Postmortem elevation in extracellular glutamate in the rat hippocampus when brain temperature is maintained at physiological levels: implications for the use of human brain autopsy tissues.

Postmortem alterations in the neuronal cytoskeleton resemble some aspects of the cytoskeletal disruption associated with neurodegenerative disorders, and are also similar to those observed following ischemia and produced by excitotoxins in vivo and in vitro. This suggests the involvement of excitotoxic mechanisms during the postmortem interval. The purpose of this study was to determine if extracellular levels of glutamate are elevated postmortem. Extracellular levels of GABA and taurine were also monitored using in vivo microdialysis. These three amino acids were analyzed using high-performance liquid chromatography. When postmortem rat brain temperature cooled rapidly to near room temperature, dialysate concentrations of glutamate were not increased in the hippocampal CA1 region during a 2-h postmortem interval, although increased extracellular levels of GABA and taurine were observed. In contrast, maintenance of brain temperature at 37 degrees C resulted in a 12-to-40 fold elevation in extracellular glutamate levels 20-120 min postmortem. In addition, the elevation in dialysate taurine concentration was greater than that observed in rats in which postmortem brain temperature was not maintained. Excitatory amino acid antagonists, NBQX (2, 3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline) and MK-801 (dizocilpine, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cylohepten-5, 10-imine hydrogen maleate blocked the additional elevation in taurine associated with maintaining brain at 37 degrees C, but had less robust effects against glutamate and GABA release. The results indicate that extracellular concentrations of glutamate, taurine and GABA increase in postmortem rat brain when physiologic temperatures are maintained, but that these increases are blunted when brain temperature decreases. After death, the human brain cools much more slowly than does the rat brain. Therefore, extracellular glutamate levels are likely to increase in the postmortem human brain and may contribute to excitotoxic neuronal damage occurring in the interval between death and autopsy.

Correlation between suckling-induced changes in the ultrastructure of mammotrophs and prolactin release.

Effects of suckling on the structure of mammotrophs and the release of prolactin, were studied in rats on the 10th day of lactation with the use of electron microscopy and radioimmunoassay techniques. Nursing animals were separated from their young for 8 hr and subsequently united and permitted to nurse for 1, 5, 15, 30 min; or 1, 2 and 4 hr. Blood samples were obtained prior to and throughout the suckling interval and pituitary glands were processed for electron microscopy. Control animals consisted of normal lactating females and animals separated from their young for 8 hr. Normally lactating controls had high prolactin serum levels (501 +/- 95 ng/ml) and synthetically active appearing mammotrophs. An 8 hr separation from the pups induced a dramatic lowering of serum prolactin (32 +/- 5 ng/ml), an increase in secretory granule storage, and a great dilation of rough endoplasmic reticulum (RER) cisternae. Five min of renewed suckling resulted in a rise of plasma prolactin levels (605 +/- 183 ng/ml) which remained high thereafter. The major ultrastructural changes observed during the first 30 min of suckling were as follows: 1) at 1 min, the RER became cmone?); 2) AT 5 MIN, AND MUCH MORE OBVIOUSLY AT 15 AND 30 MIn, a massive discharge of secretory granules was observed; and 3) at 15 min, the collapsed RER underwent transformation for 1,2 and 4 hr) induced new hormone synthesis as suggested by the presence of hypertrophied Golgi elements and numerous immature granules. This was accompanied by a new transformation of the RER from the vesicular into a lamellar form now consisting of very slender cisternae lined with numerous ribosomes, presumably involved in the renewal of the synthetic process. The morphologic findings described correlate well with the time table of prolactin release. In addition, the dramatic early changes in the structure of the RER suggest a possible involvement of this organelle in the storage and release of a proposed rapidly releasable pool of prolactin.

The interrelation of LH-RH, ovarian steroids and the adenohypophysis.

In this presentation experimental evidence is reviewed and new information is presented concerning the question of cyclic fluctuations in pituitary responsiveness to endogenous and exogenous hypothalamic-releasing hormones as well as the direct role of steroids in modulating pituitary sensitivity. Experiments are described which show that four-day and five-day cyclic rats respond differently to LH-RH, presumably because of differences in secretion patterns of ovarian steroids. The direct role of these steroids in this process was investigated both from the morphological (light and electron microscopy) and physiological (radioimmunoassay measurement of LH) viewpoint in hypophysectomized rats bearing pituitary homografts. It was found that steroids may directly alter both the morphology and the secretory function of pituitary gonadotrophs. Estradiol was found to depress the amount of LH that was secreted while progesterone was seen to influence primarily the time of LH release following LH-RH injection. Thus, it appears that direct feedback effects of ovarian steroids at the level of the pituitary are important regulators of the sensitivity of pituitary gonadotrophs to the LH- releaseing action of hypothalamic LH-RH.