Identification of alpha-tubulin as an hsp105alpha-binding protein by the yeast two-hybrid system

Hsp105alpha is a mammalian stress protein that belongs to the HSP105/110 family. Hsp105alpha prevents stress-induced apoptosis in neuronal cells and binds to Hsp70/Hsc70 and suppresses the Hsp70 chaperone activity in vitro. In this study, to further elucidate the function of Hsp105alpha, we searched for Hsp105alpha-binding proteins by screening a mouse FM3A cell cDNA library with full-length Hsp105alpha using the yeast two-hybrid system and obtained alpha-tubulin as an Hsp105alpha-binding protein. Hsp105alpha bound directly to alpha-tubulin both in vitro and in vivo. Indirect immunofluorescence analysis with anti-Hsp105 and anti-alpha-tubulin antibodies indicated that Hsp105alpha was colocalized with microtubules. Furthermore, the disorganization of microtubules induced by heat shock was prevented in Hsp105alpha-overexpressing COS-7 cells. These findings suggested that Hsp105alpha associates with alpha-tubulin and microtubules in cells and plays a role in protection of microtubules under conditions of stress.     

Circadian communication between unicells?

Populations ofGonyaulax polyedra, in two different phases, about 11 h apart, were mixed, and the intensity of their spontaneous bioluminescence glow recorded for about 2 wk under conditions of constant dim (35±3 μE/m²/s) white light and constant temperature (19.0±0.3°C). The phases and amplitudes of glow signals recorded from mixed cultures were compared with those obtained from the arithmetic sum of the intensity data from two control vials. Peaks in control cultures generally remained separate, but there was a spontaneous increase in the period beginning 6–11 d after the onset of constant conditions. This did not occur in cultures in which the medium was exchanged with fresh medium every 2 d. In the actual mixes of two cultures there was a merging of the two subpeaks in the signal, which did not occur when the medium was exchanged. The results indicate that conditioning of the medium by cells may affect the period of the circadian rhythm and that this might result in a type of communication. Supported by the Deutsche Forschungsgemeinschaft; present address     

Evaluation of adriamycin nephropathy by an in vivo electron paramagnetic resonance

A rat model for human minimal change nephropathy was obtained by the intravenous injection of adriamycin (ADR) at 5 mg/kg. By using an in vivo electron paramagnetic resonance (EPR) spectrometer operating at 700 MHz, the temporal changes in signal intensities of a nitroxide radical, 4-hydroxyl-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL), in the kidneys of rats with ADR nephropathy were investigated. The decay rate of the EPR signal intensity obtained in the kidney is indicative of the renal reducing ability. It was found that the reducing ability in the kidney declined on the 7th day after ADR administration and recovered after the 14th day. Impairment of the reducing ability occurred before the appearance of continuous urinary protein. The in vitro EPR study showed that this impairment of in vivo renal reducing ability is related to impairment of the reducing ability in the mitochondria.     

Porcine brain natriuretic peptide-like immunoreactivity in rat tissues

The presence of immunoreactive porcine brain natriuretic peptide in rat tissues was studied with a specific radioimmunoassay for porcine brain natriuretic peptide-26. The cross-reactivity of the antiserum used was less than 0.001% with rat atrial natriuretic peptide, rat brain natriuretic peptide-32 and rat brain natriuretic peptide-45. Immunoreactive porcine brain natriuretic peptide was detectable in various tissues of the rat, and high concentrations of immunoreactive porcine brain natriuretic peptide were found in the brain and cardiac atrium, with the highest level in the hypothalamus (159±30 fmol/gram wet tissue, mean±SEM, n=4). Reverse phase high performance liquid chromatography showed that the immunoreactive porcine brain natriuretic peptide of the whole brain and heart extracts eluted mainly at an identical position to synthetic porcine brain natriuretic peptide-26. These findings indicate that porcine brain natriuretic peptide-like substance, distinct from rat brain natriuretic peptide, is present in high concentrations in the rat brain and cardiac atrium.     

Notes on some plant-inhabiting fungi collected from the Nansei Islands (2)

In this paper, four fungi collected in Nansei Islands are reported. One is Strasseria garciniae, which must be transferred to the genus Phyllosticta. This fungus was collected in Okinawa Main Island and Iriomote Island. The other three fungi are newly added to the Japanese fungal flora. Phyllosticta ghaesembillae on Codiaeum variegatum and Cercospora asplenii on Asplenium antiquum were collected in Yoron Island. The last one, Coniella australiensis on Eucalyptus robusta was collected in Okinawa Main Island. Their morphology and symptoms on the host plant are described, with some mycological notes.     

Distribution of the electrophoretic variants of serum alpha1-antitrypsin in six species of the macaques

Individual variations of ₁-antitrypsin of the macaques were investigated by means of starch gel electrophoresis. The material comprised a total of 1,084 plasma samples taken from six species, namely,Macaca irus, mulatta, cyclopis, nemestrina, speciosa, andfuscata, including several geographical groups. At least ten phenotypes which were assumed in analogy to human Pi-system to be under genetic control of five codominant alleles tentatively denoted byPi _Mac ^{A, B, C, D, E} were identified. It was considered that these alleles are commonly possessed by different macaque species. A marked difference in the distribution of allele frequencies was found both within and between species groups. Several aspects of this new polymorphic variation in the macaques were discussed with special reference to the geographical distribution of the alleles and the origins of the Japanese macaque,M. fuscata. This study was carried out as part of the Special Project of the Japan-U.S. Cooperative Scientific Program: Blood Macromolecules and the Genetic Origins of the Japanese Macaques (Chief investigators:T. Miki andM. Goodman)     

Vasotocin/isotocin-immunoreactive neurons in the medaka fish brain are sexually dimorphic and their numbers decrease after spawning in the female

In teleosts, the distribution of neurons in the preoptic-hypothalamic region and their associated neurohypophysial hormones, such as vasotocin (VT), appears to be different among species. This differential distribution is thought to reflect the social and/or sexual status of individuals within a species. In the present study, we analyzed the number, size and the distribution of vasotocin/isotocin (VT/IT) neurons in the brains of both male and female medaka (Oryzias latipes) using immunohistochemistry. VT/IT neurons were similarly located in an inverted L-shape in the nucleus preopticus in both gender, as has been already reported in salmonids. However, computer-assisted image analysis revealed sexual dimorphism in the number of VT/IT-immunoreactive (ir) neurons, with greater numbers found in males as compared to females. Further, in the female brain, the number of VT/IT-ir neurons decreased significantly after spawning. In pre-spawning compared to post-spawning females, the small-sized VT/IT-ir neurons dominated. Sexual differentiation of the medaka is fully dependent upon the steroid status during the early developmental stages and steroids are also known to trigger gender-specific behavior in the adult medaka. Our findings strongly suggest that VT and/or IT neurons may be functionally related to ovulation and/or the reproductive axes through connections to their steroidal status.