Histochemical localization of cholinesterases in the neural tissue of the pineal in the rhesus monkey.

The neural tissue of the monkey pineal contains both acetyl and butyryl cholinesterases. Acetylcholinesterase was localized in the cisternae of the nuclear membrane, rough endoplasmic reticulum, on the plasma membrane of the neurones, and on the axolemma of both non-myelinated and myelinated fibres. The enzyme was not found in the axosomatic or axo-dendritic synapses. It is therefore suggested that the pineal neurones have a cholinergic function rather than a cholinoceptive one.     

Histochemical localization of cholinesterases in the neural tissue of the pineal in the rhesus monkey

Summary The neural tissue of the monkey pineal contains both acetyl and butyryl cholinesterases. Acetylcholinesterase was localized in the cisternae of the nuclear membrane, rough endoplasmic reticulum, on the plasma membrane of the neurones, and on the axolemma of both non-myelinated and myelinated fibres. The enzyme was not found in the axosomatic or axo-dendritic synapses. It is therefore suggested that the pineal neurones have a cholinergic function rather than a cholinoceptive one.This study was supported by the WHO.     

Neither prolactin nor growth hormone restore the nocturnal rise in pineal N-acetyltransferase activity or melatonin content in hypophysectomized rats

Hypophysectomy in adult male rats greatly attenuated the nocturnal rise in both pineal N-acetyltransferase (NAT) activity and melatonin content. High nighttime levels of NAT and melatonin were not restored by treating the animals with either prolactin or growth hormone, alone or in combination. Treating intact rats with bromocriptine, which depresses circulating prolactin levels, also was without effect on pineal melatonin synthesis. It appears that neither prolactin nor growth hormone are of major importance in determining pineal melatonin production.     

Neither prolactin nor growth hormone restore the nocturnal rise in pineal N-acetyltransferase activity or melatonin content in hypophysectomized rats

Summary Hypophysectomy in adult male rats greatly attenuated the nocturnal rise in both pineal N-acetyltransferase (NAT) activity and melatonin content. High nighttime levels of NAT and melatonin were not restored by treating the animals with either prolactin or growth hormone, alone or in combination. Treating intact rats with bromocriptine, which depresses circulating prolactin levels, also was without effect on pineal melatonin synthesis. It appears that neither prolactin nor growth hormone are of major importance in determining pineal melatonin production.     

Vasoactive intestinal peptide (VIP) occurs in nerves of the pineal gland

Summary Nerves staining with antibodies against vasoactive intestinal peptide (VIP) were detected in the pineal gland of the rabbit, cat and pig. VIP nerves were numerous in the cat but few in the rabbit and pig. A particularly rich VIP nerve supply was noted in the pineal stalk of the cat. The nerves were predominantly located around small blood vessels. Occasionally, nerve fibres were seen in the glandular parenchyma without obvious relation to blood vessels.Grant support from the Swedish Medical Research Council (04X-4499).     

Influence of pinealectomy on corticotropin (ACTH)

Sensory deprivation produced by removing the eyes and olfactory bulbs in male rats allowed pinealectomy to markedly augment the post-adrenalectomy elevation of ACTH levels. Pineal removal or sensory deprivation separately did not have this effect. Thus, intact sensory input and an intact pineal gland are independently capable of restricting the post-adrenalectomy rise in ACTH levels.     

Effects of sham-pinealectomy,performed under white and red light,on the melatonin content of rat pineal glands

Summary Sham-pinealectomy, performed under different light conditions in newborn and adult rats, is followed by changes of pineal activity resulting in variations of melatonin content. The pineal glands of rats sham-operated under white light produce significantly less melatonin. In contrast, glands of rats operated on under red light show a melatonin content corresponding to that of intact rats. This result implies that normal white light causes a disturbance in melatonin production by a non-retinal pathway.     

Estimation of the methylating capacity of the pineal gland. With special reference to indole metabolism

In order to obtain more information on the methylating capacity of the pineal gland, a method determining the formation of different 5-methoxyindoles in the pineal gland was developed. The method depends on measuring the incorporation of labelled methyl groups into the various hydroxyindoles present in the pineal gland, after incorporation of pineal tissue with labelled S-adenosyl methionine. Hydroxyindoles were not added to the incubation medium. After incubation thin-layer chromatography was performed with pineal tissue together with the incubation medium; the spots were scraped and counted.     

Influence of pinealectomy on corticotropin (ACTH)

Summary Sensory deprivation produced by removing the eyes and olfactory bulbs in male rats allowed pinealectomy to markedly augment the post-adrenalectomy elevation of ACTH levels. Pineal removal or sensory deprivation separately did not have this effect. Thus, intact sensory input and an intact pineal gland are independently capable of restricting the post-adrenalectomy rise in ACTH levels.This work was supported in part by NIH Grant No. P30 HD 10202. We thank F. Lynd and R.J. Reiter for advice, P. Starr and J. Sackman for technical assistance, and Bess Mitchell for typing.     

The process of survival of denervated and freely autotransplantated skeletal muscle.

The study shows that two distinct processes occur in free autografts of denervated intact muscle during the early critical phase of the transplantation: survival of transplanted fibres at the periphery of the graft, and regeneration of new muscle fibres following breakdown of the originally transplanted fibres in the central areas.     

Estimation of the methylating capacity of the pineal gland. With special reference to indole metabolism

Summary In order to obtain more information on the methylating capacity of the pineal gland, a method determining the formation of different 5-methoxyindoles in the pineal gland was developed. The method depends on measuring the incorporation of labelled methyl groups into the various hydroxyindoles present in the pineal gland, after incorporation of pineal tissue with labelled S-adenosyl methionine. Hydroxyindoles were not added to the incubation medium. After incubation thin-layer chromatography was performed with pineal tissue together with the incubation medium; the spots were scraped and counted.The authors wish to express their gratitude to Prof. Dr. J.C. van de Kamer and Dr F.C.G. van de Veerdonk for their stimulating interest in these studies.     

Ovarian response of pinealectomized and intact white-footed mice kept under naturally short photoperiods

Summary Pinealectomized and intact adult female white-footed mice were maintained outdoors from 28 November to 2 January. At the termination of the experiment, ovarian weight was significantly greater (p<0.02) in the pinealectomized group compared with that of the intact animals. Microscopic examination of the ovaries revealed that those of the intact animals had fewer and smaller follicles than those from animals which had had their pineal glands removed.Supported by NSF grant PCM 77-05734 and NIH grant HD 10202 through the Center for Research in Reproductive Biology Morphology Core.     

Variation in the number and size of myelinated nerve fibers regenerated after repeated localized freezing of the sciatic nerve of rats]

The number and size of myelinated fibres have been determined in the nerve to medial head of the gastrocnemius muscle of Rats in which the sciatic nerve had been frozen locally 1 to 5 times at monthly intervals. The contralateral nerve was used as a control. When the measurements were made one month after the last freezing, the number of fibres increased progressively until the 3rd freezing, reaching about 220% of the normal value. No higher values were observed after 4 or 5 freezings. The mean diameter of fibres decreased with the number of operations. When the measurements were made 3, 6, 12 or 18 months after the 3rd freezing, the number of fibres decreased by about 30% between the 1st and the 3rd month and then stabilized. The mean diameter of fibres increased progressively. However, at the 18th month, the size of the myelinated fibres had not reached the normal value.     

Evidence for no relationship of the pineal activity with the neonatal shrinkage of Leydig cells in the rat

Summary The collective volume of Leydig cells in prenatally pinealectomized newborn rats declined as sharply as in intact newborn rats. Also, the collective volume in pups born in the dark declined in a fashion similar to that in pups born in the light. The results indicate that neither the pineal gland nor the light is responsible for the neonatal shrinkage of Leydig cells.This study was supported in part by a Grant-in Aid for Scientific Research (No. 548067) from the Ministry of Education, Science and Culture of Japan.     

Development of post-hatching melatonin rhythm in zebra finches (Poephila guttata)

We examined levels of melatonin in the pineal, eyes and plasma over a 24 h period during development in the altricial zebra finch. Beginning as early as 2 days after hatching there was a distinct 24 h rhythm in melatonin in the pineal and plasma. Beginning at day seven after hatching there was also a 24 h rhythm present in the eyes. In the pineal and eyes the amplitude of the 24 h rhythm increased with age. In contrast, the amplitude of the plasma melatonin rhythm at 2 days was already within the range of adults and did not increase with age. These results confirm and expand earlier findings in the European starling and parallel those from precocial birds indicating that the circadian system is already competent at or shortly after hatching even in atricial birds.     

Ultrastructure and biochemistry of the pineal organ in deep-sea lanternfishes (Myctophidae)

Summary Pineal structural and biochemical adaptations in lanternfishes included: 1) few photoreceptor outer segment discs; 2) conventional synapses between photoreceptors and pineal neurons; and 3) low levels (0–60 pg/pineal) of serotonin compared to those (>1.0 ng/pineal) in the goldfish pineal organ. These findings suggest reduced photosensory and/or neuroendocrine functions in these deep-sea fishes.     

Ultrastructure and biochemistry of the pineal organ in deep-sea lanternfishes (Myctophidae)

Pineal structural and biochemical adaptations in lanternfishes included: 1) few photoreceptor outer segment discs; 2) conventional synapses between photoreceptors and pineal neurons; and 3) low levels (0-60 pg/pineal) of serotonin compared to those (greater than 1.0 ng/pineal) in the goldfish pineal organ. These findings suggest reduced photosensory and/or neuroendocrine functions in these deep-sea fishes.     

Effects of a bovine pineal peptidic fraction (E5) on plasma and pituitary levels of LH,FSH and prolactin

Summary Pituitary levels of prolactin were significantly diminished after 5 s.c. injections spaced 12 h apart of 5 g of a partially purified bovine pineal peptide fraction (E₅) in both intact and castrated rats. E₅ treatment did not significantly affect the castration-induced changes in plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH) or pituitary LH but did partially block the pituitary fall in FSH in castrated animals.Supported by NSF grant PCM 8003441 and NIH Center for Reproductive Biology (Bioassay Core) No.P30 HD 10202. Many thanks are due A. Moreno for his excellent technical assistance.     

Similarity between the effects of suprachiasmatic nuclei lesions and of pinealectomy on gonadotropin release in ovariectomized,sulpiride-treated and melatonin-replaced rats

The aim of this study was to compare the effects of pineal indole treatments on LH and FSH release in pinealectomized and suprachiasmatic lesioned and ovariectomized rats rendered hyperprolactinemic by acute sulpiride treatment. pinealectomy or suprachiasmatic nuclei lesions in female rats both decreased plasma LH and FHS at 10, but not at 20 d after surgery, whereas the daily afternoon administration of melatonin effectively restored levels of both gonadotropins to control values. In ovariectomized rats, pinealectomy or suprachiasmatic nuclei lesions were ineffective in counteracting the high plasma levels of LH and FSH. However, sulpiride treatment in both pinealectomized and suprachiasmatic nuclei lesioned and castrated female rats significantly decreased the levels of LH and FSH, an effect which was counteracted by daily afternoon melatonin administration. Other pineal indoles tested, i.e., 5-hydroxy- and 5-methoxytryptophol, were ineffective in regulating gonadotropin levels. The results suggest that the pineal gland, through its hormone melatonin, can modulate gonadotropin secretion by acting on a dopamine mechanism independent of hypothalamic suprachiasmatic areas.     

Elaunin fibres in the basement membrane of sweat gland secretory coil are rich in disulfide-groups.

Disulfide-groups of elaunin fibres of sweat gland basement membrane are demonstrated a) by thiosulfation/aldehyde-fuchsin staining or thiosulfation/Alcian Blue + 0.8 M MgCl2 staining, and b) by identifying SH-groups after reduction with sodiumthioglycollate. Elaunin fibres share this staining behaviour with "elastic fibre microfibrils" and with oxytalan fibres.