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Biomedical subjects

S D Wainwright

Publications and source records attributed to S D Wainwright.

At least 37 records · Page 2Linked to original sources
PubMed1989

On thymidine incorporation by the cultured chick pineal gland.

Explanted chick pineal glands exhibited a cycle in thymidine incorporation when cultured either under a cycle of illumination or in constant darkness. This cycle appeared to be entrained to the light cycle under which the birds were maintained. The incorporation reflected gene replication in a small fraction of the cell population that was largely, but not entirely, located in the stroma of the gland. Glands cultured with colchicine for 28 h contained a very small number of cells showing metaphase chromosomes.

Animals↗
PubMed1989

Rhythmic incorporation of thymidine by chick pineal glands in vitro.

Cultured chick pineal glands showed a cycle in the cumulative incorporation of thymidine into DNA. In undisturbed cultures the rate of thymidine incorporation, amount of thymidine incorporated per 1-day cycle, and persistence of the incorporation process were all markedly affected by the concentration of exogenous precursor. However, more than two full cycles of incorporation were found when culture medium of low thymidine content was renewed daily, or when medium of intermediate concentration was replaced on the 3rd day of culture. At a high thymidine concentration the second cycle of incorporation sometimes appeared to be impaired. At all concentrations tested, less than 2% of the available thymidine was incorporated.

Animals↗
PubMed1986

Effects of some inhibitors of DNA synthesis and repair upon the cycle of serotonin N-acetyltransferase activity in cultured chick pineal glands.

When chick pineal glands were cultured in the dark with aphidicolin from midphotoperiod, the increase of serotonin N-acetyltransferase (NAT) activity was stimulated and the time of peak NAT activity was advanced. The peak level of NAT activity was also reached sooner on the 2nd day of culture. The increase of NAT activity was also stimulated in glands cultured under diurnal illumination, but the time of peak activity was not advanced. Effects with glands explanted into culture in the dark at other times were smaller and the time of peak NAT activity was not changed. Cytosine arabinoside and dideoxythymidine also stimulated the increase of NAT activity and advanced the time of peak activity with glands cultured in the dark from midphotoperiod. 3-Aminobenzamide markedly stimulated the increase of NAT activity both in the dark and under diurnal lighting when pineal glands were explanted into culture at mid- or late photoperiod. In contrast, with glands in culture from earlier in the photoperiod, aminobenzamide had no effect upon the increase of NAT activity up to the peak level found with control glands. Thereafter results were variable. Effects of cordycepin upon development of NAT activity were similar to those of 3-aminobenzamide but less marked. Incorporation of thymidine into acid-insoluble material in the dark was very markedly inhibited by aphidicolin, cytosine arabinoside, and dideoxythymidine, but only slightly by cordycepin. Aminobenzamide strongly inhibited incorporation by glands cultured from midphotoperiod, but had little effect with glands in culture from near the end of the photoperiod. We adopt the working hypothesis that excision repair of DNA may be a major component in the mechanism of the chick pineal clock.

Acetyltransferases↗
PubMed1985

Interdependent effects of the ionophore A23187 and serum on the serotonin N-acetyltransferase activity of cultured chick pineal glands.

Marked effects of the ionophore A23187 on the cycle of N-acetyltransferase (NAT) activity in cultured chick pineal glands were observed under three conditions of illumination. However, the effects were qualitatively and quantitatively dependent on the batch of fetal calf serum used in the medium and time of explanation into culture. Ionophore increased the level of NAT activity remaining in glands exposed prematurely to light regardless of the serum used. The ionophore suppressed the "spike" in cyclic GMP content of glands cultured in the dark, and extended the period of maximum cyclic GMP content of glands under diurnal illumination.

Acetyltransferases↗
PubMed1985

Effects of some fatty acids on the serotonin N-acetyltransferase activity in cultured chick pineal glands.

Low concentrations of arachidonate, oleate, or palmitate significantly affected the cycle of NAT activity of cultured chick pineal glands in different ways. The effects observed were altered by change of the lighting conditions of culture. Effects of arachidonate were also shown to be altered by change of the serum component of the culture medium. Effects of premature exposure to light of glands cultured under diurnal conditions of illumination were changed markedly by substitution of the serum component of the medium or a supplement of ionophore A23187 and less markedly by supplements of fatty acids.

Acetyltransferases↗
PubMed1984

Relationship between cycles in level of serotonin N-acetyltransferase activity and cyclic GMP content of cultured chick pineal glands.

We examined effects of supplements on cycles in cyclic GMP content and serotonin N-acetyltransferase (NAT) activity in cultured chick pineal glands. Increases in cyclic GMP content and NAT activity were stimulated by 1-ethyl-4(isopropylidene-hydrazino)-1H-pyrazolo[3,4-b]pyridene-5-c arboxylic acid, ethyl ester, hydrochloride and isobutylmethyl xanthine under diurnal illumination and in constant darkness, but subsequent decreases were not inhibited. Hypoxanthine had little effect on NAT activity under all lighting conditions, or on the content of cyclic GMP in glands cultured in the dark. However, it markedly stimulated accumulation of cyclic GMP in illuminated cultures. EGTA or additional Ca2+ had no effect on pineal NAT activity. However, EGTA markedly stimulated accumulation of cyclic GMP both in the light and in the dark. Supplementary Ca2+ slightly retarded accumulation of cyclic GMP in the dark but stimulated slightly in the light.

1-Methyl-3-isobutylxanthine↗
PubMed1984

Phase-shifting of cycles in level of serotonin N-acetyltransferase activity and cyclic GMP content of cultured chick pineal glands by methotrexate.

Methotrexate at 1 microM stimulated increase of serotonin N-acetyltransferase (NAT) activity in chick pineal glands cultured under each of three conditions of illumination. The peak of the circadian rhythm in NAT activity and the "spike" in content of cyclic GMP were both advanced in pineal glands cultured in the dark from midphotoperiod. In contrast, the time of peak NAT activity in glands cultured in the dark from late photoperiod was unaffected. In addition, methotrexate did not affect times of reaching maximum NAT activities in glands cultured from midphotoperiod in the light or under diurnal illumination. Doubling the concentration of methotrexate also eliminated the lag phase in increase of NAT activity in glands cultured in the dark. However, at a concentration of 5 microM methotrexate the curve depicting increase of NAT activity was biphasic, and neither time nor level of peak NAT activity differed from those of control glands. Results of attempts to demonstrate persistent effects of exposure to methotrexate were inconclusive.

Acetyltransferases↗
PubMed1984

Neopterin retards loss of serotonin N-acetyltransferase activity from cultured chick pineal glands.

D-Neopterin at 10 microM delayed start of the decline of serotonin N-acetyltransferase (NAT) activity from the peak level in the cycle exhibited by chick pineal glands cultured under standard conditions in the dark. A less marked retardation of decline of NAT activity was found with glands cultured under diurnal illumination or those exposed prematurely to light. There were no significant effects of neopterin on the increases of NAT activity or peak levels of activity developed. The pteridine also retarded loss of NAT activity from the peak level developed in the dark when the time of explanting into culture was later in the (solar) day, but not when it was earlier. Neopterin had no effect on the cycle in cyclic GMP content of cultured chick pineal glands.

Acetyltransferases↗
PubMed1980

Diurnal cycles in serotonin acetyltransferase activity and cyclic GMP content of cultured chick pineal glands.

Levels of serotonin N-acetyltransferase (NAT: acetul CoA:arylamine N-acetyltransferase; EC 2.1.1.5.) activity in the chick pineal gland exhibit a marked diurnal variation in birds kept under a diurnal cycle of ilumination. Activity begins to rise rapidly at the start of the dark phase of the cycle and reaches maximum levels at mid-dark phase about 25-fold greater than the minimum basal level at mid-light phase. Thereafter, the level of activity declines to the basal level about the start of the light phase. This diurnal cycle in chick pineal NAT activity found in vivo has recently been reproduced in vitro with intact glands incubated in organ culture. The mechanism of the 'biological clock' which regulates these variations in level of chick pineal NAT activity is unknown. However, I now report that chick pineal glands cultured under a diurnal cycle of illumination exhibit a diurnal cycle in content of cyclic GMP which roughly parallels the cycles in NAT activity. In contrast, there was no correlation between variations in pineal content of cyclic AMP and in level of NAT activity.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗
PubMed1980

Regulation of the cycle in chick pineal serotonin N-acetyltransferase activity in vitro by light.

The diurnal cycle in vivo in the level of serotonin N-acetyltransferase (NAT) activity found in the pineal gland of chicks kept under diurnal lighting was reproduced in vitro with glands incubated in organ culture under the same diurnal lighting conditions. We have examined the effects of varying the lighting conditions of culture upon development of NAT activity in vitro with pineal glands from birds which were killed at different times during the photoperiod in vivo. The ability of chick pineal glands to develop increased NAT activity in culture during the dark period of a diurnal cycle of illumination was determined primarily by the proportion of the photoperiod in vivo elapsed at the time of sacrifice of the birds. The time of development of maximal NAT activity in cultured chick pineal glands could be advanced or delayed by corresponding changes in time of start of the dark period. However, the 'settings' of the pineal 'biological clock' remained unchanged. Glands from birds sacrificed in the final 30 min of the photoperiod in vivo developed increased NAT activity without lag when cultured in the light. Similarly, pineal glands cultured under continuous illumination developed an increased photostable NAT activity beginning at the time of the "subjective" end of the photoperiod in vitro. Ability to develop additional NAT activity in the dark was low rapidly at the "subjective" time in vitro of start of the next photoperoid. The maximal NAT activity developed in the dark consisted of both photolabile and photostable components.

Acetyltransferases↗
PubMed1979

Chick pineal serotonin acetyltransferase: a diurnal cycle maintained in vitro and its regulation by light.

We have reproduced in vitro the diurnal cycles in levels of serotonin acetyltransferase activity found in the chick pineal gland in vivo. The more closely the lighting conditions of culture matched those under which the birds were raised, the closer was the similarity between cycles in levels of enzyme activity in vitro and in vivo. Repetitive cycles in levels of acetyltransferase activity persisted in culture for at least 4 days under a diurnal cycle of illumination, and at least 2 days in continuous darkness. When glands were explanted into culture in the light phase of a cycle, short periods of further exposure to light markedly stimulated subsequent increase of acetyltransferase in the dark (after a short lag). Prolonged exposure to light in culture markedly inhibited increase of enzyme activity. Cycles in the levels of enzyme activity in glands cultured under altered light cycles were regulated primarily by changes in illumination. However, the endogenous biological 'clock' remained at least partly entrained to the original light cycle. Increase of acetyltransferase activity in vitro was markedly stimulated by theophylline plus compound Ro. 20.1724 (4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone) under all lighting conditions. Kinetics (to the time of attaining maximum levels in situ) of the increase under diurnal lighting and in constant darkness were indistinguishable from those in vivo. A high concentration of dl-propranolol markedly stimulated an increase in acetyltransferase activity in glands cultured in constant darkness but had little effect on glands under diurnal lighting or continuous illumination.

Acetyltransferases↗
PubMed1978

Preparation of cell populations with stabilized erythropoietic potential from the primitive streak chick blastodisc: some implications for control of gastrulation.

Improved methods for preparation from primitive streak chick blastodiscs of cell suspensions capable of forming erythroid cells in culture have been developed. When blastodiscs were preincubayed with hyaluronidase in the absence of collagenase before cell dispersion and a high concentration of methyl-alpha-mannoside was present in all media, the yields of cells were some 10-fold higher than those obtained by former procedures. Cell suspensions obtained consisted almost entirely of viable cells, yielded large numbers of free mature erythrocytes in liquid culture, and formed erythroid colonies and bursts in solidified medium. The capacity to form differentiated cells after resedimenrtation through Ficoll density gradients was partly stabilized. Addition of gee yolk homogenate to the blastodiscs immediately following treatment with hyaluronidase and to all media used thereafter largely stabilized the capacity to form erythroid cells during resedimentation through Ficoll density gradients. Possible relevance of observations made during development of the procedures to the control of onset of cell migration in the process of gastrulation is indicated.

Animals↗