Involvement of alpha-adrenoceptors in norepinephrine-induced prostaglandin E2 release by rat pineal gland in vitro

Rat pineal glands incubated in vitro with 10 or 100 microM norepinephrine (NE) released 51% and 415% more prostaglandin E2 (PGE2) to the medium than in the absence of NE. Phentolamine (10 microM) prevented fully the effect of NE at both concentrations, whereas propranolol failed to affect it significantly. After superior cervical ganglionectomy (SCGx), NE-induced PGE2 release was significantly higher than in sham-operated controls--an effect also blocked by phentolamine but not by propranolol. These results suggest that NE releases PGE2 in rat pineals via alpha-adrenoceptors, acting at a post-synaptic site, and that SCGx induces alpha-adrenergic supersensitivity in the pinealocytes.     

Seasonal differences in effect of thyroid hormone deficiency on indoleamine metabolism in the rat pineal gland

Laboratory of Biochemistry of Endocrine Diseases, Khar'kov Research Institute of Endocrinology and Hormone Chemistry, (Presented by Academician of the Academy of Medical Sciences of the USSR A. A. Korzh'.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 111, No. 1, pp. 69–70, January, 1991.     

Application of the physical disector to the central nervous system: Estimation of the total number of neurons in subdivisions of the rat hippocampus

Stereology is a group of mathematical and statistical methods that allows the extrapolation of three-dimensional structural information from two-dimensional sections (or slices). This allows researchers to derive important quantitative structural information, such as the volume, surface area or numbers of particular particles (e.g. cells) within defined regional boundaries. The need for such quantitative information in biology is of particular importance when evaluating the influence of various experimental treatments on specific organs, tissues and cells in the body. Knowledge of such changes has given important insights into the neural substrates that may be responsible for the functional and behavioral consequences of a disparate range of experimental treatments. Here, we describe some of these methods as applied to quantifying the total numbers of cells in defined regions of the hippocampal formation. The methods used for this evaluation were, first, the Cavalieri principle, which was used to determine the volumes of the various subdivisions of the rat hippocampus, and, second, the 'physical disector' method, which was used to estimate the numerical density of neurons within each subdivision. Once these values were derived, it was but a simple task to multiply them together to obtain estimates for the total numbers of cells in the given hippocampal region. We found that 16-and 30-day-old normal male rats had 176 800 and 152 700 pyramidal cells in the CA1 region, respectively. This decrease in the neuronal number was statistically significant. However, in the CA2 + CA3 region, there were approximately 169 300 and 149 600 pyramidal cells in 16- and 30-day-old normal male rats, respectively, which was not significantly different. In the dentate gyrus, there were approximately 36 700 neurons in the hilus region and 483 000 granule cells in the granule cell layer, irrespective of the age of the rats. There were no significant differences between these estimates of hilus neurons and granule cells.     

Modifying effect of pineal peptide hormones on the reactivity of rat thyroid gland to cooling

The study explores the effect of pineal peptide hormones (epitalamin) on the pituitarythyroid system in cold exposure. It is shown that pineal hormones block hyperreactivity of the thyroid to acute cold stress, while in chronic cooling they modulate the adaptive reaction of the thyroid gland to cold. Thus, pineal peptides can modulate the thyroid reaction to cooling. Epitalamin modulates the intensity of peripheral deiodination of thyroxine. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 12, pp. 652–654, December, 1997     

Acute neuroprotection by the synaptic blocker botulinum neurotoxin E in a rat model of focal cerebral ischaemia

Evidence indicates that accumulation of excitotoxic mediators, such as glutamate, contributes to neuronal damage after an ischaemic insult. It is not clear, however, whether this accumulation is due to excess synaptic release or to impaired uptake. To test a role for synaptic release, here we investigated the neuroprotective potential of the synaptic blocker botulinum neurotoxin E (BoNT/E), that prevents vesicle fusion via the cleavage of the SNARE (soluble NSF-attachment receptor) protein SNAP-25 (synaptosomal-associated protein of 25 kDa). Focal ischaemia was induced in vivo by infusing the potent vasoconstricting peptide endothelin-1 (ET-1) into the CA1 area of the hippocampus in adult rats; BoNT/E or vehicle were administered into the same site 20 min later. Injection of ET-1 was found to produce a transient and massive increase in glutamate release that was potently antagonized by BoNT/E. To assess whether blocking transmitter release translates into neuroprotection, the extent of the ischaemic damage was determined 24 h and 6 weeks after the insult. We found that BoNT/E administration consistently reduced the loss of CA1 pyramidal neurons at 24 h. The neuroprotective effect of BoNT/E, however, was no longer significant at 6 weeks. These data provide evidence that blockade of synaptic transmitter release delays neuronal cell death following focal brain ischaemia, and underline the importance of assessing long-term neuroprotection in experimental stroke studies.     

Evidence for the presence of two 24-h rhythms 180° out of phase in the pineal gland of male Pirbright-White guinea pigs as monitored by counting “synaptic” ribbons and spherules

Summary Previous studies have shown that the synaptic bodies of mammalian pinealocytes are a heterogeneous group of organelles. Whereas synaptic ribbons (SR) exhibit a day/night rhythm with small numbers during daytime and high numbers at night, the so-called synaptic spherules (SS) show species-specific differences in their rhythmicity. In the present study the numbers of SR and SS were monitored in male guinea pigs over a full 24-h period at 4-hourly intervals (LD 12:12, lights on at 7:00). The results obtained show that the rhythms of SR numbers and SS numbers are out of phase by 180°. SR numbers are small during daytime and high at night, whereas SS numbers are high during daytime and low at night. As there are no indications that SR are transformed into SS and vice versa and as SR and SS lie in different parenchymal areas it is suggested that they characterize different types of pinealocytes with differing rhythmicities. — Serum melatonin levels were low during daytime (30 pg/ml) and increased at night to reach a peak (84 pg/ml) at 24:00 h.Supported by the Deutsche Forschungsgemeinschaft within the Schwerpunktprogramm Neuroendokrinologie     

Inverse behaviour of "synaptic" ribbon and spherule numbers in the pineal gland of male guinea-pigs exposed to continuous illumination

Summary There is increasing evidence that pineal synaptic ribbons are a heterogeneous population of organelles. In addition to synaptic ribbons (SR) sensu stricto, which consist of an electron-dense rod surrounded by electronlucent vesicles, synaptic spherules (SS) exist, the electrondense core of which is round and much wider than that of the SR. In the guinea-pig SR and SS numbers exhibit an inverse day/night rhythmicity. To gain more insight into the functional significance of SR and SS, guinea-pigs were exposed to continuous illumination for approximately 4 months (LL) and the respective structures in the pineal gland were quantitated under the electron microscope and compared with control animals kept under a lighting regiment of 12 h light and 12 h dark. It was found that SR numbers increase following LL whereas SS numbers decrease. The proximal, intermediate and distal parts of the dumbbell-shaped organ respond differently. The increase in SR numbers is significant in the distal and intermediate regions only, whereas the decrease in SS numbers is significant in the proximal and the intermediate regions only. Within each pineal region analyses of parenchymal subareas measuring 65 m by 65 m exhibit an inverse correlation of SR and SS numbers indicating that there are parenchymal domains in which either SR or SS predominate. Morphometric analyses of a number of pinealocytic parameters reveal minor differences between different pineal regions and that exposure to LL does not strikingly affect the pinealocyte perikarya. By contrast, the numbers of pinealocyte processes increase significantly after LL in the distal and intermediate, but not the proximal region of the pineal gland. These observations suggest structural and functional differences between different parts of the guinea-pig pineal gland.     

Effects of electrical stimulation of the superior cervical ganglia on the number of “synaptic” ribbons and the activity of melatonin-forming enzymes in the rat pineal gland

Summary Melatonin metabolism in the mammalian pineal gland is under the clear influence of sympathetic fibers originating in the superior cervical ganglia (SCG). Previous studies suggested that pineal synaptic ribbons (SR) as well are regulated by the gland's sympathetic innervation. To gain more insight into the mechanisms involved, we examined the effects of sympathetic stimulation on SR number and on the activity of melatonin forming enzymes, serotonin N-acetyltransferase (NAT) and hydoxyindole-O-methyltransferase (HIOMT). The SCG in adult male rats were stimulated electrically during daytime for either 15 or 120 min. Immediately following stimulation, the glands were removed and processed for electron microscopy and for the determination of NAT and HIOMT activities. No differences in pineal SR number, size or location were found in rats stimulated with either parameters when compared with sham-stimulated or control animals. While the activity of HIOMT remained unchanged, the activity of NAT was also unaltered following 15 min of stimulation, but was augmented approximately three-fold in animals stimulated for 120 min. It is concluded that if SR in the rat pineal gland are under sympathetic control, the regulation is different from that involved in melatonin formation.     

Cholinergic synaptic activation due to HCO 3 − in the superior cervical ganglion of the rat

Influence of acid-base change on synaptic transmission was studied in the isolated superior cervical ganglion of the rat. Effects of changes inP _CO2 [HCO ₃ ^– ], or pH of the superfusing solution were studied, using as an index of synaptic excitation the amplitude of the initial negative deflection of surface potential induced by preganglionic stimulation. An increase or decrease in the extracellular fluid (ECF) pH by changing [HCO ₃ ^– ] at a normalP _CO2 elicited respectively augmentation or suppression of the negative deflection. Similar shifts in the ECF pH with varyingP _CO2 at a normal [HCO ₃ ^– ] had small or almost negligible effects on the negative deflection. Simultaneous increase in both theP _CO2 and [HCO ₃ ^– ], which compensated for the pH change in the ECF, induced a consistent increase in the amplitude of the negative deflection. The amplitude of negative deflection in various acid-base conditions was positively correlated with the ECF [HCO ₃ ^– ] but not with ECF pH orP _CO2. These results suggest that an increase in the ECF [HCO ₃ ^– ] activates cholinergic (nicotinic) synaptic transmission in the ganglion.     

大鼠三叉神经节垂体腺苷环化酶激活肽免疫反应神经元对松果体的神经支配

目的　证实大鼠松果体的垂体腺苷环化酶激活肽 (PACAP)免疫反应神经纤维来源于三叉神经节神经元。方法　采用颞下窝入路切断大鼠眼 上颌神经 ,存活 3d～ 1周后 ,观察松果体的PACAP免疫反应神经纤维并计数 ,与未经手术的对照组动物比较。结果　在切断了眼 上颌神经的大鼠 ,其松果体的PACAP免疫反应神经纤维明显减少。结论　大鼠三叉神经节是松果体PACAP能神经纤维的主要来源 ,该类神经纤维可能参与调节松果体腺细胞分泌褪黑素     

Research on bodies of the executed in German anatomy: an accepted method that changed during the Third Reich. Study of anatomical journals from 1924 to 1951

While it is known that bodies of the executed were used for anatomical research in Germany during the Third Reich, it is unclear whether this type of work was unique to the time period or more common in Germany than elsewhere. The dissected persons and the anatomists involved have not been fully investigated. This study of anatomical journals from 1924 to 1951 shows that 166 out of 7,438 [2.2%] German language articles mentioned the use of "material" from the bodies of executed persons. In comparison, only 2 out of 4,702 English language articles explicitly mentioned bodies of the executed. From 1924 to1932, 33 of a total of 3,734 [1%] German articles listed the use of the executed. From 1933 to 1938 the number rose to 46 out of 2,265 [2%], and increased again from 1939 to 1945 to 73 out of 984 [7%]. After the war 15 out of 455 [3%] still dealt with "material" from the executed. German anatomists' familiarity with the use of the executed as a standard for healthy tissues even before 1933 may have contributed to the ease with which they accepted the "opportunities" (large-scale studies and research on women) presented to them by unlimited access to bodies of the executed provided by the abusive National Socialist (NS) legislation and continued using them for some years after the war. German postwar anatomy was built in part on the bodies of NS victims. Information given in some publications will help with further identification of these victims.     

Comparison of primary lung tumor incidences in the rat evaluated by the standard microscopy method and by multiple step sections

The data presented in this paper have been derived from a carcinogenicity experiment with rats as part of a comprehensive research project focused on experimental studies on the toxicity and carcinogenicity of intratracheally instilled granular dusts [Ernst H, Rittinghausen S, Bartsch W, Creutzenberg O, Dasenbrock C, G?rlitz B-D et al. Pulmonary inflammation in rats after intratracheal instillation of quartz, amorphous SiO(2), carbon black, and coal dust and the influence of poly-2-vinylpyridine-N-oxide (PVNO). Exp Toxicol Pathol 2002; 54: 109-26; Ernst H, Kolling A, Bellmann B, Rittinghausen S, Heinrich U, Pott F. Pathogenetische und immunbiologische Untersuchungen zur Frage: Ist die Extrapolation der Staubkanzerogenit?t von der Ratte auf den Menschen gerechtfertigt? Teil II: Histologie. Abschlussbericht. Umweltforschungsplan des Bundesministeriums für Umwelt, Naturschutz und Reaktorsicherheit. November 2005. http://www.umweltdaten.de/publikationen/fpdf-l/3033.pdf]. The results of the standard approach to histological sampling in rodent carcinogenicity inhalation studies were compared to those obtained after supplemental evaluation of step sections at intervals of 250microm through the entire lung. Seven lung tissue specimens (six sections) each of 251 rats (55 rats of the control group, 53 rats of the group treated with quartz DQ 12, 56 rats of the group treated with quartz DQ 12 and PVNO (poly-2-vinylpyridine-N-oxide), 53 rats of the group treated with amorphous SiO(2), and 17 rats each of the groups treated with coal dust and carbon black) were evaluated by light microscopy. At least 60 hematoxylin and eosin (H&E)-stained sections per lung were evaluated of 99 female rats (30 rats of the control group, 7 rats each of the groups treated with quartz, quartz and PVNO, and carbon black, 31 rats of the group treated with amorphous SiO(2), and 17 rats treated with coal dust). For the neoplastic and pre-neoplastic lesions detected in the serial slides an approximate value of the whole tumor volume was calculated. The detection of tumors with a diameter of 0.25mm was possible. Based on the size distribution of 75 tumor volumes, the probability of detecting a tumor was 86% when using 12 sections. The addition of step sections enhanced the tumor detection rate from 17 to a total number of 44 lung tumors in the quartz-treated rats, from 6 to 10 in the quartz- and PVNO-treated rats, from 4 to 11 in the amorphous SiO(2)-treated rats, and from 4 to 10 in the carbon black-treated rats. Both the tumor multiplicity and the number of rats with pre-neoplastic lesions increased. These additional data corroborated the initial findings in all experimental groups and provided statistically significant results confirming the equivocal evidence of carcinogenic activity of amorphous SiO(2) in female Wistar rats. This technique offered accurate information on the incidence, histological type, size, and location of proliferative lesions in the entire lung, but the benefit must be balanced against the extra financial effort.     

Prenatal morphine alters the synaptic complex of postsynaptic density 95 with N-methyl-d-aspartate receptor subunit in hippocampal CA1 subregion of rat offspring leading to long-term cognitive deficits

Infants who are passively exposed to morphine or heroin through their addicted mothers usually develop neurobiological changes. The postsynaptic density 95 (PSD-95) protein, a submembranous cytoskeletal specialization, is dynamically linked with N-methyl-d-aspartate receptors (NMDARs) to form a synaptic complex in postsynaptic neurons. This complex serves important neurobiological functions, including mammalian learning and memory. However, the effects of prenatal morphine exposure on this synaptic complex are not well understood. In this study, we determined whether prenatal morphine exposure altered the synaptic complex association between PSD-95 and three major NMDAR subunits (NR1, NR2A, and NR2B), at the mRNA and protein levels, within the hippocampal CA1 subregion (an important integration area for mammalian learning and memory) of rat offspring along with the performance of long-term cognitive functions. Sprague–Dawley rat offspring from morphine-addicted mothers were studied at a younger age (postnatal day 14; P14) and at an older age (P45). Subsequently, an eight-arm radial maze task was applied to analyze the working and cued reference memory in such offspring (P45). The real-time polymerase chain reaction results showed that prenatal morphine exposure caused significant decreases in mRNA levels of the PSD-95 and three NMDAR subunits (NR1, NR2A, and NR2B) in offspring (P14 and P45). Similarly, at the protein level, immunoblotting showed that decreased whole levels of PSD-95 and NMDAR subunits were seen in offspring subjected with prenatal morphine. Furthermore, the protein interaction of the synaptic complex between the PSD-95 and NMDAR subunit, as indicated by coimmunoprecipitation, was less in prenatal morphine samples than in vehicle controls (P14 and P45). The prenatal morphine group also showed poorer performance for an eight-arm radial maze task than the vehicle-control group. These results are particularly important for a better understanding of certain opioid-mediated neurobehavioral cognitive changes in offspring associated with altered protein interaction between PSD-95 and NMDAR subunits within the developing brain.