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C González Solveyra

Publications and source records attributed to C González Solveyra.

7 recordsLinked to original sources

Oligobodies: bench made synthetic antibodies.

Using synthetic peptides and a combinatorial library of 56 mer random oligonucleotides, we have developed reagents that behave as "synthetic antibodies". The results obtained with the protein phosphatase 2A as a model system are shown here. The specificity of these reagents, named "oligobodies", has been demonstrated by Western blot analysis and immunohistochemistry. The oligobodies have enormous advantages compared to antibodies: their production is independent of the immune system, they can be prepared in a few days and there is no need for a purified target protein. These reagents can be produced even if the corresponding protein was never isolated or purified, since only a partial DNA sequence from a database provides enough information to make them.

Animals↗

GABA and its neural regulation in rat brown adipose tissue.

By using a radioreceptor assay GABA was detectable in rat interscapular brown adipose tissue (IBAT), the levels being 1% those of CNS and 10-fold those of peripheral plasma. Injection of the glutamic acid decarboxylase (GAD) inhibitor 3-mercaptopropionic acid lowered IBAT GABA levels by about half while injection of the GABA transaminase inhibitor gamma-acetylenic GABA increased them by 230%. Rats kept at 4 degrees C for 14 days exhibited IBAT GABA levels that were about half those found at 22 degrees C. Accumulation of IBAT GABA after gamma-acetylenic GABA increased by 2-fold in cold-exposed rats. Sympathetic denervation of IBAT prevented the effect of the cold environment on GABA content and impaired that on GABA accumulation. GAD activity was detectable in IBAT homogenates and isolated brown adipocytes. Exposure of rats to cold increased Vmax of GAD without modifying its Km, regardless of intactness of innervation. In binding studies with 3H-GABA as a ligand, two types of sites were uncovered of KD = 14 and 146 nM, respectively. In the presence of 2.5 mM Ca2+ bicuculline and baclofen were 57 and 46% as effective as GABA to displace 3H-GABA from IBAT binding sites. The results indicate existence, possible synthesis and type A and B receptors of GABA in rat IBAT.

Adipose Tissue↗

Origins of the sympathetic projections to rat thyroid and parathyroid glands.

The present study aimed to characterize the localization and pathways of sympathetic neurons innervating the thyroid and parathyroid glands. In rats subjected to unilateral superior cervical ganglionectomy or unilateral decentralization of the superior cervical ganglion 7 days earlier, ipsilateral depression of thyroid norepinephrine (NE) and epinephrine content to 6-16% of the contralateral intact lobe was observed. In both groups of animals neuronal [3H]NE uptake by the ipsilateral thyroid lobe was suppressed. In unilaterally decentralized rats pineal catecholamine levels remained within normal values whereas in unilaterally ganglionectomized rats a 74% decrease of pineal NE and E content was found. Unilateral section of the external carotid nerve abolished, as did unilateral superior cervical ganglionectomy, neuronal [3H]NE uptake in the ipsilateral thyroid lobe. In contrast external carotid nerve section did not modify the neuronal uptake of [3H]NE in the pineal gland. Either unilateral superior cervical ganglionectomy or external carotid nerve section resulted in similar involution of ipsilateral thyroid lobes of hypophysectomized rats. These results indicate that postganglionic sympathetic perikarya innervating the thyroid-parathyroid territory are located in the middle and/or inferior cervical ganglia and send their axons through the SCG and the external carotid nerve to these glands.

Animals↗

Benzodiazepines decrease norepinephrine release from rat pineal nerves by acting on peripheral type binding sites.

In 3 000g-supernatants of rat pineal homogenates a single population of benzodiazepine (BZP) binding sites with dissociation constant= 97-102 nM and maximal number of sites= 6.5-9 pmoles/mg protein was detected by employing 3H-flunitrazepam (FNZP) as a radioligand. The following order of affinity for several BZP was found (Ki, nM): Ro 5-4864 (8), FNZP (99), clonazepam (7,900) Ro 15-1788 (10,000). Two weeks after bilateral superior cervical ganglionectomy (SCGx) a 18-28% reduction of site number without significant changes in affinity of 3H-FNZP binding was detectable in rat pineal glands. In pineal explants priorly incubated with 3H-norepinephrine, exposure to 0.1-10 microM of Ro 5-4864 or diazepam decreased significantly transmitter release elicited by 80 mM K+, whereas clonazepam did not affect it significantly. At 10 microM-concentrations, Ro 5-4864, diazepam or clonazepam increased pineal melatonin content of explants incubated for 6 h with the drug. In pineal explants of rats subjected to SCGx 14 days earlier, only 10 microM of Ro 5-4864 increased melatonin content significantly to about half of the percent increase detected in innervated glands. These results suggest that BZP decrease transmitter release from pineal sympathetic nerves by acting on peripheral BZP binding sites, an effect which is about 2 orders of magnitude greater than the postsynaptic stimulation of pineal melatonin synthesis.

Animals↗

Effect of naloxone on the nocturnal rise of rat pineal melatonin content.

Rats exposed to 14 light: 10 h dark photoperiods were given naloxone hydrochloride (10 mg/kg) s.c. 2 h before and 5 h after the lights were turned off. A smaller nocturnal pineal melatonin peak, as well as a slower decrease from peak values were observed in naloxone-treated rats. Naloxone (up to 10(-4) M) did not affect either in vitro rat pineal melatonin content or 3H-labeled transmitter release in glands previously incubated with [3H]norepinephrine. Hence a central, rather than a peripheral opioid synapse appears to be involved in naloxone effect on nocturnal melatonin increase.

Animals↗

In vitro uptake of benzodiazepines by rat pineal gland.

As a part of a study aiming to characterize the physiological and pharmacological significance of the high affinity pineal benzodiazepine (BZP) binding sites reported previously, we examined the uptake of the BZP derivative 3H-flunitrazepam (FNZP) by rat pineal glands in vitro. At 37 degrees C, 3H-radioactivity was taken up by tissue up to a pineal/medium concentration of about 12, while at 0 degrees C the uptake amounted to only one-third that at 37 degrees C. Reciprocal of uptake analyzed by Lineweaver-Burk plots indicated apparent Km's of 1.74 and 1.45 microM, and Vmax's of 1.32 and 1.04 pmol per min per mg tissue, for control and superior cervical ganglionectomized rats, respectively, suggesting that the neural compartment does not participate significantly in 3H-FNZP uptake. Cerebral cortex explants of similar size and weight as the pineal ones took up 3H-FNZP to a maximum tissue/medium concentration of about 2. Neither pineal nor cerebral cortex 3H-radioactivity uptake exhibited significant changes as a function of time of day. A number of agents, including several BZP analogues, cocaine, desipramine, melatonin, fluoxetine, nomifensine, and dipiridamol, as well as changes in the ionic environment or metabolic inhibitors, did not affect 3H-FNZP uptake significantly. Other tissues, such as liver, muscle, kidney, adrenal gland, or anterior pituitary, took up 3H-radioactivity to tissue concentrations slightly lower than those of the cerebral cortex, suggesting that drug liposolubility accounted only to a limited extent for the high in vitro uptake detected in incubated pineals.

Animals↗