Morphological assessment in pinealectomy and foetal pineal gland transplantation in rats: Part I

Summary The effects of pineal gland, an endocrine organ known to affect the physiology of various organs and systems including the hypothalamo-hypophyseal axis, on the morphological characteristics of target organs were investigated in rats after pinealectomy and foetal pineal gland transplantation to a subpial cortical area close to the pinealectomized region.It was demonstrated that weight gain was significantly slower in pinealectomized male rats (p<0.01); pituitary gland weight was lower in the pinealectomized group (p<0.01) and transplantation had no effect on the weight of this organ; weight of adrenal gland and testis were lower after pinealectomy (p<0.01) but restored back to control levels after transplantation; the mitotic activity in seminiferous tubules increased with pinealectomy and returned back to control indexes after transplantation.Research on hypothalamic catecholaminergic content revealed a diminished histofluorescence in pinealectomized rats which showed a potentiated restoration after transplantation.The innervation of the pineal gland was studied by anterograde and retrograde injections of Wheat Germ Agglutinin Horse Radish Peroxidase from superior cervical ganglion and pineal gland, respectively. It was demonstrated that cell bodies were traced both at the transplanted tissue and hypothalamus.In view of these data, the multifunctional and a probable high level homeostatic harmony regulator essence of pineal gland is discussed.The paper has been awarded the 1993 Upjohn prize for Neurosurgical Research of the European Association of Neurosurgical Societies.     

Effects of pinealectomy and exogenous melatonin on rat hearts

The effects of pinealectomy and administration of melatonin, the major secretory product of the pineal gland, which is a direct free radical scavenger and an indirect antioxidant, were studied in rat hearts on the basis of cardiac morphology and biochemical findings. Three groups of Wistar rats were used: one group was the sham-operated control, one group consisted of pinealectomized rats and one group consisted of pinealectomized rats that were treated with melatonin. Serum cholesterol, tissue levels of malondialdehyde (MDA) and reduced glutathione (GSH), and heart weight were determined. Histochemical staining with the Van Gieson, PAS/Alcian blue at pH 2.5 and Masson's trichrome methods were performed in addition to hematoxylin-eosin staining. Levels of serum cholesterol and tissue MDA, and heart weight were increased in pinealectomized rats whereas GSH levels did not change. Melatonin administration reversed these effects. Microscopically, myocardial fibrosis and myxomatous degeneration of cardiac valves were detected in all pinealectomized rats. It can be concluded that pinealectomy of rats causes morphological changes in rat hearts, and short-term application of melatonin does not reverse these changes.     

Androgenic control of porphyrin in the harderian glands of the male syrian hamster is modulated by the photoperiod,which suggests that the sexual differences in porphyrin concentrations in this gland are important functionally

Background: The porphyrin concentrations of the Harderian glands of Syrian hamsters show marked sexual differences, with male levels being much lower than those of females. Porphyrinogenesis is inhibited by androgens, so orchidectomy leads to elevated male porphyrin concentrations; however, a number of other procedures (some of which also lower androgen levels) prevent this. We studied the effects of short-day photoperiods and melatonin on Harderian porphyrin concentrations. Methods: Intact, castrated, or pinealectomized hamsters of both sexes were exposed to long-day or short-day photoperiods. Intact or castrated hamsters were given melatonin injections in the morning or the afternoon, or were given beeswax pellets containing melatonin. After a variable period, Harderian glands were dissected and porphyrins were measured. Results: Prolonged short-day exposure (13 weeks) led to increased Harderian porphyrin concentrations and this rise was prevented by pinealectomy. The rise in Harderian porphyrins following short-day exposure was small, compared with that following castration. Short-day photoperiods also prevented the rise in porphyrin levels associated with castration and this effect was prevented by removal of the pineal. Melatonin injections, whether given in the morning or in the afternoon, had no effect on Harderian porphyrin concentration of castrated male hamsters. Continuous release melatonin pellets reduced the postcastrational rise in porphyrin levels in one experiment, while having no effect in another. In female hamster, neither short photoperiods nor melatonin pellets influenced Harderian porphyrin concentrations. Conclusions: These results suggested that a factor from the pineal gland helps maintain the low levels of porphyrin which are characteristic of male Harderian glands, despite the decrease in androgen levels which typically results from exposure to short days. Morning and afternoon injections of melatonin and continuous release melatonin pellets failed to resolve the question of whether this pineal factor is melatonin. Our results demonstrated that low male and high female porphyrin levels are maintained in Syrian hamsters, despite seasonal variations in the hormonal milieu, suggesting that these sexual differences are important for the (still unestablished) function of the Harderian glands in this species. © 1994 Wiley-Liss, Inc.     

Influence of pinealectomy and photoperiod on courtship and nest-building in male doves

Sham-operated male doves displayed greater nesting activity under long days (16L:8D) than under short days (8L:16D). This difference was abolished by pinealectomy—the activity of long-day pinealectomized birds dropped to a level comparable to that seen on short days. Removal of the pineal had no effect under the short photoperiod. These results are not due to changes in peripheral androgen production as castrated, testosterone-implanted males also exhibited higher levels of nest-building on long days if sham-operated, but not if they were pinealectomized. Neither treatment had a significant effect on courtship or copulatory behavior. It is suggested that the pineal mediates the stimulatory effect of long photoperiods on hormonally-induced nest-building in doves.     

The pineal gland is not essential for circadian expression of rat period homologue (rper2) mRNA in the suprachiasmatic nucleus and peripheral tissues

To investigate the functional involvement of the pineal gland in circadian expression of the rat period homolog gene (rPer2) in the suprachiasmatic nucleus (SCN) and peripheral tissues, we performed Northern blot analysis in tissues from pinealectomized rats. The ectomy did not have any significant effects on rPer2 mRNA expression patterns both in a daily light-dark condition and in a constant darkness. These results suggest that the rhythmic secretion of pineal melatonin is not essential for the circadian expression of clock genes in the SCN and other peripheral tissues of rats.     

Pinealectomy Does Not Affect the Healing of Experimental Colonic Anastomoses

Gastrointestinal system anastomoses, especially colonic anastomoses, have significant morbidity and mortality despite recent technical improvements. Besides regulating the circadian rhythm, the pineal gland and its main neurohormone product melatonin have widespread actions in the organism. The purpose of this study was to investigate the effects of pinealectomy on the healing of colonic anastomoses. One hundred male albino Wistar rats were used in this study. The rats were separated into three groups: control, pinealectomy, and sham groups. In the control group, only colonic resection and anastomoses were performed. Following pinealectomy, colonic anastomosis was performed 2 weeks later on one half and 2 months later on the other half of the pinealectomy group. Only craniotomy was performed on the sham group, and the rats were separated and evaluated like the pinealectomy group. Colonic anastomoses were evaluated on postanastomotic day 3 and 7 by measuring the bursting pressure and the hydroxyproline levels in the anastomotic segments. There was no difference in the bursting pressure measurements between the groups on both postoperative day 3 and 7. Although hydroxyproline levels were different between groups on both postanastomotic days 3 and 7, it has been observed that neither normal nor anastomotic hydroxyproline levels influenced the anastomotic bursting pressure measurements. The percent deviation from the normal values was compared in the anastomotic segments, and no differences were found regarding the bursting pressure and hydroxyproline levels. It was concluded that pinealectomy has no effect on the healing of colonic anastomoses.     

Surgical pinealectomy accelerates intervertebral disc degeneration process in chicken

Despite the importance of intervertebral disc (IVD) degeneration both in research and clinical practice, the underlying biological mechanism of this phenomenon remains obscure. The current study investigated the effects of neonatal pinealectomy on the development of IVD degeneration process in chicken. Thirty chicks (3 days of age) were divided into two equal groups: unoperated controls (Group X) and pinealectomized chicks (Group Y). Pinealectomies were performed at the age of 3 days. At the age of 8 weeks, magnetic resonance imaging examination of one animal in each experimental group was taken. At the end of the study, serum melatonin level was determined by using ELISA method and histopathological or biochemical examination of specimens from all subjects was done. The results of biochemical analyses were compared using Mann–Whitney U test, whereas The Chi-square test was adopted for the histological findings. In this study, the serum melatonin levels in Group Y were significantly lower than those in Group X (P < 0.001). Similarly, scoliosis was developed in 14 out of 15 (93%) in Group Y. Hydroxyproline content of IVD tissue was high in Group Y compared with the values in Group X, although there was no significant difference. Histologically, an appearance of normal IVD was observed in Group X, while the presence of a degenerated IVD was observed in Group Y. From the results of the current study, it is evident that surgical pinealectomy in new-hatched Hybro Broiler chicks has a significant effect on serum melatonin level as well as on the development of IVD degeneration and spinal malformation. In the light of these results from present animal study, melatonin may play a role in the development of IVD degeneration in human beings, but this suggestion need to be validated in the human setting.     

Effect of melatonin and pinealectomy on avoidance and exploratory activity in the rat

Melatonin in a daily dose of 50 μg/rat facilitated the extinction of active avoidance reflex and decreased the intertrial activity during extinction. However, it had no effect on learning and intertrial activity during acquisition. Pinealectomy was ineffective on acquisition, extinction and intertrial activity. Melatonin (100 μg/rat) given in 2 consecutive days facilitated the passive avoidance behavior in water deprived animals in two different experimental situations. Fifty μg of melatonin or pinealectomy was without effect on passive avoidance behavior. Neither melatonin nor pinealectomy had any influence on water intake or on exploratory activity.     

Effects of pinealectomy and melatonin supplementation on endometrial explants in a rat model

Objective

To determine the effects of pinealectomy on endometrial explants in rats and evaluate the activity of superoxide dismutase (SOD) and catalase (CAT) and the levels of malondialdehyde (MDA) in the rat endometriosis model.

Study design

Rats with experimentally induced endometriosis were randomly divided into three groups after second-look laparotomies. Group 1 (pinealectomy, n = 8) and Group 2 (pinealectomy + melatonin, n = 8) underwent pinealectomies after the second-look laparotomies. Group 3 was presented as control group (vehicle solution + without pinealectomy (n = 6)). Melatonin was administered intraperitoneally for 4 weeks in Group 2, whereas an equal volume of vehicle solution was given to Groups 1 and 3. Evaluation of the volume of the endometrial explants, histopathological examination and preservation of explant epitheliums according to the scoring system were undertaken.

Results

There was a statistically significant increase in spherical explant volumes of Group 1 compared to Groups 2 and 3. In Group 1, the level of MDA was significantly higher and SOD and CAT activity was significantly lower compared to Groups 2 and 3. A statistically significant increase in the epithelial lining scores of explants was noted in Group 1 compared to Groups 2 and 3.

Conclusion

The effects of pinealectomy on the progression of endometriosis explants were reversed by melatonin.