Localization of acidic fibroblast growth factor within the mouse brain using biochemical and immunocytochemical techniques.

The localization of acidic fibroblast growth factor (aFGF) in the male mouse brain was studied with biochemical and immunocytochemical techniques. Using two peptide-based aFGF antisera directed against independent epitopes, Western gel analysis of dissected brain demonstrated significant levels of aFGF immunoreactivity in the pons-medulla, hypothalamus and cerebellum. The cortex contained much less immunoreactivity. Consistent with the biochemical data, immunocytochemical analysis with the same two antisera demonstrated that aFGF immunoreactivity is localized in neuronal cell bodies in these regions. Numerous immunoreactive neurons were observed in the reticular formation of the pons and medulla, as well as in several other brainstem nuclei and areas. Immunoreactive neurons were also present in the lateral and medial hypothalamus, and some thalamic, subthalamic and epithalamic nuclei. In the basal ganglia, immunoreactive neurons were present in the amygdala and septum. Few intensely stained immunoreactive neurons were observed in the striatum, pallidum and neocortex. Limbic cortices contained more numerous immunoreactive neurons than neocortex. These results support the concept that aFGF is present in the brain, where it is heterogeneously distributed in neuronal cell bodies in regions involved in sensory, extrapyramidal motor, limbic and autonomic functions. The results are consistent with various neurotrophic, mitogenic, and neuromodulatory functions associated with aFGF in the mammalian central nervous system.

Distribution and regulation by estrogen of progesterone receptor in the hypothalamus of the cat.

The diencephalon is critically involved in the estrogen-dependent receptor-mediated stimulation of respiration by progesterone in cats. To identify a neuroanatomic basis for this effect of progesterone, the diencephalon of the ovariectomized cat was examined immunohistochemically with an antiprogesterone receptor (anti-PR) monoclonal antibody. No immunostaining was found in ovariectomized animals pretreated with sesame oil alone. In contrast, numerous cells in the ventromedial aspect of the hypothalamus from cats pretreated with estradiol benzoate were PR immunoreactive. Thus, PR is induced by estrogen in hypothalamic neurons of cats. In animals pretreated with estradiol benzoate, the highest density of immunostained neurons was found throughout the infundibular nucleus, especially in the region of the mammillary recess of the third ventricle. PR-immunoreactive cells were also distributed throughout the periventricular nucleus, with the highest density located rostrally and immediately above the suprachiasmatic nucleus. Notably and in contrast to a number of other species (e.g. rat and guinea pig), only very few weakly stained PR-containing cells were found in the hypothalamic ventromedial nucleus. This latter finding could reflect the progesterone independence of sexual behaviors in cat. Overall, we have identified hypothalamic areas that may subserve estrogen-dependent receptor-mediated effects of progesterone in the cat, such as the stimulation of respiration.

Ventral mesencephalic neurons containing both cholecystokinin- and tyrosine hydroxylase-like immunoreactivities project to forebrain regions.

The coexistence of cholecystokinin- and tyrosine hydroxylase-like immunoreactivities within neurons of the rat ventral mesencephalon was analyzed by using an indirect immunofluorescence technique for the simultaneous demonstration of two antigens in the same tissue section. A high degree of colocalization was observed in the substantia nigra pars compacta, in which 80-90% of all labeled neurons at rostral and up to 70% at intermediate levels contained both cholecystokinin and tyrosine hydroxylase. At caudal levels, the incidence of colocalization declined to approximately 30-50%. All of the immunoreactive perikarya in the substantia nigra pars lateralis were labeled with both substances. Other areas of the ventral midbrain that exhibited a moderate proportion of neurons immunoreactive for both cholecystokinin and tyrosine hydroxylase included the ventral tegmental area, interfascicular nucleus, and rostral and caudal linear nuclei. In addition, coexistence was occasionally observed within neurons of the central and ventral periaqueductal gray matter, supramammillary region, peripeduncular region, retrorubral field, and extremely rarely, within the substantia nigra pars reticulata. Cell bodies containing tyrosine hydroxylase-like immunoreactivity (indicative of dopamine) usually outnumbered those containing the peptide except in the supramammillary region and in the ventral periaqueductal gray matter, where the cholecystokinin perikarya were present in higher numbers. The double-labeling colocalization technique was combined with fluorescence retrograde tracing to determine some of the forebrain projections of these neurons. Ventral midbrain neurons containing both cholecystokinin and tyrosine hydroxylase were found to project to the caudate-putamen, nucleus-accumbens, prefrontal cortex, and amygdala. These projections originated from neurons located predominantly in the substantia nigra pars compacta and the ventral tegmental area. Thus, cholecystokinin occurs within the well-known dopaminergic nigrostriatal pathway in the rat. Overall, these results demonstrate that a significant proportion of the dopamine neurons giving rise to the ascending mesotelencephalic projections also contain the peptide cholecystokinin.

Cholecystokinin innervation of rat thalamus, including fibers to ventroposterolateral nucleus from dorsal column nuclei.

The distribution of cholecystokinin octapeptide immunoreactive fibers and puncta in the adult rat thalamus was studied using immunocytochemical methods. Small to moderate numbers of immunoreactive fibers were present in the lateral habenular nucleus, ventral lateral geniculate nucleus, zona incerta, parataenial, mediodorsal, medioventral, and submedial nuclei, the rhomboid, paracentral, central lateral and parafascicular nuclei, and in the medial geniculate and dorsal lateral geniculate nuclei. Moderate to large numbers of cholecystokinin (CCK)-positive fibers were present in the paraventricular nuclei, the reticular nucleus, the anteroventral, anteromedial, and central medial nuclei, and in the rostral extension of the internal medullary lamina between the parataenial and anteroventral nuclei. Dense concentrations of immunoreactive fibers were also found in a principal sensory relay nucleus, the ventroposterolateral nucleus (VPL), of the ventrobasal complex. The number of CCK-positive fibers in VPL showed a marked unilateral decrease in rats which had received lesions of the contralateral gracile and cuneate nuclei. The results of this study demonstrate that CCK-immunoreactive fibers and puncta are widely distributed in the rat thalamus, and that the source of these fibers in VPL is probably the dorsal column nuclei.