The effect of PGF2 alpha on in vivo cerebral arteriolar diameter in cats and rats

We compared the effect of topical application of PGF2 alpha on cerebral arterioles in cats and rats equipped with an acutely implanted cranial window. Arterial diameter was measured using a microscope and image splitting device. PGF2 alpha in a concentration ranging from 10(-7) to 10(-5) M had no effect on large (greater than or equal to 100 microns) or small (less than 100 microns) cat pial arterioles, but induced a dose dependent constriction of rat pial arterioles with a maximum constriction to 76% of control diameter. Dilation of cat large cerebral arterioles by topically applied PGE2 was not affected by simultaneous application of PGF2 alpha and PGE2 induced dilation of small arterioles was decreased 3% by PGF2 alpha. While we and others have previously shown that both cat and rat brain can synthesize PGF2 alpha, it appears that PGF2 alpha is not likely to normally be a major modulator of cerebral arteriolar resistance in all species.     

Effect of 15-HETE on cerebral arterioles of newborn pigs

Effects of topical application of 15-HETE on pial arteriolar diameter and cortical perirachnoid cerebrospinal fluid (CSF) prostanoid concentrations were investigated in chloralose-anesthetized newborn pigs. Pial arteriolar diameters were measured using a closed cranial window, and CSF samples from under the window were collected for prostanoid analysis after applying artificial CSF without drug and CSF containing 15-HETE (1, 10, 100, 1000 ng/ml). 15-HETE caused significant dose-related constriction from 162 ± 17.0 μm (control diameter) to 136 ± 14.5 and 129 ± 18.7 μm (100 and 1000 ng/ml, respectively). The concentration of PGE₂ (but not of PGF_2α or 6-keto-PGF_1α increased in CSF at 100 and 1000 ng/ml of 15-HETE. Pial arteriolar responses to 15-HETE were determined before and after indomethacin treatment (5 mg/kg, i.v.). 15-HETE (100 ng/ml) constricted pial arterioles before indomethacin (diameter change, −15 ± 10%); after indomethacin, constriction was potentiated in response to the same dose (diameter change, −26 ± 7%). These data support the hypothesis thet, in newborn piglets, 15-HETE exerts a vasoconstrictor effect on pial arterioles, which appears to be attenuated by 15-HETE-induced stimulation of dilator prostanoids.     

Prostaglandin levels in isolated brain microvessels and in normal and norepinephrine-stimulated cat brain homogenates

The levels of PGD₂, PGE₂, PGF_2α and 6-keto-PGF_1α (6KF_1α) produced from endogenous archidonic acid (AA) were quantitated in cat cerebral cortical homogenates and microvessels isolated from cat cerebral cortex using gas chromatography/mass spectrometry (GC/MS). There was a six-fold enrichment of 6KF_1α levels in isolated microvessels, compared to homogenates, suggesting that 6KF_1α is of vascular, rather than neuronal origin. In order to further understand any possible role that norepinephrine (NE)_might have on modulation of PG synthesis, we studied the effects of 0.5 mM NE on PG synthesis from endogenous AA and from ³H-PGG₂, the endoperoxide precursor of PGs. In cat cortical homogenates NE induced a 74% increase in PGD₂ and PGF_2α, a 62% increase in PGE₂, and a 36% increase in 6KF_1α, as measured by GC/MS. NE caused a twofold increase in the conversion of ³H-PGG₂ to ³h-PGG_2α, with a concomitant decrease in ³H-PGE₂ and ³H-6KF_1α formation, and no change in ³H-PGD₂ synthesis. NE had no effect on the total conversiob of ³H-PGG₂ to ³H-PGs, nor on the breakdown of ³H-PGG₂ in the absence of brain tissue. We conclude that NE stimulates extravascular synthesis of PGD₂, PGE₂ and PGF_2α by stimulation of the prostaglandin synthetase complex, in addition to NE's stimulatory effect on the conversion of PGG₂ to PGF_2α, and that the lack of effect of NE on 6KF_1α synthesis reflects either a failure to achieve an adequate concentration at the vascular tissue, or an absence of the mechanism whereby NE stimulates PG synthetase.     

Bone resorption and prostaglandin production by mouse calvaria in vitro: Response to exogenous prostaglandins and their precursor fatty acids

Mouse calvaria were maintained in organ culture for 96 h and endogenous prostaglandin production and active bone resorption (⁴⁵ Ca release) measured. After a lag phase of 12 h, active resorption increased over the 96 h period. The amounts of prostaglandins released into the culture medium (measured by radioimmunoassay) were highest in the first 24 h of culture. Unless these were removed by preculturing for 24 h, or suppressed by indomethacin, no response to exogenous PGE₂, PGF_2α or prostaglandin precursors could be demonstrated. Bone resorption was stimulated after preculture by both PGE₂ and PGF_2α in a dose-dependent manner (10^?18M – 10^?5M), with PGE₂ being the more potent. Collagen synthesis was unaffected by PGF_2α, whereas PGE₂ (10^?5M) had an inhibitory effect. Eicosatrienoic acid did not stimulate bone resorption at lower concentrations (10^?7M – 10^?5M_, but was inhibitory at 10^?4M. Arachidonic acid also inhibited resorption at 10^?4M, but at lower concentrations (10^?7M – 10^?5M0 increased active resorption. This was concomitant with a rise in PGE₂ and PGF_2α levels, PGE₂ production being significantly higher than PGF_2α. The effects of PGE₂ (10^?8M) and PGF_2α (10^∞M appeared additive: there was no evidence of synergistic or antagonistic effects when varying ratios of PGE₂ : PGF_2α were employed.     

Effects of cerebral ventricular, systemic, and local administration of prostaglandin F2a on canine cerebral hemodynamics

The effects of Prostaglandin F_2a (PGF_2a) on cerebral blood flow, cerebral vascular resistance, and cerebrospinal and systemic arterial pressures were determined in anesthetized dogs. Flow was measured from the cannulated sinus confluens after occlusion of the transverse canals. Infusion of 1 to 100 ug/ml of PGF_2a into the cerebral ventricular system did not affect cerebral venous outflow but increased cerebral vascular resistance, arterial blood pressure, and cerebrospinal fluid pressure at the higher concentrations. Systemic, intra-aortic arch infusion of PGF_2a from 50 to 200 ug/min decreased cerebral venous outflow and increased cerebral vascular resistance slightly. Bilateral, intra-carotid artery infusion of PGF_2a at 20 to 80 ug/min produced effects similar in magnitude and direction to systemic, intra-aortic infusion. PGF_2a appears to increase cerebral vascular resistance by active vasomotion, dependent upon the route of administration. However, the magnitude of this constriction is not great considering the dose used. Also, PGF_2a can increase systemic arterial blood pressure via a central effect.     

Regional and species differences in endogenous prostaglandin biosynthesis by brain homogenates

Endogenously formed prostaglandins (PGs) D₂, E₂ and F_2α were determined in homogenates of brain regions from rat, guinea-pig, rabbit and cat, using gas-chromatography-mass spectrometry. The main PGs formed in the brain regions of the rat were PGD₂, in the guinea-pig PGD₂ and PGF_2α, in the rabbit PGF_2α and in the cat PGE₂. Brain regions from the same animal species showed the same pattern of PG formation. They varied, however, in the amount of total PGs formed, the limbic system and the cerebral cortex being highest and cerebellum lowest.     

Sodium-potassium-ATPase electrogenicity in cerebral precapillary arterioles

Electrogenicity of the Na(+)/K(+) pump has the capability to generate a large negative membrane potential independently of ion-channel current. The high background membrane resistance of arterioles may make them susceptible to such an effect. Pump current was detected by patch-clamp recording from smooth muscle cells in fragments of arterioles (diameter 24-58 microm) isolated from pial membrane of rabbit cerebral cortex. The current was 20 pA at -60 mV, and the extrapolated zero current potential was -160 mV. Two methods of estimating the effect of pump electrogenicity on resting potential indicated an average contribution of -35 mV. In 20% of the recordings, block of inward rectifier K(+) channels by 10-100 microM Ba(2+) led to a small depolarization, but hyperpolarization was a more common response. Ba(2+) also inhibited depolarization evoked by 20 mM K(+). In arterioles within intact pial membrane, Ba(2+) failed to evoke constriction but inhibited K(+)-induced constriction. The data suggest that cerebral arterioles are vulnerable to the hyperpolarizing effect of the Na(+)/K(+) pump, excessive effects of which are prevented by depolarizing inward rectifier K(+) current     

Prostaglandin modulation of the mechanism of ACTH action in the human adrenal.

PGE₁ and PGE₂ significantly increased human adrenal cAMP levels $\text{in}$$\text{vitro}$; cortisol output was also increased in a dose related fashion. In contrast, PGF_1a and PGF_2a depressed adrenal cAMP (except PGF_2a at 100 μg/ml). PGF_1a and PGF_2a depressed cortisol levels at all doses. Indomethacin or 7-oxa-13-prostynoic acid did not affect these parameters. However, when applied in conjunction with ACTH they inhibited or enhanced hormonal action depending upon the temporal sequence of application. The findings indicate that prostaglandins modulate ACTH-adrenocortical cell interaction bidirectionally, initially potentiating and subsequently depressing ACTH stimulated events.     

The presence of discrete receptors for prostaglandin F2 alpha in the cell membranes of bovine corpora lutea.

The cell membranes isolated from bovine corpora lutea bound ³H-prostaglandin (PG) F₂α with high affinity and specificity. The specific binding of ³H-PGF₂α was detectable at 10^?10M added ³H-PGF₂α and reached saturation at 10^?7M to 10^?6M. Unlabeled PGF₂α, as low as 10^?9M, inhibited the binding of ³H-PGF₂α with complete inhibition occurring at 10^?6M. The Scatchard analysis of equilibrium binding data revealed that the PGF₂α receptors are heterogeneous: Kd₁?5.1 × 10^?9M, n?289 fmoles/mg protein; Kd₂?1.8 × 10^?8M, n?780 fmoles/mg protein. The relative affinities of various other PGs for binding to PGF₂α receptors were (PGF₂α?100%): PGF₁α?17.5; PGE₁?0.8; PGE₂?22.4; PGA₁?0.007; PGB₁?0.01. The specificity and affinity of ³H-PGF₂α binding is consistent with the possibility that this receptor interaction may reflect an initial event in the action of PGF₂α as a luteolytic agent.     

Alteration in the reactivity of hamster cheek pouch arterioles to prostaglandin E2 and noradrenaline during pregnancy or sex steroid treatment

Dioestrous and pregnant, or ovariectomized hamsters treated with sunflower oil, oestradiol or progesterone were anesthetized with pentobarbital and the arterioles of the cheek pouch membrane were prepared for microcirculatory study. Prostaglandin E₂ (PGE₂) or noradrenaline (NA) were topically applied and changes of the arteriolar diameter were measured on the screen of a closed-circuit television.PGE₂ induced arteriolar dilatation in dioestrous or ovariectomized hamsters, but induced vasoconstriction in pregnant or in oestrogen-treated animals. Vasoconstriction induced by PGE₂ in oestrogen-treated animals disappeared after administration of alpha-adrenergic blockers. In this situation, PGE₂ induced vasodilatation once again. NA elicited arteriolar constriction in each experimental group. In ovariectomized hamsters treated with oestrogen the constriction was more pronounced than in progesterone treated animals. In pregnant animals it was significantly greater on day 14 than in dioestrous animals.Progesterone treatment blunted the vascular effect of both PGE₂ and NA.It was concluded that the reverse effects of PGE₂ and the increased sensitivity to NA induced by high oestradiol levels may have roles in the regulation of local blood flow during the ovarian cycle and pregnancy.     

Influence of prostaglandins (PG) E2 and F2α on the inflammatory process

PGF_2^α, but not PGE₂, induces a slight pedal edema when given alone. Both compounds were equipotent in the carrageenin-induced rat paw edema. Locally administered, PGE₂ and PGF_2^α did not exacerbate, but rather inhibited inflammations induced by various agents such as 1% carrageenin or 1% egg white. The administration of PGE₂ directly into cotton pellets or into the rat's hind paw in combination with M. butyricum significantly inhibited, respectively, granuloma formation and the polyarthritis. Subcutaneously, both prostaglandins inhibited the adjuvant induced polyarthritis. Neither PGE₂ nor PGF_2^α inhibited the anti-edema properties of non-steroidal or steroidal anti-inflammatory standards. A greater anti-edema activity was observed with the combination treatment than with the anti-inflammatory standards alone. We were unable to decrease the anti-inflammatory activity of the steroidal and non-steroidal standards or increase the inflammatory potential of the phlogistic agents.     

Cigarette smoke ventilation decreases prostaglandin inactivation in rat and hamster lungs

The effects of cigarette smoke on the metabolism of exogenous PGE₂ and PGF_2α were investigated in isolated rat and hamster lungs. When isolated lungs from animals were ventilated with cigarette smoke during pulmonary infusion of 100 nmol of PGE₂ or PGF_2α, the amounts of the 15-keto-metabolites in the perfusion effluent were decreased. Pre-exposure of animals to cigarette smoke daily for 3 weeks did not change the metabolism of PGE₂ when the lungs were ventilated with air. Cigarette smoke ventilation of lungs from pre-exposed animals caused, however, a similar decrease in the metabolism of PGE₂ as in animals not previously exposed to smoke. After pulmonary injection of 10 nmol of ¹⁴C-PGE₂ the radioactivity appeared more rapidly in the effluent during cigarette smoke ventilation suggesting inhibition of the PGE₂ uptake mechanism. In rat lungs pulmonary vascular pressor responses to PGE₂ and PGF_2α were inhibited by smoke ventilation.     

Prostaglandins in rainbow trout (Oncorhynchus mykiss Walbaum, 1792) sperm biology – searching for answers

The purpose of this study was to determine the concentrations of prostaglandins E₂ and F_2α (PGE₂ and PGF_2α) in the blood, testis and seminal plasma of mature male rainbow trout and in the ovarian fluid to assess the effects of these prostaglandins on sperm motility parameters when present in activation media. Also prolonged incubation with prostaglandins on sperm motility and calcium influx were studied. The profile of PGE₂ and PGF_2α differed in concentration between blood, testicular supernatant and seminal plasma. PGE₂ was predominant in the blood sample (0.29 ng ml^?1) and testicular supernatant (3.1 ng ml^?1) whereas their level in seminal plasma was lower than PGF_2α (0.23 ng ml^?1). The concentrations of PGF_2α in blood, testis and seminal plasma were 0.04, 0.99, 1.3 ng ml^?1, respectively. In the ovarian fluid the concentrations of both prostaglandins were higher than in the male reproductive tract. Adding both prostaglandins to activation buffer (at concentrations 15 and 70 ng ml^?1) had no effect on any CASA parameters. Calcium influx related to rainbow trout sperm incubations with PGE₂, and PGF_2α was not detected. After 24 h incubation of sperm in artificial seminal plasma solution without and with prostaglandins all sperm samples increased their motility potential and intracellular calcium concentration. Therefore, this effect was not related to the presence of prostaglandins. In summary PGE₂, and PGF_2α were present in the rainbow trout male reproductive tract, and their profile varies from that of blood, testis and seminal plasma. The specific role of both prostaglandins in salmonid sperm biology remains unclear.     

Stimulatory effect of prostaglandin E1 on thyroliberin-induced α-melanotropin release from perifused neuro-intermediate lobes of frog pituitary gland

A possible direct effect of prostaglandins on α-melanotropin (α-MSH) release at the level of the intermediate lobe of the frog pituitary was investigated $\text{in vitro}$ using a perifusion system technique. The effect of prostaglandins was studied on both spontaneous and TRH-stimulated α-MSH secretion. No significant effect of PGE₁, PGE₂, PGF_1α or PGF_2α on basal release of α-MSH could be detected. Indomethacin did not alter the α-MSH release induced by TRH. Conversely a significant increase in TRH-induced α-MSH secretion was observed in the presence of 1 x 10^?6M PGE₁. This magnifying effect was directly related to the concentration of TRH for doses ranging from 1 x 10^?8M to 1 x 10^?6M.     

Prostaglandin endoperoxides IV. Effects on smooth muscle

The effect on smooth muscle of the endoperoxides PGG₂ and PGH₂, which are intermediates in prostaglandin biosynthesis, was studied in different systems $\text{in vitro}$ and $\text{in vivo}$. On gastrointestinal smooth muscle (gerbil colon, rat stomach) PGG₂ and PGH₂ produced contractions comparable to those of PGE₂ and PGF_2a whereas contractions elicited on vascular (rabbit aorta) and airway (guinea-pig trachea) smooth muscle were considerably greater than those of PGE₂ and PGF_2a respectively. On intravenous injection into guinea-pigs PGG₂ and PGH₂ caused a triphasic change in blood pressure and were 8–10 times more effective than PGF_2a in producing an increase in tracheal insufflation pressure. When given as aerosols the unstable endoperoxides were less effective than PGF_2a. It is concluded that the endoperoxides are potent smooth muscle stimulants and that they are more effective than their degradation products (PGD₂, PGE₂, PGF_2a) in some systems.     

The effects of prostaglandins E2 and F2α on synaptosomal accumulation and release of 3H-norepinephrine

Prostaglandins (PG) of both the E and F series may serve as modulators of norepinephrine (NE) release from peripheral sympathetic neurons. We have studied the effects of PGE₂ and PGF_2α on the accumulation and release of ³H-NE in the CNS using synaptosomes isolated from rat hypothalami.The release of ³H-NE from synaptosomes superfused with Krebs-Ringer bicarbonate buffer was multiphasic with an initial fast release phase followed by a slower release. Raising KC1 concentration of the superfusion medium to 56mM during the slow release phase is known to stimulate ³H-NE release. PGE₂ (1 × 10^?6M) attenuated ³H-NE release during the fast phase and reduced the amount of ³H-NE released due to KC1 stimulation. At lower concentrations of PGE₂ there was no change in the release profile. PGF_2α was without effect on ³H-NE release at all concentrations tested.The accumulation of ³H-NE was significantly diminished by PGE₂ at a concentration of 1 × 10^?6M, while a lower concentration (1 × 10^?7M) was ineffective. PGF_2α had no effect on ³H-NE accumulation at all concentrations investigated.     

Production of prostaglandins by cells in vitro: radioimmunoassay measurement of the conversion of arachidonic acid to PGE2 and PGF2 alpha

A specific radioimmunoassay has been applied to the measurement of the conversion of arachidonic acid to PGE₂ and PGF_2α. PGE₂ and PGF_2α biosynthesis was linearly related to the amount of arachidonic acid added and was significantly inhibited by indomethacin in concentrations as low as 10^?10 M. Sonicated Hela, L, and HEp-2 cells synthesized 244.0, 42.3, and 22.6 ng PGE₂ per mg of protein, but made substantially less PGF_2α.     

Different responsiveness of a variety of isolated dog arteries to prostaglandin D3

The addition of prostaglandin (PG) D₂ contracted helical strips of dog cerebral, coronary, renal and femoral arteries; the contraction was greatest in cerebral arteries. The contractile response of cerebral arteries was potentiated by aspirin and attenuated by polyphloretin phosphate. In the arterial strips contracted with PGF_2α, PGD₂ elicited a concetration-related relaxation; the relaxation was greatest in mesenteric arteries. In mesenteric arterial strips contracted with norepinephrine, a lesser degree of relaxation was induced, and in the K⁺-contracted arteries, only a contraction was induced. Treatment with PGD₂ attenuated the contractile responses of cerebral and mesentric arteries to PGF_2α or PGE₂; this inhibitory effect was approximately 10 times greater in mesenteric arteries. However, the response to serotonin (for cerebral arteries) or norepinephrine (for mesenteric) was unaffected. It may be concluded that the heterogeneity of response to PGD₂ of a variety of dog arteries is due to different contributions of vasoconstrictor and vasodilator mechanisms. PGD₂ appears top share the mechanism underlying arterial contraction with PGF_2α and PGE₂, and interferes with the effect of these PG's possibly on receptor sites.     

Cerebral endothelial cell culture II. Adenylate cyclase response to prostaglandins and their interaction with the adrenergic system

The response of endothelial adenylate cyclase (AC) to prostaglandins (PGE₁, PGE₂, PGF_1α, PGF_2α, PGD₂ and PGI₂) and the relationship of PGE₂ to adrenergic systems were investigated in cerebrovascular endothelial cultures. E-type prostaglandins and PGI₂ were more effective in stimulating endothelial AC (EC₅₀ = 3 × 10^?7M, and 3 × 10^?7M, respectively) than prostaglandins of the F-series and PGD₂ which activated AC at high doses only. A modulation of endothelial AC response to either PGE₂ or norepinephrine (NE) was observed in the presence of both agents in the system. It was manifested by a dose-dependent NE inhibition of the PGE₂-stimulated formation of cAMP, which was partially restored by phentolamine. Alpha and β-adrenergic agonists (α, clonidine and 6-fluoronorepinephrine; β, isoproterenol) also partly blocked while forskolin and PGE₂ synergistically stimulated the production of cAMP in the endothelial cultures. These findings strongly suggest that the interaction of prostaglandins and α- and β-adrenergic agonists with the AC system in cerebrovascular endothelium may play a role in the regulation of the cerebral microcirculation and/or blood pressure.     

Differential effects of detergents and dimethylsulfoxide on membrane prostaglandin E1 and F2alpha receptors.

Pretreatment of membranes for 1 hr at 4° with up to 0.1% Triton X-100 (TX-100) and sodium desoxycholate (SDC), resulted in a greater loss of [³H] prostaglandin (PG)F₂α binding compared to E₁ binding. Lubrol WX (LWX) tended to cause a greater loss of [³H]PGF₂α than E₁ binding. However, the differential loss was not as marked as with TX-100 or SDC. Triton X-305 was relatively ineffective, but loss of [³H]PGE₁ binding was greater than for PGF₂α. Increasing concentrations of dimethylsulfoxide (DMSO) progressively inhibited PGF₂α binding without affecting PGE₁ binding. The detergent, but not DMSO, induced losses of membrane PG binding were due to solubilization of the receptors. Greater amounts of membrane protein and phospholipids were solubilized at detergent (TX-100 and SDC) concentrations that solubilized 100% of PGE₁ receptors compared to 100% solubilization of F₂α receptors. Neither the duration of preincubation nor the amount of membrane protein chosen were responsible for differential PGE₁ and F₂α receptor losses. These differential membrane PG receptor losses raise the possibility of differences in PGE₁ and F₂α receptors association with the membrane structure.