Biomedical subjects

Raquel E Rodríguez

Publications and source records attributed to Raquel E Rodríguez.

8 recordsLinked to original sources

PubMed2006

New kappa opioid receptor from zebrafish Danio rerio.

A cDNA that encodes a kappa opioid receptor like from zebrafish (ZFOR3) has been cloned and characterized. The encoded protein is 377 residues long and presents 70% identity with the mammalian kappa receptors, although less homology is found in the amino- and carboxyl-terminus as well as in the extracellular loops. In situ hybridization studies have revealed that ZFOR3 mRNA is highly expressed in particular brain areas that coincide with the expression of the kappa opioid receptor in other species. When ZFOR3 is stably expressed in HEK293 cells, [(3)H]-diprenorphine binds with high affinity (K(D)=1.05+/-0.26 nM), being this value on the same range as those reported for mammalian kappa opioid receptors. On the other hand, the selective agonist for mammalian kappa receptors U69,593 does not bind to ZFOR3. [(3)H]-diprenorphine binding is readily displaced by the peptidic ligand dynorphin A and by the non-endogenous compounds bremazocine, naloxone and morphine, although with different affinities. Our results demonstrate that ZFOR3 is a unique model to study the kappa opioid receptor functionality.

Amino Acid Sequence↗

PubMed2005

Characterization of opioid-binding sites in zebrafish brain.

The pharmacological profile of opioid-binding sites in zebrafish brain homogenates has been studied using radiolabeled binding techniques. The nonselective antagonist [(3)H]diprenorphine binds with high affinity (K(D) = 0.27 +/- 0.08 nM and a B(max) = 212 +/- 14.3 fmol/mg protein), displaying two different binding sites with affinities of K(D1) = 0.08 +/- 0.02 nM and K(D2) = 17.8 +/- 9.18 nM. The nonselective agonist [(3)H]bremazocine also binds with high affinity to zebrafish brain membranes but only displays one single binding site with a K(D) = 1.1 +/- 0.09 nM and a B(max) = 705 +/- 19.3 fmol/mg protein. Competition binding assays using [(3)H]diprenorphine and several unlabeled ligands were performed. The synthetic selective agonists for mammalian opioid receptors DPDPE ([DPen(2),D-Pen(5)]-enkephalin), DAMGO ([D-Ala(2),NMe-Phe(4),Gly(5)-ol]-enkephalin), and U69,593 [(5alpha,7alpha,8beta)-(+)-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro[4.5]dec-8-yl]-benzeneacetamide] failed to effectively displace [(3)H]diprenorphine binding, whereas nonselective ligands and the endogenous opioid peptides such as dynorphin A showed good affinities in the nanomolar range, although several of the endogenous peptides only displaced approximately 50% of the specifically bound [(3)H]diprenorphine. Our results provide evidence that, although the selective synthetic compounds for mammalian receptors do not fully recognize the opioid-binding sites in zebrafish brain, the activity of the endogenous zebrafish opioid system might not significantly differ from that displayed by the mammalian opioid system. Hence, the study of zebrafish opioid activity may contribute to an understanding of endogenous opioid systems in higher vertebrates.

Animals↗

PubMed2005

Evidence of involvement of the nNOS and the kappa-opioid receptor in the same intracellular network of the rat periaqueductal gray that controls morphine tolerance and dependence.

Tolerance and dependence are the most important side effects of opioid-mediated pain therapies. However, the mechanisms through which these phenomena are produced still remain unknown. Among the opioid receptors, the kappa-opioid receptor has been the focus of strong research efforts, since it contributes to the reversal of morphine-induced tolerance and dependence. Parallel to this, neuronal nitric oxide synthase has been shown to play a key role in the development of these unwanted effects. Both the kappa-opioid receptor and neuronal nitric oxide synthase are abundantly located in the CNS. One of the areas where these cellular agents are best represented is a key encephalic nucleus in the development of tolerance to the analgesic action of opioid drugs, the periaqueductal gray. In this work, we studied whether morphine-induced tolerance and dependence causes changes (a) in the activity of neuronal nitric oxide synthase and (b) in kappa-opioid receptor expression in the rat periaqueductal gray. Besides, we examined the colocalization of both molecules. Our results point to an involvement of KOR and nNOS in the same intracellular network that controls the development of morphine tolerance and dependence.

Animals↗

PubMed2005

Kappa opioid receptor is expressed in the rat cerebellar cortex.

Classically, the cerebellum has been shown to be involved in motor and visual functions, although recent evidence point to new roles of this organ. Pain processing is one of the recently described functions of the cerebellum. According to the importance of the opioid system in nociception, a detailed characterization of the expression pattern of opioid peptides in the cerebellum is the first step towards understanding the precise involvement of this organ in pain management. By using two different approaches (reverse transcription/polymerase chain reaction and in situ hybridization), we have detected, for the first time, expression of the kappa opioid receptor (KOR) gene in the cerebellar cortex of the rat. Expression is found in the molecular and granular layers in all the lobules of the cerebellum. Approximately 34% of the cells present in the molecular layer express KOR mRNA. This work contributes to the deeper knowledge of the mechanisms that are involved in cerebellar function and may lead to a better understanding of the relationships between nociceptive activity and drug abuse potential.

Animals↗

PubMed2005

COMT (Val158Met) polymorphism is not associated to neuropathic pain in a Spanish population.

It is well known that the response to painful stimuli varies between individuals and this could be consequence of individual differences to pain sensitivity that may be related to genetic factors. Catechol-O-methyltransferase (COMT) is one of the enzymes that metabolize catecholamine neurotransmitters. Differences in the activity of COMT influence the functions of these neurotransmitters. A single nucleotide polymorphism (Val158Met) of COMT leads to a three to four fold reduction in the activity of the enzyme and has been associated to modifications in the response to a pain stressor. Neuropathic pain is a progressive nervous system disease due to an alteration of the peripheral or central nervous system. To elucidate the possible role of COMT polymorphism in the susceptibility to neuropathic pain, we have performed a case-control study in a Spanish population. Analysis of the (Val158Met) COMT polymorphism was performed by PCR amplification and DNA digestion with restriction enzymes. Our study concludes that functional Val158Met polymorphism of COMT gene is not associated to increased susceptibility to neuropathic pain.

Case-Control Studies↗

PubMed2004

A highly toxic morphine-3-glucuronide derivative.

By the coupling of octylamine to the uronic acid function of morphine-3-glucuronide (M3G) a new glycoconjugate (morphine-3-octylglucuronamide, M3GOAM) was prepared. When assayed in both rats and mice up to ng/kg (i.p.) doses none of the animals survived. The aliphatic octyl chain may be the lethal factor since a closely related derivative (M3GNH2), was not toxic and showed similar opioid antagonist properties than naloxone.

Animals↗

PubMed2003

Identification of two proopiomelanocortin genes in zebrafish (Danio rerio).

Characterization of a newly cloned proopiomelanocortin (POMC) gene in the teleost zebrafish, Danio rerio, is reported. This gene is formed by three exons and two introns, and its complete cDNA codes for a polypeptide of 222 amino acids. Zebrafish proopiomelanocortin (zfPOMC) contains the consensus sequences for ACTH, gamma-LPH, beta-MSH and beta-endorphin (beta-END). RT-PCR expression studies indicate that zfPOMC is selectively expressed in nervous tissue and in the pituitary gland. An homologous sequence to zfPOMC in the zebrafish genome is also presented. It is possible that this sequence represents part of a duplicate POMC, which might have appeared as a result of an extra genome duplication that have taken place in the cyprinidae family or even in all teleosts. Comparisons between the two zebrafish beta-endorphins and among these peptides and its homologues in other species are also presented.

Adrenocorticotropic Hormone↗

PubMed2003

Differential expression and cellular localization of somatolactin-1 and -2 during early development in the gilthead sea bream.

The patterns of expression of the somatolactin 1 and 2 (SL1 and SL2) transcripts were studied during the early development of the gilthead sea bream (Sparus aurata). Gene expression of SL1 and SL2 were detected in embryos and in larvae, although both transcripts presented different levels of expression. The SL1 transcripts in contrast to the SL2 transcripts presented high expression levels in embryos and younger larvae. Moreover, the SL2 transcripts were slightly present or absence in embryonic stage and the newly hatched larvae, respectively. The differences in the expression levels of SL1 and SL2 in embryos and larvae may be due to the fact that two distinct genes express both isoforms of the protein. Thus, both SLs may play different physiological roles throughout development. Moreover, the hybridization signals for SL1- and SL2-mRNAs were detected in 4-day-old larvae. Both in larvae and adults the somatolactotroph cells co-expressed both transcripts of SL and were located bordering the neurohypophysis in the pars intermedia.

Aging↗