Neuroanatomical connections between corticotropin-releasing factor (CRF) and somatostatin (SRIF) nerve endings and thyrotropin-releasing hormone (TRH) neurons in the paraventricular nucleus of rat hypothalamus.

In order to investigate the possible involvement of corticotropin-releasing factor (CRF) and somatostatin (SRIF) on thyrotropin-releasing hormone (TRH) neuronal cell activity in the rat hypothalamic paraventricular nucleus, we have proceeded to the simultaneous localization of CRF or SRIF and TRH. For this purpose, we used a dual immunostaining procedure that employed antibodies to CRF and SRIF and peroxidase-labeled goat anti-rabbit IgG as a first sequence, and antibodies to a cryptic fragment (Phe178-Glu199) of pro-TRH (to label TRH neurons) and alkaline phosphatase-labeled goat anti-rabbit IgG as the second sequence. A rich innervation of the paraventricular nucleus by immunoreactive CRF and SRIF fibers was observed. A large number of CRF and SRIF nerve endings were seen intimate anatomic proximity and often appeared to surround TRH-containing cell bodies. These results strongly suggest that TRH neurons might be regulated by both CRF and SRIF. These interactions might be the neuroanatomical basis for the already observed inhibitory effects of CRF and SRIF on TRH release.     

Morphological dispersion of introduced Hawaiian finches: evidence for competition and a Narcissus effect

Summary Morphological analyses were conducted on finch species introduced to Oahu, Hawaii. As many as 25 species of finches representing four families (Emberizidae, Fringillidae, Ploecidae, Estrildidae) have been introduced to Oahu. Of these, 15 species currently have established wild populations. When compared with pools of 25 and 23 species introduced to Oahu, the 15 surviving species are morphologically overdispersed. This result is consistent with the hypothesis that competition has influenced the outcome of these introductions. We also tested the hypothesis that surviving introduced finches in one habitat (sugarcane fields) were morphologically overdispersed. When the seven surviving finch species found in sugarcane were compared with the pools of 25 and 23 species, they too were morphologically overdispersed. However, when the seven sugarcane species were compared with a species pool consisting of only the surviving 15 species found across Oahu, they were not morphologically overdispersed. This result suggests that morphological analyses of community structure based only on comparisons of assemblages of surviving species may be biased by a Narcissus effect.     

1H and15N resonance assignments and secondary structure of the human thioredoxin C62A,C69A,C73A mutant

Summary The complete assignment of¹H and¹⁵N backbone resonances and near-complete¹H side-chain resonance assignments have been obtained for the reduced form of a mutant of human thioredoxin (105 residues) in which the three non-active site cysteines have been substituted by alanines: C62A, C69A, C73A. The assignments were made primarily on the basis of three-dimensional.¹⁵N-separated nuclear Overhauser and Hartmann-Hahn spectroscopy, in conjunction with two-dimensional homonuclear and heteronuclear correlation experiments. Based on comparisons of short-range and interstrand nuclear Overhauser effects, patterns of amide exchange, and chemical-shift differences, the structure appears essentially unchanged from that of the previously determined solution structure of the native protein [Forman-Kay. J.D. et al. (1991)Biochemistry, 30, 2685–2698). An assay for thioredoxin shows that the C62A, C69A, C73A mutant retains activity. The assignment of the spectrum for this mutant of human thioredoxin constitutes the basis for future studies aimed at comparing the details of the active-site conformation in the reduced and oxidized forms of the protein.     

Nuclear genome characterization of the carrageenophyteAgardhiella subulata (Rhodophyta)

DNA reassociation kinetics were used to determine nuclear genome organization and complexity inAgardhiella subulata (Gigartinales, Rhodophyta). Results indicate the presence of three second-order components corresponding to fast (22%), intermediate (68%) and slow (10%) fractions. Thus, the genome consists of 90% repetitive sequences. Microspectrophotoometry with the DNA-localizing fluorochrome DAPI was used to confirm ploidy level differences in the gametophytic and tetrasporophytic phases. Results indicate that meiosis occurs during tetrasporogenesis. Comparison of mean nuclear DNA (I_f) values to chicken erythrocytes (RBC) resulted in an estimate of 0.9 pg/2C genome forAgardhiella. Karyological studies using aceto-orcein revealed a chromosome complement of 2N = 44 in carposporangia and the presence of 22 bivalents during diakinesis of tetraspore mother cells.     

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     

The malmö polymorphism of coagulation factor IX, an immunologic polymorphism due to dimorphism of residue 148 that is in linkage disequilibrium with two other F.IX polymorphisms

A mouse monoclonal antibody (MAB 9.9) to coagulation factor IX (F.IX) detects a polymorphism in the plasma of normal people. Its epitope has been narrowed down to <6 amino acids in the activation peptide of the X-linked F.IX protein. The activation peptide contains a dimorphism—Thr:Ala—at position 148 of the protein. Using synthetic oligonucleotides, we have demonstrated that (1) the F.IX which reacts with 9.9 has Thr at position 148 and (2) that which does not has Ala. Positive reactors (148^thr) are designated Malmö A, and negative reactors (148^ala) are designated Malmö B. The plasma levels of AA women are indistinguishable from those of A men, and both B men and BB women are null against MAB 9.9. The plasma level of Malmö A in AB women is approximately half that of AA women, and “lyonization” is clearly operating in the heterozygotes. The dimorphism is in strong linkage disequilibrium with two other intragenic RFLPs, TaqI and XmnI. Furthermore, intragenic crossing-over—including double crossing-over—appears to have occurred between the three sites. Seven of the eight possible haplotypes have been identified, five in men and two others in women. The immunoassay that identifies ～50% of the AB women in the pool of Malmö A females with 95% confidence identifies men unambiguously as A or B. The assay would be very useful for population-genetic studies of the Malmö epitope if the studies were limited to men.