5′-Nucleotidase in spinal meningeal compartments in the rat

Summary The distribution of 5-nucleotidase (5-Nu) is reported in spinal meninges of the rat on the basis of an immunohistochemical and enzyme histochemical investigation. Strong immunoreactivity was found in the arachnoid membrane and in the sheaths of the spinal roots as well as in septa subdividing the roots. Also the superficial layer of the ligamentum denticulatum showed enzyme staining. No immunoreactivity could be detected in the pia mater or along the spinal nerve roots outside the subarachnoid space. Within the arachnoid mater the immunoreactivity was concentrated in the basal zone of the arachnoid membrane, thus appearing as a narrow fluorescent band near the border of the dura. An accentuation of immunoreactivity could be observed in areas where small dural blood vessels approach the subarachnoid space. It is well known that adenine nucleotides released from neural and glial cells of the central nervous system finally reach the cerebrospinal fluid. We presume that 5-Nu in the arachnoid membrane and spinal root sheaths is responsible for the conversion of adenine nucleotides into adenosine and that this conversion is associated with the reabsorption process of cerebrospinal fluid which most probably also takes place in spinal meninges. Adenosine, the product of 5-nucleotidase, could play a role in the reabsorption process by its vasodilatatory effect on dural and epidural vessels.     

Über die interspezifische Wirkung des Sexuallockstoffes von Apis mellifica und Apis cerana

Zusammenfassung In Feldversuchen an Drohnensammelplätzen wurden Königinnen von Apis mellifica und A. cerana von mellifica-Drohnen angeflogen. Die arteigene Königin wurde bei gleichzeitiger Darbietung der artfremden Königin in manchen Fällen (an Tagen mit geringer sexueller Appetenz der Drohnen) gesichert bevorzugt.Elektrophysiologisch konnten von den Porenplatten der Drohnenantennen beider Arten langsame Potentiale bei Reizung mit 9-Oxo-trans-2-decensäure (Queen substance) abgeleitet werden. Auf dieselben Riechzellen wirkt auch das Mandibeldrüsensekret von A. cerana. Bei beiden Arten standen die Wirkungen von Queen substance-Proben und von Cerana-Mandibeldrüsen zueinander im gleichen Verhältnis.Riechzellen des gleichen Typs finden sich auch auf den Antennen von Arbeiterinnen und Königinnen beider Arten. Ein anderer Riechzelltyp bei allen Kasten beider Arten reagiert auf den Sterzelduft von A. mellifica.

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On the interspecific effect of the sex attractant of Apis mellifica and Apis cerana

Summary In field experiments drones of Apis mellifica were attracted by queens of Apis mellifica and of A. cerana. On days of low sexual appetite the drones showed a significant preference for their own queen.In both species we recorded spikes and slow potentials from the poreplates of the drones antennae. The same type of olfactory sense cells responds to 9-oxotrans-2-decenoic acid (queen substance) and to the secretion of the mandibular gland of Apis cerana. The quantitative relationship of the effects of the queen substance probes and of the cerana-mandibular glands were the same in both species.Olfactory cells of the same type were found on the queens' and workers' antennae of both species. Another cell type common to all castes of both species responds to the scent of the Nasanov gland of A. mellifica.

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Für die freundliche Überlassung von Proben synthetischer 9-Oxo-trans-2-decensäure danken wir Herrn R. K. Callow (Rothamsted) und H. Rembold (München).     

Cellular components of the immune barrier in the spinal meninges and dorsal root ganglia of the normal rat: immunohistochemical (MHC class II) and electron-microscopic observations

This report deals with the distribution, morphology and specific topical relationships of bone-marrow-derived cells (free cells) in the spinal meninges and dorsal root ganglia of the normal rat. The morphology of these cells has been studied by transmission and scanning electron microscopy. Cells expressing the major histocompatibility complex (MHC) class II gene product have been recognized by immunofluorescence. At the level of the transmission electron microscope, free cells are found in all layers of the meninges. Many of them display characteristic ultrastructural features of macrophages, whereas others show a highly vacuolated cytoplasm and are endowed with many processes. These elements lack a conspicuous lysosomal system and might represent dendritic cells. Scanning electron microscopy has revealed that free cells contact the cerebrospinal fluid via abundant cytoplasmic processes that cross the cell layers of the pia mater and of the arachnoid. Cells expressing the MHC class II antigen are also found in all layers of the meninges. They are particularly abundant in the layers immediately adjacent to the subarachnoid space, in the neighbourhood of dural vessels, along the spinal roots and in the dural funnels. In addition to the meninges, strong immunoreactivity for MHC class II antigen is observed in the dorsal root ganglia. The ultrastructural and immunohistochemical findings of this study suggest the existence of a well-developed system of immunological surveillance of the subarachnoid space and of the dorsal root ganglia.     

The transition of the thick ascending limb of Henle's loop into the distal convoluted tubule in the nephron of the rat kidney

Summary Examination of serial semithin sections of rat kidney cortex and a subsequent electron microscopic study of selected areas revealed that the characteristic epithelium of the cortical part of the thick ascending limb of Henle extends for a varying distance beyond the macula densa. The transition from the relatively thin epithelium of the thick ascending limb at this site to the three -or even four-fold thicker epithelium of the convoluted part of the distal tubule is sharply defined and occurs without the interposition of an intermediate cell type.The position of the macula densa at the end but still clearly within the ascending limb of Henle's loop is functionally interpreted to guarantee the separation of the sensor point macula densa from disturbing influences which might arise from the secretory activity of the subsequent tubular portion.Investigations supported by the Deutsche Forschungsgemeinschaft. The skillful technical assistance of Mrs. Saliha Sabanovic is gratefully acknowledged     

Zahl und Verteilung antennaler Sensillen bei der Honigbiene (Apis mellifera L.)

Zusammenfassung An Totalpräparaten der Antennengeißel von Arbeiterin und Drohne vonApis mellifera carnica wurden Zahl und Verteilung aller Sensillen und Setae ermittelt. Dabei ließen sich anhand des cuticularen Baues folgende Sensillentypen unterscheiden: S. placodeum, S. ampullaceum, S. coeloconicum, S. campaniforme und 5 Haarsensillen S. trichodeum A, B1, B2, C, D, sowie 4 Setatypen (A 1–3, B), die wahrscheinlich nicht innerviert sind. Die Benennungen der Sensillen wurde den bisher gebrauchten Bezeichnungen gegenübergestellt. Sensillenzahl und -Verteilung, Sinneszellzahl und Funktion der Sensillen wurden anhand von Literaturangaben zusammengestellt und diskutiert. Bemerkenswert ist der starke Dimorphismus zwischen Arbeiterin und Drohne in der relativen Sensillenzahl für die einzelnen Sensillentypen und in der Gesamtzahl der Sinneszellen. So sind bei der Arbeiterin die wahrscheinlich olfaktorischen S. trichodea A und die mechanorezeptorischen S. trichodea B 1 wesentlich stärker vertreten. Die Drohne hat keine S. basiconica und im übrigen wesentlich mehr S. placodea als die Arbeiterin. Insgesamt hat die Drohne eine ca. 2-fach größere Geißeloberfläche und etwa 5-mal soviele Sinneszellen wie die Arbeiterin. Die Arbeiterinnengeißel hat auf ihrer Rückseite eine porenplattenfreie Zone, die dicht mit nichtinnervierten Setae besetzt ist. Bei der Drohne findet man stattdessen eine porenplattenärmere Zone mit einer geringeren Zahl von Setae. Charakteristische Verteilungsmuster bestehen auch für alle anderen Sensillen und Setae.

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Number and distribution of the sensilla on the antennal flagellum of the honeybee (Apis mellifera L.)

Summary Number and distribution of sensilla and setae on the antennal flagellum of the honeybeeApis mellifera carnica were determined on whole antennal preparations. The following types of sensilla were distinguished according to their cuticular structure: Sensillum placodeum, S. ampullaceum, S. coeloconicum, S. basiconicum, S. campaniforme and 5 hair sensilla S.trichodeum A, B1, B2, C, D, as well as 4 types of probably non-innervated setae (A1–3, B). The names used here for the different types were compared with the previously used terms. Number and distribution of sensilla, the number of sensory cells and the function of the sensilla were discussed with respect to the data available from the literature. There is a notable dimorphism between the worker and drone with respect to the relative number of sensilla of each type and to the total number of sensory cells. The worker has far more of the presumably olfactory S. trichodea A and of the mechanoreceptive S. trichodea B1. The drone lacks the S. basiconica and has far more S. placodea than the worker. The flagellum surfarce of the drone is twice as large as that of the worker and has 5 times as many sensory cells. The worker flagellum has a poreplate-free zone on the side facing the head which is densely packed with non-innervated setae. In the corresponding zone the drone has a lower density of poreplates than elsewhere on its antennal flagellum. All other sensilla and setae are also characteristically distributed.

     

Sex pheromone of the moth,Antheraea polyphemus

The sex pheromone system of Antheraea polyphemus was characterized from female abdominal tips by classical and electroantennogram techniques as rans-6,cis-11-hexadecadienyl acetate and trans-6,cis-11-hexadecadienal. A 90 : 10 mixture of acetate and aldehyde was highly attractive to wild males in the field. The synthetic pheromone and A. polyphemus females were not attractive to released Antheraea pernyi males.     

Proliferation of acid-secretory cells in the kidney during adaptive remodelling of the collecting duct

The renal collecting duct adapts to changes in acid-base metabolism by remodelling and altering the relative number of acid or alkali secreting cells, a phenomenon termed plasticity. Acid secretory A intercalated cells (A-IC) express apical H(+)-ATPases and basolateral bicarbonate exchanger AE1 whereas bicarbonate secretory B intercalated cells (B-IC) express basolateral (and apical) H(+)-ATPases and the apical bicarbonate exchanger pendrin. Intercalated cells were thought to be terminally differentiated and unable to proliferate. However, a recent report in mouse kidney suggested that intercalated cells may proliferate and that this process is in part dependent on GDF-15. Here we extend these observations to rat kidney and provide a detailed analysis of regional differences and demonstrate that differentiated A-IC proliferate massively during adaptation to systemic acidosis. We used markers of proliferation (PCNA, Ki67, BrdU incorporation) and cell-specific markers for A-IC (AE1) and B-IC (pendrin). Induction of remodelling in rats with metabolic acidosis (with NH(4)Cl for 12 hrs, 4 and 7 days) or treatment with acetazolamide for 10 days resulted in a larger fraction of AE1 positive cells in the cortical collecting duct. A large number of AE1 expressing A-IC was labelled with proliferative markers in the cortical and outer medullary collecting duct whereas no labeling was found in B-IC. In addition, chronic acidosis also increased the rate of proliferation of principal collecting duct cells. The fact that both NH(4)Cl as well as acetazolamide stimulated proliferation suggests that systemic but not urinary pH triggers this response. Thus, during chronic acidosis proliferation of AE1 containing acid-secretory cells occurs and may contribute to the remodelling of the collecting duct or replace A-IC due to a shortened life span under these conditions.     

The antennal benzoic acid receptor cell of the female silk moth <Emphasis Type="Italic">Bombyx mori</Emphasis> L.: structure–activity relationship studies with halogen substitutes

Studies on structure–activity relationships were carried out to characterize the response specificity of the benzoic acid cell of the female of the moth Bombyx mori by means of single sensillum electrophysiological recordings. We demonstrated that this cell type responds best to a natural key substance (benzoic acid) and has similar response profiles for less effective compounds, including various halogen substitutes of benzoic acid, benzaldehyde and other derivates of the key compound. Using different halogen substitutes (F, Cl, Br, I), we showed that the cellular response decreases with increasing atomic size of the substitute and that halogen substitutes were most effective in the meta-position. Thus, m-fluor benzoic acid was even more effective than benzoic acid. These results indicate that a critical feature of the stimulus molecule is the inductive effect generated by the halogen substitutes. Increasing the atomic size of the halogen substitute impairs the recognition of the molecule by the receptor cell, possibly due to steric effects. Decreasing the electron density in the aromatic ring improves the receptor response. The benzoic acid receptor cell can be considered as specialist despite not being involved in pheromone detection as it responds maximally to a key substance and has similar response profiles for less effective compounds.     

Characterization of renal interstitial fibroblast-specific protein 1/S100A4-positive cells in healthy and inflamed rodent kidneys

Fibrosis is considered as a central factor in the loss of renal function in chronic kidney diseases. The origin of fibroblasts and myofibroblasts that accumulate in the interstitium of the diseased kidney is still a matter of debate. It has been shown that accumulation of myofibroblasts in inflamed and fibrotic kidneys is associated with upregulation of fibroblast-specific protein 1 (FSP1, S100A4), not only in the renal interstitium but also in the injured renal epithelia. The tubular expression of FSP1 has been taken as evidence of myofibroblast formation by epithelial–mesenchymal transition (EMT). The identity of FSP1/S100A4 cells has not been defined in detail. We originally intended to use FSP1/S100A4 as a marker of putative EMT in a model of distal tubular injury. However, since the immunoreactivity of FSP1 did not seem to fit with the distribution and shape of fibroblasts or myofibroblasts, we undertook the characterization of FSP1/S100A4-expressing cells in the interstitium of rodent kidneys. We performed immunolabeling for FSP1/S100A4 on thin cryostat sections of perfusion-fixed rat and mouse kidneys with peritubular inflammation, induced by thiazides and glomerulonephritis, respectively, in combination with ecto-5-nucleotidase (5NT), recognizing local cortical peritubular fibroblasts, with CD45, MHC class II, CD3, CD4 and Thy 1, recognizing mononuclear cells, with alpha smooth muscle actin (SMA), as marker for myofibroblasts, and vimentin for intracellular intermediate filaments in cells of mesenchymal origin. In the healthy interstitium of rodents the rare FSP1/S100A4+ cells consistently co-expressed CD45 or lymphocyte surface molecules. Around the injured distal tubules of rats treated for 3–4 days with thiazides, FSP1+/S100A4+, 5NT+, SMA+, CD45+ and MHC class II+ cells accumulated. FSP1+/S100A4+ cells consistently co-expressed CD45. In the inflamed regions, SMA was co-expressed by 5NT+ cells. In glomerulonephritic mice, FSP1+/S100A4+ cells co-expressed Thy 1, CD4 or CD3. Thus, in the inflamed interstitium around distal tubules of rats and of glomerulonephritic mice, the majority of FSP1+ cells express markers of mononuclear cells. Consequently, the usefulness of FSP1/S100A4 as a tool for detection of (myo)fibroblasts in inflamed kidneys and of EMT in vivo is put into question. In the given rat model the consistent co-expression of SMA and 5NT suggests that myofibroblasts originate from resident peritubular fibroblasts.Ivan Hegyi and Michel Le Hir contributed equally to the study     

In vivo protein biotinylation for identification of organ-specific antigens accessible from the vasculature

We describe a new methodology, based on terminal perfusion of rodents with a reactive ester derivative of biotin that enables the covalent modification of proteins readily accessible from the bloodstream. Biotinylated proteins from total organ extracts can be purified on streptavidin resin in the presence of strong detergents, digested on the resin and subjected to liquid chromatography-tandem mass spectrometry for identification. In the present study, in vivo biotinylation procedure led to the identification of hundreds of proteins in different mouse organs, including some showing a restricted pattern of expression in certain body tissues. Furthermore, biotinylation of mice with F9 subcutaneous tumors or orthotopic kidney tumors revealed both quantitative and qualitative differences in the recovery of biotinylated proteins, as compared to normal tissues. This technology is applicable to proteomic investigations of the differential expression of accessible proteins in physiological and pathological processes in animal models, and to human surgical specimens using ex vivo perfusion procedures.