inositol depletion. In opaque lenses, ouabain-sensitive Rb influx, an index of Na-K-ATPase activity, decreased to only 53.8% of mean values in age-matched controls; the ouabain-insensitive component increased by 63.6%. Zopolrestat treatment prevented these diabetic-induced changes and maintained ouabain-sensitive and ouabain-insensitive Rb influx. Collectively, these results suggest that Zopolrestat exerts a protective effect on the slowly developing diabetic cataract, as well as reducing albuminuria and proteinuria.

Albuminuria↗
PubMed1995

Kinetics of myo-inositol transport in rat ocular lens.

Myo-inositol (MI) influx as a function of concentration in rat lens consisted of a saturable component, fit by a rectangular hyperbola, and a linear component which was more distinct at high myo-inositol concentrations suggesting passive diffusion. The hyperbolic component was half-maximally saturated (Kt) at 61.3 microM and had a maximal transport rate (Jmax) of 44.6 mumol/kg wet wt/h. The linear component had an apparent permeability coefficient of 1.44 x 10(-6) s-1. Sorbitol, which distributed rapidly in the extracellular space (6.83 ml/100 g wet wt), also appeared to enter the intracellular space with a permeability coefficient of 1.37 x 10(-6) s-1, similar to that of myo-inositol. The influx of myo-inositol was critically dependent on the concentration of extracellular sodium consistent with a sodium-myo-inositol cotransport. The kinetics of influx activation by sodium suggested an apparent 2:1 coupling ratio for sodium and myo-inositol. When potassium was used as sodium substitute, a significantly stronger influx inhibition was observed than with nondepolarizing sodium substitutes, indicating that myo-inositol was driven by the electrochemical gradient of sodium rather than the chemical gradient only. Reducing the extracellular Na concentration increased the MI concentration at which transport was half-maximally activated, suggesting an ordered binding sequence of Na followed by MI. Myo-inositol influx was competitively inhibited by phlorizin with an inhibitory coefficient (Ki) of 35 microM. Phloretin also was capable of inhibition but with a much lesser efficacy. Myo-inositol desaturates from the lens at a rate of 0.00862 h-1. Approximately 19% of the efflux can be inhibited with phlorizin, suggesting that it represents carrier-mediated flux. The phlorizin insensitive flux has a rate of 0.00695 h-1 or 1.93 x 10(-6) s-1, similar to the Na-independent passive influx. MI influx is due to a Na-dependent, phlorizin-sensitive active transport while the efflux consists largely of a phlorizin-independent passive leakage.

Animals↗
PubMed1993

Mechanisms for D-glucose inhibition of myo-inositol influx into rat lens.

Myo-Inositol depletion as a result of hyperglycemia is considered one of the leading contributors to chronic diabetic complications. We investigated the possible mechanisms through which elevated extracellular glucose levels affect the loss of intracellular myo-inositol in rat lens. Short-term incubation (up to 4 h) in solutions with elevated glucose concentrations revealed a concentration-dependent inhibition of myo-inositol influx. This inhibition was caused by both an increase of the transport coefficient and a decrease of maximal flux and thus was a mixed competitive and noncompetitive inhibition. If polyol accumulation was prevented with sorbinil, an aldose reductase inhibitor, the inhibition of myo-inositol influx was partially reduced. The remaining inhibition was the result of an increased transport coefficient without a change in maximal flux and therefore represents a strictly competitive inhibition. A similar competitive inhibition was observed with the nonmetabolizable glucose analogue L-glucose, which cannot be converted to polyol. Longer exposure (16 h) to solutions with high glucose concentrations resulted in an inhibition that correlated with high lens polyol levels. This inhibition persisted after the lenses were returned to solutions with normal glucose concentrations and was the result of a decrease of maximal flux without a significant change in transport coefficient, a strictly noncompetitive inhibition. The noncompetitive inhibition associated with polyol accumulation and the competitive inhibition due to extracellular glucose were additive. Lens myo-inositol depletion after exposure to elevated glucose concentrations thus resulted from a competitive inhibition caused by the interaction of extracellular glucose with the myo-inositol carrier and a noncompetitive inhibition associated with polyol accumulation.

3-O-Methylglucose↗
PubMed1992

Myo-inositol transport in the lens of galactose-maintained rats.

Lens myo-inositol (MI) content is regulated by a pump-leak system consisting of an active Na-dependent MI transport and its passive permeability through the membrane. We measured the active MI uptake and membrane permeability in lenses of rats maintained on a 50% galactose diet for 1, 3 and 7 days. After only 1 day of galactose feeding, active MI uptake in the lens was reduced dramatically by 74% compared to age-matched control lenses; by day 3, active MI transport was decreased by 89% and it was undetectable by day 7. The passive membrane permeability was determined by measuring (a) the passive MI influx and (b) the 3H-sorbitol flux. After 1 day of galactose feeding, the membrane permeability increased such that within 3 days it increased to 5-6 fold. Galactose feeding also led to a rapid increase in lens polyol content. After 1 day, lens polyol increased to 53 mumol/g wet wt compared to a control value of 0.35 mumol/g wet wt and increased further to 65 and 72 mumol/g wet wt after 3 and 7 days of galactose feeding respectively. Lens galactose accumulation was low (3 mumol/g wet wt) up to 7 days; however, it was rapidly increased after 7 days. Our results indicate that galactose feeding rapidly interfered with MI homeostasis by a severe depression of active MI transport and a rapid increase in membrane permeability. These interferences of MI homeostasis correlate with the appearance of high polyol levels.

Animals↗
PubMed1991

Cyclic GMP modulates release of norepinephrine from adrenergic nerves innervating canine arteries.

Evidence is presented that compounds which stimulate the soluble form of the enzyme guanylate cyclase or which inhibit the enzyme cGMP phosphodiesterase (PDE), responsible for the degradation of cGMP (including endothelium-derived relaxing factor) are inhibitors of sympathetic neurotransmission to vascular smooth muscle and inhibit the efflux of norepinephrine from sympathetic nerves. Moreover, prostacyclin, papaverine, iloprost, and forskolin, compounds which stimulate the enzyme adenylate cyclase, and rolipram (neural specific) and milrinone, enoximone, and piroximone (muscle specific) inhibitors of Type III cAMP PDE and degradation of cAMP, do not inhibit nerve stimulation to most blood vessels. The data support the concept that cGMP may act as a negative feedback modulator of physiologic frequencies of sympathetic nerve activity to blood vessels. cAMP does not appear to modulate adrenergic neurotransmission to vascular smooth muscle at physiologic frequencies of neural stimulation.

Acetylcholine↗
PubMed1990

Inhibition of sympathetic neurotransmitter release by modulators of cyclic GMP in canine vascular smooth muscle.

The contractile response to neurally released norepinephrine (NE) from sympathetic nerve endings innervating vascular smooth muscle are inhibited by substances which raise either cyclic AMP and cyclic GMP concentrations in smooth muscle. However, cyclic AMP is believed to facilitate NE release from sympathetic nerves whereas the role of cyclic GMP in this process is undefined. We examined the effects of presumed modulation of the intraneuronal concentration of cyclic AMP and cyclic GMP on sympathetic neurotransmission to isolated canine mesenteric artery by measurement of the efflux of [2-14C]NE during transmural nerve stimulation (calcium dependent release of NE) and administration of tyramine (calcium independent release of NE) and measurement of the contractions to exogenous NE and tyramine. Stimulation of adenylate cyclase with forskolin, prostacyclin and iloprost, a stable prostacyclin analog, and inhibition of Type III cyclic AMP phosphodiesterase with neural specific rolipram, 'non-specific pelrinone and milrinone and isobutylmethylxanthine did not enhance the efflux of [2-14C]NE from sympathetic nerves innervating the blood vessels. Isoproterenol enhanced the efflux of [2-14C]NE. The effect was inhibited by propranolol but not affected by milrinone, amrinone or rolipram. Activators of guanylate cyclase (SIN-1a an active metabolic of molsidomine, nitroglycerin and sodium nitroprusside) and inhibitors of Type II cyclic GMP phosphodiesterase (M&B-22948 and verofyllin) inhibited the efflux of NE released by transmural nerve stimulation but not by tyramine. These data support the conclusion that cyclic GMP may be an inhibitory modulator of calcium and depolarization dependent NE release from sympathetic nerves, whereas neuronal cyclic AMP may not be a primary modulator of neurotransmission to vascular smooth muscle.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1990

Release of norepinephrine from adrenergic nerve endings of blood vessels is modulated by endothelium-derived relaxing factor.

The release of norepinephrine from adrenergic nerve endings is inhibited by substances which raise cyclic 3',5'-guanosine monophosphate (cGMP) in neural tissue. Endothelium-derived relaxing factor (EDRF) elevates cGMP in vascular smooth muscle. Thus, EDRF may also modulate the release of norepinephrine (NE) from adrenergic nerves. We tested this postulate in isolated canine pulmonary arteries and veins using the technique of superfusion and measurement of the efflux of radiolabeled NE during transmural nerve stimulation at 1, 2, 4, 8, 16 and 32 Hz for 10 min. In segments of artery and vein with intact endothelium the contractile responses to low frequency nerve stimulation were decreased when compared to endothelium rubbed blood vessels. Electrical stimulation of arteries and veins with intact endothelium for 10 min released less 2-[14C]-NE than rubbed blood vessels, especially at the lower frequencies of 1, 2 and 4 Hz, with lesser effects at frequencies of 16 and 32 Hz. Using the technique of bioassay, EDRF from porcine thoracic aorta inhibited the efflux of 2-[14C]-NE from the pulmonary artery and vein. The findings support the conclusion that the endothelium can inhibit release of NE from sympathetic nerve innervating canine pulmonary artery and vein. The endothelium, in part through EDRF, can act as an endogenous inhibitor or sympathetic neurotransmitter release.

Acetylcholine↗
PubMed1989

The endothelium modulates adrenergic neurotransmission to canine pulmonary arteries and veins.

We tested the postulate that endothelium-derived relaxing factor (EDRF) modulates adrenergic neuroeffector transmission in isolated canine pulmonary arteries and veins, using the technique of superfusion and measurement of the efflux of [2-14C]NE during transmural nerve stimulation at 1, 2, 4, 8, 16 and 32 Hz for 10 and 30 min. In endothelium-rubbed artery and vein the contractile responses to low frequency nerve stimulation were enhanced, when compared to those from endothelium rubbed blood vessels. Transmural nerve stimulation of endothelium competent arteries and veins for 10 min released less [2-14C]NE than denuded arteries and veins, especially at 1, 2 and 4 Hz, with smaller differences evident at higher frequencies (16 and 32 Hz) of stimulation. Superfusion of endothelium rubbed blood vessels with effluent from canine thoracic aorta decreased the release of [2-14C]NE during nerve stimulation. These findings suggest that the endothelium and EDRF can inhibit release of adrenergic neurotransmitter from canine pulmonary arteries and veins. The endothelium may act as an endogenous modulator of adrenergic neurotransmission to canine vascular smooth muscle.

Acetylcholine↗