Effect of selective midbrain and diencephalic 5,7-dihydroxytryptamine lesions on serotonin content in individual preopticohypothalamic nuclei and on serum luteinizing hormone level.

Injection of the serotonin neurotoxin, 5,7-dihydroxytryptamine (5,7-DHT), into the midbrain dorsal (DR) or median (MR) raphe nucleus of castrated and normal male rats was followed by measurement of serum luteinizing hormone (LH) level and the 5-hydroxytryptamine (5-HT) content of several hypothalamic and amygdaloid nuclei. Only the DR lesions lead to a significant decrease (42%) in serum LH level in normal rats. The elevated LH level in castrated animals was not affected by either lesion. The DR lesions were followed by 5-HT reductions only in the medial preoptic area, the arcuate and ventromedial hypothalamic nuclei, and in the basal and central amygdaloid nuclei. In contrast, the 5-HT reductions produced by MR lesions were much more widespread, being found in all nuclei assayed with the exception of the dorso- and ventromedial hypothalamic nuclei. In a second experiment, degeneration of serotonergic terminals in the ventromedial region of the hypothalamus following intradiencephalic injections of 5,7-DHT led to a significant decrease in serum LH level and a 5-HT reduction in the arcuate, ventromedial and dorsomedial hypothalamic nuclei. 5,7-DHT injections into the medial preoptic area and the anterior hypothalamic area did not affect serum LH level. These results suggest that a serotonergic pathway originating in the midbrain dorsal raphe nucleus and innervating the mediobasal hypothalamus has a stimulatory influence on LH secretion.

Modification of ethanol intoxication by dopamine agonists and antagonists.

The effects of ethanol on locomotor activity and rotarod performance were measured in the rat. Apomorphine (4 mg/kg s.c.) antagonized and pimozide (0.75 - 1.50 mg/kg s.c.) enhanced the effects of a 2 g/kg dose of ethanol on rotarod performance. No such interaction was seen with pentobarbital, 25 mg/kg, a dose sufficiency to produce an equivalent motor performance deficit. Animals with 6-hydroxydopamine lesions of the substantia nigra were more sensitive to the ethanol-induced impairment of rotarod performance and suppression of locomotor activity than shamoperated controls. The pattern of dopamine depletion in lesioned animals indicated a degree of selectivity of the lesion for the nigrostriatal pathway. The results are compatible with the hypothesis that ethanol interferes with dopaminergic transmission, and that such an interference may be involved in the behavioral effects of ethanol.

The effects of subhypnotic doses of ethanol on regional catecholamine turnover.

A 2 g/kg dose of ethanol given intraperitoneally to rats significantly reduced the turnover of dopamine in the substantia nigra and caudate nucleus, increased dopamine turnover in the olfactory tubercle and had no effect on dopamine turnover in the nucleus accumbens, amygdala and hypothalamus. The same dose of ethanol decreased the probenecid-induced homovanillic acid accumulation in the caudate nucleus. The turnover of norepinephrine was also decreased in hypothalamus and increased in the pons medulla region. No change in norepinephrine turnover was observed in frontal cortex, parietal cortex, cerebellum, amygdala, hippocampus and locus ceruleus region. The distribution of ethanol was similar in cortex, caudate nucleus, hypothalamus and pons-medulla. Catecholamine turnover in different brain regions seems to be differentially sensitive to the effects of ethanol, with most regions being unaffected by ethanol.

Histochemical and biochemical characterization of the rat paracervical ganglion.

The nature and identity of the catecholamines in the paracervical ganglion and superior cervical ganglion small, intensely fluorescent (SIF) cells were investigated using fluorescence histochemical and immunohistochemical techniques. The paracervical ganglion SIF cells were found to contain norepinephrine and the superior cervical ganglion SIF cells, dopamine. The norepinephrine content of the paracervical ganglion SIF cells averaged about 72 ng/ganglion and did not change during the rat estrus cycle. The activity of the enzyme tyrosine hydroxylase in the PCG was very low (about 0.48 nmoles DOPA formed/h/mg protein) and was about 1/50th of the activity of the enzyme in the SCG, where it averaged about 23.90 nmoles DOPA formed/h/mg protein. These experiments suggested that the paracervical ganglion has large numbers of norepinephrine containing SIF cells with a relatively slow turnover of their catecholamine content.

Dopaminergic activity of some apomorphine analogs.

The dopaminergic activity of ten apomorphine analogs was studied in rats lesioned unilaterally with 6-hydroxydopamine in the nigro-striatal system. Of these ten compounds, N,N-dimethyl-5,6-dihydroxy-2-amino-tetralin (M-7), N-methyl-5,6-dihydroxy-2-aminotetralin (M-8) and N,N-dimethyl-4,5-dihydroxy-2-aminoindan (DDAI) exhibited potent dopaminergic stimulant activity by causing the rat to turn to the unoperated side. The turning behavior of apomorphine, M-7, DDAI and d-amphetamine were antagonized by haloperidol. M-7 and DDAI also induced pecking in pigeons and their effects were also blocked by haloperidol. It is concluded that M-7, M-8 and DDAI are direct acting central dopaminergic agents.

Adrenergic neuronal degeneration induced in portal vein and caudal artery by 6-hydroxydopamine in vitro.

Fluorescence histochemistry and electron microscopy were used to study the structural consequences of in vitro exposure to the sympatholytic agent 6-hydroxydopamine on two blood vessels, the portal mesenteric vein and caudal artery of the rat. The results showed depletion of catecholamines to indetectable levels associated with clear signs of adrenergic nerve degeneration, such as cytoplasmic shrinking, virtual absence of dense core vesicles and swelling of mitochondria. All of the changes observed occurred within 2 hours in the caudal arteries and 3.5 hours in the portal veins. Comparison of electron and fluorescence micrographs of incubated control specimens with those of unincubated, fresh specimens showed that the nerve endings of the incubated controls were well preserved for at least 3.5 hours. With destruction of nerve endings in such a short period of time, the processes of specific neuronal degeneration could be clearly demonstrated in isolated blood vessels.

A modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagon and growth hormone.

The requirement of using homologous antisera (primary antiserum and peroxidase-antiperoxidase (PAP) complex raised in the same species) in the unlabeled antibody enzyme method has been investigated at the light and electron microscopic level using the localization of insulin, glucagon and growth hormone as model systems. Optimum immunocytochemical staining for all three antigens was observed when sheep or goat antirabbit gamma-globulin (S-ARgammaG or G-ARgammaG) were used to couple rabbit peroxidase-antiperoxidase complex with either guinea pig antisera to insulin (GP-AIS) or glucagon (GP-AGS), or monkey antisera to rat growth hormone (M-ARGH). The cross-reactivity between S-ARgammaG or G-ARgammaG and immunoglobulins in these primary antisera were substantiated by immunoelectrophoresis and radioimmunoassay. S-ARgammaG was shown to produce precipitation arcs with GP-AIS and M-ARGH that were similar to those seen when the latter were reacted with rabbit antiguinea pig gamma-globulin antiserum and goat antimonkey gamma-globulin antiserum, respectively. Radioimmunoassay results revealed that immunoprecipitation of 6-10% as compared to homologous antisera controls yielded excellent staining localization when S-ARgammaG was used for immunocytochemistry. Thus, heterologous antisera (primary antiserum and PAP complex raised in different species) may be used in the unlabeled antibody enzyme method as long as the coupling antiserum shows cross-reactivity with immunoglobulins of the primary antiserum and the PAP complex.