The analyses of antigen and DNA polymorphism within the bovine major histocompatibility complex: 1. The class I antigens

Summary. Serology, isoelectric focusing (IEF) of expressed antigens, and restriction fragment length polymorphisms (RFLP) were compared for the identification of BoLA class I haplotypes. Expressed antigens identified as bands by IEF correlated well with serological definition confirming and extending our earlier findings (Joosten et al. 1988). Comparison of serology and isoelectric focusing bands with restriction fragments was more complicated; fragments were found which correlated both with broadly reacting and antigen specific sera. We also found correlation of fragments with two or more sera which showed no cross-reactivity. Fragments unique to particular haplotypes were also observed.
Serology remains the simplest method of typing BoLA class I antigens. Isoelectric focusing generally agrees with serological definition of antigens and detects antigens not yet defined by serology. It may also be useful in defining the products of other expressed BoLA class I loci. In order to identify RFLPs which could be used for typing, comparison with serology or IEF is essential. Haplotype specific RFLPs could be useful in identifying genes linked to the MHC.     

In vitro Capacitation of Ejaculated Rabbit Spermatozoa

Prior to fertilization mammalian spermatozoa undergo physiological changes in the female reproductive tract. These changes are collectively known as capacitation. In essence capacitation is a further differentiation that the sperm cell must acquire beyond the maturational changes that it undergoes in the epididymis. Although capacitation in vitro has been easily achieved in rodents, its accomplishment in the rabbit is inconsistent and difficult. We report here successful in vitro capacitation of ejaculated rabbit sperm, assessed by in vitro and in vivo fertilization of rabbit ova. Sperm were used from pooled ejaculates collected from bucks of proven fertility since sperm collected from individual bucks resulted in significant differences in fertilization levels. Conditions favoring in vitro capacitation were: (1) extended incubation time of 12 h, (2) addition of 20% heated rabbit serum to incubation medium, and (3) an atmosphere of 5% CO2, 8% O2, and 87% N2 during incubation. In vitro capacitation of sperm under these conditions resulted in 67% fertilization compared to 89% for control sperm capacitated for 13 h in the uterus.     

Effects of Temperature, Light, Nutrients and Carbon Dioxide on the Strength of the Self-Incompatibility Response in Detached Flowers of Lycopersicon peruvianum

The effects of temperature, light, nutrients and CO₂on the gametophyticself-incompatibility response in a clone of Lycopersicon peruvianumhave been quantified using fluorescence microscopy of stylesquashes prepared from detached flowers held under experimentalconditions for 48 h after self pollination. Self-incompatibilitywas significantly weakened by raising the temperature from 15°C to 22 or 25°C, and by incubating flowers withoutan energy source - i.e. in the dark and/or without externallysupplied sucrose. The sucrose effect was not duplicated by equimolarmannitol or a full mineral nutrient solution plus vitamins.CO₂ applied at 5% in air for 8 h after pollination had no detectableeffect on pollen tube arrest. Observed weakening effects werenot sufficient for use in production of selfed seed. They indicate,however, that self-incompatibility in this clone of L. peruvianumis a temperature-sensitive, energy-dependent process, and supportthe oppositional model of self-incompatibility in which incompatiblepollen tubes are actively inhibited. Lycopersicon, Solanaceae, carbon dioxide, light, nutrients, self-incompatibility, temperature     

Inter-strain Variability of Structural Proteins in Tetrahymena*

SYNOPSIS The high molecular weight proteins found in isolated pellicles of Tetrahymena have been compared in several individual strains within the genus using SDS polyacrylamide gel electrophoresis. Three variants of the B-protein of epiplasm (MW 174,000; 155,000; 145,000) and 2 of the C-protein (MW 140,000; 122,000) were found among the strains examined. No variation was observed in the major kinetodesmal fiber protein (MW 250,000). The variation found between strains in the proteins of a structure which is (as far as we know) the same in all strains indicates a disjunction between evolutionary change at the 2 levels of organization. The taxonomic implications of the observed variation in structural proteins in Tetrahymena are discussed.