Perfusion method for preparing pig brain cortex for Golgi-Cox impregnation

The perfusion procedure described in this paper produces high quality impregnation of pig visual and somatosensory cortical neurons with a Golgi-Cox solution. Starting within 30 min after death, pig heads were perfused with a fixative solution composed of a mixture (v/v) of liquid phenol, 5%; formalin, 14%; ethylene glycol, 25%; methanol, 28%; and water, 28% for two periods of 4 hr each. After perfusion, the heads were chilled for at least 18 hr. The entire brain was removed from the skull and then placed in 10% buffered formalin, where it remained for at least 10 days before taking the blocks that were to be immersed in the Golgi-Cox solution. Three weeks spent in the Golgi-Cox solution typically produced uniform neuron impregnation. The tissue blocks were then embedded in celloidin and sectioned at 120 micron. This procedure avoids the following difficulties: Golgi-Cox methods that produced excellent results with rodent or primate tissue were unsuccessful with pig tissue, placing fresh tissue in Golgi-Cox solution resulted in incomplete neuron impregnation, and immersion fixation in 10% buffered formalin without perfusion resulted in excessive staining of glia.     

Characterization of circadian oscillations in alanine dehydrogenase activity in non-dividing populations of Euglena gracilis (z)

Possible factors that could generate the circadian oscillations in alanine dehydrogenase (EC 1.4.1.1.) activity observed in cultures of non-dividing Euglena gracilis (Z) have been examined in an effort to learn more about the basic timekeeping mechanism of biological clocks. No differences in K_m, pH optimum or electrophoretic mobility could be demonstrated between enzyme extracted from the minimum part of the 24-h oscillation in activity and that extracted from the maximum part. Also, no evidence for the presence of activators or inhibitors was found in mixing experiments. The effect of cycloheximide on the rhythm was examined; it was shown that the oscillation ceases in the presence of the inhibitor, but that if the inhibitor is removed after 12 h, the rhythm resumes with no apparent change in phase. Analyses of gel scans of enzyme preparations partially purified by (NH₄)₂SO₄ fractionation and polyacrylamide gel electrophoresis indicated that there was more alanine dehydrogenase protein present at the maximum part of the cycle than there was at the minimum part. In view of these and other data, an operational model of a circadian biological clock is discussed.     

Effects of pralidoxime applied locally into the midbrain reticular formation on the hippocampal theta rhythm induced by acetylcholinesterase inhibition

After local administration into the midbrain reticular formation of an acetylcholinesterase reactivator--Pralidoxime, a significant decrease of intensity of hippocampal theta rhythm induced by previous inhibition of acetylcholinesterase by DFP was observed already after 10 min. This result suggests that cholinergic structures are localized in midbrain reticular formation and that they play a role in the origin of hippocampal theta rhythm.     

Effects of temperature and infection with Eimeria ochrogasteri on digestive organs of the prairie vole, Microtus ochrogaster

Prairie voles, Microtus ochrogaster, were infected with Eimeria ochrogasteri and exposed to 2 environmental temperatures, 5 and 22 C. Dry weights of the small and large intestines increased by 33% and 19%, respectively, in infected animals. Infected animals also exhibited a 14% decrease in cecal length compared to uninfected animals. The interaction between temperature and infection affected the length of the small intestine. Infected animals maintained at 5 C had longer small intestines than both infected animals housed at 22 C, and uninfected animals at 22 or 5 C. Furthermore, the dry weight of the small intestine was affected by a 3-way interaction (infection, temperature, and sex). Temperature affected stomach and liver dry weights, as well as lengths of the small intestine and cecum. Stomach and liver dry weights, as well as small intestine lengths, were greater in those animals held at 5 C, whereas cecum lengths decreased. Prepatency, patency, and total oocyst production were not affected by temperature; however, infected animals held at 5 C exhibited diarrhea during the patent period.