Browsing by Author "Fuchs, E."

Although it has been reported that hypothalamic 5HT1A-receptor functioning is modulated by oestrogen and that this modulation contributes to the regulation of female sexual behaviour, there have been no reports up to now showing changes in numbers of these receptors during the oestrus cycle and after oestrogen treatment. We therefore analysed 5HT1A-receptors in eight brain areas of female rats at different stages of the oestrus cycle, and in ovariectomized (OVX) females without and with oestrogen replacement. In-vitro receptor autoradiography with the agonist 3H-8-OH-DPAT(3H-8-hydroxy-2-[di-n-propylamino]tetralin) was used to determine numbers and affinities of 5HTA1A-receptors. To evaluate the hormonal state of the animals, serum concentrations of oestradiol, progesterone, luteinizing hormone (LH), and prolactin were also measured. Hormone determinations confirmed the expected endocrine states of the animals. In the ventromedial hypothalamic nucleus, the number of 3H-8-OH-DPAT binding sites (Bmax-value) during oestrus was increased compared to dioestrus yielding significant differences when using ANOVA statistics. In OVX females, the number of binding sites was decreased compared to pro-oestrus and oestrus, and after oestrogen replacement, it was as high as during oestrus. All other brain areas analysed (medial preoptic area, bed nucleus of the stria terminalis, lateral septum, cingulate cortex, amygdala, hippocampal region CA1, and layers V and VI of the occipital cortex) showed no significant changes in 3H-8-OH-DPAT binding site numbers. Also the affinity of 3H-8-OH-DPAT binding sites did not change during the oestrus cycle, but in the medial preoptic area, oestradiol-treated OVX animals showed a tendency for increased affinity compared to untreated OVX females. This was indicated by a change in Kd which appeared to be significant when groups were compared with the t-test. We conclude from our data, that in the ventromedial hypothalamic nucleus, which is involved in the regulation of sexual function, 5HT1A-receptors are up-regulated during oestrus, that ovariectomy reduces the receptor numbers, and that oestradiol replacement counteracts the effect of ovariectomy. Since the ventromedial hypothalamic nucleus contains a high number of oestrogen receptive cells, our data indicate that oestrogen up-regulates 5HT1A-receptor expression in this nucleus.

We have shown previously that repeated laboratory restraint stress or daily corticosterone administration affects the structure of CA3 hippocampal neurons in rats. In the present study, we investigated the effect of repeated daily psychosocial stress on the structure of hippocampal CA3 pyramidal neurons in male tree shrews (Tupaia belangeri). Male tree shrews develop social hierarchies in which subordinates show characteristic changes in physiological and behavioral parameters when confronted with a dominant. In the present experiments, subordinate animals lost body weight soon after starting the daily social conflict, and urinary excretion of cortisol was elevated throughout the experiment as compared with the control period. Golgi-impregnated brain tissue from subordinates exposed to 28 d (1 hr/d) of social confrontations was compared with that from control nonstressed animals. The apical dendrites of the CA3 pyramidal cells from subordinates had a decreased number of branch points and total dendritic length as compared with controls. No differences were observed in apical dendritic spine density or in the basal dendritic tree morphology. The stress-induced CA3 apical dendritic atrophy in subordinates was prevented by administering daily oral doses of the antiepileptic drug phenytoin (Dilantin, Sigma, St. Louis, MO) (200 mg/kg), which interferes with excitatory amino acid (EAA) action. These results suggest that the naturalistic stressor psychosocial stress induces specific structural changes in hippocampal neurons of subordinate male tree shrews. These changes, like those in the rat after glucocorticoid treatment or restraint stress, probably are mediated by activation of the hypothalamo-pituitary-adrenal-axis acting in concert with endogenous EAAs from mossy fiber input.

Intestinal trichomoniasis was observed in 156 of 202 Tupaia belangeri (77.2%). The parasites were located principally in the cecum (75%) and were far less common in the proximal portion of the colon (19%) or terminal portion of the ileum (6%). Advanced trichomoniasis was associated with liquid cecal contents but not diarrhea. The trichomonads had a tendency to penetrate the mucosal epithelial layer, causing desquamation of entire crypts. They never penetrated the epithelial basement membrane and never triggered inflammatory responses. The trichomonads were characterized by three anterior flagella and one trailing flagellum that extended over the entire parasite body, connected to it by an undulating membrane. The capitulum of the straight axostyle formed a small but well defined pelta, and the stout costa had distinct banding. The parasites thereby matched the description of Tritrichomonas mobilensis.

From brain, heart and muscle tissue of the tree shrew (Tupaia belangeri), a higher order mammal, cDNA clones were isolated that encoded two functional splice variants of the corticotropin-releasing factor (CRF) type 2 receptor (CRF-R2). The first, full-length splice variant, amplified from brain and heart tissue, encoded a CRF receptor protein that is 410 amino acids in length and approximately 96% homologous to human CRF-R2alpha. The second, full-length splice variant, derived from skeletal muscle tissue, encoded a 437-amino acid CRF receptor protein that is approximately 92% homologous to human CRF-R2beta. Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) amplifications and RNase protection analyses, showed that tree shrew CRF-R2alpha (tCRF-R2alpha) and tree shrew CRF-R2beta (tCRF-R2beta) were coexpressed in brain tissue but not in heart and skeletal muscle tissue. Finally, human embryonic kidney 293 (HEK293) cells stably transfected with tCRF-R2alpha and tCRF-R2beta were used to demonstrate that the CRF analogs urocortin and sauvagine bind with significantly greater affinity (21- to 140-fold) to these two CRF-R2 splice variants than do human/rat and ovine CRF analogs. In keeping with these results of our CRF binding studies, EC50 values were substantially lower for urocortin-and sauvagine-stimulated than for h/rCRF-and oCRF-stimulated cyclic AMP accumulation in HEK293 cells stably transfected with tCRF-R2alpha or tCRF-R2beta cDNAs. The tree shrew therefore constitutes an important animal model in which to investigate the role of CRF receptor subtypes in the stress response.

Human insulin-like growth factor I (IGF-I) is a growth and differentiating factor produced by various adult and fetal tissues. In the kidney, it has been linked to the proliferative response of renal tubular and glomerular cells, following unilateral or partial nephrectomy, in acromegaly, in diabetes mellitus and in glomerulonephritis. To gain insight into the potential effects of IGF-I in human kidney, a quantitative analysis of IGF-I-binding sites was performed in fetal and adult tissue using 125I-IGF-I. The ligand consistently labelled renal cortex, medulla, and glomeruli while renal vessels were not uniformly marked. The highest affinity of binding sites was found in glomeruli (adult kidneys: Kd 24.7 +/- 5.1 pM; Bmax 5.2 +/- 0.5 fmol/mg tissue equivalent (TE); n = 4; fetal kidneys: Kd 17.0 +/- 2.5 pM; Bmax 4.5 +/- 0.7 fmol/mg TE; n = 4) and cortical tubules, while vessels and renal medulla (adult kidneys: kd 47 +/- 3.9 pM, Bmax 2.6 +/- 0.3 fmol/mg TE; n = 4; fetal kidneys: kd 41.6 +/- 9.2 pM, Bmax 3.5 +/- 0.4 fmol/mg TE; n = 3) had only about half the affinity of binding and a significantly reduced maximal capacity. The strong binding of 125I-IGF-I to glomeruli supports the view that IGF-I may be involved in modulating glomerular structure and function. Fetal renal growth may depend on the action of IGF-I on glomerular cells and tubular epithelia of the kidney.

These studies were designed to determine whether adult neurogenesis occurs in the dentate gyrus of the tree shrew, an animal phylogenetically between insectivores and primates, and to explore the possibility that this process is regulated by stressful experiences and NMDA receptor activation. We performed immunohistochemistry for cell-specific markers and the thymidine analog bromodeoxyuridine (BrdU), a marker of DNA synthesis that labels proliferating cells and their progeny, on the brains of adult tree shrews subjected to psychosocial stress or NMDA receptor antagonist treatment. Cells that incorporated BrdU in the dentate gyrus of adult tree shrews were primarily located in the subgranular zone, had morphological characteristics of granule neuron precursors, and appeared to divide within 24 hr after BrdU injection. Three weeks after BrdU injection, BrdU-labeled cells had neuronal morphology, expressed the neuronal marker neuron specific enolase, and were incorporated into the granule cell layer. Vimentin-immunoreactive radial glia were observed in the dentate gyrus with cell bodies in the subgranular zone and processes extending into the granule cell layer. Exposure to acute psychosocial stress resulted in a rapid decrease in the number of BrdU-labeled cells in the dentate gyrus. In contrast, blockade of NMDA receptors, with the NMDA receptor antagonist MK-801, resulted in an increase in the number of BrdU-labeled cells in the dentate gyrus. These results indicate that adult neurogenesis occurs in the tree shrew dentate gyrus and is regulated by a stressful experience and NMDA receptor activation. Furthermore, we suggest that these characteristics may be common to most mammalian species.

Noradrenaline has been shown to play an important role within the visual system of the brain. To analyze the postnatal development of alpha2-noradrenergic receptors in the visual system of tree shrews, we localized and quantified binding sites for the antagonist [3H]-rauwolscine by in vitro-autoradiography in the dorsal lateral geniculate nucleus and the striate cortex at different postnatal ages. At birth, the dorsal lateral geniculate nucleus is only slightly labeled by [3H]-rauwolscine. During the postnatal period, the number of binding sites increases to reach a maximum around postnatal day 20. Since the young tree shrews open their eyes at approximately day 19, it appears that this high concentration of alpha2-adrenoceptors is related to eye opening. In the adult animal, [3H]-rauwolscine labeling shows a laminated pattern in the dorsal lateral geniculate nucleus. Laminae 1, 2, and 3 are more strongly labeled than laminae 4, 5, and 6. In the striate cortex, the pattern of [3H]-rauwolscine-binding sites changes dramatically during the early postnatal period. Immediately after birth, there is only one layer, located within the subplate zone, which is labeled. From postnatal day 5 onwards, all cortical layers which can be distinguished on histologically stained sections reveal [3H]-rauwolscine-binding sites, but in layer IV, which is known to receive major inputs from the dorsal lateral geniculate nucleus, there is very little labeling during the first two postnatal weeks. In this layer, a large number of [3H]-rauwolscine-binding sites occurs between postnatal day 15 and 20, that is slightly before and around the time of eye opening.(ABSTRACT TRUNCATED AT 250 WORDS)

Affective disorders are accompanied by central nervous changes that may lead to diseases of brain and peripheral organs. To gain an insight into neurobiological mechanisms that underlie such diseases we are studying tree shrews (Tupaia belangeri). This animal model is based on the fact that male tree shrews are very territorial and that under laboratory conditions, two males establish a clear social rank order with a dominant and a subordinate animal. In the visual presence of the dominant, the subordinate shows all typical signs of stress with pronounced activation of the hypothalamic-pituitary-adrenal axis and of the sympathetic nervous system. If there are daily confrontations with the dominant during a time period of several weeks, the subordinate experiences chronic psychosocial stress. Tree shrews can be regarded as a suitable animal model to investigate the neurobiological basis of affective disorders since (1) behavioral and endocrine symptoms of subordinates resemble those of depressive patients, (2) antidepressant treatments lead to an improvement of symptoms, and (3) also in humans chronic stress can lead to depression. Using this model we showed that chronic stress induces changes in the morphology of hippocampal pyramidal neurons, affects neurogenesis in the hippocampal formation, and changes the expression of glucocorticoid, serotonergic and noradrenergic receptors in the brain. These changes depend on the duration of the stress period with some of the alterations being reversible whereas others persist during a longer time period. Since the above receptors modulate neuronal activity, the stress induced alterations lead to an impairment of neuronal activity in distinct brain regions.

Granulomatous lesions similar to those of pulmonary histiocytosis in rats developed spontaneously in the lungs of captive tree shrews. Incidence peaked in 3-year-old tree shrews. Sex dependency was not observed, and development of the granulomas was unrelated to experimental procedures because the lesions were observed in animals from the breeding stock as well. The granulomas consisted of amorphous material, foam cells, and a few foreign body-type multinuclear giant cells; they also contained acicular clefts, often with some fibrous material. Alveolar septa within and adjacent to the granulomas were thickened in most instances, but did not contain inflammatory cells in appreciable numbers or amyloid. Only traces of cholesterol and calcium were detected in the amorphous material; neutral fat was stored in the foam cells and the amorphous masses. Fibers without birefringency were documented by polarization and scanning electron microscopy in the vicinity of granulomas, which in energy-dispersive X-ray microanalysis consisted mostly of calcium, but lacked silicon.

Biopterin, 6-hydroxymethyl-pterin, isoxanthopterin, neopterin and, pterin were quantified in stress-free collected spontaneous morning urine samples from Callithrix jacchus, Saguinus fuscicollis, Saguinus labiatus, Saimiri sciureus, Presbytis entellus, Cercopithecus albogularis, Cercocebus torquatus, Macaca fascicularis, Hylobates concolor, Pongo pygmaeus, and Gorilla gorilla. In most species, biopterin was the most frequent urinary pteridine followed by neopterin. Sex differences in biopterin and neopterin excretion were observed in Gorilla gorilla and Pongo pygmaeus. Pterin and isoxanthopterin were only present in minor concentrations. 6-hydroxymethyl-pterin was barely detectable and not present in the urine of Saguinus labiatus, Saimiri sciureus, and both male Gorilla gorilla and Pongo pygmaeus.

Vascular endothelial growth factor (VEGF) has an important function in renal vascular ontogenesis and is constitutively expressed in podocytes of the adult kidney. The ability of VEGF to be chemotactic for monocytes and to increase the activity of collagenase and plasminogen activator may have implications for renal development and renal disease. In humans, the cellular actions of VEGF depend on binding to two specific receptors: Flt-1 and KDR. The aims of this study were: (1) to localize VEGF receptor proteins in human renal ontogenesis; (2) to quantify VEGF binding in human fetal and adult kidney; and (3) to dissect the binding into its two known components: the KDR and Flt-1 receptors. The latter aim was achieved by competitive binding of VEGF and placenta growth factor-2, which only binds to Flt-1. Quantification of 125I-VEGF binding sites was performed by autoradiography and computerized densitometry. By double-label immunohistochemistry, VEGF receptor proteins were localized solely to endothelial cells of preglomerular vessels, glomeruli, and postglomerular vessels. In developing glomeruli, VEGF receptor protein appeared as soon as endothelial cells were positive for von Willebrand factor. Specific 125I-VEGF binding could be localized to renal arteries and veins, glomeruli, and the tubulointerstitial capillary network in different developmental stages. Affinity (Kd) of adult (aK) and fetal (fK) kidneys was: Kd: glomeruli 38.6 +/- 11.2 (aK, n = 5), 36.3 +/- 7.1 (fK, n = 5); cortical tubulointerstitium 19.4 +/- 2.6 (aK, n = 5), 11.6 +/- 7.0 (fK, n = 5) pmol. Placenta growth factor-2 displaced VEGF binding in all renal structures by approximately 60%. VEGF receptor proteins thus were found only in renal endothelial cells. A coexpression of both VEGF binding sites could be shown, with Flt-1 demonstrating the most abundant VEGF receptor binding sites in the kidney. These studies support the hypothesis of a function for VEGF in adult kidney that is independent of angiogenesis.

Push-pull cannulae were implanted into the preoptic area and into the mediobasal hypothalamic median eminence complex of ovariectomized rhesus monkeys. After recovery, perfusion of the implanted areas was performed over a period of 56 h before and following estradiol benzoate treatment. This treatment results in a drop of LH levels followed by an increase. Catecholamine (norepinephrine, epinephrine and dopamine) concentrations in perfusates collected at 15 min intervals fluctuated tremendously prior to treatment with the estrogen. These fluctuations were largely reduced in perfusates of both structures following the estrogen treatment. They reoccurred at the time of increasing LH levels. beta-endorphin and GABA concentrations were also measured in the perfusates of both structures. Occasional secretory bursts were observed without any obvious relation to the estrogen treatment. It is concluded that catecholamine release in the preoptic area and in the mediobasal hypothalamic median eminence complex is of a pulsatile nature in ovariectomized rhesus monkeys. This pulsatility is largely reduced or abolished following estrogen treatment. The reduced pulsatility may bear a signal character for the release of LH.