Thomas S. King

The influence of different light spectra on the suppression of pineal melatonin content in the Syrian hamster.

The purpose of this study was to test the capacity of different visible wavelengths of light to suppress nocturnal levels of pineal melatonin in hamsters. It was found that the visible wavelengths vary in their ability to perturb pineal melatonin. During the period of peak pineal melatonin production, animals were exposed to fluorescent light sources having half-peak bandwidths of 339-371 nm (near-ultraviolet), 435-500 nm (blue), 510-550 nm (green), 558-636 nm (yellow) and 653-668 nm (red). In each experiment, animals were exposed to equal irradiances of each light source. The different irradiances used were 0.928, 0.200, 0.186, 0.074 and 0.019 microW/cm2. The resultant data demonstrated that blue fluorescent light was the most efficient in suppressing pineal melatonin. Green fluorescent light was found to be the next most efficient light for inhibiting pineal melatonin followed by yellow fluorescent light. Near-ultraviolet and red light were the least capable of suppressing pineal melatonin. These observations suggest that the retinal photopigment responsible for mediating the pineal glands response to light in the hamster may be either rhodopsin or another blue-sensitive chromophore.

The transfer of cocaine and its metabolites across the term human placenta

This study defines human placental transport of cocaine and its two minor, but pharmacologically active, metabolites—norcocaine and cocaethylene. The experimental system was the single, isolated perfused cotyledon of a normal term human placenta, and antipyrine served as a freely diffusible marker. Cocaine was transferred rapidly by the placenta at a rate about 80% that of antipyrine. The transfer had characteristics of passive transport consistent with the high lipid solubility of the drug. We found no evidence of significant placental metabolism of cocaine during its rapid placental transfer. Ethanol did not alter the cocaine transfer rate. Norcocaine and cocaethylene were equally as rapidly transferred. Thus the placenta is no barrier to the transfer of cocaine and its derivatives to the fetus.

Differential response of pineal melatonin levels to light at night in laboratory-raised and wild-captured 13-lined ground squirrels (Spermophilus Tridecemlineatus)

Pineal melatonin levels were compared in laboratory-raised or wild-captured 13-lined ground squirrels (Spermophilus tridecemlineatus) that were either exposed to 10 h of darkness at night or to light which had an irradiance of 400 microW/cm2. In laboratory-born squirrels the period of darkness was associated with a gradual rise in pineal melatonin levels with peak values being reached at 0200 h, 6 h after darkness onset. Thereafter, melatonin levels decreased and were back to low daytime levels by 0800 h, 2 h after light onset. The exposure of laboratory-raised animals to an irradiance of 400 microW/cm2 during the night totally prevented the nocturnal rise in pineal melatonin levels in these animals. In wild-captured ground squirrels the period of darkness at night was associated with a rapid rise in pineal melatonin such that by 2200 h, 2 h after lights out, peak melatonin values were already attained; additionally, melatonin levels remained high throughout the period of darkness but returned to daytime values by 0800 h. Exposure of wild-captured squirrels to a light irradiance of 400 microW/cm2 during the normal dark period was completely incapable of suppressing pineal melatonin levels. The difference in the sensitivity of the pineal gland of laboratory-raised and wild-captured ground squirrels may relate to their previous lighting history.

Azidothymidine (Zidovudine) Transport by the Human Placenta

The diagnosis of Acquired Immunodeficiency Syndrome (AIDS) is increasingly made in pregnant women, and the disease may be transmitted to the fetus. Azidothymidine (AZT, Zidovudine) is the one therapeutic agent of some promise in this condition. As there is no information on the transport of this drug by the human placenta, such studies were carried out using the single cotyledon placental perfusion system and human placental vesicles. AZT crossed the placenta readily and bidirectionally. The transfer rate was about 70% that of a freely diffusible reference marker, antipyrine, and was comparable in both directions. There was no evidence of active or carrier-mediated transport and no glucuronidated metabolites of the drug were identified in either maternal or fetal compartments. The authors believe that the drug crosses the placenta by diffusion, consistent with its lipophilicity and transport into various blood cells.

Cocaine Disrupts Estrous Cyclicity and Alters the Reproductive Neuroendocrine Axis in the Rat

Although a common drug of abuse, cocaines effects on cyclic reproductive functions and the neuroendocrine systems regulating these functions have not been studied. Here, we report the effects of cocaine on (1) estrous cyclicity and ovulation rates and (2) the stimulated in vitro release of hypothalamic GnRH and aminergic neurotransmitters directly involved in regulating or modulating GnRH release. Within 7 days of treatment with 10 mg kg-1 day-1 of cocaine HCl subcutaneously, rats demonstrated significant estrous cycle irregularity including repetitive days of estrus and prolonged periods of diestrus. After 6 weeks of treatment, cocaine-treated rats exhibited a 44.3% decrease in ovulation rates. For the in vitro studies, bilaterally ovariectomized rats were injected with cocaine (10 mg kg-1 day-1) or with saline for 2 weeks. Each rat received estradiol benzoate (50 mg kg-1 day-1 s.c.) for 2 days before sacrifice. Hypothalamic slices were prepared, placed in 0.1 ml microchambers and perfused with modified Krebs buffer (pH 7.4) using a programmable perfusion system. Basal release of norepinephrine (NE) and serotonin (5HT) was significantly increased in the cocaine-treated group versus controls. Ten-minute pulses of 10(-7)M progesterone (P4) increase NE and 5HT, but not dopamine (DA), release in the saline-treated group. In contrast, pulses of P4 increased NE, but not 5HT or DA, in the cocaine-treated rats. Ten-minute pulses of 0.1 microM NE increased GnRH release in both saline- and cocaine-treated rats. However, the response to pulsed NE was significantly attenuated in the cocaine-treated group.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Ovarian Steroids to Stimulate Region-Specific Hypothalamic 5-Hydroxytryptamine Synthesis in Ovariectomized Rats

Accumulations of 5-hydroxytryptophan (5HTP) and L-dihydroxyphenylalanine (L-DOPA) following decarboxylase inhibition as indices for 5-hydroxytryptamine (5HT) and catecholamine (dopamine and/or norepinephrine) synthesis, respectively, were both increased in the preoptic area-anterior hypothalamus (POA-AH) of ovariectomized rats treated with a combination of 17 beta-estradiol benzoate (E2) and progesterone (P). Similarly, an increased accumulation of L-DOPA was seen in the mediobasal hypothalamus (MBH) and median eminence (ME) of these animals although no change was observed in 5HTP accumulation in the MBH or ME of these rats. Hypophysectomy negated these steroid-induced effects on L-DOPA accumulation. However, hypophysectomy had no apparent effect on steroid-stimulated 5HTP accumulation in the POA-AH of these rats. Under the experimental conditions of our study, the results suggest that the stimulatory effect of ovarian steroids on hypothalamic catecholamine synthesis is dependent on an intact pituitary gland, that the stimulatory effect of ovarian steroids on 5HT synthesis in the POA-AH is not dependent on an intact pituitary gland, and (3) that ovarian steroids do not seem to influence 5HT synthesis in the ME or MBH. The significance of these results may lie in the function of these hypothalamic monoaminergic neurotransmitter systems to regulate gonadotrophin release and subsequent steroid feedback modulation of such central regulatory mechanisms.

Comparison of the Effects of β‐Adrenergic Agents on Pineal Serotonin N‐Acetyltransferase Activity and Melatonin Content in Two Species of Hamsters

The nighttime rise in pineal melatonin levels can be blocked by administration of the β‐adrenergic receptor antagonist, propranolol, in both Syrian hamsters and rats. Although the administration of β‐adrenergic receptor agonists such as norepinephrine or isoproterenol stimulates pineal melatonin production in the rat, these drugs are without apparent effect on indole production in the Syrian hamster. To determine whether this lack of stimulatory effect in the Syrian hamster is characteristic of this species, a comparison of the effects of norepinephrine and isoproterenol on pineal serotonin N‐acetyltransferase activity and melatonin content was conducted. In contrast to their lack of effect in the Syrian hamster, norepinephrine and isoproterenol stimulated pineal serotonin N‐acetyltransferase activity and melatonin content in the Djungarian hamster. Hourly injection of norepinephrine during a continuation of light into the normal dark period stimulated increases in the activity of serotonin N‐acetyltransferase and melatonin content in the Djungarian hamster but was without effect on these pineal parameters in the Syrian hamster.

Pharmacological studies on the regulation of N-acetyltransferase activity and melatonin content of the pineal gland of the Syrian hamster

Thus far, all attempts to stimulate melatonin synthesis by β‐adrenergic receptor agonists in the Syrian hamster pineal gland have failed. Neither a wide range of dosages of isoproterenol (0.5 mg/kg to 24 mg/kg), nor prolonged treatment with norepinephrine, the natural neurotransmitter, increased N‐acetyltransferase (NAT) activity or melatonin production. In the present study, the administration of isoproterenol at night was likewise ineffective in advancing or enhancing the normal nightly melatonin peak. Also, we did not find a delayed effect 7 or 8 h after the administration of the drug. Furthermore, we tested the idea of coneurotransmitters such as octopamine or dopamine being possibly necessary for stimulation, but could not find any effect of these substances on melatonin synthesis. In addition, a parasympatholytic agent, atropine, did not increase the responsiveness to sympathomimetic agents. Administration of a phosphodiesterase inhibitor was also ineffective in stimulating NAT activity. On the other hand, isoproterenol did retard the drop in NAT and melatonin after lights‐on at night, indicating that β‐receptors are involved in maintaining elevated melatonin levels.

Studies on pineal melatonin levels in a diurnal species, the Eastern chipmunk (Tamias striatus): Effects of light at night, propranolol administration or superior cervical ganglionectomy

Five experiments were carried out on the control of melatonin levels in the pineal gland of a diurnal species, the Eastern chipmunk (Tamias striatus). We confirmed that the exposure of chipmunks to fluorescent white light of 3,981–4,304 lux during the normal dark period does not prevent the rise in pineal melatonin levels normally associated with darkness. Also, the administration of propranolol (20mg/kg) at 8 p.m. did not block the rise in pineal melatonin in animals exposed to either dark or light at night. Similarly, if chipmunks received propranolol 4 hours into the dark phase, pineal melatonin levels were not depressed 2 hours later. When animals were superior cervical ganglionectomized, however, the pineal content of melatonin remained low regardless of whether the animals were exposed to darkness or light at night. The exposure of chipmunks acutely to light at midnight (4 hours after darkness onset) had only a slight depressive effect on pineal melatonin 30 min later; by comparison, when chipmunks were acutely exposed to light at 3 a.m. (7 hours after darkness onset) daytime pineal melatonin levels were reached within 15 min after light onset. These findings in a diurnal species, the Eastern chipmunk, differ markedly when compared to previously reported observations on nocturnal laboratory rodents.

Tryptophan administration inhibits nocturnal N-acetyltransferase activity and melatonin content in the rat pineal gland. Evidence that serotonin modulates melatonin production via a receptor-mediated mechanism.

In the rat pineal gland, the activity of serotonin N-acetyltransferase (NAT) and the concentration of melatonin are normally high at night; conversely, the concentration of serotonin (5-HT), the precursor of melatonin, is low. Since tryptophan administration increases the concentration of pineal 5-HT at night, we examined its effect of melatonin production. Nighttime tryptophan loading led to substantial increases in pineal 5-hydroxytryptophan, 5-hydroxyindole acetic acid (5-HIAA), and 5-HT but a highly significant reduction in NAT activity in comparison to saline-injected controls. In contrast to other measured indoles, melatonin levels also were significantly diminished by tryptophan loading. Nocturnally high pineal norepinephrine levels were unaltered by tryptophan administration. The idea that high concentrations of 5-HT could lead to substrate inhibition of NAT activity was not supported by kinetic analysis of control NAT levels versus tryptophan-inhibited NAT activity under varied substrate concentrations. Hypotheses to explain these results include the possibility that tryptophan inhibition of melatonin synthesis is mediated by the release of 5-HT from the pinealocyte and its subsequent autocrine action on melatonin production.