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PubMed1980

Comparative localization of prolactin-binding sites in different rat tissues by immunohistochemistry, radioautography and radioreceptor assay.

Prolactin was localized in various rat tissues, using two different histological approaches: immunohistochemistry and autoradiography. Following injection of ovine prolactin (oPRL) to intact rats, oPRL was localized in the proximal convoluted tubules (PCT) of the kidney using an immunoperoxidase technique. Diffuse and weak staining was observed in the liver while no staining was observed on sections of adrenal, testis and brain. Incubation of [125I]oPRL on frozen cryostat tissue sections followed by radioautographic processing revealed specific radioautographic grains on cells of convoluted tubules of the kidney. Specific grains were also seen on hepatocytes of the liver, cells of the adrenal cortex, interstitial cells of the testis and epithelial cells of the choroid plexus of the brain. The amount of specific radioautographic grains seen per tissue agreed well with in vitro binding values of [125I]oPRL to crude membrane preparations in the same tissues. Our observations clearly show that in the rat specific binding sites for prolactin are present on cells of the proximal convoluted tubules of the kidney, hepatocytes of the liver, glandular cells of the zona reticularis, zona fasiculata and zona glomerulosa of the adrenal, interstitial cells of the testis and epithelial cells of the choroid plexus. These studies demonstrate the necessity of using more than one morphological approach in the identification of prolactin-binding sites in different tissues at the cellular level.

Adrenal Glands↗
PubMed1985

Glutamate decarboxylase-immunoreactive boutons in synaptic contacts with hypothalamic dopaminergic cells: a light and electron microscopy study combining immunocytochemistry and radioautography.

Double post-embedding immunolabeling of both tyrosine hydroxylase and glutamate decarboxylase on 1-micron semi-thin sections allowed the visualization of numerous endings that use gamma-aminobutyrate as a transmitter apposed to dopaminergic cell bodies in the periventricular-arcuate hypothalamic complex. Up to fifteen glutamate decarboxylase-positive contacts per tyrosine hydroxylase-positive cell profile could be observed. In some favourable planes of section glutamate decarboxylase-positive endings were also seen in close apposition to proximal dopaminergic dendrites. About 250 tyrosine hydroxylase-positive cell profiles, whose diameter approached the maximum diameter of the dopaminergic cells, were surveyed. An average of 7.4 glutamate decarboxylase-positive contacts were counted on these profiles. From these figures it was estimated that a dopaminergic cell body was contacted on average by 75-175 terminals that use gamma-aminobutyrate as a transmitter. At the electron-microscopic level, the nature of these contacts was investigated by a method combining radioautographic detection of cell bodies having taken up tritiated dopamine and pre-embedding immunostaining of glutamate decarboxylase containing endings. Glutamate decarboxylase-positive axon terminals were seen apposed to somatic and dendritic elements. On some favorable planes of section, they were found to be engaged in morphologically defined synaptic complexes of the symmetrical or asymmetrical type. A number of the postsynaptic perikarya were labelled by tritiated dopamine and, in agreement with the light microscopic observations, they were frequently seen in contact with more than one immunopositive ending. The present findings provide a morphological substratum for a direct gamma-aminobutyrate control of the tuberoinfundibular dopaminergic neurons. Such a control could account more particularly for the central, stimulatory effects of gamma-aminobutyrate on prolactin secretion.

Animals↗