Immunolocalization of choline acetyltransferase in 2 types of efferent synapses of the organ of Corti

The efferent (olivo-cochlear) innervation of the organ of Corti was studied using a monoclonal antibody against choline acetyltransferase (ChAT). In the inner spiral bundle (ISB), below the inner hair cells (IHCs), the anti-ChAT immunoreactivity was observed within unvesiculated fibers and vesiculated varicosities. Unreactive varicosities, at least as numerous as the immunoreactive ones, were also detected. Both types of vesiculated varicosities synapsed with the dendrites of the primary auditory neurons (afferent fibers) connected to the IHCs. At the outer hair cell (OHC) level, nearly all the vesiculated terminals making axo-somatic synapses with the OHCs were anti-ChAT immunoreactive. Only few terminals synapsing with the OHCs were unreactive. These findings allowed the differentiation of at least three types of efferent synapses in the organ of Corti. In the ISB, a first population of axo-dendritic synapses seems to be cholinergic whereas a second population might use another neurotransmitter. At the OHC level, our results support the hypothesis that acetylcholine is the neurotransmitter of nearly all the large axo-somatic synapses. The rare unreactive axo-somatic synapses could constitute a fourth and minor type of efferent synapse. Thus, it would be helpful to subclassify the efferent innervations of the organ of Corti according to their neurochemical nature. A re-evaluation of the whole body of available electrophysiological data would be also necessary, as until now, acetylcholine was considered as being the only efferent cochlear neurotransmitter.     

Effect of historical factors on genetic variation in three terrestrial Cephalanthera species (Orchidaceae) with different breeding system on the Korean Peninsula

Previous studies have shown that levels of genetic diversity in species of the genus Cephalanthera covary with the breeding system. In the southern part of the Korean Peninsula, the three self‐compatible terrestrial orchids Cephalanthera erecta, C. falcata and C. longibracteata flower synchronously in sympatric populations. The food‐deceptive C. falcata with bright yellow flowers is predominantly outcrossing, whereas autogamy is the dominant strategy in both C. erecta and C. longibracteata, whose white flowers do not open fully. We examined genetic diversity (by means of allozymes) of the three species in sympatric populations (600 × 600 m area) in the Yeonwhasan Provincial Park (YPP) and in non‐sympatric populations outside YPP, South Korea. Thirteen out of 20 putative loci were variable across the three species, but there was a complete lack of allozyme variation within each species and we found no evidence of hybridisation. Our results suggest that historical factors, i.e. the Quaternary climate oscillations, have played a major role in determining levels of genetic diversity in the three Cephalanthera species. The Korean populations of C. erecta (a warm‐temperate/temperate element) and C. falcata (a warm‐temperate element) may have been established by a single introduction from a genetically depauperate ancestral population, likely located outside the Korean Peninsula. On the other hand, since C. longibracteata is a boreal/temperate element, it may have survived the Last Glacial Maximum in microrefugia located in low elevation regions within the Peninsula where it has been subjected to population bottlenecks reducing its genetic diversity.     

p-Chlorophenylalanine-Induced Alteration of Somatodendritic Levels of Tryptophan Hydroxylase Within the Rat Mesencephalic Raphe Nuclei

Abstract: Tryptophan hydroxylase distribution was examined across the nuclei raphe dorsalis, medianus, and pontis of the adult rat, under basal conditions and 2 days after a single injection of p -chlorophenylalanine, an irreversible tryptophan hydroxylase inhibitor. Tryptophan hydroxylase-expressing cells were numbered in transverse sections processed for immunohistochemistry, and the area of tryptophan hydroxylase distribution was delineated in adjacent sections transferred onto nitrocellulose and processed for immunoautoradiography. Two distinct areas were visualized: an inner zone, corresponding to the area displaying tryptophan hydroxylase-immunoreactive cells (so-called somatic area), and an outer zone, here called perisomatic, devoid of perikarya yet rich in tryptophan hydroxylase-positive neuropil in the histological sections. After treatment with p -chlorophenylalanine, a significant decrease in the number of tryptophan hydroxylase-immunoreactive cells could be observed only in the rostral raphe dorsalis, particularly within its ventromedian and dorsomedian subdivisions. In all raphe nuclei, the topological reconstruction of the somatic area was not modified. Based on the densitometric measurements in the immunoautoradiographs, however, a dramatic decrease in the content, concentration, and volume of expression of tryptophan hydroxylase could be documented in the three raphe nuclei. Detailed analysis of these results led to the conclusion that (a) tryptophan hydroxylase expression is differentially regulated in different serotoninergic cell body subpopulations of the raphe, some of which are more sensitive to p -chlorophenylalanine, and (b) distribution of tryptophan hydroxylase protein is modified also in the somatodendritic area in all raphe nuclei.     

Pharmacological effects of GABA on serotonin metabolism in the rat brain

The pharmacological effects of GABA-related drugs were studied on the serotonin (5-HT) and 5-hydroxyindole-acetic acid (5-HIAA) contents of various regions of the rat brain. These effects were examined in the nuclei raphe dorsalis, magnus and centralis and in structures receiving a dense serotonin innervation such as the habenula complex and subcommissural organ. The GABA agonist, muscimol, increased the 5-HT contents and reduced 5-HIAA levels in structures containing serotoninergic terminals suggesting an inhibitory effect of GABA on the firing of serotoninergic neurons with concomitant reduction of 5-HT utilisation. In contrast, the GABA antagonist, bicuculline, probably stimulated 5-HT turnover since its intraperitoneally administration produced significant increase of 5-HT and/or 5-HIAA levels in the same brain regions. These data are in agreement with a transsynaptic inhibitory control of GABA on serotoninergic neurons. Drugs which inhibit the GABA catabolism such as amino-oxyacetic acid or gamma-vinyl-GABA and which should elevate GABA levels in the synaptic gap were capable of increasing or decreasing the 5-HT and the 5-HIAA levels depending on the experimental conditions. These results suggest that several processes are probably involved in the control of serotoninergic neurons by GABA in the rat brain. Among them, an intracellular effect of GABA on 5-HT metabolism might well occur in cells containing both GABA and 5-HT.     

Nucleolar structures in chromosome and SC preparations from human oocytes at first meiotic prophase

Summary We describe a comparative study of the behavior of nucleolar structures and their relationship with nucleolar chromosomes and synaptonemal complexes at first meiotic prophase of human oocytes in an attempt to elucidate the nature of this cellular organization and to learn more about maternal nondisjunction. The number of main nucleoli varies along the different stages of prophase I and is usually low. It shows an increase from leptotene to pachytene and a decrease from pachytene to diplotene related to a decrease and an increase of main nucleoli volume, respectively. The methodology employed has enabled us to analyze in detail dark bodies, round bodies, dense bodies, and main nucleoli in chromosome or synaptonemal complex spreads. The relationship between nucleolar chromosomes or synaptonemal complexes and the nucleoli implies the existence, in a very reduced space, of chromosomal regions that contain homologous sequences and that are often unpaired. This situation may facilitate the production of heterologous pairing and chromosomal exchanges between nonhomologous chromosomes and finally result in aneuploidy. THus, the situation explained above together with the differences between the oocyte and spermatocyte NOR cycles could be one of the reasons for the higher incidence of aneuploidies of maternal origin at meiosis I.     

Endothelial cell junctions

In the course of a freeze-cleave study on intercellular junctions in the regenerating rat liver, we observed an unusual array of intramembranous particles located in regions of contact between endothelial cells lining the hepatic sinusoids. These arrays were characterized by an accumulation of particles which resembled a zonula occludens in their linear deployment but differed in that the contact regions were composed of individual particles which remained separated from each other by regular particle-free intervals.     

Le collagène du byssus de Mytilus edulis L.

Résumé L'autoradiographie révèle, au niveau du pied, une incorporation massive et sélective de la ³H-Proline dans la glande blanche de Mytilus edulis. Cette étude a permis de suivre le processus qui mène de la synthèse de la sécrétion dans la partie basale des cellules jusqu'a son émission dans le sillon pédieux où elle participe à la formation du filament. La collagénase détruit la presque totalité du marquage, attestant ainsi la nature collagénique du produit sécrété. Les autres glandes pédieuses ainsi que la glande du byssus proprement dite, située à la base du pied, montrent une incorporation très faible, sans commune mesure avec celle de la glande blanche. Ceci démontre de façon définitive que le collagène présent dans le filament prend naissance dans cette glande et justifie la dénomination de glande du collagène. Des contrôles réalisés dans différentes régions (bords du manteau, manteau, branchies) montrent que l'injection du précurseur dans le bord palléal constitue une méthode satisfaisante pour marquer de façon relativement rapide et différentielle le collagène de la glande.

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The collagen of the byssus in Mytilus edulis L.II. Autoradiographic study on the incorporation of ³H-Proline

Summary Autoradiographic studies reveal a strong specific incorporation of ³H-Proline in the white gland in the foot of Mytilus edulis. The author could trace the radioactive secretory product from its synthesis in the basal part of the cells down to its outflow into the pedial groove where it takes part in the formation of the filament. Purified collagenase takes out radioactivity from the sections. This observation confirms the collagenous nature of the secretion.The other foot-glands as well as the main byssus gland located at the base of the foot show but a very weak labelling not comparable with that of the white gland. This clearly evidences that the collagen occuring in the filament originates from the latter. The white gland may be properly called: collagen gland.Control sections through different parts of the body (mantle-edge, mantle, gills) confirm that our injection technique of the precursor into the palleal margin is a suitable method for a rather quick and specific labelling of the glandular collagen.

Cette note fait partie d'un travail pour l'obtention d'une thèse de doctorat.