Axonal sub-populations in the central nervous system demonstrated using monoclonal antibodies against alpha-tubulin

Using two monoclonal antibodies that specifically recognise alpha-tubulin we describe differences in their light and electron microscopic immunoperoxidase staining of axons in cerebellum, hippocampus, and cerebral cortex. In the molecular layer of the cerebellar cortex at the light microscopic level, one of the antibodies (YOL/34) labelled parallel fibre axons (in an identical manner to a beta-tubulin monoclonal antibody) while the other antibody (YL1/2) failed to label them. Extending these studies to the electron microscopic level in the cerebellum we have determined the sub-cellular localisation of alpha-tubulin in microtubules and the postsynaptic density, and also demonstrated a sub-population of parallel fibres and myelinated axons labelled with antibody YL1/2. The monoclonal antibodies were further characterised using immunoblotting against alpha-tubulin separated by isoelectric focusing gels. The results suggest that the contrasting staining patterns between the alpha-tubulin antibodies may reflect axonal sub-populations containing different isotypes of alpha-tubulin.     

Specific-locus mutation rates in the mouse following inhalation of ethylene oxide, and application of the results to estimation of human genetic risk

Male (101 × C3H)F₁ mice were exposed in an inhalation chamber to ethylene oxide (EtO) in air at a concentration of (generally) 255 ppm. After accumulating total exposures of 101 000 or 150 000 ppm.h in 16–23 weeks, the males were mated to T-stock females for a standard specific-locus mutation-rate study in which 71 387 offspring were observed. The spermatogonial stem-cell mutation rate at each exposure level, as well as the combined result, does not differ significantly from the historical control frequency. At the lower and higher exposure levels, the results rule out (at the 5% significance level) an induced frequency that is, respectively, 0.97 and 6.33 times the spontaneous rate; the combined results rule out a multiple of 1.64.

The relationship between mouse spermatogonial stem-cell mutation rates and EtO-induced testis ethylations was compared with the relationship between Drosophila post-stem-cell mutation rates and sperm ethylations (Lee, 1980). The comparison does not rule out equal mutability per ethylation; but it cannot prove parallelism. An assessment of the mouse-Drosophila relationship will require a more efficient alkylator than EtO and the use of comparable germ-cell stages.

More meaningful conclusions may be drawn by utilizing the data for direct estimation of human risk by expressing the induced mutation frequency that is ruled out (at the 5% significance level) as a multiple of control rate and extrapolating to human exposure levels. The probable absence of major stem-cell killing (and thus, possibly, cell selection) by EtO indicates that such extrapolation probably does not produce an underestimate. For a human exposure concentration of 0.1 ppm on working days during the reproductive lifespan, the mouse experimental results rule out (at the 5% significance level) an induced spermatogonial stem-cell gene mutation rate greater than 8% of the spontaneous rate; for 1.0 ppm, they rule out an induced rate roughly equal to the spontaneous rate. The induced rate for any one poststem-cell stage would have to be about 3 orders of magnitude higher than that for stem cells to constitute an equivalent risk.     

Interactions between dietary protein,ovulation rate and follicle stimulating hormone level in the ewe

Ovulation rate (OR) was studied in two experiments using mature Border Leicester × Merino ewes in which oestrous cycles were synchronized using a prostaglandin analogue. In both experiments a basal ration of 500 g of lucerne/barley pellets was provided. In the first experiment, ewes were fed individually iso-energy supplements of 500 g of either peas, lupins, soybean pellets or lucerne/barley pellets. The ovulation studied at laparoscopy occurred approximately 34 days after starting the supplementary feeding. Ewes fed lupins or soybean pellets had higher (P < 0.05) OR's than the ewes fed the other diets.In the second experiment, ewes were fed either iso-protein supplements of peas or lupins or casein supplement (170 or 100 g of protein) either formalin treated or untreated. The ovulations studied at laparoscopy occurred approximately either 17 or 34 days after the first feeding of the supplement. Follicle stimulating hormone (FSH) levels in plasma were measured over the 8 days prior to the second ovulation. There were no differences (P < 0.05) in OR's at the first ovulation. However, by the second, ewes fed peas had the highest (P < 0.05) OR while those fed lupins or protected casein had similar OR's. These tended to be higher than in the ewes fed untreated casein. FSH levels were generally higher from 8 days to 3 days prior to ovulation in ewes which were to have twin ovulations compared to those having single ovulations.The results confirm that feeding high energy or high protein will increase OR. There are independent effects of energy and protein. The results suggested that the ovulation rate may be related to FSH levels.     

Material Legacies and Environmental Constraints Underlie Fire Resilience of a Dominant Boreal Forest Type

Ecosystems - Resilience of plant communities to disturbance is supported by multiple mechanisms, including ecological legacies affecting propagule availability, species’ environmental...