Immunohistochemical demonstration of the distribution of serotonin neurons in the brainstem of the rat and cat

The morphological characteristics and distribution of the somata of serotonin-containing neurons in the brainstem of rats and cats were studied by use of the peroxidase-anti peroxidase (PAP) immunohistochemical method employing highly specific antibodies to serotonin. Antibodies were raised in rabbits against an antigen prepared by coupling serotonin to bovine thyroglobulin and using formaldehyde as the coupling reagent. The distribution pattern of serotonin neurons observed in the present material is essentially in agreement with that described by other investigators who used the Falck-Hillarp method. In addition, this immunohistochemical technique revealed serotonin-containing perikarya in the following regions: 1) the periaqueductal gray, especially lateral to the nucleus raphe dorsalis, 2) the nucleus interpeduncularis, 3) the nucleus parabrachialis ventralis and dorsalis, 4) the field of the lemniscus lateralis, and 5) the reticular formation of the pons and medulla oblongata. The described immunohistochemical procedure makes it possible to study central serotonin neurons in detail without pharmacological pretreatment. The wide distribution of serotonin neurons demonstrated in this study should be considered when interpreting experiments dealing with the serotonin system.     

Subcellualr localizations of guanylate cyclase and 3′,5′-cyclic nucleotide phophodiesterase in sea urchin sperm

The subcellular localizations of guanylate cyclase and 3′,5′-cyclic nucleotide phophodiesterase in sea urchin sperm were examined. Both the specific and total activities of these two enzymes were much higher in sperm flagella (tails) than in the heads. In addition to the observation that guanylate cyclase in the flagella was particulate-bound and solubilized by Triton X-100, more than 980% of the cyclase activity in the flagella was found in the plasma membrane fraction, whereas the activity of cyclic nucleotide phosphodiesterase was observed in both the axonemal and plasma membrane fractions. The observations indicated that the cyclase in the flagella appeared to be associated with the plasma membrane. Cyclic nucleotide phosphodiesterase in the plasma membrane fraction as well as the axonemal fraction hydrolyzed both cyclic GMP and cyclic AMP; however, the rates of hydrolysis for cyclic GMP were obviously higher than those for cyclic AMP. The enzymic properties of guanylate cyclase and cyclic nucelotide phosphodiesterase in sperm flagella were also briefly described.     

Immunohistochemical demonstration of serotonin-containing nerve fibers in the mammalian hippocampal formation

The distributional pattern of serotonin-immunoreactive nerve fibers in the hippocampal formation of six different mammalian species (rat, chipmunk, hamster, cat, dog and monkey) was studied in detail by means of a modified peroxidase-antiperoxidase immunohistochemical method, using a specific serotonin antiserum. Furthermore, the density of varicosities distributed in each layer of the hippocampus was semiquantitatively analyzed. In all species investigated, the routes of serotonin fibers entering the hippocampal formation were found to be almost the same. These fibers were extensively distributed throughout the hippocampal formation, and had a characteristic arrangement corresponding to the laminar structure of this region. A dense innervation by varicose serotonin fibers was found in the stratum lacunosum-moleculare, but a few serotonin fibers were also distributed in the stratum lucidum of the CA2 and CA3 fields. The stratum pyramidale and the granule cell layer of the dentate gyrus contained a small number of serotonin fibers. The concentration and the direction of serotonin fibers were different in each area of each animal. Two peculiar observations should be stressed: (1) in the rat, the strata oriens and radiatum of CA2 and CA3 as well as the stratum lacunosum-moleculare displayed abundant serotonin fibers; (2) in the dog, abundant serotonin fibers were diffusely distributed in the CA1 field except for the stratum pyramidale and the most dense concentration of serotonin fibers was seen in the stratum oriens of CA3. The present study provides a morphological basis for further study of the functional significance of serotonin in the limbic system.