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Biomedical subjects

L Vollrath

Publications and source records attributed to L Vollrath.

At least 127 records · Page 7Linked to original sources

The ultrastructure of the nerve fibers and pinealocytes in the rat pineal stalk.

In view of the increasing interest in the central innervation of the mammalian pineal gland, this aspect was studied in depth in the rat. This species is especially suited since the nerve fibers in question form a distinct bundle running from the deep to the superficial pineal gland through the pineal stalk. The axons were counted and analysed ultrastructurally in the pineal stalks cut transversely at three levels (proximal, intermediate, and distal) relative to the neural axis and in longitudinal sections. The number of nerve fibers was highly variable, ranging from 551 to 1,132 proximally and from 110 to 448 distally, indicating that many fibers terminate in the stalk or leave the stalk after forming a loop. Large myelinated axons, which are abundant proximally, appear to lose their sheaths along their course through the stalk. Most of the axons were small and unmyelinated. A few of these had the appearance of sympathetic fibers and disappeared after sympathectomy. Others contained abundant neurosecretory granules, and, according to the literature, may originate in the hypothalamic paraventricular nuclei. The majority of the small axons which are apparently devoid of granules and dense-cored vesicles may come from the habenular nuclei and the stria medullaris. In addition to axons, the stalk contains astrocytes, a few oligodendrocytes and Schwann cells, as well as pinealocytes identical to those of the superficial pineal gland.

Acid Phosphatase↗

Different types of magnetically sensitive cells in the rat pineal gland.

Extracellular recordings from rat pinealocytes reveal different cell responses following experimental changes in the horizontal component of the ambient magnetic field. While two-thirds of the units recorded did not respond at all, one group is activated, the activation continuing after switching off the magnetic stimuli. These cells were not further activated by a second stimulus. Another group comprises cells characterized by a sustained inhibition or excitation, respectively, during magnetic stimulation. These cells could be influenced by a second stimulus. After guinea pigs and homing pigeons the rat is now the third species in which the pineal gland has been shown electrophysiologically to respond to changes of the ambient magnetic field.

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"Synaptic" ribbons and spherules of the guinea pig pineal gland: inverse day/night differences in number.

The present study deals with the functionally enigmatic "synaptic" ribbons and spherules of guinea pig pinealocytes. Whereas the ribbons have been shown to exhibit a 24-hr rhythmicity with low numbers during the day and high numbers at night, very little of a definitive nature is known about the spherules. Sixteen male guinea pigs of the Hartley strain were perfusion fixed, 8 between 0900-1100 hr, and 8 between 2100-2300 hr. The ribbons and spherules were counted in the pineal parenchyma of the proximal, intermediate, and distal regions. In confirmation of earlier studies, it was found that "synaptic" ribbons are equally abundant in the proximal, intermediate, and distal regions of the gland, during both the day and the night, and that they increase significantly in number at night when compared with daytime values. The spherules, by contrast, are more abundant proximally and are present in greater numbers during the day than at night. As ribbons and spherules are usually not found in one and the same pinealocytic profile, and based on previous electrophysiological studies, it is proposed that the ribbons are morphological markers of nocturnally active pinealocytes, whereas spherules characterize diurnally active pinealocytes.

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Depressive effect of LHRH on the numbers of "synaptic" ribbons and spherules in the pineal gland of diestrous rats.

Previous studies have shown that LHRH or LHRH-like substances are present in the pineal gland. In order to investigate whether exogenous LHRH may affect the pineal gland, in the present study the effects of a single dose of LHRH (1 microgram, i.p.) on pineal "synaptic" ribbons and spherules as well as serum melatonin levels were examined in diestrous Wistar rats. One hour after the injection both ribbons and spherules exhibited a statistically significant decrease in number. Serum melatonin levels were not affected. It is concluded that humoral feedback mechanisms may exist between the hypothalamus and the pineal gland.

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Effects of an artificial magnetic field on serotonin N-acetyltransferase activity and melatonin content of the rat pineal gland.

In the present study the effects of artificial magnetic fields on pineal serotonin-N-acetyltransferase (NAT) activity and melatonin content in male Sprague-Dawley rats were investigated to study the secretory activity of the pineal gland. Experimental inversion of the horizontal component of the natural magnetic field, performed at night-time, led to a significant decrease of both parameters investigated. During day-time, this effect was less conspicuous. During night-time, inversion of the horizontal component is followed by a reduced pineal secretory activity for about 2 h. After 24 h exposure to the inverted horizontal component, return to the natural condition was followed by a renewed clear depression of pineal NAT activity and melatonin content, indicating that the main stimulus is not the inverted magnetic field itself but rather its change. Changing the inclination of the local magnetic field from 63 degrees to 58 degrees, 68 degrees or 78 degrees, respectively also decreased the secretory activity of the rat pineal gland.

Acetyltransferases↗

Pineal "synaptic" ribbons and spherules during the estrous cycle in rats.

In previous studies pineal "synaptic" ribbons have been shown to undergo striking numerical changes under various physiological and experimental conditions and to be regulated by beta-adrenergic mechanisms. The aim of the present investigation was to study the numbers of pineal "synaptic" ribbons and spherules in Wistar rats throughout the estrous cycle and to compare them with those in males. There were no statistically significant differences in the numbers of ribbons and spherules between males and females and in the females at the different stages of the estrous cycle, indicating that the structures in question, in vivo, do not appear to be regulated by naturally occurring changes of sex steroid hormones and gonadotrophins.

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24-hour-variation of pineal gland volume, pinealocyte nuclear volume and mitotic activity in male Sprague-Dawley rats.

In two experiments carried out on two alternate days, the 24-h-rhythmicity of pineal gland volume, pinealocyte nuclear size in cortex and medulla and mitotic activity were studied in male Sprague-Dawley rats, to assess to what extent morphological parameters reflect the pronounced day/night differences in pineal melatonin formation. Pineal volume exhibited statistically significant changes in the second experiment only, with a distinct trough at 6 p.m. Karyometry revealed highly variable patterns. In the first experiment, pinealocyte nuclear changes lacked parallelism in cortex and medulla. The cortex exhibited a bimodal curve with peaks at noon of the first day and at 6 a.m. of the second day, and two troughs at 6 a.m. and midnight respectively of the first day. The medulla showed no clear-cut rhythmicity. In the second experiment, cortex and medulla reacted similarly, nuclear size decreasing from 6 a.m. to 6 p.m., remaining low thereafter. Mitotic activity of pinealocytes is low (on average 23 mitotic figures/gland). In both experiments statistically significant differences existed between certain times, pointing in the direction of 24-h-rhythmicity, but whereas the curve exhibited a peak at midnight in the first experiment, mitotic activity in the second experiment showed a trough at midnight. It is concluded that for as yet unexplained reasons morphological parameters do not appear to accurately reflect circadian rhythmicity of pineal melatonin formation.

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Mitotic activity and its 24-hour rhythm in the rat pineal gland.

Previous studies have yielded equivocal results concerning the 24-hour rhythmicity of mitotic activity in the rat pineal. The aim of the present study was to re-investigate this problem by carrying out three separate 24-hour experiments on alternate days. The results obtained confirm previous findings showing that in the pineal gland of adults mitotic activity is low. On average 22.3 mitotic figures of pinealocytes are seen per pineal gland, corresponding to a mitotic index of 0.2-0.6/1,000 pinealocytes. Mitotic activity is distinctly higher at daytime than at night. The timing of the peaks and troughs differs slightly from experiment to experiment. The majority of observations now indicate that in the rat pineal gland mitotic activity is higher at day time than at night.

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Pineal complex of rats: effects of superficial pinealectomy on the deep pineal.

Previous studies have shown that in rodents the pineal complex may consist of two parts, i.e. the small deep pineal and the large superficial pineal. As commonly used techniques of pinealectomy remove the superficial pineal only, it was decided to examine what effect this operation has on the deep pineal in male Sprague-Dawley rats. 3 and 6 weeks after superficial pinealectomy, the deep pineal exhibited a clear decrease in volume, due to an atrophy of the intrinsic pinealocytes. An atrophy was still noticeable after 6 or 12 months. It is concluded that after superficial pinealectomy the deep pineal is not an important factor with respect to melatonin formation, in particular, as there are extrapineal sites of melatonin formation.

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Morphological findings relating to the problem of cortex and medulla in the pineal glands of rat and hamster.

Because, in previous investigations on the rat pineal gland, karyometric studies of pinealocytes from cortical and medullary regions had yielded contradictory results, experiments were carried out to resolve this problem. In immersion-fixed, paraffin-embedded pineal glands, nuclear size of cortical regions was invariably larger than that in the medulla, the nuclear size clearly depending on the plane of sectioning. The differences between cortex and medulla were abolished in (a) pineal glands sagittally bisected prior to immersion fixation and (b) perfusion-fixed, epoxy resin-embedded pineal tissue, suggesting that unequal pinealocyte nuclear size in cortex and medulla is artefactual. In the hamster pineal gland, cortical and medullary regions are separated by a narrow band of interstitial cells, indicating that different structural features account for this subdivision in different species.

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Anomalous occurrence of immunoreactive calcitonin cells in the thymus of the rat.

In a study of the effect of pinealectomy on thyroid C-cell number, 8 animals out of 66 were found to have thymic tissue in close association with the thyroid. Cells containing immunoreactive calcitonin were found in all of the thyroids but in only one of the 8 pieces of thymus. These cells found in a piece of thymic tissue associated with the right thyroid lobe were located immediately under the capsule and did not form or associate with follicles. Unlike the other animals the rat with thymic calcitonin cells had an unequal distribution of C-cells between the left and right thyroid lobes, but the total number of thyroidal C-cells was the same as that of the other rats. Since the thymus proper was not examined in these 66 animals, ten additional rats were taken for such a study. Thyroid-associated thymic tissue was found in three of these, but none of these thymi showed any immunoreactive cells.

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"Synaptic" ribbons and spherules of the rat pineal gland: day/night changes in vitro?

In the present study pineal glands of rats aged 69-71 days were studied in vivo and in vitro with respect to day/night changes of "synaptic" ribbons and spherules. It was found that ribbons outnumber spherules by a factor of 3. In vivo, both ribbons and spherules show a roughly 3-fold increase in number at 1 a.m. when compared to 1 p.m. Up to 39 h in vitro, the two structures in question did not reveal day/night differences in amount, suggesting that diurnal rhythmicity of the gland did apparently not persist in organ culture. After 3 h in organ culture, the spherules, but not the ribbons, showed a striking increase in number, showing that ribbons and spherules may be governed by different mechanisms.

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Structural dissimilarities in different regions of the pineal gland of Pirbright white guinea-pigs.

In the present study, the proximal, intermediate and distal parts of dumbbell-shaped pineal glands of Pirbright White guinea-pigs were investigated with respect to structural dissimilarities. The most conspicuous difference was that while in the proximal part the pinealocytes were arranged in a random fashion, the intermediate and distal regions, except a small ventral area in the latter, exhibited a prominent follicular arrangement of the pinealocytes. Concretions were restricted to the subcapsular region of the intermediate part, whereas cells with hyperchromatic nuclei increased in number in an anterior-posterior direction. In males, pinealocyte nuclear size did not differ significantly in the regions investigated, whereas in females the karyometric results were much more heterogeneous. Exposure of male guinea-pigs to continuous illumination for 45 days resulted in (i) a 25% decrease of pineal volume, the intermediate area being relatively little affected, (ii) an increase in the number of follicles and (iii) a decrease in pinealocyte nuclear size, especially in a ventral zone of the proximal part of the pineal gland.

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Alterations in the spontaneous activity of cells in the guinea pig pineal gland and visual system produced by pineal indoles.

The indoles serotonin (SER), melatonin (MEL), 5-methoxytryptophol (5-MTL) and 5-hydroxytryptophol (5-HTL) were administered during daytime microelectrophoretically to 240 cells in the pineal gland of the guniea-pig. The action of SER and 5-HTL was predominantly depressant on the electrical activity, MEL and 5-MTL caused an excitation in most of the units. Although MEL and 5-MTL caused fairly similar reactions on average, they appear to act on different cells. The effects of microelectrophoretically applied MEL and 5-MTL on the spontaneous or evoked activity in the visual system (retinal ganglion cells, optic tract, lateral lateral geniculate body, superior colliculus) of the guinea-pig were also investigated. Of the 76 cells tested in the visual system 25 of the ON- and OFF-cells increased the rate of discharge when the two indoles were applied. Cells in the optic layer of the superior colliculus showed no measurable response to the application of the two substances. 5-HTL caused no effect on cells in the visual system.

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Ultrastructural observations on the central innervation of the guinea-pig pineal gland.

In the present study the central innervation of the guinea-pig pineal gland was investigated. The habenulae and the pineal stalk contain myelinated and non-myelinated nerve fibres with few dense-cored and electron-lucent vesicles. Some myelinated fibres leave the main nerve fibre bundles, lose their myelin-sheaths and terminate in the pineal gland. Although direct proof is lacking, the non-myelinated fibres appear to end near the site where the bulk of the myelinated fibres are located. Here a neuropil area exists where synapses between non-myelinated fibre elements are abundant. Neurosecretory fibres were also seen. The results support the concept of functional interrelationships between hypothalamus, epithalamus and the pineal gland.

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Electrical responses of pineal cells to melatonin and putative transmitters. Evidence for circadian changes in sensitivity.

The effects of microelectrophoretically applied melatonin (N-acetyl-5-methoxy-tryptamine), noradrenaline (NOR) or acetylcholine (ACH) on the electrical activity of pineal cells during both the day- and night-time were studied in urethane anesthetised guinea-pigs. A total of 288 cells were tested with melatonin, and in addition with either NOR (120 cells) or ACH (138 cells). Of the 206 cells responding to melatonin application, 139 were excited and 67 inhibited. A total of 85 cells responded to the application of NOR, 45 being excited and 40 inhibited. Responses to ACH application were observed in 75 pineal cells, 49 units being inhibited and 26 excited. It was possible to observe excitation in 18 units following electrical stimulation of the lateral habenular nucleus, and of these cells, nine units were additionally tested with ACH. Application of the cholinergic transmitter caused enhanced activity in eight of these cells, the remaining unit being inhibited. The responses to melatonin and NOR varied significantly depending on whether the cells were tested during the day or night. It is apparent from these studies that (1) melatonin, noradrenaline and acetylcholine can bring about direct changes in the firing frequency of pineals cells, and (2) pineal cells do not respond to these substances in the same way. Furthermore, since melatonin application had no effect when nerve cells in brain regions surrounding the pineal organ were tested, the results suggest that melatonin, either produced in the pineal or reaching the gland in the general circulation may be capable of directly influencing the cells engaged in its own synthesis.

Acetylcholine↗

The effects of sex hormones, prolactin, and chorionic gonadotropin on pineal electrical activity in guinea pigs.

Microelectrophoretic application of sex hormones onto pineal cells in guinea pigs has shown different responses in pregnant females as compared to males. In pregnant females estrone caused excitation in 74% of the cells tested, while progesterone and testosterone, prolactin, and HCG were inhibitory in a majority of the cells tested, while progesterone and testosterone, prolactin, and HCG were inhibitory in a majority of the cells. In contrast, in males estrone caused excitation of only 19% but inhibition of 37%. A smaller percentage of cells was inhibited by progesterone, while the predominant response to testosterone was excitation. These results suggest that the pineal gland may be under a feedback control.

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Morphological and electrophysiological evidence for habenular influence on the guinea-pig pineal gland.

The central innervation of the guinea-pig pineal gland was investigated by histological and electrophysiological methods: Staining the pineal gland and the epithalamus, a double route of central innervation could be shown in the anterior part of the organ: (a) Fibres from the habenular nuclei, mainly from the lateral part, penetrate the organ via the pineal stalk. (b) Other fibres join the striae medullares and running in the habenulae reach the organ more dorsally. The fibres end in the intercellular space where they form a dense network. In 15 male guinea-pigs under urethane anesthesia, two series of unit recording experiments were performed: (a) Recordings were made from 128 units in the posterior and anterior part of the pineal gland and the effects of electrical stimulation of the habenular nuclei were observed. Lateral habenular stimulation influenced 44% of the units. 80% of these were excited and 20% were inhibited. (b) Recordings were made from 42 units in the lateral habenular nucleus. Twelve units (29%) responded with an augmentation of spontaneous activity following pineal gland stimulation. No inhibition response was observed.

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