Serotonin- and Dopamine-Related Gene Expression in db/db Mice Islets and in MIN6 ß-Cells Treated with Palmitate and Oleate.

High circulating nonesterified fatty acids (NEFAs) concentration, often reported in diabetes, leads to impaired glucose-stimulated insulin secretion (GSIS) through not yet well-defined mechanisms. Serotonin and dopamine might contribute to NEFA-dependent ß-cell dysfunction, since extracellular signal of these monoamines decreases GSIS. Moreover, palmitate-treated ß-cells may enhance the expression of the serotonin receptor Htr2c, affecting insulin secretion. Additionally, the expression of monoamine-oxidase type B (Maob) seems to be lower in islets from humans and mice with diabetes compared to nondiabetic islets, which may lead to increased monoamine concentrations. We assessed the expression of serotonin- and dopamine-related genes in islets from db/db and wild-type (WT) mice. In addition, the effect of palmitate and oleate on the expression of such genes, 5HT content, and GSIS in MIN6 ß-cell was determined. Lower Maob expression was found in islets from db/db versus WT mice and in MIN6 ß-cells in response to palmitate and oleate treatment compared to vehicle. Reduced 5HT content and impaired GSIS in response to palmitate (-25%; p < 0.0001) and oleate (-43%; p < 0.0001) were detected in MIN6 ß-cells. In conclusion, known defects of GSIS in islets from db/db mice and MIN6 ß-cells treated with NEFAs are accompanied by reduced Maob expression and reduced 5HT content.

Fluoxetine impairs insulin secretion without modifying extracellular serotonin levels in MIN6 ß-cells.

INTRODUCTION: Pancreatic ß-cells synthetize and store Serotonin (5-Hydroxytriptamine, 5HT) which is co-released with insulin. It has been proposed that extracellular 5HT binds to specific cell surface receptors and modulate insulin secretion. On the other hand, Selective Serotonin Reuptake Inhibitor (SSRI) fluoxetine seems to reduce Glucose-Stimulated Insulin Secretion (GSIS). However, it is unknown whether this effect results from changes in extracellular 5HT concentration owed to the blockade of 5HT transporter (SERT) or from non-5HT dependent actions. The aims of this work were: 1) to quantify extracellular 5HT levels and GSIS in ß-cell lines, 2) to determine whether extracellular 5HT levels and GSIS are changed by fluoxetine or 5-Hydroxytryptophan (5HTP, the immediate 5HT biosynthetic precursor), and 3) to quantify the expression of Slc6a4 gene (encoding SERT) in ß-cell lines in relation to other genes involved in 5HT system. MATERIAL AND METHODS: ß-cell lines MIN6 and RINm5f were subjected to GSIS protocols, after treatment with fluoxetine, 5HTP or 5HT. Insulin and 5HT were quantified by ELISA and HPLC, respectively. Relative mRNA expression was quantified by RT-qPCR. RESULTS: MIN6 ß-cells secretes 5HT in response to glucose, showing a sharp increase in 5HT release when cells were preloaded with 5HTP. Treatment with 5HT or fluoxetine reduces GSIS. Fluoxetine fails to further increases 5HTP-induced elevation of secreted 5HT. MIN6 ß-cells express both isoforms of Tryptophan Hydroxylase (Tph1 and Tph2), and have high expression levels of L-Dopa decarboxylase (Ddc), both enzymes involved in 5HT biosynthetic pathway, but do not express the 5HT transporters Slc6a4 or Slc6a3 (the Dopamine-5HT transporter) genes. CONCLUSION: The inhibitory effect of fluoxetine on ß-cell glucose stimulated insulin secretion is not mediated by blockage of 5HT transporter through SERT.

Teste de cornea e exame do humor aquoso no diagnostico da raiva natural em caes. / Cornea test and examination of the aqueous humor in the natural rabies diagnosis in dogs

O teste de cornea foi comparado ao exame do humor aquoso por inoculacao em camundongos para o diagnostico da raiva natural em 30 caes. Os dois testes foram positivos em 66,66% dos casos estudados e negativos em 3,33% deles. Nos demais pares de amostras, os resultados foram positivos apenas em um, 13,33% e somente no outro teste, 16,66%. Em relacao ao total de resultados positivos, o teste de cornea (80%) foi comparavel ao exame do humor aquoso por inoculacao em camundongos (83,33%). A associacao dos dois metodos aumentou a taxa de positividade do diagnostico para 96,66%

Diagnostico da raiva em caes a partir da saliva por imunofluorescencia direta. / Direct immunofluorescence rabies diagnosis from dog saliva samples

Um total de 30 amostras de saliva obtidas de 30 caes raivosos foi examinado para o diagnostico da raiva por imunofluorescencia direta. A identificacao de estruturas corpusculares e puntiformes fluorescentes em 134 (55,83%) das 240 impressoes examinadas foi satisfatoria para concluir o diagnostico em 29 (96,66%) dos 30 caes estudados. Uma amostra negativa por imunofluorescencia direta (3,33%) e positiva por inoculacao em camundongo sugere o uso associado das duas tecnicas para o exame da saliva. A possibilidade para o diagnostico precoce da raiva foi observada na maioria dos casos