Biomedical subjects

J D DeCristofaro

Publications and source records attributed to J D DeCristofaro.

5 recordsLinked to original sources

PubMed1992

Tissue- and treatment-specific usage of multiple preproenkephalin transcriptional start sites.

The significance of the 5' heterogeneity of the transmitter gene ppEnk was evaluated by comparing start site usage (E1-E4) between 12 tissues from untreated rats, using primer extension analysis. In the basal state, we found that E3- and E4-initiated transcripts accounted for 80% of the total striatal RNA present compared with a preferential usage of the E2 start site in all other tissues. To determine whether this selective expression could be modified by biologically relevant pathways, rats were made hypoglycemic. After insulin shock, only E3 + E4-initiated transcripts increased (16-fold at 1 day) in the adrenal medulla but were unaffected in the striatum. As the effects of insulin shock on the adrenal medulla are mediated by cholinergic pathways and the striatum also receives cholinergic inputs, we also compared the effects of cholinergic drug treatments on start site usage in these two tissues. Rats were treated with cholinergic agonists (nicotine + oxotremorine) which induced adrenomedullary E2 and E3 + E4 transcripts (5- and 80-fold, respectively). This effect peaked at 2 days. In contrast, in the same animals, striatal ppEnk RNA (E3 + E4) increased only 10-15-fold after drug treatment. Hence it appears that biologically relevant whole animal stimuli (insulin shock or cholinergic agents) activate biochemical pathways, which affect start site usage in a tissue-specific fashion. Selective RNA start site usage suggests a biological significance, which may be important in the widespread tissue expression of this gene.

Adrenal Medulla↗

PubMed1992

Regulation of adrenomedullary preproenkephalin mRNA: effects of hypoglycemia during development.

To further evaluate whether transsynaptic mechanisms account for stress-induced changes in adrenomedullary preproenkephalin mRNA (ppEnk mRNA), neonatal rats were made hypoglycemic at a time when synapses are non-functional (less than 10 days postnatal age). While ppEnk mRNA in medullae from adult rats increased as much as 60-fold in this paradigm (insulin 10 U/kg), ppEnk mRNA levels in the newborn increased only 1.6-fold (insulin 20 U/kg). To evaluate whether postsynaptic cholinergic pathways of the neonatal adrenal medulla were functional, we treated 5-day-old pups with cholinergic agonists (nicotine [1 mg/kg, s.c., q 12 h] + carbachol [1.7 mumol/kg, s.c., q 12 h x 4 days]). Combined cholinergic agonist treatment augmented enkephalin prohormone and peptide levels up to 3-fold (P less than 0.05). To determine whether the blunted response to hypoglycemia in the newborn resulted from a deficiency in functional transsynaptic activity, synapses were matured using thyroid hormone pretreatment (postnatal days 2 and 3) before hypoglycemic stress. Hypoglycemia now caused a 40-fold increase in adrenomedullary ppEnk mRNA levels only in the T3/insulin treated group. To exclude other secondary effects of hypoglycemia (eg. hormonal, or insulin treatment-dependent), intracellular glycopenia was produced in the presence of secondary hyperglycemia by injecting adult rats or pups with 2-deoxyglucose (500 mg/kg). Similar to the insulin-hypoglycemia group, a large increase in adrenomedullary ppEnk mRNA resulted in the adult but not in the 5-day-old neonatal adrenal medullae. We conclude that enkephalin biosynthesis, like co-stored catecholamines, is induced by a transsynaptic process.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Medulla↗

PubMed1990

Multiple preproenkephalin transcriptional start sites are induced by stress and cholinergic pathways.

A major control of gene expression occurs at the level of initiation of RNA transcription. In the nervous system this is reflected in part by 5' end RNA heterogeneity of neural transcripts. We now report the characterization of four preproenkephalin (ppEnk) RNA initiation sites in the rat brain striatum. In the adrenal medulla two ppEnk transcriptional start sites were detected. Moreover, cholinergic induction of ppEnk RNA initiation was observed in the adrenal medulla, but not in the striatum. The converse was true following handling stress. Our observations suggest that selective start site usage and stimulus evoked induction of specific RNA initiation is tissue-specific. We speculate that start site usage and induction may provide separate mechanisms through which neuronal gene expression is controlled in anatomically, as well as functionally distinct neurohumoral structures.

Adrenal Medulla↗

PubMed1990

Bimodal regulation of adrenal opiate peptides by cholinergic mechanisms.

Physiologic stressors increase trans-synaptic impulse activity and result in adrenal catecholamine release and biosynthesis. To determine the effects of stress on the co-localized opiate peptide system, rats were cold stressed at 4 degrees C. While cold stress slightly decreased enkephalin levels, a more severe stress (wetting and cold) increased enkephalin levels by 95%. Examining trans-synaptic-cholinergic mechanisms, treatment with either nicotinic or muscarinic agonists alone resulted in no change in adrenal enkephalin content. However, treatment with both nicotinic and muscarinic agonists together resulted in a three-fold rise in enkephalin levels. To further examine cellular mechanisms, medullae were explanted in the presence of agents that increase second messenger cyclic nucleotide levels. Treatments that increase the levels of cAMP, the cyclic nucleotide associated with nicotinic receptor activation, prevented the rise in medullary enkephalin relative to control explants. In contrast, treatments that increased cGMP levels, the cyclic nucleotide associated with muscarinic receptor activation, had no effect on enkephalin content compared to control explants. However, in the presence of both forskolin (10 microM) plus db-cGMP (5 mM), enkephalin content rose three-fold over control explants. These data suggest that, distinct from catecholamine pathways, enkephalin levels can be positively or negatively regulated by the severity of a stressful stimulus, by cholinergic receptor mechanisms and by an interaction of cyclic nucleotide second-messenger pathways.

Adrenal Medulla↗

PubMed1986

Calcium.

Disorders of calcium homeostasis are frequent and seldom diagnosed in the Emergency Department. In both the adult and pediatric population, the causes of increased or decreased calcium are many, and the index of suspicion needs to be raised for these disorders. Selective ordering is warranted for patients with altered mental status, seizures, neuromuscular irritability, or in patients with diseases known to have abnormalities of calcium as a complicating factor. These include renal failure, multiple myeloma, metastatic cancer, granulomatous disease, and endocrinopathies.

Adult↗