Differential accumulation of hydroxyproline-rich glycoproteins in bean root nodule cells infected with a wild-type strain or a C4-dicarboxylic acid mutant of Rhizobium leguminosarum bv. phaseoli

An antiserum raised against deglycosylated hydroxyproline-rich glycoproteins (HPGPs) from melon (Cucumis melo L.) was used to study the relationship between Rhizobium infection and induction of HRGPs in bean (Phaseolus vulgaris L.) root nodule cells infected with either the wild-type or a C₄-dicarboxylic acid mutant strain of Rhizobium leguminosarum bv. phaseoli. In effective nodules, where fixation of atmospheric dinitrogen is taking place, HRGPs were found to accumulate mainly in the walls of infected cells and in peribacteroid membranes surrounding groups of bacteroids. Internal ramifications of the peribacteroid membrane were also enriched in HRGPs whereas the peribacteroid space as well as the bacteroids themselves were free of these glycoproteins. In mutant-induced root nodules, HRGPs were specifically associated with the electron-dense, laminated structures formed in plastids as a reaction to infection by this mutant. The presence of HRGPs was also detected in the host cytoplasm. The aberrant distribution of HRGPs in infected cells of mutant-induced nodules likely reflects one aspect of the altered host metabolism in relation to peribacteroid-membrane breakdown. The possibility that the antiserum used for HRGP localization may have cross-reacted with ENOD 2 gene products is discussed in relation to amino-acid sequences and sites of accumulation.     

Carbocyclic phosphonate analogs of 2′,3′-dideoxyadenosine-5′-monophosphate as substrates of 5-phosphoribosyl-1-pyrophosphate (PRPP) synthetate

Several carbocyclic phosphonate analogs of 2′,3′-dideoxyadenosine-5′-monophosphate (ddAMP) were pyrophosphorylated by E. coli 5-phosphoribosyl-1-pyrophosphate (PRPP) synthetase in the presence of PRPP. Structure-activity relationships are discussed.     

Co-distribution of annexin VI and actin in secretory ameloblasts and odontoblasts of rat incisor

Summary Annexin VI and actin were detected by immunoblot analysis in the enamel- and dentin-related portions of dental tissues. Annexin VI was found mainly in the particulate fraction whereas actin was detected in both the soluble and particulate fractions. By immunoelectron microscopy, annexin VI antibodies conjugated with colloidal gold were seen to label the mitochondria, the cytosol and the nucleus of secretory ameloblasts and odontoblasts of rat incisor. In the processes of these cell, the plasmalemmal undercoat was labeled. Antiactin antibodies labeled the desmosome-like junctions, the cytosol, and the mitochondria of the cell bodies. Extensive labeling was seen at the periphery of the Tomes' processes and odontoblast processes. These results suggest that annexin VI may play a role in Ca²⁺-regulation in the cell bodies, especially as a calcium receptor protein in the mitochondria. Moreover, annexin VI and actin seem to be co-distributed in secretory processes. Thus, these proteins might be both involved in exocytotic and endocytotic events.     

Localization of neuropeptide Y-immunoreactive cells and fibres in the brain of the Japanese quail

Summary In the present study, we have demonstrated, by means of the biotin-avidin method, the widespread distribution of neuropeptide Y (NPY)-immunoreactive structures throughout the whole brain of the Japanese quail (Coturnix coturnix japonica). The prosencephalic region contained the highest concentration of both NPY-containing fibres and perikarya. Immunoreactive fibres were observed throughout, particularly within the paraolfactory lobe, the lateral septum, the nucleus taeniae, the preoptic area, the periventricular hypothalamic regions, the tuberal complex, and the ventrolateral thalamus. NPY-immunoreactive cells were represented by: a) small scattered perikarya in the telencephalic portion (i.e. archistriatal, neostriatal and hyperstriatal regions, hippocampus, piriform cortex); b) medium-sized cell bodies located around the nucleus rotundus, ventrolateral, and lateral anterior thalamic nuclei; c) small clustered cells within the periventricular and medial preoptic nuclei. The brainstem showed a less diffuse innervation, although a dense network of immunopositive fibres was observed within the optic tectum, the periaqueductal region, and the Edinger-Westphal, linearis caudalis and raphes nuclei. Two populations of large NPY-containing perikarya were detected: one located in the isthmic region, the other at the boundaries of the pons with the medulla. The wide distribution of NPY-immunoreactive structures within regions that have been demonstrated to play a role in the control of vegetative, endocrine and sensory activities suggests that, in birds, this neuropeptide is involved in the regulation of several aspects of cerebral functions.Abbreviations AA archistriatum anterius - AC nucleus accumbens - AM nucleus anterior medialis - APP avian pancreatic polypeptide - CNS centrai nervous system - CO chiasma opticum - CP commissura posterior - CPi cortex piriformis - DIC differential interferential contrast - DLAl nucleus dorsolateralis anterior thalami, pars lateralis - DLAm nucleus dorsolateralis anterior thalami, pars medialis - E ectostriatum - EW nucleus of Edinger-Westphal - FLM fasciculus longitudinalis medialis - GCt substantia grisea centralis - GLv nucleus geniculatus lateralis, pars ventralis - HA hyperstriatum accessorium - Hp hippocampus - HPLC high performance liquid chromatography - HV hyperstriatum ventrale - IF nucleus infundibularis - IO nucleus isthmo-opticus - IP nucleus interpeduncularis - IR immunoreactive - LA nucleus lateralis anterior thalami - LC nucleus linearis caudalis - LFS lamina frontalis superior - LH lamina hyperstriatica - LHRH luteinizing hormone-releasing hormone - LoC locus coeruleus - LPO lobus paraolfactorius - ME eminentia mediana - N neostriatum - NC neostriatum caudale - NPY neuropeptide Y - NIII nervus oculomotorius - NV nervus trigeminus - NVI nervus facialis - NVIIIc nervus octavus, pars cochlearis - nIV nucleus nervi oculomotorii - nIX nucleus nervi glossopharyngei - nBOR nucleus opticus basalis (ectomamilaris) - nCPa nucleus commissurae pallii - nST nucleus striae terminalis - OM tractus occipitomesencephalicus - OS nucleus olivaris superior - PA palaeostriatum augmentatum - PBS phosphate-buffered saline - POA nucleus praeopticus anterior - POM nucleus praeopticus medialis - POP nucleus praeopticus periventricularis - PP pancreatic polypeptide - PYY polypeptide YY - PVN nucleus paraventricularis magnocellularis - PVO organum paraventriculare - R nucleus raphes - ROT nucleus rotundus - RP nucleus reticularis pontis caudalis - Rpc nucleus reticularis parvocellularis - RPgc nucleus reticularis pontis caudalis, pars gigantocellularis - RPO nucleus reticularis pontis oralis - SCd nucleus subcoeruleus dorsalis - SCv nucleus subcoeruleus ventralis - SCNm nucleus suprachiasmaticus, pars medialis - SCNl nucleus suprachiasmaticus, pars lateralis - SL nucleus septalis lateralis - SM nucleus septalis medialis - Ta nucleus tangentialis - TeO tectum opticum - Tn nucleus taeniae - TPc nucleus tegmenti pedunculo-pontinus, pars compacta - TSM tractus septo-mesencephalicus - TV nueleus tegmenti ventralis - VeL nucleus vestibularis lateralis - VLT nucleus ventrolateralis thalami - VMN nucleus ventromedialis hypothalami A preliminary report of this study was presented at the 15th Conference of European Comparative Endocrinologists, Leuven, Belgium, September 1990     

Spectrum of phenylketonuria mutations in Western Europe and North Africa,and their relation to polymorphic DNA haplotypes at the phenylalanine hydroxylase locus

Summary A total of 252 chromosomes from 126 patients with phenylalanine hydroxylase (PAH) deficiencies were analyzed for both mutant genotypes and restriction fragment length polymorphism (RFLP) haplotypes at the PAH locus. The mutant genes studied originated either from Western Europe (116 alleles) or from Mediterranean countries (136 alleles). Only 27% of all mutant alleles were found to carry identified mutations, particularly mutations at codon 252 (2.3%), 261 (7.5%), 280 (6.3%), 408 (3.5%) and at the splice donor site of intron 12 (6.3%). The mutant genotypes were associated with RFLP haplotypes 7, 1, 38, 2 and 3 at the PAH locus respectively. Except for the splice mutation of intron 12, these associations were preferential, but not exclusive, since the other four mutations were found on the background of at least two RFLP haplotypes. These results, together with the observation that 85% of PAH deficient patients are heterozygotes for their mutant genotypes, emphasize the great heterogeneity of PAH deficiencies in Mediterranean countries and hamper systematic DNA testing for carrier status in this population.     

Characterization of kinetochores in multicentric chromosomes

Long-term cultures of certain rat and mouse cell lines carry several dicentric and some multicentric chromosomes. Using antikinetochore antibodies obtainable from serum of scleroderma (var. CREST) patients we studied the number of kinetochores formed along the length of these chromosomes. The rat cells displayed as many kinetochores as there were centromeres. However, mouse cells showed the synthesis of only one kinetochore in dicentric and multicentric chromosomes which had been in the culture for a period of 1 year or more. When translocations were induced by bleomycin in mouse L cells, the newly formed dicentric chromosomes showed the formation of two kinetochores. It is not known when the accessory centromeres lose their capacity to assemble kinetochore proteins. Possibly, in the rat the latent kinetochores lack a specific component which renders them ineffective for microtubule binding. The reason for the formation of only one kinetochore in mouse multicentric chromosomes is not clear. It may be due to the accumulation of mutations, modification of the kinetochore protein so that it lacks the antibody binding component, or a more effective regulatory gene than in the rat.     

Chromosomal distribution of P and I transposable elements in a natural population of Drosophila melanogaster

The chromosomal distribution of P and I transposable elements was studied, by in situ hybridisation, in 25 isofemale lines of Drosophila melanogaster collected at Nasr'Allah in Tunisia. An important interline variability for the number of copies of both elements was revealed. The mean number of copies per line was 31.3 for P and 21.0 for I. Certain chromosome arms had a higher frequency of copies than others: arm 3R had the highest frequency of I elements; the X chromosome had the highest frequency of P elements and the lowest frequency of I elements. For both P and I elements the number of copies on the different chromosome arms is independent. Furthermore, there is no significant correlation between the number of copies of P and the number of copies of I for a given line. A study of the localisation of hybridisation sites on the X chromosome revealed the existence of preferred regions for each family. The population studied was of type M' in the P-M system of hybrid dysgenesis. There is no direct relationship between the M potential of an isofemale line and its number of copies of P elements. These results are compared with those of other investigators and the consequences for cytotype determination are discussed.