Knowledge about sites and mechanisms of action of vitamin D and its analogs has been greatly advanced by histochemical approaches. High resolution and high sensitivity, combined with the integrative potential of relatively intact histochemical tissue preparations, contributed information that is difficult or impossible to obtain otherwise. In in vivo distribution studies with conventional biochemical assays, target cell populations associated with non-target tissues frequently remain unrecognized without the resolution achieved by cellular autoradiography. Autoradiography, alone or combined with immunohistochemistry when applied to in vivo drug targeting and target characterization, has provided information on cellular-subcellular receptor distribution in over 50 tissues. These discoveries, importantly, contribute to a new understanding of the biological role of vitamin D and challenge the concept of "the calcium homeostatic steroid hormone" as being too narrow. While some of the outstanding effects of vitamin D deficiency and toxicity relate to calcium homeostasis, the vast majority of the target tissues appear not to be primarily related to calcium metabolism, but rather to the activation and regulation of exo- and endocrine secretory and somatotrophic processes such as cell differentiation and proliferation. Also, several highly calcium-dependent tissues such as striated and smooth muscles are not genomic targets for vitamin D. The reviewed data on the diverse and extensive presence of target tissues forecast a high therapeutic potential for vitamin D and especially its low-calcemic analogs, far beyond that which is presently utilized. The evidence provided for vitamin D also testifies to the utility and need to include in vivo cytopharmacology in any target evaluation of bioactive compounds to further the understanding of their mechanisms of action, and to identify preferential targets and their differential therapeutic and toxic potentials.

The presence of c-fos, a marker for cell activation, was investigated in cerebral neurons actively expressing ecdysteroid receptors during larval-pupal development in the tobacco hornworm, Manduca sexta. Colocalization was accomplished by ecdysteroid autoradiography using the tritiated high affinity 20-hydroxyecdysone agonist ponasterone A and immunocytochemistry with an antibody to a peptide sequence which is highly conserved in both human and murine c-fos. Immunoreactivity to a c-fos-like protein(s) was present in nuclei of many neurons of all the developmental stages examined. However, with the exception of the optic lobe, cells expressing nuclear ecdysteroid receptors were more immunoreactive than non-ecdysteroid-binding neurons. These data suggest that ecdysteroid-induced gene activation and translation may involve c-fos expression.

The present study examined the number and distribution of progestin receptor cells in the 8-day-old male and female cortex and compared cortical labeling with that in the preoptic area and central hypothalamus. Eight-day postnatal mice (four males and four females), treated with estradiol, were each sc injected with 0.32 micrograms/100 g BW [125I]progestin (SA, 2200 Ci/mM). Brains were frozen 2 h after injection of [125I]progestin, sectioned, and processed for thaw-mount autoradiography. Cells with a nuclear concentration of radioactivity were localized in lamina VI of the lateral cortical regions of the male and female brain, while only a few cortical cells were seen in laminae II, III, and V of the suprarhinal, lateral, and cingulate/paracingulate regions. Comparison of the number of labeled cells revealed that the female cortex contained significantly more labeled cells than the male at three of the four levels investigated. Similarly, the number of target cells was higher in the female medial preoptic nucleus, but not in the arcuate nucleus and ventromedial hypothalamic nucleus, while the distributions of labeled cells in the male and female preoptic/hypothalamic regions were comparable. Injection of unlabeled progesterone or R5020 1 h before [125I]progestin reduced the nuclear concentration of radioactivity in all target regions and verified the specificity of [125I]progestin for the progestin receptor. The results of these studies indicate that mouse 8-day-old cortex and preoptic area in the female animal have more progestin receptor cells than those in the male and demonstrate that progestin receptor cells are localized in a region of the cortex known to contain few estrogen target cells. These results further suggest that a sexual dimorphism in progestin cell number may result in a differential effect of progestin on the cortex and preoptic area of the mouse, perhaps establishing a dimorphism in development and function.

Atrial myoendocrine cells of rat were investigated regarding estradiol uptake. It was found that, in addition to their specific endocrine function of producing cardiac polypeptides of the cardiodilatin/atrial natriuretic peptide (CDD/ANP) family, these cells also specifically accumulate radiolabeled estradiol. This co-localization supports the view that steroid hormones play an important role in the regulation of the CDD/ANP gene.

Immunohistochemical investigations with different antisera against cardiodilatin 99–126 or alpha atrial natriuretic polypeptide revealed the presence of cardiac hormones not only in the atria of rats but also in strands of myoendocrine cells located in subendocardial regions of the ventricular septum. The localization of CDD-IR (cardiodilatin immunoreactivity) in the ventricle is associated with the location of the conduction system in the rat. The significance of the morphological relationship between cardiodilatin and the conduction system of the rat heart is discussed.

The topographical distribution of enkephalin in the central nervous system of the lizard, Anolis carolinensis, has been studied by the immunoperoxidase technique with antiserum to leucine‐enkephalin. Immunoreactive enkephalin perikarya, fibers and probably terminals are widely distributed throughout the central nervous system, which agrees well with the distribution of enkephalins in the mammalian brain. Enkephalin‐containing perikarya are found in the subpallium (septum, nucleus accumbens, striatum, amydgala), preoptic and hypothalamic region, ventromedial nucleus and ventromedial area of thalamus, pretectal geniculate nucleus and posterodorsal nucleus of pretectum, nucleus of the lateral lemniscus, locus ceruleus, spinal trigeminal nucleus, nucleus of the solitary tract, medial parvocelluar nucleus, and dorsal horn of the spinal cord. Enkephalinergic fibers and terminals are found in the above–mentioned areas as well as in the pallium (medial and dorsal cortex, dorsal ventricular ridge), dorsomedial and anterior dorsolateral nucleus of the thalamus, habenua, nucleus of the stria medullaris, torus semicircularis, mesencephalic tegmental area, interpeducular nucleus, mesencephalic trigeminal nucleus, central gray, reticular formation, raphe nucleus, substantia nigra, isthmus region, and nucleus of the trapezoid body. Enkephalinergic pathways appear to exist between the septum and the medial cortex, nucleus accumbens and nucleus of the lateral olfactory tract, striatum and certain mesencephalic structures, hypothalamus and tegmentum, and between nucleus of the lateral lemniscus and torus semicirculais. In the pituitary, cells of the pars intermedia, and certain cells of the rostral pars distalis also show immunoreactivity to enkephalin antiserum. The distribution of enkephalin immunoreactivity throughout the hypothalamus and in the median eminence suggests involvement in neuroendocrine regulation. Presence of enkephalin in many extrahypothalamic brain areas indicates its important role in various sensory functions and in behavioral and autonomic integration.