Contribution of synapses in the medial supramammillary nucleus to the frequency of hippocampal theta rhythm in freely moving rats.

We have previously shown that in urethane-anesthetized rats the frequency of rhythmical slow activity in the hippocampus ("theta") is controlled by the medial supramammillary nucleus (SuM). In particular, injections of procaine into SuM in urethane-anesthetized animals reduce the frequency of theta. However, it has been reported that, in freely moving animals, lesions of SuM do not affect theta. The present experiments were designed to resolve this anomaly. Injections of procaine or chlordiazepoxide into SuM in urethane-anesthetized animals reduced the frequency of theta elicited by reticular stimulation. Mapping showed that procaine injections in freely moving animals were effective in the same locations as under urethane anesthesia. Injections of chlordiazepoxide were effective in a more restricted area than procaine, consistent with an action on synapses in SuM and sparing fibers afferent to SuM. Analysis of the functional spread indicated an effective radius of diffusion of the drugs of 500 microns. With optimal placements, this implied an action on at least 80% of SuM. However, in contrast to the results under urethane, the maximal frequency reductions obtained were less than 50% of the theoretical maximum. In a number of animals receiving repeated injections into SuM, lesions developed which encompassed the whole of SuM. As previously reported, theta was largely intact in SuM-lesioned animals. However, the frequency of theta produced by reticular stimulation was reduced after lesion by approximately the same amount as by procaine injections before lesion. These results suggest that in freely moving animals SuM is only one of two or more nuclei which jointly control the frequency of reticular-elicited theta.

Developmental exposure of mice to pulsed ultrasound.

Exposure of pregnant mice on gestation day (gd) 8 to 1 MHz continuous-wave ultrasound (0, 0.05, 0.50, or 1.00 W/cm2) was reported previously to result in a slight (nonsignificant) increase in malformations. The present study was conducted in a similar fashion using pulsed ultrasound but was designed to maximize the likelihood of finding effects of gd 8 ultrasound exposure on prenatal development. Pregnant ICR mice (approximately 60 animals/group) were exposed on gd 8 to pulsed ultrasound with a center frequency of 1 MHz at levels of 0 (sham control), 0.05, 0.50, or 1.00 W/cm2 (spatial average, temporal average intensities; ISATA) with a spatial peak, pulse average intensity (ISPPA) of 90 W/cm2 and pulse duration of 6.5 microseconds. Anesthetized animals were placed in a degassed water bath (30 degrees C) and exposed for two 10 min intervals during which the beam was centered 1 cm on either side of the abdominal midline. On gd 17, dams were killed; the uterus and its contents were weighed and examined; and live fetuses were weighed and examined for external, visceral, and skeletal malformations. Although one female in the 0.50 W/cm2 group and seven animals in the 1.00 W/cm2 group died following exposure, no other significant change from controls was seen in any maternal or fetal parameter evaluated. Thus the results of this study indicate that there was no detectable effect on prenatal development of mice following exposure to ultrasound on gd 8 (a time of maximal sensitivity), even at exposure intensities that were lethal to some maternal animals.

Cytotoxicity and mutagenicity of ophthalmic solution preservatives and UVA radiation in L5178Y cells.

Four chemical preservatives commonly used in ophthalmic solutions were tested for their toxic and mutagenic potential in mouse lymphoma cells with and without exposure of the cells to ultraviolet A (UVA) radiation. The preservatives tested were benzalkonium chloride (BAK), chlorhexidine, thimerosal and ethylenediaminetetraacetic acid (EDTA). Cell survival and mutagenesis were measured using the L5178Y mouse lymphoma (TK +/-) system. Cells were exposed to varying amounts of preservatives for 1 h at 37 degrees C, and then aliquots were irradiated with UVA radiation (during the exposure to preservative). Cells were then assayed for survival, and for mutagenesis at the thymidine kinase (TK) locus. In concentrations commonly found in ophthalmic solutions, BAK, chlorhexidine, and thimerosal were toxic to cells, and thimerosal was slightly mutagenic. When cells were exposed to preservative and UVA radiation, chlorhexidine was mutagenic and the mutagenic activity of thimerosal was enhanced.

Behavioral effects of erythrosine following light exposure.

Three studies were conducted to examine the effects of erythrosine on the activity level of mice in a figure 8 maze. Results of the first study show that activity in a dark maze is not influenced by intraperitoneal doses as high as 1.25 mg/kg. In two additional studies, subjects were subjected to combinations of dye and pre-exposure to blue (Experiment II) or blue and green (Experiment III) light. Light exposure consistently produced increased activity levels. However, there was little evidence of a dye-light interaction effect.