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The studies described here were undertaken to characterize the hepatic insulin and glucagon receptors of control (C), pinealectomized (Pn), and melatonin-treated pinealectomized (Pn + Mel) rats. Compared with C rats, an increase in plasma glucose and glucagon levels and a reduction in circulating concentrations of insulin in Pn animals were observed. Melatonin treatment of Pn rats reverses all three parameters toward the normal values. In liver membranes, insulin binding was lower in Pn than in C rats, and glucagon binding was greater in Pn than in C animals; in Pn + Mel rats both insulin and glucagon binding reverse toward the normal values that were observed in C rats. The modifications in hormone binding reflect changes in the number of receptors but not in the affinity constants. The time courses of hormone association and dissociation from liver membranes were similar in all three experimental groups. The degradation of both hormones by liver membranes was similar in all three groups. Insulin receptor degradation also was similar in the three groups, while glucagon receptor degradation was similar in the liver membranes of C and Pn rats but smaller in Pn + Mel animals. These findings suggest that the pineal gland may modulate the circulating levels and liver receptor concentrations of insulin and glucagon. In addition, our results indicate that insulin and glucagon did not induce a down-regulation of liver receptors in Pn rats.

It has traditionally been accepted that, in the process of cellular differentiation, developmental options are progressively restricted until commitment to a specific fate is established and then only terminal differentiation along this lineage is possible. Although this is usually the case in normal physiological development, the latest experimental evidences indicate that the differentiated state of mature cells is not always as stable and durable as it was thought to be. In fact, recently, a hidden plasticity has been revealed in differentiated cells which allows them to deviate to other cell types that might be, functionally, very far away in other developmental pathways. This plasticity has biological significance since it is necessary for normal development to occur, but it also makes possible the emergence of aberrant lineages when interferences with the normal transcriptional and epigenetic mechanisms in charge of maintaining cellular identity do appear. Cancer is one of the possible outcomes of this aberrant reprogramming. The plasticity of the initial cell suffering the first oncogenic alteration plays an essential role in cancer development, since only if this cell possesses enough plasticity a tumoral reprogramming will be possible and a full-blown tumor will develop. Also, plasticity makes it possible for differentiated cells to acquire cancer stem cell properties in the presence of the appropriate oncogenic insults. In this review we discuss the role of cellular plasticity in the normal development of adult tissues and how cellular susceptibility to reprogramming plays an essential part in cancer development.

Glucagon-like peptide-1 does not have specific, high-affinity receptors on rat liver membranes, does not displace glucagon from glucagon receptors on these membranes and does not stimulate the production of cyclic AMP by isolated rat hepatocytes. In the presence of glucagon, high concentrations of glucagon-like peptide-1 do not significantly alter the production of cyclic AMP. Thus, glucagon-like peptide-1 appears unlikely to have a direct action on hepatic carbohydrate metabolism.

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The studies described in this paper were undertaken to characterize the circulating and hepatic insulin and glucagon receptor concentrations of control (C), diabetic (Db), and pinealectomized-diabetic (Pn + Db) rats. Compared with C rats, an increase in plasma glucose and glucagon levels and a reduction in circulating insulin concentrations in Db animals was observed; these differences were greater in Pn + Db rats. In liver membranes, insulin binding was lower in Db and in Pn + Db than in C rats, and glucagon binding was greater in Db and in Pn + Db than in C rats. The modifications in hormone binding did not reflect changes in the affinity constants. The time courses of hormone association and dissociation from liver membranes were similar in all three experimental groups. The degradation of both hormones and their receptors was similar in all three groups. These findings indicate that in either pinealectomized-diabetic or diabetic rats there were significant changes in the circulating and liver insulin and glucagon receptor concentrations and that the changes in the circulating levels of both pancreatic hormones were more pronounced in pinealectomized-diabetic animals. In addition, the absence in Db and in Pn + Db rats of the insulin and glucagon down-regulation of their own receptors could further modify metabolic activities in these animals.

The trkA proto-oncogene encodes a high-a nity NGF receptor that is essential for the survival, di erentiation and maintenance of many neural and non-neural cell types. Altered expression of the trkA gene or trkA receptor malfunction have been implicated in neurodegeneration, tumor progression and oncogenesis. We have cloned and characterized the 5' region of the mouse trkA gene and have identi®ed its promoter. trkA promoter sequences are GC-rich, lack genuine TATA or CAAT boxes, and are contained within a CpG island which extends over the entire ®rst coding exon. The mouse trkA transcription start site is located 70/71 bp upstream to the AUG translation initiation codon. Sequence analysis showed that the gene encoding the insulin receptor-related receptor, IRR, is located just 1.6 kbp upstream to the trkA gene and is transcribed in the opposite direction. We have used trkA-CAT transcriptional fusions to study trkA promoter function in transient transfection experiments. RNase protection assays and CAT protein ELISA analyses showed that a 150 bp long DNA segment, immediately upstream to the start site, is su cient to direct accurate transcription in trkA-expressing cells. Dissection of this fragment allowed us to identify a 13 bp cis-regulatory element essential for both promoter activity and cell-type speci®c expression. Deletion of this 13 bp segment as well as modi®cation of its sequence by site-directed mutagenesis led to a dramatic decline in promoter activity. Gel mobility shift assays carried out with double-stranded oligonucleotides containing the 13 bp element revealed several speci®c DNA-protein complexes when nuclear extracts from trkA-expressing cells were used. Supershift experiments showed that the Sp1 transcription factor was a component of one of these complexes. Our results identify a minimal trkA gene promoter, located very close to the transcription start site, and de®ne a 13 bp enhancer within this promoter sequence.

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