Biomedical subjects

S Reuss

Publications and source records attributed to S Reuss.

At least 55 records · Page 3Linked to original sources

PubMed1994

Substance P-like immunoreactivity in the hypothalamic suprachiasmatic nucleus of Phodopus sungorus--relation to daytime, photoperiod, sex and age.

The immunohistochemical distribution of substance P (SP) in the hypothalamic suprachiasmatic nucleus (SCN) was studied in adult male and female Djungarian hamsters (Phodopus sungorus) held under either long or short photoperiods. Intact animals were killed by perfusion with a fixative at the middle of the light or dark periods, respectively. The tissue was processed by routine immunohistochemical methods. Perikarya exhibiting SP-like immunoreactivity (LI) were found in the SCN of animals of all groups. These cell bodies predominantly were restricted to a distinct portion of the nucleus extending less than 150 microns rostrocaudally and were often concentrated in its lateral aspect. SP-LI fibers were rarely observed in the SCN, however, other hypothalamic parts, e.g. anterior and paraventricular regions, exhibit strong SP-LI innervation patterns. Sex-related differences were not observed. Long-term exposure to short days decreased the number of neurons exhibiting SP-LI by approximately 60% when compared to long-day animals at both day- and nighttime. At night, SP-LI neurons were augmented in number by 34% (long-day group) and 56% (short-day group). Further, the numbers of SP-LI perikarya in the SCN of aged hamsters at day- and nighttime were augmented 3- to 4-fold when compared to adult animals. These results suggest that substance P in the SCN is involved in the regulation of circadian and seasonal mechanisms in this highly photoperiodic rodent species.

Aging↗

PubMed1994

Clorgyline effect on pineal melatonin biosynthesis in adrenalectomized rats pretreated with 6-hydroxydopamine.

The response to administration of the specific monoamine oxidase A (MAO-A) blocker clorgyline was investigated in adult male Sprague-Dawley rats which were adrenalectomized four days prior to treatment or were additionally sympathectomized as newborns by injection of 6-hydroxydopamine. In both groups, the contents of pineal indoles melatonin and N-acetylserotonin were augmented, and the contents of 5-hydroxyindoleacetic acid and 5-hydroxyindoletryptophol decreased 90 min following clorgyline injections when compared to rats receiving saline. The observed responses were less pronounced in rats both adrenalectomized and sympathectomized. The results are in line with the hypothesis that preservation from oxidation of both MAO-A substrates, noradrenaline and serotonin, upon clorgyline administration contributes to the observed increase in melatonin biosynthesis thought to be associated with the anti-depressant effects of MAO inhibition.

Adrenalectomy↗

PubMed1994

Expression of nicotinic acetylcholine receptors in the rat superior cervical ganglion on mRNA and protein level.

The expression of nicotinic acetylcholine receptors (nAChR) in the rat superior cervical ganglion was investigated by Western blotting, immunohistochemistry and non-radioactive in situ hybridization applying probes for the alpha 4-1 and beta 2 subunit mRNA. Immunoblot analysis of homogenized ganglia using the anti-nAChRs antibody WF6 revealed a labeled protein band of apparent molecular weight of 40 kDa which is typical for the alpha subunit of nAChRs. Applying double-labeling immunofluorescence with antibodies against tyrosine hydroxylase, nAChR-like molecules were identified in most postganglionic neurons and in a subpopulation of small intensely fluorescent (SIF) cells. alpha 4-1 and beta 2 subunit mRNAs were detected in all perikarya of postganglionic sympathetic neurons but not in SIF cells. These results suggest that antibodies raised against purified Torpedo AChR bind to nAChR in sympathetic ganglia and indicate that alpha 4-1 and beta 2 subunits are constituents of nAChRs in sympathetic postganglionic neurons but not of SIF cells.

Animals↗

PubMed1994

NADPH-diaphorase activity of nitric oxide synthase in the olfactory bulb: co-factor specificity and characterization regarding the interrelation to NO formation.

The neuronal form of the enzyme nitric oxide synthase (nNOS) synthesizes the messenger molecule nitric oxide (NO). In addition to NO formation, nNOS exhibits a so-called NADPH-diaphorase (NADPH-d) activity. This study focused on the characterization of NADPH-d activity with regard to NO formation in the rat olfactory bulb. In this area of the brain pronounced staining is localized in discrete populations of neuronal somata and in olfactory glomeruli. Diaphorase staining combined with demonstration of nNOS by polyclonal antibodies revealed that NADPH-d activity of neuron somata is associated with nNOS immunoreactivity. It is concluded that neuron somata exhibit NADPH-d activity of nNOS. NADPH-d activity of nNOS did not utilize beta-NADH or alpha-NADPH. Moreover, NADPH-d activity was inhibited in the presence of alpha-NADPH. Dichlorophenolindophenol (DPIP), an artificial electron acceptor and an inhibitor of NO formation, totally suppressed NADPH-d staining of neurons, supporting the concept that the NADPH-d of neuron somata is due to nNOS. Cytochrome C, miconazole, EGTA, and trifluoperazine, which have been reported to inhibit cytochrome P450 reductase activity of NOS, did not affect NADPH-d staining. Hence, NADPH-d activity of NOS does not involve cytochrome P450 reductase activity as required for NO formation. Contrary to NADPH-d activity of neuron somata, staining of olfactory glomeruli was not co-localized with nNOS immunoreactivity. Glomerular staining was also observed in the presence of beta-NADH and alpha-NADPH. Further, it was unchanged in the presence of the NO formation inhibitor DPIP. Hence, the glomerular staining in the presence of NADPH is not due to the NADPH-d activity of NOS. We conclude that staining of neuronal structures in the presence of NADPH does not necessarily represent NADPH-d activity of NOS.

2,6-Dichloroindophenol↗

PubMed1994

An improved technique for visually controlled pinealectomy in the rat.

An improved method for pinealectomizing rats and other small laboratory rodents is described which avoids bleeding and allows excision of the organ under exact visual control. After removing a small piece of the skull dorsal to the confluence of the transverse and dorsal sagittal sinuses, the latter is ligated and cut. The underlying pineal gland is then exposed and may be excised reliably without damage to the gland tissue or to adjacent brain structures. Intra- and post-operative complications are extremely rare.

Anesthesia, General↗

PubMed1993

Lack of effect of oxytocin on the numbers of "synaptic" ribbons, cyclic guanosine monophosphate and serotonin N-acetyltransferase activity in organ-cultured pineals of three strains of rats.

In addition to the stimulating influence of the sympathetic system on the function of the mammalian pineal gland, neuropeptides such as neuropeptide Y, vasoactive intestinal polypeptide and arginine-vasopressin (AVP) are thought to function as modulators. Since AVP has been shown to influence pineal melatonin synthesis, the aim of the present study was to investigate the possible effects of the second hypothalamic nonapeptide oxytocin (OT), which likewise has been detected in the pineal gland. We therefore studied "synaptic" ribbon (SR) numbers, N-acetyltransferase (NAT) activity and the intracellular concentration of cyclic guanosine monophosphate (cGMP) following in vitro incubation of rat pineals in media containing OT (10(-5) M), noradrenaline (NA, 10(-5) M) or both NA and OT. Pineal glands were derived from rats of three different strains (Sprague-Dawley, Long-Evans and the AVP-deficient strain Brattleboro). Neither morphological nor biochemical analyses showed a difference between control and OT-incubated organs in any of the strains tested. In Brattleboro rats, but not in the other strains, noradrenaline slightly increased the number of SR which was not observed when NA and OT were combined. The addition of NA resulted in distinct augmentation of NAT activity and cGMP content, which were not affected by additional OT application. These results suggest that oxytocin is not crucially involved in the regulation of pineal gland function.

Animals↗

PubMed1993

[Work of the inner clock. Neuroanatomy of circadian systems of mammals].

Many aspects of mammalian life exhibit distinct alterations throughout the 24-h cycle. Morphological, physiological, and biochemical parameters display circadian rhythms which are thought to be generated by an endogenous pacemaker and regulated by environmental factors. The morphological substrates of the endogenous circadian system have been studied extensively during the last two decades. Although knowledge is far from complete, there is general agreement that the pathways involved consist mainly of retina, hypothalamus, spinal cord, sympathetic trunk, and pineal gland. This review characterizes the anatomical structures and tracts responsible for generation and maintenance of circadian rhythmicity and discusses functional implications of neurotransmitter involvement and the selectivity of connections.

Animals↗

PubMed1993

Strain differences in the ratio of synaptic body types in photoreceptors of the rat retina.

In the retinal outer plexiform layer of seven different rat strains, synaptic bodies (SB) were counted and, according to their morphology, characterized as synaptic ribbons (SR), synaptic spheres (SS) or intermediate structures. It was found that absolute SB numbers showed relatively small variations while SR/SS ratios differed considerably between the strains investigated. These results are discussed with respect to retinal pigmentation and to formation and degradation, respectively, of synaptic ribbons.

Animals↗

PubMed1993

The Harderian gland of the Djungarian hamster (Phodopus sungorus): light- and electron-microscopical investigations.

The Harderian gland of the Djungarian hamster Phodopus sungorus was investigated by light and electron microscopy in adult animals of both sexes, held under either long or short photoperiods. These glands have a tubulo-alveolar structure. Epithelial cells were seen as small cylindric cells with a large, round nucleus located basally, many small vacuoles distributed throughout the cytoplasm (type I cells) or as rather broad cells with larger vacuoles (type II cells). The ratio of both cell types differed from 1:1 to 2:1 (type I:type II), regardless of the animal's sex. In the electron microscope, abundant smooth endoplasmic reticulum of the vesicular type and rod-like or oval-shaped mitochondria of the crista type were very numerous; while Golgi complexes were rarely seen. 'Membranous bodies' were about nine times higher in amount in type II cells and were further augmented fourfold, in number, in glands of female animals. The few bundles of poly-tubular structures that were observed, were predominantly in type I cells. Myoepithelial cells were linked to the basal part of acinar cells by interdigitations. Many microvilli were observed at the apical portion of cellular membranes. Acinar cells predominantly showed signs of merocrine secretion, although apocrine secretion was seen in some cases. Under epifluorescence, red porphyrin fluorescence was seen predominantly in type I cells, where it is mainly found apically. The comparison of animals held under either long or short photoperiods did not reveal any differences in Harderian gland structure.

Animals↗

PubMed1992

Nicotinic cholinoceptors in the rat pineal gland as analyzed by western blot, light- and electron microscopy.

The monoclonal antibody WF6, raised against purified Torpedo nicotinic acetylcholine receptor (nAChR) was used to study the distribution of cholinoceptors in the rat pineal gland by means of Western blot analysis, light- and electron microscopy. The immunoblot analysis using homogenized pineal gland revealed a labeled protein band of apparent molecular weight 40 kDa which was identified as alpha-subunit of a nAChR. In the light microscope, approximately one-fourth of the pinealocytes exhibited cytoplasmic immunoreactivity (IR) of varying density. In the electron microscope, IR was seen as patchy staining of cell membranes of pinealocyte somata and processes. Presynaptic IR material was not found. Distribution and intensity of the observed IR was not significantly different in pineal sections from ganglionectomized rats, nor were any alterations found that would relate to the animals' sex or to the time of killing (day vs night). Our results provide further evidence for the existence of cholinergic receptors in the mammalian pineal. They may be important for the understanding of the gland's regulation.

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↗

PubMed1991

Photoperiod effects on bombesin- and cholecystokinin-like immunoreactivity in the suprachiasmatic nuclei of the Djungarian hamster (Phodopus sungorus).

The immunocytochemical distribution of the putative satiety peptides bombesin (BBS) and cholecystokinin (CCK) were studied in the hypothalamic suprachiasmatic nuclei (SCN) of male and female Djungarian hamsters (Phodopus sungorus) held under either long (light/dark, LD 16:8 h) or short (LD 8:16) photoperiod. The animals were killed by perfusion with a fixative at the middle of the light period and the tissue was processed by routine immunohistochemical methods. Perikarya exhibiting BBS- or CCK-like immunoreactivity (LI) were found in the SCN of animals of all groups. Sex-related differences were not observed. In contrast, long-term exposure to short days decreased the number of neurons exhibiting CCK-LI dramatically when compared to long-day animals, while BBS-LI was not affected. The results suggest that CCK in the SCN is involved in the regulation of differential food intake in this highly photoperiodic rodent species.

Animals↗

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↗

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

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

Neuropeptide Y localization in telencephalic and diencephalic structures of the ground squirrel brain.

The distribution of neuropeptide Y-immunoreactive (NPY-IR) perikarya, fibers, and terminals was investigated in the brain of two species of hibernatory ground squirrels, Spermophilus tridecemlineatus and S. richardsonii, by means of immunohistochemistry. In the telencephalic and diencephalic structures studied, distinct patterns of NPY-IR were observed which were essentially identical in male and female animals of both species. No differences in amount or distribution of NPY-IR structures were observed between animals which had been in induced hibernation for several months before sacrifice in March/April and those sacrificed one week after their capture in May. In some brain structures (e.g., the hypothalamic arcuate nucleus), IR cell bodies were observed only after pretreatment with colchicine. NPY-IR perikarya and fibers were found in the cerebral cortex, caudate nucleus-putamen, and dorsal part of the lateral septal nucleus. Dense fiber plexuses were seen in the lateral and medial parts of the bed nucleus of the stria terminalis. The numbers of IR perikarya observed in the medial part of the nucleus increased following intraventricular colchicine injections. The accumbens nucleus exhibited few IR cells and many fibers. Claustrum and endopiriform nuclei showed a considerable number of stained cells and fibers that increased in number and staining intensity in colchicine-treated ground squirrels. The induseum griseum showed a small band of IR cell bodies and varicose fibers. Bipolar of multipolar IR cells and varicose fibers were found in the basal nucleus of the amygdala. Dense fiber plexuses as well as IR terminals were seen in the median, medial, and lateral preoptic areas of the hypothalamus. Terminals and relatively few fibers were located in the periventricular, paraventricular, and supraoptic nuclei. The anterior, lateral, dorsomedial, and ventromedial hypothalamic nuclei contained relatively large numbers of terminals and fibers. In the suprachiasmatic nuclei, dense terminals were distributed mainly in the ventromedial subdivision. In the median eminence, immunoreactive terminals were concentrated in the external layer, with fibers predominant in the internal layer. NPY-IR perikarya were observed only in the arcuate nucleus of the hypothalamus and only following colchicine treatment. In the epithalamus (superficial part of the pineal gland and habenular nuclei), varicose fibers appeared mainly in perivascular locations (pineal) or as a dense plexus (habenular nuclei). These results from ground squirrels are discussed in comparison to those obtained in other species and with regard to considerations of the physiological role of NPY.

Animals↗

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↗