Brain and erythrocyte microtubules from chicken contain different beta-tubulin polypeptides

beta-Tubulin subunits isolated from chicken brain tissue and erythrocytes are distinguishable as unique biochemical species by electrophoretic and peptide mapping procedures. 1) The subunits of beta-tubulin exhibit major differences in electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gels that vary according to the pH and ionic strength of the gel. 2) The isoelectric points of urea-denatured beta subunits from brain tissue and erythrocytes are pH 5.1 and 5.4, respectively, whereas those of both alpha subunits are approximately pH 5.2.3) Two-dimensional peptide maps prepared with alpha-chymotrypsin or V8 protease show that alpha-tubulin peptides are indistinguishable, whereas beta-tubulin peptides are very different. Only one-third of the 15 major tyrosine-containing beta-tubulin peptides prepared with alpha-chymotrypsin are common to both beta-tubulin species. The data indicate that the beta-tubulin subunits of brain tissue and erythrocytes are biochemically distinct and may be different gene products. The presence of tubulin variants in brain tissue and erythrocytes may indicate special requirements for microtubule assembly and function in different cell types.     

Identity and origin of the ATPase activity associated with neuronal microtubules. II. Identification of a 50,000-dalton polypeptide with ATPase activity similar to F-1 ATPase from mitochondria

We determined that the ATPase activity contained in preparations of neuronal microtubules is associated with a 50,000-dalton polypeptide by four different methods: (a) photoaffinity labeling of the pelletable ATPase fraction with [gamma-32P]-8-azido-ATP; (b) analysis of two- dimensional gels (native gel X SDS slab gel) of an ATPase fraction solubilized by treatment with dichloromethane; (c) ATPase purification by glycerol gradient sedimentation and gel filtration chromatography of a solvent-released ATPase fraction, (d) demonstration of the binding of affinity-purified antibody to the 50-kdalton polypeptide to ATPase activity in vitro. Beginning with preparations of microtubules we have purified the ATPase activity greater than 700-fold and estimate that the purified enzyme has a specific activity of 20 mumol Pi x mg-1 x min- 1 and comprises 80-90% of the total ATPase activity associated with neuronal microtubules. With affinity-purified antibody we also demonstrate cross-reactivity to the 50-kdalton subunits of mitochondrial F-1 ATPase and show that the antibody specifically labels mitochondria in PtK-2 cells. Biochemical comparisons of the enzymes reveal similar but not identical subunit composition and sensitivity to mitochondrial ATPase inhibitors. These studies indicate that the principal ATPase activity associated with microtubules is not contained in high molecular weight proteins such as dynein or MAPs and support the hypothesis that the 50-kdalton ATPase is a membrane protein and may be derived from mitochondria or membrane vesicles with F-1-like ATPase activity.     

Food and feeding habits of the lesser spotted dogfish, Scyliorhinus canicula (L.), in Isle of Man waters

Stomach contents of the lesser spotted dogfish, Scyliorhinus canicula , from Isle of Man waters were investigated. Dogfish fed opportunistically on a wide range of macrobenthic fauna with hermit crabs, cockles and whelks the dominant prey. Items of secondary importance included various crabs, callianassid shrimps, bivalve molluscs, holothurians, polychaetes and, when locally abundant, herring, Clupea harengus . Dogfish predated more heavily on molluscs than reported in previous studies, reflecting the abundance of molluscs in the associated benthic community.
Composition of the diet altered gradually with dogfish size, reliance on small crustaceans declined and consumption of hermit crabs and molluscs increased with growth. Seasonal changes in the composition of the diet were observed. Feeding intensity was greatest during summer, related in part to increased prey availability, and was least during autumn.     

Sexual behavior and reproduction ofCercocebus albigena johnstonii in Kibale forest,Western Uganda

Gray-cheeked mangabeys live in multimale social groups. Two groups of these monkeys were studied extensively over a period of 22 months, and a further eight groups were observed opportunistically. Data on the occurrence of sexual swelling infernales were collected and the different phases of swelling and deflation are described in detail together with data on their duration. The data were found to agree broadly with those presented by other workers on captive animals. Pre and postpartum swellings are described. The majority of copulations was observed to occur at peak swelling and a specific gesture, the head flag, was noted as a solicitation. The data on initiation of copulations show that the males initiated more copulations than the females. However, female-initiated copulations ended in ejaculation more often than male-initiated copulations. The behaviors of the mangabeys during precopulation and copulation were found to be broadly similar to those of the macaque. Copulations were observed most often on the day of peak swelling;however, adult males were the only animals to copulate prior to peak swelling. Subadult male copulations were most often after peak swelling. Masturbation was observed to ejaculation on two occasions. The data show gestation periods of between 184 and 189 days. A mean interbirth interval was calculated to be 33.33 ± 15.87 months.     

Virus Aggregation as the Cause of the Non-neutralizable Persistent Fraction

The non-neutralizable or persistent fraction of virus populations has been found to be caused by aggregated virus. Detailed investigation was performed with the prototype strain of echovirus type 4 (Pesascek), as this virus is notorious for its large non-neutralizable fraction. When Pesascek virus was clarified by low-speed centrifugation, homologous antiserum hardly neutralized the virus. However, when the virus was filtered through membranes having a porosity only twice the diameter of the virus, monodispersed virus was obtained which was efficiently neutralized. Serum titers were up to 1,000 times higher if the neutralization test was carried out with monodispersed virus. Virus in non-neutralizable aggregates was found to constitute 30% of the infective units of unfiltered Pesascek virus but only 0.1% of the antigenically related DuToit strain. This explains why DuToit strain has been a more satisfactory indicator strain for detecting type 4 antibodies, regardless of the echo 4 strain used for inducing the antibodies. Clarified suspensions and ultrafiltrates of viruses belonging to the picorna-, reo-, myxo-, adeno-, herpes-, and poxvirus groups were studied. Clarified suspensions yielded persistent fractions of 0.005% for poliovirus, of 0.1% for reovirus, of 0.6% for influenza virus, of <0.001% for adenovirus, of 0.06% for herpesvirus, and of 10 to 30% for vaccinia virus. In all cases the persistent fractions were removed by membrane filters which had a pore diameter no larger than twice that of the virus under test, and the high concentration of virus in each ultrafiltrate was completely neutralized by antiserum.     

Thermostabilization and thermosensitization of herpesvirus

Wallis, Craig (Baylor University College of Medicine, Houston, Tex.), and Joseph L. Melnick. Thermostabilization and thermosensitization of herpesvirus. J. Bacteriol. 90:1632-1637. 1965.-Herpesvirus, long considered as one of the most thermolabile of viruses, was stabilized by 1 m Na(2)SO(4) or Na(2)HPO(4) so that it withstood heating at 50 C, but the virus was not protected by 1 m MgCl(2), MgSO(4), or KH(2)PO(4), or 2 m KCl or NaCl; 1 m Na(2)SO(4) also stabilized herpesvirus at 25 and 37 C. In contrast, herpesvirus was made extremely thermosensitive in the presence of isotonic salt concentrations or of isotonic tris(hydroxymethyl)aminomethane buffer, especially at pH 7.2 or above. Partially purified virus was relatively thermostable when suspended in distilled water at pH 7.2, but in Earle's salt solution the virus immediately became thermosensitive. As found in tissue culture harvests, herpesvirus was thermolabile, but the virus was rendered stable at 50 C by simple dilution in distilled water. Protection by proteins or amino acids, generally accepted as virus-stabilizing agents, did not seem to be the result of a direct effect upon herpesvirus. The present data suggest that the added proteins counteract in part thermosensitizing effects of the salts contained in the virus harvest.     

SSCP and segregation analysis of the human type X collagen gene (COL10A1) in heritable forms of chondrodysplasia.

Type X collagen is a homotrimeric, short chain, nonfibrillar collagen that is expressed exclusively by hypertrophic chondrocytes at the sites of endochondral ossification. The distribution and pattern of expression of the type X collagen gene (COL10A1) suggests that mutations altering the structure and synthesis of the protein may be responsible for causing heritable forms of chondrodysplasia. We investigated whether mutations within the human COL10A1 gene were responsible for causing the disorders achondroplasia, hypochondroplasia, pseudoachondroplasia, and thanatophoric dysplasia, by analyzing the coding regions of the gene by using PCR and the single-stranded conformational polymorphism technique. By this approach, seven sequence changes were identified within and flanking the coding regions of the gene of the affected persons. We demonstrated that six of these sequence changes were not responsible for causing these forms of chondrodysplasia but were polymorphic in nature. The sequence changes were used to demonstrate discordant segregation between the COL10A1 locus and achondroplasia and pseudoachondroplasia, in nuclear families. This lack of segregation suggests that mutations within or near the COL10A1 locus are not responsible for these disorders. The seventh sequence change resulted in a valine-to-methionine substitution in the carboxyl-terminal domain of the molecule and was identified in only two hypochondroplasic individuals from a single family. Segregation analysis in this family was inconclusive, and the significance of this substitution remains uncertain.     

Isozyme analysis of Galaxias species (Teleostei: Galaxiidae) from the Taieri River, South Island, New Zealand: a species complex revealed

We examined genetic differentiation among 23 samples of non-migratory river galaxias from 17 streams in the Taieri River system, South Island, New Zealand. Four major genetic types were found, two of which occur in narrow sympatry in one location. These were compared with topotypical material representing Galaxias anomalus from the Clutha system (Otago) and G. vulgaris from the Waimakariri system (Canterbury) in order to establish identity. Morphological examination of these four major genetic types revealed consistent concomitant differences. The results suggest that there are at least three species of river galaxias in the Taieri system: G. anomalus, G. vulgaris and at least one previously undescribed species. We propose that the genetic structuring and subsequent speciation of this group has been promoted by the absence of the marine juvenile phase that is found in five other members of the genus native to New Zealand. This structuring may be exacerbated by population fragmentation over the last century owing to the negative influence of introduced trout. The phylogenetic diversity within the river system mirrors the diverse flora and invertebrate fauna of the region, and has conservation implications that parallel those resulting from our improved knowledge of the New Zealand herpetofauna through the application of genetic analysis.