scholarly journals THE EFFECTS OF CAFFEINE ON OXYGEN CONSUMPTION AND CELL DIVISION IN THE FERTILIZED EGG OF THE SEA URCHIN, ARBACIA PUNCTULATA

1945 ◽  
Vol 29 (2) ◽  
pp. 63-72 ◽  
Author(s):  
Ralph Holt Cheney

1. By means of the Warburg-Barcroft microrespirometer apparatus and the Warburg direct method, the relative effect of caffeine upon the O2 consumption of the fertilized egg of Arbacia punctulata was shown for the following concentrations in sea water: 0.002 per cent (M/10,000), 0.004 per cent (M/5,000), 0.02 per cent (M/1,000), 0.1 per cent (M/200), 0.2 per cent (M/100), 0.5 per cent (M/40), and 2 per cent (M/10). 2. In comparison with the normal eggs (uninhibited, non-caffeine-treated controls), caffeine in concentrations including and greater than 0.1 per cent (M/200) depressed the average uptake from approximately 25 to 61 per cent over the 3 hour period. In a number of instances, as typified by Experiment 10, the effective inhibitory concentration ranged from 0.02 per cent (M/1,000) upward and the degree of depression of the O2 consumption ranged from 10.6 per cent to 60.6 per cent. 3. All caffeine concentrations including and above 0.02 per cent (M/1,000) in the series used, resulted in decreasing the normal rate of cleavage division in the fertilized Arbacia eggs. 4. The higher concentrations (0.5 and 2 per cent) produced a complete blockage of the cleavage process. 5. Complete cleavage inhibition was noted only when the O2 uptake had been depressed to 50 per cent or more of the normal controls. 6. O2 consumption-time relationship data indicate an average depression, in O2 consumption over a 3 hour period, ranging from 25 per cent with a caffeine concentration of 0.1 per cent to a 61 per cent inhibition with a concentration of 2 per cent. 7. Concentrations of less than 0.1 per cent (certainly of less than 0.02 per cent) give variable results and indicate no significant effect. 8. It is inferred from the respiration data presented that it is probable that the inhibition of the O2 consumption in fertilized Arbacia eggs is due to the influence of caffeine upon the main (activity or primary) pathway. It will be observed that there are certain similarities of the caffeine data to the degree of inhibition accomplished by sodium cyanide. Moreover, it has been demonstrated that the cyanide probably acts on the cytochrome oxidase step in the cytochrome oxidase-cytochrome chain of reactions constituting the O2 uptake phase of respiratory metabolism. It is not improbable, therefore, that caffeine also may act upon the cytochrome oxidase enzyme. 9. From the viewpoint of environmental conditions influencing reproductive phenomena, it is of interest that caffeine can affect the normal metabolism of the zygote.

1961 ◽  
Vol 38 (2) ◽  
pp. 249-257
Author(s):  
H. MOHRI ◽  
K. HORIUCHI

1. The respiratory quotient of sea-urchin spermatozoa has been determined with Pseudocentrotus depressus, Anthocidaris crassispina and Hemicentrotus pulcherrimus. 2. The R.Q. of sea-urchin spermatozoa, measured by the Warburg direct method, has been reported to be near unity. This was also the case with the present material when the suspending medium was sea water, the R.Q. being 0.8-1.0. It was found, however, that the pH of sperm suspensions was markedly different in the presence and absence of alkali to absorb C02. 3. When the pH of the suspension was fixed by such buffers as 0.025 M-glycyl glycine, the R.Q. measured by the above method was about 0.7. This is in accord with the results of earlier metabolic studies, which indicated that endogenous phospholipids are the main substrates for the respiration of sea-urchin spermatozoa. 4. The O2 uptake of the present material, however, was found to be little affected by variation in pH. The difference in the R.Q. values obtained with ordinary sea water and buffered sea water, therefore, cannot be explained in terms of pH. 5. When the spermatozoa were suspended in ordinary sea water, the utilization of endogenous phospholipids was much reduced in the absence of alkali, while in buffered sea water the change in phospholipids was almost the same, with and without the absorption of CO2. 6. Determination of the R.Q. by the first method of Dickens & Šimer, in which the O2 uptake and the CO2 output were measured with one and the same sperm suspension, gave a value of about 0.7 with both ordinary and buffered sea water.


1948 ◽  
Vol 25 (4) ◽  
pp. 353-368
Author(s):  
LORD ROTHSCHILD

1. An investigation has been made into the causes of the interrelated phenomena of Senescence and the Dilution Effect in the spermatozoa of Echinus esculentus. The Dilution Effect is the increase in O2 uptake and in movement observed when a concentrated sperm suspension, obtained by partial dilution of semen, is diluted with sea water. 2. The supernatant sea water obtained by centrifuging partially metabolized sperm suspensions contains no substances which depress the respiration or motility of the spermatozoa of the same species. This supernatant sea water has a small but definite protective action on spermatozoa in maintaining but not increasing their O2 consumption. The effect is enhanced by agitating the suspensions before centrifugation. 3. E. esculentus spermatozoa do not glycolyse aerobically, anaerobically, or in the presence of 5% O2 in CO. The absence of aerobic acid production proves that the reduced O2 uptake per sperm of dense suspensions is not due to a lowering of the pH of the medium. 4. As the Dilution Effect is observed in manometric experiments in which CO2 is continuously and adequately removed, neither accumulation of CO2 nor a reduction in pH resulting from the accumulation of CO2 are responsible for the reduced O2 uptake of dense suspensions. 5. The O2 uptake of dense sperm suspensions (> 109 sperm/ml.) is altered by changes in O2 tension. O2 consumption in 100% oxygen is nearly double that in 10% O2 in N2. An increase in O2 tension has no stimulating action on dilute suspensions. 6. An atmosphere of 100% O2 has a gradual toxic effect on dense and dilute sperm suspensions. 7. Carbon monoxide has a greater inhibitory action on dilute than on concentrated sperm suspensions. 8. The difference between the effects of varying O2 tensions or CO on dense and dilute suspensions is partly, or perhaps wholly, due to gaseous diffusion being a limiting factor. This may partly explain the Dilution Effect. 9. Apart from photo-reversibly inhibiting respiration, CO exerts an irreversible toxic influence on sea-urchin spermatozoa. 10. Illumination of spermatozoa with a wave-length of light outside the region of photo-chemical absorption by CO cytochrome oxidase enables visual examination to be made in the presence of CO. The O2 uptake of a suitable concentration of spermatozoa is 70% inhibited by 5% O2 in CO. The same inhibition occurs when the suspension is illuminated by light of wave-length 548 mµ, but there is no equivalent decrease in motility. 11. After motility and respiration have ceased sea-urchin spermatozoa still contain oxidizable substrates and the appropriate dehydrogenases as evidenced by their ability to reduce methylene blue in an evacuated Thunberg tube.


1983 ◽  
Vol 245 (5) ◽  
pp. G697-G702 ◽  
Author(s):  
P. T. Nowicki ◽  
B. S. Stonestreet ◽  
N. B. Hansen ◽  
A. C. Yao ◽  
W. Oh

Regional and total gastrointestinal (GI) blood flow, O2 delivery, and whole-gut O2 extraction and O2 consumption were measured before and 30, 60, and 120 min after feeding in nonanesthetized, awake 2-day-old piglets. Cardiac output and blood flow to kidneys, heart, brain, and liver were also determined. Blood flow was measured using the radiolabeled microsphere technique. In the preprandial condition, total GI blood flow was 106 +/- 9 ml X min-1 X 100 g-1, while O2 extraction was 17.2 +/- 0.9% and O2 consumption was 1.99 +/- 0.19 ml O2 X min-1 X 100 g-1. Thirty minutes after slow gavage feeding with 30 ml/kg artificial pig milk, O2 delivery to the GI tract and O2 extraction rose significantly (P less than 0.05) by 35 +/- 2 and 33 +/- 2%, respectively. The increase in O2 delivery was effected by a significant increase in GI blood flow, which was localized to the mucosal-submucosal layer of the small intestine. O2 uptake by the GI tract increased 72 +/- 4% 30 min after feeding. Cardiac output and blood flow to non-GI organs did not change significantly with feeding, whereas arterial hepatic blood flow decreased significantly 60 and 120 min after feeding. The piglet GI tract thus meets the oxidative demands of digestion and absorption by increasing local blood flow and tissue O2 extraction.


1986 ◽  
Vol 235 (2) ◽  
pp. 337-342 ◽  
Author(s):  
D G Clark ◽  
M Brinkman ◽  
S D Neville

The effects of the sequential addition of glucose, noradrenaline, propranolol and oleic acid on the rates of O2 consumption and heat production by isolated interscapular brown adipocytes from control and cafeteria-fed rats were compared. Although the chemical agents produced very similar changes in oxidative metabolism, the actual rates of O2 uptake and heat output in adipocytes from the cafeteria-fed rats, when expressed per g dry wt. of cells, were approx. 65% less than those obtained with cells from the control rats. However, when the same results were expressed per 10(8) multiloccular brown adipocytes, rather than gravimetrically, rates of O2 consumption and heat production were equivalent. Further interpretation of these data is complicated, because the average volume of multiloccular brown adipocytes from cafeteria-fed rats was 2.5 times that for multiloccular cells from control animals.


1946 ◽  
Vol 29 (5) ◽  
pp. 267-275 ◽  
Author(s):  
Herbert Shapiro

A study was made of the diffusion of the red pigment echinochrome from the eggs of the sea urchin, Arbacia punctulata, into sea water. Unfertilized eggs retained their pigment, over periods of hours. Outward diffusion of pigment from unfertilized eggs normally is entirely negligible, or does not occur at all. Enchancing the calcium or potassium content of the artificial sea water (while retaining isosmotic conditions) did not induce pigment release. Under anaerobic conditions, unfertilized eggs release pigment in small quantities. Fertilization alone brings about echinochrome release. Fertilized eggs invariably released pigment, whether in normal sea water, or sea water with increased calcium or potassium. This diffusion of the pigment began during the first cleavage, possibly soon after fertilization. The pigment release is not a consequence solely of the cell's permeability to echinochrome (or chromoprotein, or other pigment combination) but is preceded by events leading to a release of echinochrome from the granules in which it is concentrated within the cell. These events may be initiated by activation or by anaerobiosis. The phenomenon was not due to cytolysis.


1984 ◽  
Vol 57 (2) ◽  
pp. 601-607 ◽  
Author(s):  
G. A. Ordway ◽  
D. L. Floyd ◽  
J. C. Longhurst ◽  
J. H. Mitchell

A description is given of a technique that provides a relatively simple means by which O2 consumption and hemodynamic variables can be measured in exercising dogs. We used a multistage submaximal treadmill test to study the responses of 10 foxhounds to dynamic exercise. They were also studied during maximal treadmill exercise. O2 consumption increased from 16.3 +/- 1.7 ml O2 X min-1 X kg-1 at rest to 92.9 +/- 9.7 ml O2 X min-1 X kg-1 at a work load of 6.4 km/h, 20% grade and to 111.9 +/- 9.6 ml O2 X min-1 X kg-1 during maximal exercise. Cardiac output (CO) increased from 6.11 +/- 0.45 l/min at rest to 16.91 +/- 1.46 and 17.66 +/- 0.60 l/min at 6.4 km/h, 20% grade and maximal exercise, respectively. Arteriovenous O2 difference increased from 5.8 +/- 0.3 vol% at rest to 12.0 +/- 0.4 and 13.2 +/- 0.7 vol% at 6.4 km/h, 20% grade and maximal exercise, respectively. Heart rate (HR) increased from 116 +/- 7 beats/min at rest to 250 +/- 8 beats/min at 6.4 km/h, 20% grade and to 278 +/- 6 beats/min during maximal exercise. O2 uptake, CO, and arteriovenous O2 difference increased with the onset of exercise, appeared to level at lower work intensities (6.4 km/h, 4 and 8% grade), and increased significantly at each of the higher work intensities (6.4 km/h, 12, 16, and 20% grade). Additionally, we observed linear relationships between O2 consumption and HR (HR = 1.35 X VO2 + 120.5; r = 0.87; P less than 0.001) and between O2 consumption and CO (CO = 5.91 X VO2 + 216.6; r = 0.96; P less than 0.001). Further, the linear relationship between O2 consumption and CO demonstrated in the present study is similar to that observed in humans.


1979 ◽  
Vol 57 (10) ◽  
pp. 1863-1865 ◽  
Author(s):  
Roger M. Evans

Seawater-adapted teleosts drink to offset water loss by osmosis. A direct method of monitoring drinking by implanting a fistula to drain the stomach indicated that rainbow trout began drinking from about 9 to 12 (range 1 to 22) h after being placed in 15‰ sea water. Unlike the Japanese eel (Anguilla japonica). in which the onset of drinking has been shown to be immediate and reflex-like, the onset of drinking in trout appears to occur only after appreciable water has been lost to the medium. The trout resembles the eel in that the capacity to shallow water in the absence of postingestional negative feedback exceeds the rate of drinking required to maintain normal water balance.


1985 ◽  
Vol 86 (1) ◽  
pp. 105-134 ◽  
Author(s):  
M Mahler ◽  
C Louy ◽  
E Homsher ◽  
A Peskoff

Previously we tested the validity of the one-dimensional diffusion equation for O2 in the excised frog sartorius muscle and used it to measure the diffusion coefficient (D) for O2 in this muscle and the time course of its rate of O2 consumption (Qo2) after a tetanus (Mahler, 1978, 1979, J. Gen. Physiol., 71:533-557, 559-580, 73:159-174). A transverse section of the frog sartorius is in fact well fit by a hemi-ellipse with width divided by maximum thickness averaging 5.1 +/- 0.2. Using the previous techniques with the two-dimensional diffusion equation and this hemi-elliptical boundary yields a value for D that is 30% smaller than reported previously; the revised values at 0, 10, and 22.8 degrees C are 6.2, 7.9, and 10.8 X 10(-6) cm2/s, respectively. After a tetanus at 20 degrees C, Qo2 rose quickly to a peak and then declined exponentially, with a time constant (tau) approximately 15% faster than that reported previously; tau averaged 2.1 min in Rana temporaria and 2.6 min in Rana pipiens. A technique was devised to measure the solubility (alpha) of O2 in intact, respiring muscles, and yielded alpha (muscle)/alpha (H2O) = 1.26 +/- 0.04. With these modifications, the values for O2 consumption obtained with the diffusion method were in agreement with those measured by the direct method of Kushmerick and Paul (1976, J. Physiol. [Lond.]., 254:693-709). Using results from both methods, at 20 degrees C the ratio of phosphorylcreatine split during a tetanus to O2 consumption during recovery ranged from 5.2 to 6.2 mumol/mumol, and postcontractile ATP hydrolysis was estimated to be 13.6 +/- 4.1 (n = 3) nmol/mumol total creatine.


1989 ◽  
Vol 67 (2) ◽  
pp. 871-878 ◽  
Author(s):  
M. Constantinopol ◽  
J. H. Jones ◽  
E. R. Weibel ◽  
C. R. Taylor ◽  
A. Lindholm ◽  
...  

Because the maximal rate of O2 consumption (VO2max) of the horse is 2.6 times larger than that of steers of equal size, we wondered whether their pulmonary gas exchanger is proportionately larger. Three Standardbred racehorses [body mass (Mb) = 447 kg] and three domestic steers (Mb = 474 kg) whose cardiovascular function at VO2max had been thoroughly studied (Jones et al. J. Appl. Physiol. 67: 862–870, 1989) were used to study their lungs by morphometry. The basic morphometric parameters were similar in both species. The nearly 2 times larger lung volumes of the horses caused the gas exchange surfaces and capillary blood volume to be 1.6 to 1.8 times larger. Morphometric pulmonary diffusing capacity was 2 times larger in the horse than in the steer; the 2.6-fold greater rate of O2 uptake thus required the alveolar-capillary PO2 difference to be 1.3 times larger in the horse than in the steer. Combining physiological and morphometric data, we calculated capillary transit time at VO2max to be 0.4–0.5 s. Bohr integration showed capillary blood to be equilibrated with alveolar air after 75 and 58% of transit time in horses and steers, respectively; horses maintain a smaller degree of redundancy in their pulmonary gas exchanger.


1936 ◽  
Vol 19 (4) ◽  
pp. 625-632 ◽  
Author(s):  
Kenneth S. Cole ◽  
Robert H. Cole

The alternating current resistance and capacity of suspensions of unfertilized and fertilized eggs of Arbacia punctulata have been measured at frequencies from 103 to 1.64 x 107 cycles per second. The unfertilized egg has a static plasma membrane capacity of 0.73 µf./cm.2 which is practically independent of frequency. The fertilized egg has a static membrane capacity of 3.1 µf./cm.2 at low frequencies which decreases to a value of 0.55 µf./cm.2 at high frequencies. The decrease follows closely the relaxation dispersion of the dielectric constant if the dissipation of such a system is ignored. It is considered more probable that the effect is due to a fertilization membrane of 3.1 µf./cm.2 capacity lifted 1.5 µ. from the plasma membrane, the interspace having the conductivity of sea water. The suspensions show a frequency-dependent capacity at low frequencies which may be attributable to surface conductance. The equivalent low frequency internal specific resistance of both the unfertilized and fertilized egg is about 186 ohm cm. or about 6 times that of sea water, while the high frequency data extrapolate to a value of about 4 times sea water. There is evidence at the highest frequencies that the current is penetrating the nucleus and other materials in the cytoplasm. If this effect were entirely due to the nucleus it would lead to a very approximate value of 0.1 µf./cm.2 for the capacity of the nuclear membrane. The measurements do not indicate any change in this effect on fertilization.


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