Biomedical subjects

L Vollrath

Publications and source records attributed to L Vollrath.

At least 55 records · Page 3Linked to original sources

PubMed1992

Proliferation-associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture.

The oxygen consumption rate, proliferative activity, and morphology of EMT6/Ro mouse mammary sarcoma cells in monolayer and multicellular spheroid culture have been investigated in a comparative study. During the transition of monolayer cells from the exponential into the plateau growth phase, there is a distinct decrease in the cellular volume that is associated with a corresponding decrease in the proliferative and respiratory activity of the cells. The decline in cell volume is mainly due to a decrease in the content of cytoplasm, whereas the size of the nucleus is only slightly reduced. A concomitant decrease in the number of mitochondria per cell obviously accounts for the reduction in cellular oxygen uptake. Despite a continuous decrease of cell proliferation from the surface to interior regions of EMT6 spheroids reflected by a gradient in tritiated thymidine labeling, volume-related oxygen consumption is rather uniform in viable regions of these aggregates. The finding can be explained by the results of the morphometric evaluation showing a uniform volume density of mitochondria, i.e., of oxygen-consuming sites within these spheroids.

Animals↗

PubMed1992

In vitro effects of putative neurotransmitters on synaptic ribbon numbers and N-acetyltransferase activity in the rat pineal gland.

The pineal contains a large number of classical transmitters and neuropeptides. Some of these neurochemicals are involved in the regulation of serotonin N-acetyltransferase (NAT) activity and hence in melatonin synthesis. Synaptic ribbons present in the pineal gland also exhibit a numerical day/night rhythm parallel to that of NAT activity. There is scarcity of information regarding the regulation of synaptic ribbon (SR) numbers. In the present study, we have investigated in vitro effects of a number of classical neurotransmitters and neuropeptides. NAT activity was used to monitor melatonin synthesis under the experimental conditions used. Norepinephrine (NE), Delta sleep-inducing peptide (DSIP), vasoactive intestinal polypeptide (VIP), adenosine and N-acetyl-asp-glu (NAAG) significantly increased NAT activity in rat pineal. DSIP and VIP also increase the stimulatory effect of NE on NAT activity. These neurochemicals had no effect on SR numbers. Gamma aminobutyric acid (GABA), serotonin and taurine affected neither NAT activity nor SR. Somatostatin increased SR numbers significantly, without having any effect on NAT activity. The effect of somatostatin is regarded to be pharmacologic, since rather high dosages (10(-4) M) were required to obtain a significant effect. Although somatostatin is present in the pineal and may change rhythmically, the inconsistency of the day/night rhythmicity and the lack of such a rhythm in female rats and male gerbils speaks against an important physiological role of somatostatin in regulating SR numbers.

Animals↗

PubMed1992

Synaptic ribbons, spheres and intermediate structures in the developing rat retina.

The present study was conducted to investigate the qualitative and quantitative development of synaptic bodies in retinae of Wistar rats during postnatal days 4-28. In addition, the effects of different light regimens and of eye pigmentation on SB numbers were studied. Synaptic bodies were counted and measured in the outer plexiform layer of retinal tissue fixed and processed by routine electron microscopical techniques. At postnatal days 4 and 5, retinae showed only few synaptic bodies. The main numerical development of synaptic bodies occurred between postnatal days 4 and 9, numbers remaining more or less constant thereafter. The intracellular location of synaptic ribbons changed from predominantly cytoplasmic sites to positions at the membrane. In Wistar rats of postnatal day 15 held under a light/dark regimen, synaptic ribbon numbers and lengths were found to be significantly larger at night than at daytime. This was not observed in animals kept under constant darkness. In retinae of a pigmented rat strain, Brown Norway, total numbers of synaptic bodies were similar to those of Wistar rats, whereas the relative proportions of synaptic ribbons and spheres or sphere-like structures, respectively, differed between strains. These results are discussed with regard to synaptic body formation and regulation under the influence of light and eye pigmentation.

Aging↗

PubMed1992

Substance P- and calcitonin gene-related peptide-like immunoreactive neurons in the rat trigeminal ganglion--with special reference to meningeal and pineal innervation.

The distribution of perikarya showing substance P- (SP) or calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) in the rat trigeminal ganglion (TG) were investigated by means of immunohistochemical methods. Approximately 50% of the perikarya contain CGRP while SP-LI was observed in 1/3 of the cells. IR fibres were seen to leave the ganglion via the ophthalmic, maxillary, and mandibular nerves. The combination of peptide histochemistry and retrograde labelling of cells in the ganglion following injection of a fluorescent tracer into the pineal gland reveals that few SP- or CGRP-LI trigeminal neurons innervate the pineal gland. In contrast, the vast majority of perikarya in the TG were labelled upon application of the tracer to the meningeal surface supporting the view that meninges and meningeal arteries in rodents are heavily innervated by SP- and CGRP-LI trigeminal neurons.

Animals↗

PubMed1992

Biphasic 24-hour variations in cyclic GMP accumulation in the rat pineal gland are due to corresponding changes in the activity of cytosolic and particulate guanylate cyclase.

Various parameters of the rat pineal gland display a 24-h rhythm. However, nothing is known about possible 24-h variations in cyclic GMP (cGMP) metabolism. In the present study, 24-h variations in pineal gland cGMP accumulation were investigated by determining the increase in cGMP level with and without inhibitors of phosphodiesterase at different time points over a light/dark cycle (12/12 h). Furthermore, the activity of guanylate cyclase (GC) was determined under substrate-saturated conditions regarding the cytosolic and particulate forms of the enzyme. It has been found that cGMP accumulation and GC activity display biphasic 24-h variations with two peaks--one approximately 7 h after lights "on" and the other approximately 7 h after lights "off." The activity of cytosolic GC remains unchanged in the presence of the nitric oxide (NO) synthesis inhibitor N-monomethyl-L-arginine, indicating that 24-h variations in the activity do not reflect changes in the synthesis of the GC stimulator NO.

Animals↗

PubMed1991

Acetylcholine and muscarinic agonists increase synaptic ribbon numbers in the rat pineal.

Mammalian pinealocytes possess synaptic ribbons (SR) which are commonly present in photoreceptor cells at synaptic junctions. Pineal SR numbers undergo a diurnal rhythm parallel to that of pineal N-acetyltransferase (NAT) activity and melatonin levels. Recent findings suggest that SR numbers, unlike NAT activity and melatonin synthesis and release, do not seem to be regulated by adrenergic mechanisms or neuropeptides in adult rats. Since the pineal gland also receives cholinergic nerve fibres, we have investigated in vitro effects of acetylcholine (ACh) and carbamyl-beta-methylcholine (CBMC; a specific muscarinic agonist) in the presence and absence of pirenzipine (a specific inhibitor of muscarinic M1 receptors). ACh and CBMC increased SR numbers significantly. Pirenzipine inhibited the CBMC-induced increase in SR numbers. On the basis of these findings, it is suggested that cholinergic agonists increase pineal SR numbers by acting through muscarinic M1 receptors. Hence muscarinic mechanisms may have a functional role in pineal physiology.

Acetylcholine↗

PubMed1991

The role of arginine-vasopressin for pineal melatonin synthesis in the rat: involvement of vasopressinergic receptors.

The endogenously synthesized nonapeptide arginine vasopressin (AVP) is thought to be involved in transduction of photic information to the pineal gland. The enhancement of circulating AVP leads to a suppression of the nocturnal melatonin surge the mechanisms of which are unknown so far. We therefore studied the effect of dDAVP, an AVP analog with antidiuretic but without vasopressor activity, on pineal melatonin synthesis in Sprague-Dawley and AVP-deficient Brattleboro rats. The nocturnal intra-arterial application of dDAVP mimicked the inhibitory effect of AVP on the activity of the rate-limiting enzyme for pineal melatonin synthesis, N-acetyltransferase (NAT), in both rat strains. Furthermore, since the pineal is equipped with receptors for VP4-9 (the major proteolytic AVP fragment) only, the influence of this substance on the gland's metabolic activity was investigated in vitro. Neither this peptide nor AVP alone did not affect NAT activity, but either substance potentiated the norepinephrine-induced enhancement of NAT activity. These results reveal that at least two mechanisms mediate the influence of AVP on pineal melatonin synthesis. The AVP-induced pineal inhibition in vivo is probably due to a receptor-mediated effect on pinealopetal signal transduction. This inhibition masks the potentiating effect of AVP on the pineal gland itself which is delayed by the conversion of AVP to VP4-9. The present results support the idea of a modulatory role of AVP and its metabolites in the generation and maintenance of the circadian melatonin rhythm in mammals.

Animals↗

PubMed1991

Effects of LHRH, progesterone, estradiol-17 beta and dexamethasone in vitro on pineal synaptic ribbons and serotonin N-acetyltransferase activity in diestrous rats.

Pineal glands of regularly cycling Sprague Dawley rats (180-220 g) killed on the diestrous morning (between 0900-1000 h) were incubated in appropriate media for six hours with LHRH (8.5 microM), progesterone (3.2 microM), estradiol-17 beta (370 nM) or dexamethasone (250 nM). Pineals incubated in hormone-free medium and unincubated glands served as controls. Six rats were used in each group. After incubation the glands were divided into two parts. One part was used to estimate serotonin N-acetyltransferase (NAT) activity. The other part was processed for electron microscopy to quantify synaptic ribbons (SR). The SR numbers were computed to 20,000 microns 2 area of pineal tissue. The number and distribution pattern of SR were identical in incubated as well as in the unincubated controls. In both these groups the SR located close to the cell membrane were more (23 +/- 1) than those that lay away from it (9 +/- 2). LHRH had no effect on the number of SR located close, to or distant from, the cell membrane. Incubation of pineals with progesterone significantly (p less than or equal to 0.05) depressed the number of SR present close to membranes (23 +/- 1 in controls vs 11 +/- 2 in treated group), total SR (34 +/- 3 in controls vs 21 +/- 2 in treated group) and synaptic fields (26 +/- 2 in controls vs 17 +/- 2 in treated group). Likewise, in the estradiol-17 beta group also membrane-associated SR decreased significantly. The effect of progesterone was more severe than estrogen on the SR possibly due to the differences in the doses used.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

No differences in pineal synaptic ribbon and spherule numbers in different stocks and strains of laboratory rats.

Previous studies have shown that the pineal glands of different stocks and strains of laboratory rats have different melatonin-forming capacities. In the present investigation a widely studied morphological parameter of the pineal, i.e. synaptic ribbon (SR) and synaptic spherule (SS) numbers, was explored in 6 different stocks and strains of laboratory rats, viz.:Han:WIST (albino), LEW/Han (albino), DA/Han (agouti), BN/Han (dark brown), LE/Han (black hooded) and (LEW x BN)/F1 (black with white belly). The rats were maintained under the usual laboratory conditions (lights on from 06.00-18.00 h) for 3 weeks and killed between 10.00-12.00 h, when they were 6 weeks old. The pineals were rapidly excised and processed for transmission electron microscopy. The morphology, distribution (in singles or groups, distant from, or near cell membrane etc.) and number of SR and SS per 20,000 microns2 area of pineal tissue were similar in all groups of rats studied. It is concluded that, in contrast to pineal gland size and melatonin synthesis, SR number is a fairly constant pineal parameter in different stocks and strains of laboratory rats and independent of pigmentation.

Animals↗

PubMed1990

Investigations on day-night differences of vesicle densities in synapses of the rat suprachiasmatic nucleus.

The present study was conducted to test whether the well-known circadian alterations in physiological and metabolical parameters of the hypothalamic suprachiasmatic nucleus (SCN) are accompanied by day-night differences in the number of vesicles in intrinsic synapses of the nucleus. Two groups of 5 adult male rats each were killed at mid-light or mid-dark, respectively, by perfusion with Karnovsky's fluid. The SCN were removed and processed for routine electron microscopy. In medial parts of the nucleus, synapses were characterized as being of Gray type I (asymmetrical), Gray type II (symmetrical) or of intermediate form, and the vesicles per synaptic profile (VPSP) were counted over a defined area. It was found that the ratio of different types of synapses did not differ between day and night. Median VPSP numbers were slightly augmented in animals killed at mid-dark when compared to those obtained from mid-light rats, this difference, however, lacks statistical significance. The present results suggest that vesicle density as a morphological parameter does not clearly parallel the circadian changes in SCN metabolism.

Animals↗

PubMed1990

Melatonin modulates diacylglycerol and arachidonic acid metabolism in the anterior pituitary of immature rats.

In pituitary glands of immature rats prelabeled in vitro with [3H]arachidonic acid, melatonin diminished the luteinizing hormone-releasing hormone (LHRH)-induced increase in [3H]diacylglycerol accumulation as well as [3H]arachidonic acid release from the tissue. Melatonin reduced also LHRH-stimulated incorporation of [3H]glycerol into pituitary [3H]diacylglycerol. The effect was day-time dependent: in the evening experiment melatonin was effective at 0.1 nM concentration while in the morning it had no effect even at 10 nM concentration. The effect of melatonin was also abolished by pretreatment with pertussis toxin. Diacylglycerol and/or arachidonic acid might serve as 2nd messengers transducing the effect of melatonin at the cellular level.

Animals↗

PubMed1990

A possible role for cyclic guanosine monophosphate in the rat pineal gland.

Adrenergic stimulation of pinealocytes induces an increase of both cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). However, for cGMP no biological effects have been demonstrated so far. Therefore we tested the effects of the analog 8-bromo-cGMP on synaptic ribbon numbers and on melatonin synthesis as reflected by N-acetyltransferase (NAT) activity in the rat pineal gland in vitro. Incubation for 6 h with 8-bromo-cGMP did not change the activity of serotonin NAT but it increased the number of synaptic ribbons. These results indicate that cGMP is involved as a second messenger in the regulation of synaptic ribbon numbers in the rat pineal gland.

Acetyltransferases↗

PubMed1990

The role of the hypothalamic paraventricular nuclei for the regulation of pineal melatonin synthesis: new aspects derived from the vasopressin-deficient Brattleboro rat.

There is evidence for an involvement of the hypothalamic paraventricular nuclei (PVN) in the regulation of pineal melatonin synthesis in rats. Since electrical stimulation of the PVN or the systemic administration of arginine-vasopressin (AVP) result in a depression of the nocturnal melatonin surge, this neuropeptide appears to be pivotal for the transduction of PVN-efferent, pinealopetal signals. We therefore used an AVP-deficient animal model, the Brattleboro rat, to further investigate the mechanisms responsible for pineal regulation. Anesthetized adult male animals received 2 min of bilateral electrical stimulation of the PVN either during the day or at night. Thirty min later, pineal glands were removed and pineal N-acetyltransferase (NAT) activities and melatonin contents were determined. Stimulation resulted neither during the day nor at night in any significant alterations of pineal NAT activity or melatonin content when compared to control or sham-stimulated animals. These data further support the proposed modulatory role of AVP for the regulation of melatonin synthesis in the Epiphysis cerebri of genetically intact rats.

Animals↗

PubMed1990

Prenatal development of "synaptic" ribbons in the guinea pig pineal gland.

Pineal "synaptic" ribbons are a heterogeneous population of organelles. "Synaptic" ribbons (SR) sensu stricto, "synaptic" spherules (SS), and intermediate forms (IMF) are present. Their function and origin are unknown, and a knowledge of their prenatal development is lacking. Thus the pineal glands of prenatal, neonatal, and adult guinea pigs were prepared for electron microscopy. "Synaptic" ribbons were studied morphologically and quantitatively. The three categories of "synaptic" ribbons reported in adult pineal glands were also present in prenatal pineal glands. Their structural features, distribution, grouping, and composition patterns are similar to those in adults. "Synaptic" ribbons were first detected in pinealocytes of the distal region of a 42-day postcoitus (PC) pineal gland and were comparable with those in adults. They increased in number with age and reached a peak at 63 days PC, followed by a steep decline at 66 and 67 days PC. By day 69 PC, the numbers increased again and showed a dramatic increase after birth. Several true ribbon synapses were seen at day 63 PC between pinealocyte cell processes or between pinealocyte cell process and pinealocyte cell body. Since true ribbon synapses have not been found in adult guinea pig pinealocytes, their synaptic nature could have been lost during development. No precursors for the "synaptic" ribbons were found. The endoplasmic reticulum cisternae may be the origin for the ribbon vesicles because of their close association with the "synaptic" ribbons.

Aging↗

PubMed1990

Neuronal markers in the rodent pineal gland--an immunohistochemical investigation.

Although some embryological and morphological features speak in favour of a neuronal character of rodent pinealocytes, histochemistry and ultrastructure let this issue appear controversial. Using antibodies to different neurofilaments, the neural adhesion molecule L1, synaptophysin and tubulin as neuronal markers, the pineal glands of rat and guinea-pig were studied by means of immunofluorescence. Neurofilament-immunoreactivity was present in some rat pineal nerve fibers and in the majority of guinea-pig pinealocytes, L1 decorated rat intrapineal nerve fibers, synaptophysin was almost ubiquitously distributed in the pineal of both species, while tubulin-immunofluorescence was seen in nerve fibers of rat and guinea-pig pineal and in some pinealocytes of the latter. These findings speak in favour of the neuronal character of guinea-pig pinealocytes. The lack of neurofilament- and tubulin-immunoreactivity in rat pinealocytes might be attributable to very low concentrations of these proteins or species differences as to their expression. Further studies including in situ-hybridisation of relevant mRNAs will be necessary to answer these questions definitely.

Animals↗

PubMed1990

Synaptic ribbons of the rat pineal gland: responses to in-vivo and in-vitro treatment with inhibitors of protein synthesis.

To elucidate the role of protein synthesis in the nocturnal increase of synaptic ribbons in the rat pineal gland, actinomycin-D, which inhibits transcription, and cycloheximide, an inhibitor of translation, were used. To assure that the drugs were effective and to relate morphological changes to pineal biosynthetic phenomena, the activity of N-acetyltransferase and levels of pineal indoleamine were measured. Results of in-vivo, short-term and long-term treatment with either drug suggest that transcription of proteins related to synaptic ribbon formation occurs during the first half of the light phase, whereas translation takes place during the first few hours of the dark phase. In contrast, proteins involved in enhanced melatonin synthesis are transcribed and translated during the first few hours of the dark phase. In vitro, preincubation with inhibitors of protein synthesis abolished the increase in the numbers of synaptic ribbons after stimulation with dibutyryl-adenosine-cyclic-monophosphate, indicating that the results of the in-vivo experiments are due to an interaction of the drugs with the pineal gland itself. The present study shows that, although in the rat pineal enhanced melatonin synthesis and increased numbers of synaptic ribbons occur at the same time, transcription of proteins involved in both rhythms is temporally separated.

Acetyltransferases↗

PubMed1990

Circadian variations of "synaptic" bodies in the pineal glands of Brattleboro rats.

The function of the mammalian pineal gland is regulated primarily by the sympathetic system. Arginine-vasopressin (AVP) may also be involved in the regulation of pineal melatonin synthesis under experimental conditions. The present study was conducted in the AVP-deficient rat strain, the Brattleboro rat, to investigate whether the numbers and rhythms of pineal "synaptic" bodies in this strain are different from those found in intact rats. AVP or its non-vasoconstrictive analog, deamino-D-AVP, was also injected intra-arterially in Brattleboro or Sprague-Dawley rats to test whether this procedure influences "synaptic" body numbers. Brattleboro rats were killed at different time-points throughout the 24 h-cycle in March, June and September. "Synaptic" ribbons, spherules and intermediate structures were quantified and examined with regard to their intracellular location, with or without nocturnal AVP or D-AVP treatment. Numbers of ribbons were low during the day and high during the night (as in genetically intact rats), whereas spherules and intermediate structures numbers exhibited inconstant daily patterns. Night levels of "synaptic" ribbons were highest in June, lowest in March, whereas day levels did not differ significantly. No significant alterations in pineal "synaptic" body numbers were found following administration of AVP or D-AVP. Our results therefore indicate that AVP does not play a crucial role in the regulation of pineal "synaptic" body numbers in rats.

Animals↗

PubMed1990

Quantitative analysis of "synaptic" ribbon profiles in the pineal complex of male and female Pirbright-White guinea pigs.

Previous studies have pointed in the direction of sex differences as well as regional differences in the pineal gland of guinea pigs. In the present investigation these aspects were studied at the electron-microscopic level by quantitating different types of "synaptic" bodies, intrinsic to pinealocytes. The two major types of "synaptic" organelles, ribbons and spherules, did not exhibit regional or sex differences. "Synaptic" structures intermediate in appearance to ribbons and spherules were significantly larger in number in males in the distal region of the pineal gland, compared to females. As previous studies have shown that ribbon and spherule numbers undergo characteristic changes depending on the functional state of the pineal gland, it is concluded that, as far as the "synaptic" organelles are concerned, no clear-cut sex or regional differences appear to exist in the guinea pig pineal gland.

Animals↗