Biomedical subjects

Aurea Orozco

Publications and source records attributed to Aurea Orozco.

6 recordsLinked to original sources

PubMed2005

Thyroid hormone deiodination in fish.

We review the experimental evidence accumulated within the past decade regarding the physiologic, biochemical, and molecular characterization of iodothyronine deiodinases (IDs) in piscine species. Agnathans, chondrichthyes, and teleosts express the three isotypes of IDs: ID1, ID2, and ID3, which are responsible for the peripheral fine-tuning of thyroid hormone (TH) bioactivity. At the molecular and operational level, fish IDs share properties with their corresponding vertebrate counterparts. However, fish IDs also exhibit discrete features that seem to be distinctive for piscine species. Indeed, teleostean ID1 is conspicuously resistant to propylthiouracil (PTU) inhibition, and its response to thyroidal status differs from that exhibited by other ID1s. Moreover, both the high level of ID2 activity and its expression in the liver of teleosts are unique among vertebrates. The physiologic role of iodothyronine deiodination in functions regulated by TH in fish is not entirely clear. Nevertheless, current experimental evidence suggests that IDs may coordinate and facilitate, in a tissue-specific fashion, the action of iodothyronines and other hormones involved in such processes.

Animals↗

PubMed2004

Effects of iodothyronines on the hepatic outer-ring deiodinating pathway in killifish.

Substrate availability has been thought to be a major regulator of the outer-ring deiodinating pathway (ORD) in fish. However, current information strongly suggests that while fish iodothyronine deiodinase type 2 (D2) responds to iodothyronines in the same manner as its mammalian counterpart, fish deiodinase type 1 (D1) exhibits a distinct response. Furthermore, 3,5-T2, generally considered to be an inactive product of iodothyronine metabolism, has recently been described as bioactive, but its effects upon D1 and D2 are not yet known. We examined the effect that short-term immersion in T4, T3, and 3,5-T2 (0.1 microM; 12 or 24 h) exerts on both D1 and D2 activities and on the levels of expression of D1 and D2 mRNAs in killifish liver. In agreement with previous reports in teleosts, no iodothyronine exerted a significant effect on D1 enzymatic activity. However, all three iodothyronines significantly decreased D2 activity. Furthermore, at 24 h post-immersion T4, T3, and 3,5-T2 inhibited both D1 and D2 transcription. Together, the present results confirm the differential effect of iodothyronines upon the hepatic ORD pathway in fish and show that this effect can occur at a transcriptional level. Furthermore, we provide the first evidence that 3,5-T2 can affect both activity and transcription of hepatic deiodinases in teleosts.

Animals↗

PubMed2004

Halometabolites and cellular dehalogenase systems: an evolutionary perspective.

We review the role of iodothyronine deiodinases (IDs) in the evolution of vertebrate thyroidal systems within the larger context of biological metabolism of halogens. Since the beginning of life, the ubiquity of organohalogens in the biosphere has provided a major selective pressure for the evolution and conservation of cellular mechanisms specialized in halogen metabolism. Among naturally available halogens, iodine emerged as a critical component of unique developmental and metabolic messengers. Metabolism of iodinated compounds occurs in the three major domains of life, and invertebrate deuterostomes possess several biochemical traits and molecular homologs of vertebrate thyroidal systems, including ancestral homologs of IDs identified in urochordates. The finely tuned cellular regulation of iodometabolite uptake and disposal is a remarkable event in evolution and might have been decisive for the explosive diversification of ontogenetic strategies in vertebrates.

Animals↗

PubMed2003

The liver of Fundulus heteroclitus expresses deiodinase type 1 mRNA.

The presence of a type 1 deiodinase (D1) in the liver of teleosts has been a controversial issue. Recently we characterized the deiodinase activity in rainbow trout and killifish liver and found that the liver of both species co-expresses the two enzymes (D1 and D2) that catalyze the outer ring-deiodinating pathway. We here report the cloning and characterization of an mRNA from the liver of the killifish Fundulus heteroclitus that encodes a D1 (FhD1). The cDNA amplified by RT-PCR from F. heteroclitus liver is 1314 nt long and encodes a protein of 248 aa. It contains a TGA codon in its open reading frame and a selenocysteine insertion sequence in its 3(') untranslated region, consistent with the structure of a selenoenzyme mRNA. The deduced peptide sequence is 73% identical to that encoded by the tilapia D1 cDNA cloned from kidney and 46% identical to the D1s reported in other vertebrates. Northern blot analysis shows that FhD1 mRNA is expressed in F. heteroclitus liver, consistent with prior biochemical evidence for hepatic D1 activity. Furthermore, heterologous expression of the FhD1 cDNA resulted in a protein with properties similar to the D1-like activity in F. heteroclitus liver. The cloned enzyme, like the native species, is relatively insensitive to inhibition by PTU, but mutation of Ser-159 in FhD1 to the Pro residue found in D2 and D3 isoforms increased the sensitivity to PTU. Our results show that, under basal conditions, killifish liver indeed expresses a D1 enzyme that is homologous to mammalian D1s, establishing this as a useful model in which to study the regulation of D1 and D2 concurrently.

Amino Acid Sequence↗

PubMed2002

Cloning of the gene and complete cDNA encoding a type 2 deiodinase from Fundulus heteroclitus.

Recently, we reported the cloning of a cDNA fragment from Fundulus heteroclitus liver encoding the open reading frame of type 2 deiodinase (FhD2). We here report the cloning of 14 kb of genomic sequence from F. heteroclitus that includes the previously reported coding region of the F. heteroclitus Dio2 gene (FhDio2), the 5(') and 3(') untranslated regions, and flanking regions and introns. This FhDio2 gene comprises two exons divided by a 4.8-kb intron. The position of the intron is similar to that of introns in other Dio2 genes. The analysis of approximately 1.3 kb of genomic sequence upstream of the mRNA start site revealed that, in contrast to mammalian Dio2 genes, there were no apparent TATA or CRE sequences. Nevertheless, a putative Sp1 site was found, similar to that in other F. heteroclitus TATA-less promoters. We have also cloned the complete FhD2 cDNA, which spans 4652 bp and contains a sequence adjacent to its poly(A) tail that is highly similar to the selenocysteine insertion sequence (SECIS) found in human D2 cDNA. The expression of a construct containing the FhD2 ORF plus the native SECIS resulted in a protein with deiodinase activity similar to that of the native FhD2. Analysis of the regulation of this gene, combined with ongoing studies of the F. heteroclitus D1 gene, will allow us to elucidate the functions of the colocalized deiodinases in teleost liver.

Animals↗

PubMed2002

Environmental salinity selectively modifies the outer-ring deiodinating activity of liver, kidney and gill in the rainbow trout.

We here analyzed the effect of a mild hyperosmotic challenge on the activities of deiodinases type I (D1) and II (D2) in the trout liver, and D1 in kidney and gill, two organs involved in osmoregulation. FW-adapted immature rainbow trout were transferred to 5 per thousand SW and killed 0.5, 1, 2, 4, 8 12, 24 and 48 h post-transfer (PT). Fish maintained in FW served as controls. Hepatic, renal and branchial D1 and hepatic D2 activities were assessed as well as circulating levels of T(3), T(4) and cortisol. Hyperosmotic challenge elicited significant and sustained decreases in kidney D1 and liver D2 activities at 8 h PT, which returned to control values at 48 h PT. In contrast, liver and gill D1 activities exhibited no significant change throughout the study. Also, significant increases in circulating T(4) at 2-4 and 48 h PT were observed. Circulating T(3) remained unmodified until 24-48 h PT, when it rose sharply. Simultaneously, cortisol showed a trend towards increase during the initial 4 h PT, which attained significance at 48 h PT. The present findings demonstrate that a mild hypertonic challenge is sufficient to elicit responses in the trout thyroidal axis. Hormonal changes in the circulatory compartment are in accordance with those previously described for migratory salmonids. A novel aspect of our findings is the organ-specific differential response exhibited by ORD-enzymes when trout are exposed to a mildly different osmotic environment. Our findings further establish the uniqueness of fish thyroid physiology, and can be of value in further understanding the evolutionary aspects of this ORD family of deiodinases.

Animals↗