Biomedical subjects
Publications and source records attributed to V A Alder.
We have successfully measured the intravitreal and intraretinal oxygen distribution in normal rats, during both normoxia and hyperoxia. The feasibility of such measurements in the very small rat eye has opened up the opportunity for the use of the many rat models of vascular disease in physiological experiments of retinal oxygen supply and consumption.
The mechanical, histological and pharmacological properties of the isolated cat ophthalmociliary artery are reported and compared for passively stretched ring segments from three locations on the artery. Ring segments were mounted in a modified myograph system after the method of Hogestatt, Andersson and Edvinsson (1983, Acta Physiol Scand. 117, 49-61) and comparative length tension curves were measured. The most distal segment (DOA), nearest the eye, developed by far the smallest maximal tension compared to the middle segment (MOA) and the most proximal segment (POA), despite the fact that lumen diameters were similar. All segments displayed a phasic and tonic component in response to passive stretching and during activation with K(+)-Krebs. Cumulative contractile dose-response curves were measured for nine agonists: histamine (HIS), 5-hydroxytryptamine (5-HT), dopamine (DOPA), adrenaline (A), noradrenaline (NA), tyramine (TYR), phenylephrine (PHE), isoproterenol (ISOP) and xylazine (XYL) and the maximum active tension developed at a concentration of 10(-3) M was compared with that for 0.124 M K(+)-Krebs. The order of contraction magnitude in the three segments was: POA, HIS much greater than Kmax much greater than NA greater than A greater than DOPA greater than PH = TYR greater than 5-HT = ISOP greater than XYL; MOA, Kmax much greater than NA much greater than A = HIS much greater than DOPA greater than ISOP = PHE greater than TYR = 5-HT greater than XYL; DOA, Kmax much greater than NA greater than A greater than HIS = DOPA greater than ISOP greater than PHE = 5-HT greater than XYL = TYR. Pre-activation did not produce relaxation in response to histamine and isoproterenol. Testing with specific alpha 1 and beta antagonists supported the presence of alpha 1 receptors but not beta receptors. It is concluded that functional HIS, alpha 1 and 5-HT receptors are present and that the ophthalmociliary artery is heterogeneous in its response to HIS, with the POA segment producing the largest responses. The magnitude of the contractile responses to A and NA grew with increasing passive tension, whereas the response to histamine was independent of passive tension.
This study investigates the ability of an alpha 1-adrenergic blocker, tolazoline, two Ca2+ entry channel blockers, verapamil and diltiazem, acetylcholine and Ca(2+)-free solutions to relax alpha 1-adrenergically activated cat ophthalmociliary artery ring segments. The form of a contraction elicited with adrenaline, noradrenaline or phenylephrine was shown to depend on the passive tension set. As passive tension was increased, which is equivalent to increasing blood pressure in vivo, the alpha 1-adrenergic elicited contraction became more sustained. Incubation with the Ca2+ channel blockers diltiazem and verapamil was ineffective in reducing the phasic response to the alpha 1-adrenergic agonists whereas addition of these blockers in increasing concentrations during a sustained adrenergic contraction caused a graded and significant relaxation. This implies that therapeutic use of Ca2+ channel blockers may act directly on the ophthalmociliary artery to inhibit the autonomic control of sustained tone in this vessel. Removal of exogenous Ca2+ from the bathing medium initially reduced the phasic component and eliminated the tonic component of the alpha 1-adrenergic contraction. Repeated exposure of the ring segment to Ca(2+)-free medium further reduced the phasic component elicited with alpha 1-adrenergic contraction. Pre-incubation with the alpha 1-adrenergic blocker Tolazoline reduced the alpha 1-adrenergic antagonist phasic response only at high concentrations (10(-4) M), whereas Tolazoline applied during the tonic phase was an effective relaxant at lower concentrations. Acetylcholine added to the bath during an alpha 1-adrenergic contraction induced relaxation at low concentrations and contraction for higher concentrations. The relaxation was eliminated by removal of the endothelial cell layer. These data support the notion that the phasic component of an alpha 1-adrenergic contraction is mainly dependent on intracellular Ca2+ stores whereas the tonic component relies almost exclusively on extracellular Ca2+. It is suggested that Ca2+ channel blockers and alpha 1-adrenergic antagonists may potentially play a role in relaxing the ophthalmociliary artery to improve the ocular circulation in vascular diseases in which there is considerable vascular tone present.
Hematologic values are compared for normal and streptozotocin-induced diabetic rats after 6 weeks of induced diabetes. Most hematologic parameters were the same in the two groups except for blood glucose, glycated hemoglobin, and 2,3 diphosphoglycerate, all of which were elevated in the streptozotocin group. However the P50 (the PO2 at which the oxygen-carrying capacity of blood is 50% of maximal) remained normal. We hypothesize that a left shift in the oxyhemoglobin dissociation curve caused by the glycation of a small percentage of the hemoglobin is compensated by elevation in the 2,3-diphosphoglycerate which returns the P50 to normal values. This compensatory mechanism also occurs in some stages of human diabetes.
The isometric responses of isolated human posterior ciliary artery to adrenergic agonists, histamine (HIS), and 5-hydroxytryptamine (5-HT) were studied in passively stretched ring segments mounted in a myograph bath. Cumulative dose response curves were measured for nine agonists: HIS, 5-HT, dopamine (DOPA), epinephrine (A), norepinephrine (NA), tyramine (TYR), phenylephrine (PHE), isoproterenol (ISOP), and xylazine (XYL), and the log(molar concentration) at which one half of the maximum active tension was developed (EC50) was estimated. The ring segments were unresponsive to DOPA and XYL; HIS and ISOP produced biphasic responses with a mild relaxation for low concentrations and small contractions for high concentrations of the agonist. The remaining agonists caused contractile responses of magnitude listed in the rank order following compared with the maximum active tension in response to 0.124 M K(+)-Krebs: Kmax much greater than A greater than 5-HT = PHE greater than NA greater than TYR It was concluded that functional HIS, alpha 1-adrenergic, and 5-HT receptors were present on human posterior ciliary artery but that there are no alpha 2-adrenergic receptors.
Oxygen-sensitive microelectrodes were used to measure the intraretinal oxygen distribution in the rat during both air and oxygen breathing. During air breathing the average PO2 profile obtained showed a value of 19 +/- 2.5 mmHg (n = 36) at the internal limiting membrane (ILM). As the penetration depth increased, a minimal PO2 of 8.2 +/- 2.4 mmHg was attained at 50% of the retinal depth, from which point the PO2 increased to a peak of 33 +/- 3.3 mmHg at 100% of the retinal depth at Bruch's membrane. During oxygen ventilation, all intraretinal oxygen tensions were increased and the profile became almost monotonic, increasing from 36.1 +/- 3.3 mmHg (n = 9) at the ILM to 134.3 +/- 6.7 mmHg at 100% of the retinal depth. These results are qualitatively similar to those found in the cat, and they demonstrate the importance of oxygen delivery by both the retinal and the choroidal circulation during normoxia, as well as the inability of vascular autoregulation to maintain a constant retinal PO2 during hyperoxic ventilation.
Measurements of vitreal oxygen tension have been made for the first time in the streptozotocin-induced diabetic rat eye. A total of 36 Sprague Dawley rats were divided into a control (n = 18) and streptozotocin injected group (n = 18), and after 5-6 weeks of established hyperglycaemia, an acute experiment was performed in which vitreal oxygen tension profiles were determined with oxygen sensitive microelectrodes. The control rats had significant oxygen tension gradients in the vitreous close to retinal arteries with relatively flat oxygen tension profiles close to retinal veins and intermediate regions. All control rats had a substantial arteriovenous oxygen tension difference when measurements were made on retinal arteries and veins. In contrast the oxygen tension profiles measured in the vitreous of streptozotocin rats showed markedly reduced oxygen gradients in the vicinity of retinal arteries and a smaller arteriovenous oxygen tension difference. In both groups of rats, for distances of 500 microns and greater from the retina (mid vitreous) a plateau oxygen tension value was observed. No significant difference was found in this mean mid vitreous value between the control rats and diabetic rats under the same systemic conditions. We conclude that there are significant changes in oxygen tension near retinal arteries in streptozotocin-induced diabetes before any histopathological changes are evident.
Oxygen tension measurements are presented for the first time for the vitreous body of the rat eye. Oxygen-sensitive microelectrodes were used to measure vitreal PO2 profiles for two breathing conditions: 20 and 100% O2. Results show that for ventilation with 20% O2 most of the vitreous body has a constant PO2 (mid vitreous PO2 of 22.6 +/- 1.9 mmHg (+/- S.E.; n = 6), except for a layer about 200 microns thick at the retinal surface where significant PO2 gradients exist, especially in the region of retinal arteries. Midvitreous PO2 is similar to the PO2 at retinal veins. All vitreal PO2 values increase if ventilatory oxygen is raised to 100% O2. After equilibration, the main body of the vitreous again has an almost constant PO2, a factor of 3.75 +/- 0.40 (n = 10), higher than for 20% O2 ventilation except for a region about 400 microns thick adjacent to the internal limiting membrane where the local geometry of the retinal circulation dictates PO2 distribution. When the electrode was oriented to make contact with the retina at different distances from the optic disk, venous PO2 was observed to decrease in more peripheral locations. This work opens the possibility of investigating the role of hypoxia in rat models of retinal vascular disease.
Intraocular measurements of blood flow have been made in the rat eye using hydrogen-clearance polarography. Hydrogen was delivered by a bolus injection of hydrogen-saturated saline via a cannula in the lingual artery of the anesthetized rat. An intraocularly placed hydrogen-sensitive microelectrode monitored the arrival and clearance of hydrogen preretinally adjacent to retinal arteries, veins, and intervascular areas. The effect of stepwise penetration through the retina and into the choroid was also determined, and a map of hydrogen distribution in the retina during hydrogen delivery and clearance was produced. Intraocular measurements, which have the advantage that the electrode location can be clearly visualized, together with computer control of the bolus injection and data collection, have allowed highly repeatable measurements of local blood flow to be made. The mean blood flow in retinal tissue was 373 +/- 51 ml.min-1.100 g-1 (values are means +/- SD, n = 32). The correlation between the nature of the hydrogen clearance curve and the location of the electrode helps explain data from other studies in which the position of the electrode with respect to the vasculature was unknown.
The form and magnitude of K+ Krebs-induced contractures has been measured for the dog ophthalmociliary artery. Ring segments of the vessel were mounted in a myograph after the method of Hogestatt, Andersson and Edvinsson (1983, Acta Physiol. Scand. 117, 49-61). For low passive tensions the K(-)-induced contractures show both a phasic and tonic component, with the tonic tension component being the smaller. As passive tension is increased, the phasic component grows and plateaus whilst the tonic component continues to increase. For high passive tensions the phasic component disappears and the tonic component reduces in magnitude and the contracture is not fully reversible. Comparison of the length tension curves measured during K+ Krebs-induced contractures and Ca2(+)-free Krebs-induced relaxation shows that the active tension generated by this vessel increases with increasing passive tension until a peak active tension of 3.1 mN mm-1 is reached at an effective radius of 357 microns. For greater values of passive tension the active tension decreases. The calculated transmural pressure required to maintain the vessel at this optimal radius is 51.9 mmHg, which is very similar to the only available measurements of ophthalmic artery pressure, viz. 51.8 mmHg (Grunwald, Sinclair and Riva, 1981, Invest. Ophthalmol. Vis. Sci. 20, 564-6). The optimal passive stretch to apply to these ring segments for further pharmacological or physiological work is discussed.
The effect of graded systemic hyperoxia on vitreal PO2 distribution has been determined for the rat eye. Oxygen tension profiles were measured, using oxygen-sensitive microelectrodes, as a function of distance from the internal limiting membrane as the inspired oxygen percentage was increased in 10% steps from 20-100%. Depending on the original touching location of the microelectrode on the retina, there could be substantial PO2 gradients within 500 microns of the retina; at greater distances vitreal PO2 was constant and a function of the inspired oxygen percentage. Whatever the location of the microelectrode in the vitreous, PO2 rose with increasing hyperoxia. The relationship between vitreal PO2 and inspired oxygen was nonlinear with a central relatively flat region between 50-80% inspired oxygen. The ratio between vitreal PO2 during 100% O2 breathing and air breathing was 3.42 +/- 1.08 (standard deviation, n = 7). Possible explanations for the plateau region are the maintenance of a relatively constant PO2 by vascular autoregulation and/or the buffering of capillary PO2 by hemoglobin. The rat eye, therefore, responds to hyperoxia similarly to that of the cat and monkey but differs from that of the miniature pig where there is no rise in preretinal PO2 during hyperoxia.
There is a close correlation between the sound pressure of tone burst required to affect a primary auditory neuron at its characteristic frequency and that which will produce a detectable N1 response at the same frequency. Units with thresholds from 80--0 db SPL (recorded from damaged and undamaged cochleas) were 0--20 dB , respectively, more sensitive than the action potential response.
A new approach to the guinea pig auditory nerve is presented. This approach, which has several advantages over previous methods, enables stable recordings from primary nerve fibers to be make. Entry to the auditory nerve is through the modiolus via scala tympani of the basal turn of the cochlea.