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Biomedical subjects

L J Petterborg

Publications and source records attributed to L J Petterborg.

At least 19 recordsLinked to original sources

Effect of testosterone replacement on the alteration of steroid metabolism in the hypothalamic-preoptic area of male hamsters treated with melatonin.

Adult male hamsters were maintained under 14 hours of light per day and randomly assigned to groups that received daily afternoon melatonin (25 micrograms) or vehicle injections. Animals from both groups were killed following 4, 8, and 12 weeks of treatment. By 12 weeks, the melatonin-treated hamsters had significant reductions in the weights of the testes and seminal vesicles, serum testosterone levels, and activities did not differ between groups. In a second experiment, hamsters were hypothalamic-preoptic area (HPOA) aromatase activities. Hypothalamic-preoptic area 5 alpha-reductase activities did not differ between groups. In a second experiment, hamsters were again treated with melatonin or vehicle for 12 weeks prior to being killed. After 10 weeks of treatment, groups of melatonin-treated animals received subcutaneous silastic capsules (5, 10, or 20 mm) filled with testosterone. Animals in two other groups were given blank implants or no implants at all. Two weeks later, at autopsy, reproductive organ weights, serum testosterone levels, and HPOA aromatase activities were significantly suppressed by melatonin administration. 5 alpha-Reductase activity in the HPOA was not affected. Hamsters that had been given the 10- and 20-mm testosterone implants exhibited normal seminal vesicle weights and HPOA aromatase activities. These results suggest that melatonin-induced reduction of HPOA aromatase activity is mediated by decreased circulating levels of testosterone.

Animals

Effect of melatonin replacement on serum hormone rhythms in a patient lacking endogenous melatonin.

A potentially confounding variable inherent in studies designed to examine the effect of melatonin administration in humans is the presence of an endogenous melatonin rhythm in the experimental subjects. The effects of exogenous melatonin administration on serum hormone rhythms was recently examined in a male patient who lacked detectable circulating levels of endogenous melatonin. The patient's pineal gland had been destroyed five years previously in the course of treatment for a pineal astrocytoma. On three separate occasions, over approximately a one-year period, the patient was given daily oral melatonin replacement (2 mg/day, 1 mg/day and 0.5 mg/day). These experiments were designed to assess the effects of exogenous melatonin on serum growth hormone, prolactin, cortisol and testosterone rhythms. Analysis of blood samples collected every 2-4 hours periods both before and during melatonin replacement revealed that the exogenous melatonin rhythm was associated with improvements in self-reported sleep and mood ratings. Melatonin administration produced robust nocturnal peaks in serum growth hormone and prolactin levels immediately following ingestion of the hormone, while serum cortisol and testosterone rhythms were not influenced. These results suggest that melatonin may modulate the coordination and enhancement of selected biological rhythms in man.

Adult

Effect of a 15 minute light pulse on nocturnal serum melatonin levels in human volunteers.

Monitoring the daily melatonin rhythm during the course of phototherapy for affective disorders may be beneficial in assessing the efficacy of such treatments. It is therefore of interest to study the effects of the timing, duration, and intensity of bright light pulses on melatonin levels in normal subjects. To examine the effects of a single exposure to a brief burst of bright light on serum melatonin, groups of healthy human volunteers of both sexes were treated with a 15 minute pulse of bright light (350 cd/m2) early in the evening during the winter months. Serial blood samples were collected from each person and the effect of the light pulse on serum melatonin and cortisol levels determined. Melatonin levels were significantly but only transiently suppressed by the light pulse, while cortisol levels were not affected. These results demonstrate that short duration bright light treatments can influence the melatonin rhythm generating system in humans.

Female

Effects of light and an alpha-2-adrenergic agonist on serotonin N-acetyltransferase activity in chick pineal gland.

Pineal serotonin N-acetyltransferase (NAT) is the enzyme that catalyzes the production of N-acetylserotonin from serotonin and is the rate limiting step in the biosynthesis of melatonin in the chick pineal gland. Chick pineal NAT activity is decreased by light and by noradrenergic agents that act at the alpha-2-adrenergic receptor. Light-induced inhibition of nocturnal NAT activity can be demonstrated by exposing 4-day-old chicks to light, or by exposing pineal gland explants cultured in vitro either to light or to UK 14,304 (an alpha-2-adrenergic agonist). The inhibition by either light or UK 14,304 is preceded by a transient, but consistent rise in NAT activity. The paradoxical, transient light-induced rise in enzyme activity is mimicked by UK 14,304 which suggests a similarity in mechanisms of light and adrenergic signal transduction associated with the regulation of NAT activity.

Adrenergic alpha-Agonists

Ontogeny of light-induced decrease of N-acetyltransferase activity in explanted chick pineal glands.

The ontogeny of chick pineal serotonin N-acetyltransferase (NAT) activity was investigated in explanted chick pineal glands at 4, 10 and 21 d of age. Nocturnal levels of the enzyme and the response of the enzyme to light exposure were determined in pineal glands maintained in short-term culture at each age. The results indicate that nocturnal NAT activity was increased in the glands from older birds. Nocturnal levels of NAT activity at the time of the initiation of the experiment were threefold greater in glands from 21-d-old birds as compared to that in glands from 4-day-old chicks. The response to light was similar in all three ages examined; light induced a significant decrease in NAT activity within 60 min in explanted glands from 4-d-old chicks and within 180 min in the glands from the 10- and 21-d-old chicks. A paradoxical transient increase in enzyme activity occurred immediately (within 5 min) following light exposure which was significant in the glands from the youngest chicks, and present, but more variable, in the older chicks. These data indicate that the nocturnal enzyme activity is greater in glands from older birds, but that light exposure of explanted glands initiates a transient rise followed by a decrease in NAT activity at all three ages.

Acetyltransferases

Effects of daily afternoon melatonin administration on body weight and thyroid hormones in female hamsters.

Female hamsters were treated each afternoon for 8 weeks with subcutaneous injections of 25 micrograms of either melatonin or vehicle solution. Animals were sacrificed in either the morning or afternoon of diestrus and proestrus, along with their respective melatonin-induced acyclic pairs. Melatonin-treated hamsters had significantly greater mean body weights than did the vehicle-treated hamsters (P less than 0.05). Terminal mean serum T4 levels and free thyroxine index (FT41) were significantly reduced (P less than 0.05) in melatonin-treated hamsters compared with values obtained from animals subjected to the vehicle alone. In addition, T4 levels, FT41, T3 levels, and FT31 were significantly influenced by day and time of sacrifice independently of the melatonin treatment. T3 uptake was significantly lower on the morning of proestrus in melatonin-treated animals. These results demonstrate that chronic afternoon melatonin administration in female hamsters results in the loss of estrous cyclicity, a significant gain in body weight, and the reduction of T4 levels and T3 uptake. Changes in serum T3 levels are a function of the time of sample collection and are not influenced by melatonin treatment. In addition, these data indicate that thyroid function in general changes during the estrous cycle in these animals.

Animals

Identification of alpha 2-adrenergic receptors in chicken pineal gland using [3H]rauwolscine.

The norepinephrine-induced inhibition of avian pineal N-acetyltransferase activity appears to be mediated by alpha 2-adrenergic receptors. In this study, alpha 2-adrenergic receptors in the chicken pineal gland were directly identified by radioligand binding. Membrane preparations of pineal glands from chickens from 1 to 6 weeks of age were examined using [3H]rauwolscine, a selective alpha 2-adrenergic receptor antagonist, to characterize the binding sites. The results indicate no ontological change in either the affinity (KD) or density of receptor binding sites (Bmax) during the time span examined. The binding was saturable and of high affinity with a mean KD of 0.27 +/- 0.01 nM and a mean Bmax of 242 +/- 12 fmol/mg protein. Further characterization of these binding sites indicated that the alpha 2-adrenergic receptor is of the alpha 2A subtype, since prazosin and ARC-239 bound with low affinities and oxymetazoline bound with high affinity.

Adrenergic alpha-Antagonists

Exercise reduces gonadal atrophy caused by short photoperiod or blinding of hamsters.

The effect of voluntary exercise upon several reproductive parameters was assessed in male Syrian hamsters. Forty-two animals were caged in five groups and then blinded: Ten housed five per plastic cage; eight housed individually in plastic cages; six housed individually in plastic cages with 15 cm exercise wheels in the cage; eight housed in Wahmann steel activity cages with 36 cm wheels locked to prevent rotation; ten housed in functional Wahmann running wheel cages. At 16 weeks after blinding, animals with access to functional running wheels had significantly larger testes (3.7 +/- 1.0 g) than those with no wheels or locked wheels (1.86 +/- 0.6 g; mean +/- SD; p less than 0.001). These results suggested that voluntary exercise reduces testicular atrophy caused by blinding. A second experiment was run in which the animals were subjected to a short photoperiod (LD 6:18) for 12 weeks instead of blinding. Similar results were obtained except that access to the small exercise wheels did not affect testicular atrophy: Mean testicular mass of animals in individual cages = 0.79 +/- 0.3 g, small wheels = 0.74 +/- 0.4 g, functional Wahmann wheels, 2.56 +/- 1.0 g, locked Wahmann wheels 0.52 +/- 0.1 g (p less than 0.01).

Animals

Reproductive effects of 6-chloromelatonin implants and/or injections in male and female Syrian hamsters (Mesocricetus auratus).

Adult female hamsters were injected each afternoon for 9 weeks with 2.5, 15 or 25 micrograms of melatonin or 6-chloromelatonin (CM). Each drug resulted in a significant dose-related depression in uterine, ovarian and anterior pituitary gland weights. Additionally, plasma and pituitary concentrations of prolactin fell with increasing dose of either indole whereas pituitary levels of LH and FSH were positively correlated with dose. There was no difference in effectiveness between the two drugs. Adult male hamsters treated for 10 weeks with daily afternoon injections of melatonin and a blank beeswax pellet had depressed testicular and accessory organ weights and plasma and pituitary concentrations of prolactin. Implantation of a 1 mg melatonin or CM beeswax pellet in hamsters concurrently receiving daily afternoon injections of melatonin prevented the organ weight and hormonal changes, except for plasma prolactin. Adult male hamsters treated for 10 weeks with daily afternoon injections of CM and a blank beeswax pellet had depressed reproductive organ weights and pituitary and plasma concentrations of prolactin; this depression in hormonal values and organ weights was totally prevented if the CM-injected hamsters were also bearing a beeswax-melatonin pellet. In conclusion, 6-chloromelatonin is as effective as melatonin with regards to antigonadotrophic and counter-antigonadotrophic effects in male and female Syrian hamsters.

Animals

Modification of testicular and thyroid function by chronic exposure to short photoperiod: a comparison in four rodent species.

Exposure of male Syrian hamsters (Mesocricetus auratus) for 10 weeks to short photoperiod (SP) providing 10 hr light: 14 hr darkness (10:14 LD) produced a significant reduction in the weights of the reproductive organs, plasma thyroxine (T4) levels and free T4 index (FT4I) compared to the values of animals exposed to long photoperiod (LP, 14:10 LD). C57bl male house mice (Mus musculus) kept in SP (10:14 LD) had reproductive organ weights equivalent to those of mice kept in long days (14:10 LD) and lower T3 uptake (T3U) values. Male gerbils (Meriones unguiculatus) exposed to 13 weeks of SP (10:14 LD) had lower body weights, testes and seminal vesicle weights and higher T3U values compared to LP (14:10 LD) controls. However, no effect was seen on plasma T4 and triiodothyronine (T3) values nor the FT4I and free T3 index (FT3I). White-footed male mice (Peromyscus leucopus) exposed to SP (8:16 LD) had significantly lower testes and seminal vesicle weights while plasma T4 and T3 levels were unaffected. Snell strain house mice (Mus musculus) exposed to SP (8:16 LD) had normal reproductive organ weights compared to the values of LP-exposed (16:8 LD) control animals. However, there was a significant depression in T3 and in the FT3I in the SP animals.

Animals

An immunocytochemical study of the luteinizing hormone-releasing hormone (LHRH) system in the white-footed mouse: effect of blinding and melatonin.

Previous studies have provided evidence that pineal-mediated gonadal atrophy involves the luteinizing hormone-releasing hormone (LHRH) system in some rodents. The purpose of the present study was to visualize the LHRH system in the brain of the white-footed mouse by immunocytochemical methods and to assess the influence of blinding or melatonin on the LHRH immunostaining characteristics of the median eminence. Six intact adult female mice and six adult males (two intact, two blinded, and two with subcutaneous melatonin-beeswax implants) treated for 6 wk were sacrificed by perfusion of fixative, and their brains were removed, cut into 60-micron sections and stained for LHRH by the peroxidase-antiperoxidase (PAP) method. Since the tissues were batch-processed in separate wells in a common chamber, semiquantitive comparisons may be made between similar sections from animals subjected to different treatments. The testes were also collected and prepared for microscopic examination. The distribution of LHRH-stained cell bodies and fibers was found to be essentially the same as has been described in other rodents, i.e., predominantly confined to the preoptic area and hypothalamus. The perikarya appear as unipolar and bipolar fusiform cells with "beaded fiber" type processes. In sections that included the median eminence of the blinded and melatonin-treated males, the degree of immunopositive staining appears to be greater than in the median eminence of untreated mice. The histological appearance of the testes from the treated animals revealed a profound inhibition of gametogenesis. These observations support the suggestion that pineal-mediated gonadal atrophy is the result of an inhibition of LHRH release from the median eminence.

Animals

Inclusion bodies in pinealocytes of the cotton rat (Sigmodon hispidus). An ultrastructural study and X-ray microanalysis.

Pinealocytes of the cotton rat (Sigmodon hispidus) often contain large (2-6 micron diameter) intracytoplasmic inclusions, the function of which is not known. These inclusions may represent nucleolus-like bodies, mineral deposits, secretory products or viral inclusions. In this study these inclusions were classified as type A, B or C inclusions based on the amount of electron-dense material interspersed within the finely granular material comprising the bulk of these inclusions. Each type of inclusion was analyzed by X-ray microanalysis and enzymatic proteinaceous digestion. X-ray microanalysis of these inclusions differed both quantitatively and semiquantitatively from that of human or gerbil pineal concretions, the latter two of which are extracellular deposits. Pronase, a proteolytic enzyme, digested the electron-dense material only after longer times of tissue exposure to this enzyme in contrast to the easily digested, finely granular matrix-like material of these inclusions. Such intrapinealocytic inclusions have only been observed in the cotton rat. Their functional significance remains unknown.

Animals

Effect of superior cervical ganglionectomy on the ultrastructure of the pinealocyte in the cotton rat (Sigmodon hispidus).

The ultrastructure of pinealocytes was examined morphometrically in superior cervical ganglionectomized and sham-operated cotton rats. Following denervation, the pinealocytes decreased in size. Reduced areas of the nucleoli, Golgi apparatus, granular endoplasmic reticulum, mitochondria, and vacuoles containing a flocculent material as well as a decreased number of dense-core vesicles were noted. In contrast, the numbers of "synaptic" ribbons and ribbon fields increased. The nature of ultrastructural changes observed in the cotton rat pinealocyte after sympathectomy may indicate a diminution of the presumptive secretory processes of this cell. The presence of a small number of myelinated and unmyelinated axons in the pineal gland of the cotton rat following superior cervical ganglionectomy suggests that the sympathetic nerve fibers from the superior cervical ganglia do not comprise the only source of innervation to the pineal. Additionally, the presence of striated muscle fibers has been observed in the present study in the pineal gland of the cotton rat which heretofore has not been reported in this species.

Animals

Ultrastructure of pinealocytes of the cotton rat, Sigmodon hispidus.

Fine structural features of pinealocytes of cotton rats (Sigmodon hispidus) were examined. Golgi complexes, mitochondria, endoplasmic reticulum and polysomes are usual organelles seen in the perikaryonal cytoplasm of pinealocytes. Many non-granulated vesicles (40 to 80 nm in diameter) and a few granulated vesicles (about 100 nm in diameter) are associated with the Golgi cisternae. Occasionally, the cisternae contain granular materials. The perikaryonal cytoplasm of pinealocytes is characterized by the presence of inclusion bodies. These bodies are usually round in shape, not bounded by a limiting membrane and composed of fine granular or filamentous materials of high electron-opacity, which are similar in appearance to the substance seen in the nucleolonema. Pinealocyte processes, filled with abundant non-granulated vesicles and some granulated vesicles, are mainly found within the parenchyma and occasionally in perivascular spaces.

Animals

Interaction of luteinizing hormone-releasing hormone, cyproterone acetate and arginine vasotocin on plasma levels of luteinizing hormone in intact and castrated adult male rats.

Treatment of unanesthetized castrated adult male rats every 3 h for 48 h with either 5 microgram of arginine vasotocin (AVT) and/or 1 microgram luteinizing hormone-releasing hormone (LRH) caused a significant inhibition of plasma levels of luteinizing hormone (LH) and compared to castrated control rats receiving diluent only. However, the intravenous (iv) injection of 1 microgram of AVT into urethane-anesthetized male rats which had been castrated for 0, 24 or 48 h did not affect plasma levels of LH at 10, 20 or 60 min following injection compared to their respective diluent-treated castrated control rats. Similarly, the iv injection of either 100 ng, 1 microgram or 10 microgram AVT was unable to acutely affect plasma levels of LH in intact male rats. Following the iv injection of 2 doses of 50 ng LRH spaced 1 h apart in anesthetized castrated male rats, 2 peaks of equal magnitude in plasma LH were noted. Castrated rats treated with 2 injections spaced 1 h apart of LRH + AVT had significantly higher plasma levels of LH than did rats treated with LRH alone. In subsequent studies, both AVT and arginine vasopressin were observed to augment the plasma response of LH to an injection of LRH whereas oxytocin had no effect. A single injection of AVT + LRH significantly augmented the plasma titers of LH compared to levels observed in LRH-treated control rats as did a second injection 1 h later. The administration of cyproterone acetate sc for 2 days by itself had no effect on plasma LH but in conjunction with LRH caused a marked rise in plasma LH compared to intact rats treated with LRH alone. AVT in combination with LRH and cyproterone acetate caused a significant elevation in plasma LH at 60 min post-injection when compared to plasma levels of rats treated with LRH alone or the combination of LRH and cyproterone acetate. It is concluded that acute intravenous injections of AVT augment the LH-releasing activity of LRH; chronic treatment for 48 h, however, with LRH + AVT leads to a significant depression of plasma LH perhaps due to an exhaustion of the releasable pool of LH in the anterior pituitary.

Animals