Proctolin-related material in the mouse brain as revealed by immunohistochemistry

By use of a specific antiserum against the insect peptide proctolin we were able to identify proctolin-immunoreactive neurons in the mouse brain. These nerve cells belong to the nuc. mesencephalicus n. trigemini. Furthermore, the antiserum stained very few nerve fibers with varicosities in the immediate neighborhood of the roof of the third ventricle. The chemical identity of the immunoreactive material with genuine proctolin remains to be elucidated.     

Development of the lymphatic network in the muscle coat of the rat jejunum as revealed by enzyme-histochemistry

The process of lymphangiogenesis was studied in the muscle coat of the rat small intestine by light and scanning and transmission electron microscopy; identification of lymphatic vessels was made by 5'-nucleotidase staining. Light and scanning electron microscopy demonstrated that the intramuscular lymphatic network formation, which started only postnatally, was attributable to the vascular sprouting of slender lymphatic endothelial projections and to a splitting of the vessels, causing intervascular meshes of various sizes. The growing lymphatics were consistently closed by the endothelial cells, which were characterized by an abundance of cell organelles and prominent cytoplasmic processes. The cells often revealed close contacts with the processes of developing smooth muscle cells in the jejunal muscle coat, suggesting a possible role for the latter cells in the guidance of the lymphatic extension. The present study is the first to suggest the closed nature of lymphatics persisting throughout their development, even at the initial stage of lymphangiogenesis.     

Different pathways of human T-cell activation revealed by PHA-P and PHA-M.

Antigen-specific T-cell activation is mediated via the CD3-Ti (antigen receptor) complex, and monoclonal antibodies to both CD3 and Ti cause a rapid rise in intracellular Ca2+. This calcium mobilization is not inhibited by monoclonal antibodies to CD2. The rise in calcium mobilization induced by purified PHA (PHA-P) does not occur in a cell line which lacks CD2 expression, and can be blocked in other T cells by anti-CD2 antibodies. A combination of monoclonal antibodies to different epitopes of CD2 causes calcium mobilization and mitogenesis. Reagent grade PHA (PHA-M) induces calcium moblization in cells that lack CD2, and its effects in other T cells cannot be blocked by anti-CD2 antibodies. The effects of PHA-P and PHA-M are thus mediated predominantly through different activation pathways.     

The transendothelial passage in the absorbing peripheral lymphatic vessel of the rodent diaphragm

The absorbing peripheral lymphatic vessels of rat (Wistar/Kyoto) and gerbil (Meriones unguiculatus) diaphragms were observed with SEM, TEM and three-dimensional models under normal conditions, after experimental hemorrhage and after abdominal injection of China ink and latex polystyrene particles. Superficial and deep lymphatic vessels were noted and their ultrastructural characteristics showed an endothelial wall lacking a continuous basal lamina. Moreover, "intraendothelial channels" which are dynamic, fundamental pathways for lumen migration into the lymphatic vessel for fluids, China ink and latex polystyrene particles, were also demonstrated. Concerning the transendothelial passage of the autologous erythrocytes after their migration through the stomata and submesothelium connective channel, it was observed that this passage occurs by means of a "transcellular pathway". This consists of a "migratory pore" independent from the intercellular junctions, which is formed by an active process following a probable immunological response or cellular interaction between the erythrocyte and the endothelial cell.     

The distribution and morphology of lymphatic vessels on the peritoneal surface of the adult human diaphragm,as revealed by an ink-absorption method

Application of india ink to the peritoneal and pleural surfaces of the adult human diaphragm allowed visualization of the distribution and morphology of the lymphatic vessels by light microscopy and scanning electron microscopy. The diaphragms examined had been fixed and stored in 10% formalin. Numerous lymphatic vessels were stained black with india ink, presenting reticular, radial-meshwork, ladder-like and lacy patterns. They were distributed throughout the entire sternocostal part. Analysis by light and scanning electron microscopy of the areas indicated by india ink revealed the presence of primary lymphatic vessels that formed lymphatic lacunae and stomatal openings to the peritoneal cavity. A layer of secondary collecting lymphatic vessels was located cranially with respect to the layer of primary lymphatic vessels. Thus, the peritoneum had at least two layers of lymphatic vessels. These lymphatic vessels were not tubular vessels but resembled flat cisternae, as has been suggested in the case of the mouse diaphragm. The pleura lacked lymphatic stomata and had no such double-layered lymphatic organization. This is the first report that showed distribution and morphology of the lymphatic vessels in the diaphragmatic peritoneum of the formalin-fixed, adult human diaphragm. The method and results in the present study may contribute to morphological analysis of the lymphatic system in the wall of the human body cavity.     

脑淋巴引流研究进展

大脑不存在衬以内皮的淋巴管,但同样存在淋巴引流,分为脑脊液的淋巴引流和脑实质组织间液的淋巴引流两部分。脑脊液淋巴引流的脑、脊神经根途径更为重要。而脑实质的组织间液通过血管周围间隙引流至大脑动脉的外膜内,进而引流至颈部淋巴结。阻断脑淋巴引流可引起动物以及人类脑形态结构、功能等改变,证明脑淋巴引流具有重要的生理意义。     

The ontogeny of Mauthner cells in the brain of Labeo rohita as revealed by NADPH-d and nNOS immunohistochemistry

Nitric oxide (NO) is well demonstrated to act as a neuronal messenger in neurotransmission in vertebrate animals. We are for the first time reporting nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and neuronal nitric oxide synthase (nNOS) in the Mauthner cells (MC) of the carp, Labeo rohita brain during post-embryonic development. The NADPH-d and nNOS-positive MC were seen intensely labeled in all young stages on either side of the ventricle in the hindbrain. In adult, these cells were moderately stained with NADPH-d but not with nNOS. Further, the morphometeric analysis of the staining intensities showed the decrease in the optical densities of the MC as the development progresses. The gradual increase in cell diameter was evident from hatchling to adult (18–22 μm to 98–106 μm), while the nuclear diameter remained nearly unchanged (10–12 μm to 18–20 μm). In fishes, MC are known to be involved in the control of startle response. The NADPH-d enzymatic activity seen in the MC of early development may play an important role to activate the startle response to fast escape from the predators and unfavorable environmental conditions to survive during early phase of life. As nNOS is absent in the adult MC, we assume that the adult MC express a different splice variant of nNOS which is detected by NADPH-d, but not by the currently used antibody. Hence, the presence of nNOS (enzyme required for the synthesis of NO) in the MC suggests the involvement of NO in neurotransmission to generate the startle response rapidly. Furthermore, we also suggest that NO may be involved in other aspects such as pathway finding, neuronal cell migration, signaling and refinement during the development of the brain of L. rohita.     

Decline in fertility of mouse sperm with abnormal chromatin during epididymal passage as revealed by ICSI

BACKGROUND: Recent studies showed that ICSI with cauda epididymal or ejaculated sperm of infertile mice or men, respectively, was less effective in fertilization and normal embryo development than ICSI using sperm from the testes. These studies suggested that sperm nuclear quality declined after release from the testis, but the site where this loss of fertility occurs has not been localized. METHODS: We performed ICSI with testicular, caput, and cauda epididymal sperm from infertile Tnp1-/-Tnp2+/- mutant mice, which have a minimal level of transition nuclear proteins and are sterile by natural mating. RESULTS: When the heads of motile sperm from the testis or caput epididymis of Tnp1-/-Tnp2+/- males were injected into enucleated mouse oocytes, sperm chromosomes showed no difference from those of wild-type mice, but the chromosomes from sperm taken from the cauda epididymis of mutant males showed increased abnormalities. Injection of testicular or caput epididymal sperm from Tnp1-/-Tnp2+/- males into intact oocytes resulted in normal embryonic and fetal development and yields of liveborn equivalent to wild-type, but cauda sperm from Tnp1-/-Tnp2-/- mice produced lower implantation rates and yields of liveborn than did those from wild-type mice. CONCLUSIONS: These results demonstrate that in mice with sperm chromatin abnormalities, the decline in fertility of sperm with ICSI occurs after the caput epididymis. The advantage of using caput epididymal sperm for ICSI in certain situations may be considered as an approach to be tested in human assisted reproduction.     

Mapping of benzodiazepine-like immunoreactivity in the rat brain as revealed by a monoclonal antibody to benzodiazepines.

A monoclonal antibody against benzodiazepines (21-7F9) was used to study the distribution of benzodiazepine-like immunoreactivity in the rat brain. Immunodensitometry in combination with image analysis were used for quantification. The results showed a ubiquitous distribution of benzodiazepine-like immunoreactivity throughout the brain. Very high levels of benzodiazepine-like immunoreactivity were found in the Purkinje cell layer of the cerebellum, in the primary olfactory cortex, in the stratum pyramidale of the hippocampus and in the mitral cell layer of the olfactory bulb. High densities of benzodiazepine-like immunoreactivity were found in the granule cell layer of the cerebellum, the pyramidal cell layer of the olfactory tubercle, the granule layer of the dentate gyrus, the arcuate nucleus of the hypothalamus, the mammillary bodies, the interstitial nucleus of Cajal and superficial grey layer of superior colliculus. The substantia nigra pars compacta, the islands of Calleja and layers II, III, V and VI of the cerebral cortex had moderate levels of benzodiazepine-like immunoreactivity. Lower densities were found in the internal granular layer and the external plexiform layer of the olfactory bulb, in the molecular layer of the dentate gyrus, in layers I and IV of the cerebral cortex, in the nucleus caudate-putamen and most of the thalamic nuclei. The lowest density of immunoreactivity was found in the globus pallidus, and the strata radiatum, oriens and lacunosum-moleculare of the hippocampus. The distribution of endogenous benzodiazepine-like immunoreactivity was compared with the distribution of the GABA/benzodiazepine receptor by using both immunocytochemistry and receptor autoradiography. Our studies have shown a clear mismatch between the localization of the benzodiazepine-like immunoreactivity and the GABA/benzodiazepine receptors.     

Microglia in the mature and developing quail brain as revealed by a monoclonal antibody recognizing hemopoietic cells.

The monoclonal antibody QH1, which recognizes quail endothelial and hemopoietic cells, was found to label microglia in the developing and mature brain of the quail. Forms of microglia similar to those described in mammals were labelled. Ameboid microglia predominated at embryonic stages, became less numerous in late embryonic development, and disappeared completely by day 10 post-hatch (P10). Poorly ramified microglia were present as early as day 5 of incubation (E5), and were progressively replaced by mature ramified microglia from E14 onwards. From P10 onwards, ramified microglia were the only microglial form seen in the quail brain.     

Early semantic context integration and lexical access as revealed by event-related brain potentials

A late neurophysiological response, the N400, is well known to reflect word processing and semantic context integration. Nevertheless, behavioural and neurophysiological data have demonstrated that word features and semantic sentence context influence linguistic processes already within the first 200 ms following the onset of the critical word stimulus. The lack of early word-related ERP effects in some studies might be due to large stimulus variance on relevant linguistic parameters, for example the length of written words or their frequency of usage. Here we investigated the effect of the length of words, their standardized lexical frequency and probability to occur in a given semantic context, by orthogonally varying these factors in a sentence-reading task. We found word frequency and probability to be reflected neurophysiologically already at 120 and 180 ms after written word onset. However, at these early stages frequency and probability effects were modulated by word length, as documented by significant interactions, whereas at later stages, around 300-500 ms, additive effects of these variables were found. These results indicate that semantic context integration may take place at a surprisingly early stage and near-simultaneously with the processing of information about the form of a word and its lexical properties, therefore challenging serial models of psycholinguistic information access. A methodological implication of this study is that, in order to obtain early psycholinguistic ERP effects, stimulus variance must be kept to a minimum.     

Consciousness and arousal effects on emotional face processing as revealed by brain oscillations. A gamma band analysis.

It remains an open question whether it is possible to assign a single brain operation or psychological function for facial emotion decoding to a certain type of oscillatory activity. Gamma band activity (GBA) offers an adequate tool for studying cortical activation patterns during emotional face information processing. In the present study brain oscillations were analyzed in response to facial expression of emotions. Specifically, GBA modulation was measured when twenty subjects looked at emotional (angry, fearful, happy, and sad faces) or neutral faces in two different conditions: supraliminal (10 ms) vs subliminal (150 ms) stimulation (100 target-mask pairs for each condition). The results showed that both consciousness and significance of the stimulus in terms of arousal can modulate the power synchronization (ERD decrease) during 150-350 time range: an early oscillatory event showed its peak at about 200 ms post-stimulus. GBA was enhanced by supraliminal more than subliminal elaboration, as well as more by high arousal (anger and fear) than low arousal (happiness and sadness) emotions. Finally a left-posterior dominance for conscious elaboration was found, whereas right hemisphere was discriminant in emotional processing of face in comparison with neutral face.     

The fast detection of rare auditory feature conjunctions in the human brain as revealed by cortical gamma-band electroencephalogram

Natural environments typically contain temporal scatters of sounds emitted from multiple sources. The sounds may often physically stand out from one another in their conjoined rather than simple features. This poses a particular challenge for the brain to detect which of these sounds are rare and, therefore, potentially important for survival. We recorded gamma-band (32-40 Hz) electroencephalographic (EEG) oscillations from the scalp of adult humans who passively listened to a repeated tone carrying frequent and rare conjunctions of its frequency and intensity. EEG oscillations that this tone induced, rather than evoked, differed in amplitude between the two conjunction types within the 56-ms analysis window from tone onset. Our finding suggests that, perhaps with the support of its non-phase-locked synchrony in the gamma band, the human brain is able to detect rare sounds as feature conjunctions very rapidly.     

The structure of the Graafian follicle as revealed by microdissection

Summary A study of the developing spleen in thirty-three embryos of the Gecko, Gymnodactylus kotschyi, revealed the fact that four phases occurred. In thefirst phase the spleen anlage appears as a plate of cells on the peritoneal surface of the dorsal mesentery, just above the primordial stomach. Thesecond phase shows the spleen as a brick-shaped protuberance in which the first capillaries (believed to be arterial) appear. The blood drains through the dorsal pancreas to enter the ductus venosus. Thethird phase shows the spleen as an oval with a more extensive system of capillaries which still seem to be arterial in nature. The blood still drains into the dorsal pancreas, thence to the ductus venosus. Thefourth phase persists to the time of hatching and reveals a change in the circulatory system of the spleen. The capillaries within the spleen are of two sizes. Those closest to the splenic artery are smaller in diameter than those which lead to the splenic vein. The former seem to be arterial and the latter venous in type. The splenic vein enters the ductus venosus direct. The intermediate pathway through the pancreas has now been dicontinued. The cells of the spleen are very small and almost entirely nuclear in the later embryonic stages.

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Zusammenfassung Eine Untersuchung über die Entwicklung der Milz von 33 Embryonen des Gecko (Gymnodactylus kotschyi) zeigt, daß der Vorgang in vier Zeitabschnitten stattfindet.In demersten Abschnitt erscheint die Anlage der Milz als eine Platte von Zellen an der peritonealen Oberfläche des dorsalen Mesenteriums, nahe über dem primordialen Magen.Während deszweiten Abschnittes erscheint die Milz in der Form einer ziegelsteinartigen Hervorragung in welcher die ersten Capillaren (wahrscheinlich arterielle Capillaren) zum Vorschein kommen. Das Blut geht durch die dorsale Pankreas und tritt in den Ductus venosus ein.Derdritte Abschnitt zeigt die Milz als ein Oval mit einem weitverzweitgten Capillarsystem, welches noch immer arteriell zu sein schcint. Das Blut nimmt denselben Weg im zweiten Abschnitt.Dervierte Abschnitt dauert bis zum Zeitpunkt des Ausschlüpfens und gibt einen Einblick in die Veränderung des Blutkreislaufs der Milz. Die Capillaren in der Milz sind von zweierlei Größe. Die Capillaren in der Nähe der Milzarterie sind kleiner als die, welche in die Vene führen. Die ersteren scheinen arteriell zu sein, während die letzteren veneus sind. Die Vene der Milz tritt direkt in den Ductus venosus ein. Der Umweg durch die Pankreas ist nun vermieden. Die Zellen der Milz sind sehr klein und bestehen in späteren embryonischen Stufen beinahe nur aus dem Zellkern.

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Four text-figures     

整片组织免疫染色显示淋巴管构筑

目的探讨整片组织内淋巴管免疫染色,并与间接注射法和组织切片免疫染色法比较。方法用普鲁士蓝间接注射新生儿肝被膜内淋巴管,观察肝浅淋巴管的构筑。通过火棉胶切片分析肝浅淋巴管的流向。取人大腿皮肤,作石蜡切片和冷冻切片,免疫染色后观察淋巴管的分布。取大鼠耳和背部的皮肤、膈和小肠,将整片组织作免疫染色,观察组织内淋巴管构筑。结果间接注射普鲁士蓝可显示淋巴管构筑和流向,但淋巴管染色缺乏特异性。组织切片免疫染色可显示淋巴管分布,然而不能准确地分析淋巴管密度。整片组织的免疫染色很理想,毛细淋巴管盲端、毛细淋巴管网和淋巴管丛清晰可见。结论整片组织免疫染色能够很好地显示组织内完整的淋巴管构筑。     

Neuronal pathways to the rat thyroid revealed by retrograde tracing and immunocytochemistry

The distribution and origin of the nerve fibres innervating the rat thyroid were studied by immunocytochemistry, retrograde tracing and denervation experiments. Immunocytochemistry revealed nerve fibres containing noradrenaline, neuropeptide Y, vasoactive intestinal peptide, peptide histidine-isoleucine, galanin, substance P, neurokinin A and calcitonin gene-related peptide around blood vessels and follicles. Many of these transmitter candidates were found to co-exist with each other in different combinations in different subpopulations of neurons. Sympathectomy eliminated all noradrenaline- and noradrenaline/neuropeptide Y-containing fibres in the thyroid. Cervical vagotomy eliminated about 50% of the galanin-, substance P- and calcitonin gene-related peptide-containing fibres. Local denervation (removal of the thyroid ganglion and the thyroid nerve) eliminated all galanin- and substance P-immunoreactive fibres and the majority of noradrenaline-, noradrenaline/neuropeptide Y-, vasoactive intestinal peptide- and calcitonin gene-related peptide-containing fibres in the thyroid gland. Injection of True Blue into the thyroid gland labelled cell bodies in the thyroid ganglion, the laryngeal ganglion, the superior cervical ganglion, the jugular-nodose ganglionic complex, the dorsal root ganglia (C2-C5) and the trigeminal ganglion. Judging from the number of labelled nerve cell bodies, the superior cervical ganglion and the thyroid ganglion contribute most to the thyroid innervation, while the laryngeal ganglion and the trigeminal ganglion contribute least. The True Blue-labelled ganglia were examined for the presence of various populations of nerve cell bodies (only major populations are listed). The thyroid ganglion harboured neuropeptide Y, vasoactive intestinal peptide and galanin/vasoactive intestinal peptide cell bodies (in order of predominance); the laryngeal ganglion galanin/vasoactive intestinal peptide, vasoactive intestinal peptide and calcitonin gene-related peptide cell bodies; the superior cervical ganglion noradrenaline/neuropeptide Y and noradrenaline cell bodies; the jugular ganglion calcitonin gene-related peptide, substance P/calcitonin gene-related peptide and galanin/substance P/calcitonin gene-related peptide cell bodies; the nodose ganglion vasoactive intestinal peptide and vasoactive intestinal peptide/galanin cell bodies; the dorsal root ganglia (C2-C5) and the trigeminal ganglion calcitonin gene-related peptide, substance P/calcitonin gene-related peptide and galanin/substance P/calcitonin gene-related peptide cell bodies.(ABSTRACT TRUNCATED AT 400 WORDS)