Carbonic anhydrase enzyme histochemistry of cranial nerve primary sensory afferent neurons in the rat

Summary Hansson's enzyme histochemical method for the demonstration of carbonic anhydrase has been used to examine primary sensory neurons of cranial nerves in the rat (cochlear ganglion cells excluded). Numerous carbonic anhydrase positive neurons were present in the trigeminal and geniculate ganglia as well as in the mecencephalic trigeminal nucleus. A few carbonic anhydrase positive ganglion cells were found in the nodose ganglion, but none in the petrosal and vestibular ganglia. However, in the latter ganglia, satellite cells surrounding the neurons frequently showed staining for carbonic anhydrase.     

Regional Distribution of Cytochrome P-450 in the Rat Brain: Spectral Quantitation and Contribution of P-450b,e and P-450c,d

The cytochrome P-450 (P-450) content of different regions of the rat brain was measured after partial purification of the enzyme from homogenates, and the quantitative contribution of P-450b,e and P-450c,d to brain P-450 was assessed by Western immunoblotting and immunohistochemistry using rabbit antibodies raised against purified hepatic P-450b and P-450c, respectively). P-450 could be quantitated by its reduced CO difference spectrum after chromatography of homogenates on p-chloroamphetamine-coupled Sepharose. The yield of P-450 from whole brain was 90 +/- 19 pmol/g of tissue, which is approximately 1% of the level in liver microsomes from control rats. The amount of P-450 recovered from homogenates of olfactory lobes, hypothalamus, thalamus, striatum, cerebral cortex, and brainstem varied between 40 and 100 pmol/g of tissue. The cerebellum was a region of exceptionally high P-450 content, with yields of up to 400 pmol/g whereas the substantia nigra yielded only 16-20 pmol/g. Immunohistochemical studies with anti-P-450b and anti-P-450c revealed intense staining of a limited number of cells in the cerebellum with both antibodies and in the thalamus only with anti-P-450c. In the cerebellum, both anti-P-450b and anti-P-450c stained the Bergmann glial cells together with their radial processes. Individual glial cells in the granular cell layer were also stained. There was no staining of Purkinje cells. In the thalamus, anti-P-450b gave weak staining of certain astroglia, but with anti-P-450c, there was intense staining of neuronal somata.(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantitation and tissue localization of the cellular retinoic acid-binding protein

The distribution of the cellular retinoic acid-binding protein (CRABP) in some rat tissues has been determined, and the protein has been localized by immunocytochemical techniques in sections from rat testis. In the testis CRABP was found in the seminiferous tubuli with Sertoli cells and the spermatogonia most intensely stained. All other cells of the germinal epithelium appeared largely devoid of CRABP. By use of an enzyme-linked immunosorbent assay CRABP was quantitatively estimated in several tissues and the highest levels were found in testis and eye. Comparisons of the tissue levels of CRABP and of the cellular retinol-binding protein (CRBP) did not reveal any apparent correlation.     

Developmental changes of gangliosides of the rat stomach. Appearance of a blood group B-active ganglioside

Rat stomach gangliosides were purified and their distribution in the different tissue compartments was established. Three major monosialogangliosides were found: GM3, GM1, and a ganglioheptaosylceramide carrying a blood group B determinant. This latter structure was characterized by exoglycosidase degradation, immunostaining with a monoclonal anti-blood group B antibody on thin layer chromatogram, permethylation analysis, electron-impact mass spectrometry of the permethylated-reduced and trimethylsilylated molecule, and 1H NMR spectroscopy of the native ganglioside. It was found to be (Formula: see text) i.e. a GM1 structure substituted with the blood group B determinant and was called B-GM1. A similar structure has been previously identified in precancerous rat liver and chemically induced rat hepatoma (Holmes, E. H., and Hakomori, S. (1982) J. Biol. Chem. 257, 7698-7703). Fucosyl-GM1 was also detected as a minor ganglioside in rat gastric mucosa. The ganglioside profile was modified during the postnatal development. The contribution of GM3 and GD3, which accounted for 95% of the ganglioside sialic acid at birth, decreased during the first 3 weeks of life. GM1, fucosyl-GM1, and B-GM1 were not detected at birth. The concentration of the fucogangliosides increased during the 2nd and 3rd weeks after birth, was stable during the 4th week and then decreased, whereas that of GM1 increased steadily between 6 days and 2 months of age. B-GM1, which has been defined as a tumor-associated ganglioside in the rat liver, was found to be a developmentally regulated antigen of the normal rat stomach.     

Characterization of neutral blood group B-active glycosphingolipids of rat gastric mucosa. A novel type of blood group active glycosphingolipid based on isogloboside

The blood group active glycosphingolipids of rat gastric mucosa have been investigated. Only blood group B active structures were found, two of which have been structurally characterized by monoclonal antibodies, mass spectrometry, permethylation analyses, proton NMR spectroscopy, and exoglycosidase digestions. A six-sugar compound based on a gangliotetraosylceramide core was isolated and shown to have the following structure: (Formula: see text). The same compound was recently isolated from rat bone marrow cells and characterized by Taki et al. (Taki, T., Kimura, H., Gasa, S., Nakamura, M., and Matsumoto, M. (1985) J. Biol. Chem. 260, 6219-6225). The possible precursor compounds of this structure, gangliotriaosylceramide and gangliotetraosylceramide, were also found in the gastric mucosa. A seven sugar compound, based on isogloboside, was isolated from the gastric mucosa and shown to have the following structure: (formula; see text) The latter compound is novel and extends the list of different types of core structures found for blood group glycolipids. The epithelial cells of the stomach are unique among the cells lining the gastrointestinal tract in having blood group active glycolipids based on ganglio- and isogloboseries core structures.     

Neurons in primary cultures from five defined rat brain regions--cellular composition and morphological appearance

Primary cultures from 15-17 days old fetal rat cerebral cortex, striatum, hippocampus, substantia nigra and brain stem were grown for ten days. Cell aggregates were formed one to two days after seeding. The cell bodies migrated peripherally from the clusters during development and networks of processes were formed. The cultures from the different brain regions contained predominantly neurons, stained by an antiserum against the neuron-specific enolase (NSE). There were differences in morphological appearance of the aggregates and also of the single neuronal cells cultivated from the various brain regions. On the bottom of the culture dishes a monolayer was formed of predominantly undifferentiated (mesenchymal-like) cells. Some cells of the monolayer stained for the astrocyte markers glial fibrillary acidic protein (GFAp) or S-100. The majority of the cells were, however, unstained to these markers. Very few endothelial cells and macrophages were observed.     

No Linkage between Genes Controlling Female Pheromone Production and Male Pheromone Response in the European Corn Borer, Ostrinia Nubilalis Hubner (Lepidoptera; Pyralidae)

The E and Z pheromonal strains of the European corn borer, Ostrinia nubilalis, are characterized by female production of and male preference for opposite blends of (E)-11-and (Z)-11-tetradecenyl acetate. It is known that the pheromone production is controlled by an autosomal gene and that the males' behavior is determined by a sex-linked gene. A third gene, autosomally inherited, has been shown to determine the organization of the male pheromone receptors. In the present study the linkage relationship between the autosomal genes controlling sex pheromone production and male olfactory sensilla was investigated. A recombination experiment showed unequivocally that the genes determining the variation in pheromone production and male pheromone receptors are not closely linked and are most likely inherited independently.     

Effects of 3-aminobenzamide on Chinese hamster cells treated with thymidine analogues and DNA-damaging agents. Chromosomal aberrations, mutations and cell-cycle progression

The nuclear enzyme, poly(ADP-ribose) synthetase is involved in the repair of damaged DNA. We report here the results obtained with 3-aminobenzamide (3AB), an inhibitor of this enzyme, on induced biological effects. 3AB increases the frequency of chromosomal aberrations induced by DMS, EMS, ENU, bleomycin and CldUrd. The magnitude of the effect is dependent on the type of chemical used, the combinations with DMS and EMS being the most potent ones. No potentiation was observed after treatment of cells with MMC. Mutation frequencies were determined on the HPRT locus and showed that 3AB did not increase the frequency of gene mutations induced by EMS, ENU and CldUrd. Cell-cycle progression is affected when cells are grown in medium containing CldUrd and 3AB, primarily when the inhibitor is present during the second cell cycle when substituted DNA becomes replicated. The extent of the effect depends on the amount of analogue incorporated and is independent of the presence of the analogue in the medium during the second cell cycle. Analysis of chromosomal aberrations in delayed G2 cells with the aid of the premature chromosome-condensation technique revealed numerous aberrations after incorporation of CldUrd and treatment with 3AB.     

Human arterial smooth muscle cells in culture. Effects of platelet-derived growth factor and heparin on growth in vitro

Human arterial smooth muscle cells (hASMC) were cultured from explants of the inner media of uterine arteries obtained at hysterectomy. The presence of alpha-actin and smooth muscle-specific actin isoforms and the microscopic appearance of the cells in secondary culture established their smooth muscle origin. The hASMC were diploid and had no signs of transformation. Plasma-derived serum failed to stimulate their proliferation in vitro. Their rate of proliferation was, however, proportional to the concentration of whole blood serum in the medium. Anti-PDGF IgG at high concentrations inhibited the stimulatory effect of whole blood serum on cell proliferation. This suggests that hASMC depend on exogenous PDGF for their growth. In PDS or bovine serum albumin cell numbers remained constant for 7 days in culture and the thymidine index was below 1% per 24 h. When reexposed to whole blood serum these cells started to proliferate within 2 days. This indicates that hASMC when deprived of PDGF enter a quiescent state that is fully reversible upon rexposure to the mitogen. Heparin is a powerful growth inhibitor for SMC. In our system, heparin caused a dose-dependent inhibition of cell proliferation despite optimal concentrations of whole blood serum. This inhibition was reversible upon withdrawal of heparin. At heparin concentrations which caused a half-maximal inhibition it was also competed for by increasing concentrations of whole blood serum. Quiescent hASMC expressed the PDGF receptor on their surface as judged from immunofluorescence with a monoclonal antibody. This was true irrespective of whether growth arrest was achieved by serum depletion or by the addition of heparin to serum-containing medium. Cells growing in the presence of whole blood serum did not, however, express the receptor antigen. These observations suggest that heparin may interfere with PDGF or with its binding and further processing at the level of the cell-surface receptor.     

Evolutionary consequences of climate‐induced range shifts in insects

Range shifts can rapidly create new areas of geographic overlap between formerly allopatric taxa and evidence is accumulating that this can affect species persistence. We review the emerging literature on the short‐ and long‐term consequences of these geographic range shifts. Specifically, we focus on the evolutionary consequences of novel species interactions in newly created sympatric areas by describing the potential (i) short‐term processes acting on reproductive barriers between species and (ii) long‐term consequences of range shifts on the stability of hybrid zones, introgression and ultimately speciation and extinction rates. Subsequently, we (iii) review the empirical literature on insects to evaluate which processes have been studied, and (iv) outline some areas that deserve increased attention in the future, namely the genomics of hybridisation and introgression, our ability to forecast range shifts and the impending threat from insect vectors and pests on biodiversity, human health and crop production. Our review shows that species interactions in de novo sympatric areas can be manifold, sometimes increasing and sometimes decreasing species diversity. A key issue that emerges is that climate‐induced hybridisations in insects are much more widespread than anticipated and that rising temperatures and increased anthropogenic disturbances are accelerating the process of species mixing. The existing evidence only shows the tip of the iceberg and we are likely to see many more cases of species mixing following range shifts in the near future.