Light-scattering studies of bull spermatozoa. II. Interaction and concentration effects.

The complete autocorrelation function of the intensity fluctuations of laser light scattered from motile bull spermatozoa is shown to depend upon several factors not previously considered. Samples of bull spermatozoa generally contain a substantial proportion of dead cells, which give rise to slowly decaying components of the autocorrelation function. Whereas previous work has concentrated on the form of the fast decaying autocorrelation component, we are concerned here with the relative amplitude and shape of the slow autocorrelation component and the general form of the composite function. In principle, the relative amplitudes of the fast and slow components of the autocorrelation function can be used as an assay of the proportion of swimming cells. We show that this amplitude ratio depends upon cell concentration, scattering cell geometry, and scattering angle. A simple model is developed to explain these results on the basis of the asymmetry of light scattered from these cells, motile/immotile cell interactions, wall-swimming effects, and geotactic reorientation of dead cells.     

The Scyphomedusae of the Rockall Trough, northeastern Atlantic Ocean

An extensive series of samples between the surface and 2700m depth have been taken in the Rockall Trough between 1973 and1978. The coronate medusae, in decreasing order of abundanceoccurring in the Trough are: Periphylla periphylla, Atolla wyvillei,A. parva, A. vanhoeffeni and Nausitho? globifera. The last specieswas rare and no information on its biology was obtained. Theother species breed within the Trough, the three Atolla speciesthroughout the year. Few large mature Periphylla periphyllawere caught and its breeding season remains undefined. Theirmorphology and vertical and geographical distributions are brieflydiscussed. The semaeostomid medusae, Pelagia noctiluca, Aureliaaurita and Cyanea lamarckii also occurred, the latter two speciesconfined to the Rockall Bank.     

Altered processing of integrin receptors during keratinocyte activation

We used monoclonal antibodies against specific integrin subunits to examine the role of integrin receptors in keratinocyte activation. We found that before activation, beta 1 subunits in keratinocytes showed a diffuse distribution, whereas after activation, keratinocytes organized beta 1 receptors into marginal adhesion plaques. In immunoprecipitation experiments with antibodies against beta 1 integrin subunits, we found mostly immature subunits synthesized in keratinocytes freshly harvested from skin. Moreover, integrin receptor complexes immunoprecipitated from these cells by monoclonal antibodies against alpha 2, alpha 3, or alpha 5 subunits contained only immature beta 1 subunits. With keratinocytes cultured 4-7 days, anti-beta 1 antibodies immunoprecipitated mostly mature beta 1 subunits, and integrin complexes immunoprecipitated from cultured cells by anti-alpha subunit antibodies contained mostly mature beta 1 subunits. Antibodies directed against beta 1 subunits also inhibited keratinocyte migration. Based on these results, we suggest that up-regulation of migration by activated keratinocytes depends on changes in processing of pre-beta 1 subunits to mature beta 1 subunits. We also studied the distribution of integrin subunits in skin and on keratinocytes migrating out of skin explants. Whereas beta 1, alpha 2, and alpha 3 subunits were detected in keratinocytes in skin and migrating out of explants, alpha 5 subunits were observed only in migrating cells.     

The snRNP E protein multigene family contains five pseudogenes with common mutations.

Sequence data from three previously-uncharacterized members of the snRNP E protein multigene family suggest that each is a non-transcribed processed pseudogene, even though one clone has the potential to code for a full-length protein with greater than 90% similarity to previously-characterized E protein cDNAs. Each of the newly-analyzed family members is without introns, contains a tract of polyadenylic acid residues, and is flanked by short direct repeats. In addition, the three sequences all contain point mutations that distinguish them from the E protein coding sequence. Seven point mutations are common to the three sequences described here and to two previously-described E protein pseudogenes. Although all of these mutations are transitions, only 5 of 7 could have been generated by deamination of methylated cytosines in inactive genes. Thus, the common mutations in the pseudogenes suggest an origin other than the expressed gene that we have described. Allelic variants for two of the pseudogenes were detected and repetitive elements are located near four of the five E protein pseudogenes that have been characterized.     

The rapid purification of 3-hydroxybutyrate dehydrogenase and malate dehydrogenase on triazine dye affinity matrices.

3-Hydroxybutyrate dehydrogenase (EC 1.1.1.30) and malate dehydrogenase (EC 1.1.1.37) were purified to homogeneity on a large scale involving only two sequential affinity-chromatography steps on two triazine dye-Sepharose matrices. Recoveries of both enzymes were in excess of 60%. Malate dehydrogenase could also be purified by a combination of triazine dye affinity chromatography and gel filtration on Ultrogel AcA-44, but this offered no significant advantage over the purely affinity procedure.     

The relationship between the size of mitochondria and the intensity of light that they scatter in different energetic states

The intensity of light scattered at 90° to the incident beam and the effective hydrodynamic radii of mitochondria incubated under a variety of conditions have been measured. Addition of high concentrations of uncouplers to respiring mitochondria resulted in a decrease in scatter which was not due to swelling. Addition of valinomycin to mitochondria depleted of substrate in K⁺-free medium produced an increase in scatter that was not due to shrinking. It is concluded that changes in the intensity of scattered light are not reliable indices of changes of volume of mitochondria, and that changes in conformation with changes in metabolic state dominate changes in light scatter. A molecular mechanism for the effect of metabolic state upon the scattered intensity is suggested.