Characterization of the extent of pontomesencephalic cholinergic neurons' projections to the thalamus: comparison with projections to midbrain dopaminergic groups.

We sought to determine whether pontomesencephalic cholinergic neurons which we have been shown previously to project to the substantia nigra and ventral tegmental area also contribute to the thalamic activation projection from the pedunculopontine and laterodorsal tegmental nuclei. Retrograde tracing, immunohistochemical localization of choline acetyltransferase and statistical methods were used to determine the full extent of the cholinergic projection from the pedunculopontine and laterodorsal tegmental nuclei to the thalamus. Progressively larger Fluoro-Gold injections in to the thalamus proportionally labeled increasing numbers of pontomesencephalic cholinergic cells both ipsi- and contralaterally in the pedunculopontine and laterodorsal tegmental nuclei. Multiple large thalamic injections left only a small fraction of the ipsilateral pontomesencephalic cholinergic group unlabeled. This small remainder did not correspond to the populations which project to the substantia nigra and ventral tegmental area, thereby indicating that substantia nigra- and ventral tegmental area-projecting cholinergic neurons must also project to the thalamus. We examined whether there existed any set of cholinergic neurons in the pedunculopontine and laterodorsal tegmental nuclei which did not innervate a thalamic target. The distribution of descending projections of the pedunculopontine and laterodorsal tegmental nuclei demonstrated that the unlabeled remainder cannot correspond to a purely descending group. We also show that substance P-positive cholinergic cells in the laterodorsal tegmental nucleus project to the thalamus. Further studies demonstrated that the small population of cholinergic cells left unlabeled from the thalamus were the smallest sized cholinergic cells, and included two groups of small, light-staining cholinergic cells located in the parabrachial area and central gray, adjacent to the main pedunculopontine and laterodorsal tegmental nuclei cholinergic groups. These small cells, in contrast to thalamic-projecting cholinergic cells, did not stain positively for reduced nicotinamide adenine dinucleotide phosphate-diaphorase. Taken together, these results indicated that all of the reduced nicotinamide adenine dinucleotide phosphate diaphorase-positive/choline acetyltransferase-positive neurons of the pedunculopontine/laterodorsal tegmental nuclei ascend to innervate some portion of the thalamus, in addition to the other targets they innervate. These findings indicate that the diverse physiological and behavioral effects attributed to the activity of pontomesencephalic cholinergic neurons should not be dissociated from their activating effects in the thalamus.

Concentration of chicken gonadotropin-releasing hormones I and II in microdissected areas of turkey hen brain during the reproductive cycle.

Chicken gonadotropin-releasing hormones I and II (cGnRH I and II) were measured by radioimmunoassay (RIA) in extracts of microdissected regions of turkey hen brain (preoptic area [POA], region of periventricular nuclei [PHN], septum [SEP], hippocampus [HP], dorsomedial thalamus/habenula [DMT], midbrain central gray [MCG], and caudal lateral hypothalamus [LH]) at five stages of the reproductive cycle: before photostimulation, during egg laying, during incubation, during photorefractoriness, and after return to short daylengths. The highest concentration of cGnRH I occurred in PHN, followed by POA, SEP, DMT, HP, LH, and MCG, in decreasing order, whereas the highest concentration of cGnRH II occurred in SEP, followed by POA, DMT, HP, MCG, PHN, LH. These results agree, with some exceptions, with the distribution of fibers and cells as determined by immunohistochemistry. cGnRH II was from 1.3 to 24 times as abundant as cGnRH I in different brain areas. During incubation, cGnRH I concentrations were significantly elevated in the POA and cGnRH II levels were significantly elevated in HP; few other significant differences were detected. Correlation analysis detected occasional significant positive and negative correlations between cGnRH I and II concentrations in forebrain areas and MCG of laying birds and in PHN and LH of incubating birds. These results demonstrate an approximate correspondence between hormone concentrations measured in tissue extracts by RIA and immunohistochemistry and indicate an abundance of cGnRH II as compared with cGnRH I. cGnRH I and II concentrations did not, however, change in parallel in all brain areas, suggesting that these peptides do not function in an exactly parallel fashion. Thus, an extent to which cGnRH II is involved in gonadotropin release remains unresolved.

Increase in hypothalamic cholecystokinin following acute and chronic morphine.

Recent evidence supports an antagonistic interaction between cholecystokinin (CCK) and opiate peptides. The present study determined the effects of various levels of morphine treatment on hypothalamic levels of CCK as determined by radioimmunoassay. Acute treatment with morphine sulfate (10 mg/kg) or implantation of one morphine pellet (75 mg free base) increased levels of CCK in whole hypothalamus. Increased exposure to morphine by either chronic injections or implantation of two pellets did not result in a further change in whole hypothalamic CCK levels. In samples dissected into hypothalamic subregions, the effect of morphine on CCK levels was localized to medial but not lateral or posterior regions. These experiments extend earlier in vitro findings and suggest that some of the physiological and behavioral effects of opiate peptides may result from modulation of endogenous CCK.