It is well known that animals exposed to stressful stimuli during their early life develop different neurological disorders when they become adults. In this study, we evaluated the effect of acute cold stress on gamma-aminobutyric acid (GABA) and L-Serine (L-Ser) transporters in vitro, using the uptake of [(3)H]-GABA and [(3)H]L-Ser by synaptosomes-enriched fractions isolated from rat cerebral cortex during postnatal development. GABA and L-Ser uptake studies in vitro will be used in this investigation as a colateral evidence of changes in the expression of transporters of GABA and L-Ser. We observed that the maximum velocity (V (max)) in L-Ser and GABA uptake after stress session increased in all stages studied. In contrast, K (m) values of L-Ser uptake enhancent in almost age calculated, excluding at PD21 after cold stress during development, at the same time as K (m) (uptake affinity) values of GABA increased in just about age considered but not at PD5 compared with the control group. Finally we investigated the mechanism by which cells regulate the substrate affinity of L-Ser and GABA transporters. We demonstrated a significantly increase in total PKC activity to PD5 from PD21. Pretreatment with PKC inhibitor: staurosporine (SP) led to a restoration of control uptake in several postnatal-days suggesting a relationship between amino acids system and PKC activation. These findings suggest that a single exposure to postnatal cold stress at different periods after birth modifies both GABA and L-Ser transporters and the related increase in total PKC activity could be intracellular events that participate in neuronal plasticity by early life stress, which could be relevant to function of transporters in the adult rat brain.
Postnatal stress alters stress responses for life, with serious consequences on the central nervous system (CNS), involving glutamatergic neurotransmission and development of voluntary alcohol intake. Several drugs of abuse, including alcohol and cocaine, alter glutamate transport (GluT). Here, we evaluated effects of chronic postnatal stress (CPS) on alcohol intake and brain glutamate uptake and transporters in male adolescent Wistar rats. For CPS from postnatal day (PD) 7, pups were separated from their mothers and exposed to cold stress (4 °C) for 1 h daily for 20 days; controls remained with their mothers. Then they were exposed to either voluntary ethanol (6%) or dextrose (1%) intake for 7 days (5-7 rats per group), then killed. CPS: (1) increased voluntary ethanol intake, (2) did not affect body weight gain or produce signs of toxicity with alcohol exposure, (3) increased glutamate uptake by hippocampal synaptosomes in vitro and (4) reduced protein levels (Western measurements) in hippocampus and frontal cortex of glial glutamate transporter-1 (GLT-1) and excitatory amino-acid transporter-3 (EAAT-3) but increased glutamate aspartate transporter (GLAST) levels. We propose that CPS-induced decrements in GLT-1 and EAAT-3 expression levels are opposed by activation of a compensatory mechanism to prevent excitotoxicity. A greater role for GLAST in total glutamate uptake to prevent enlarged extracellular glutamate levels is inferred. Although CPS strongly increased intake of ethanol, this had little impact on effects of CPS on brain glutamate uptake or transporters. However, the impact of early life adverse events on glutamatergic neurotransmission may underlie increased alcohol consumption in adulthood.
<p>En los últimos años se está dando una importancia creciente a las normas sobre bienestar animal debido a la confluencia de varios factores, como el mayor conocimiento en distintas disciplinas relacionadas con los animales de producción (el comportamiento animal, la fisiología del estrés y el manejo correcto de los animales), la relación directa entre estos conocimientos y los niveles de producción estables y competitivos a mediano y largo plazo y una mayor concientización social sobre las necesidades de los animales, así como el rechazo hacia los abusos De esta forma, una vez superadas las necesidades de incremento en el abastecimiento de productos, se han empezado a fijar otros parámetros para cumplir con las demandas sociales en el ámbito de la producción ganadera, uno de esos parámetros, cada día más importante, es el bienestar animal. En esta revisión se intenta destacar la importancia del bienestar de los animales, tanto por la calidad de vida de los mismos, como por su impacto sobre la producción ganadera. Se exponen los principales factores generadores de estrés en ganado, así como los efectos fisiológicos y sus impactos negativos sobre la producción. En vías de brindar una mejor vida al animal y aumentar la calidad del producto, cada vez más países y consumidores imponen exigencias legales y reglamentarias que determinan estándares de bienestar para el manejo animal. Es por todo esto que el bienestar animal adquiere cada vez mayor relevancia en todo el mundo. América Latina se encuentra actualmente atravesando un proceso de adaptación a los nuevos requerimientos internacionales.</p>
Serological assays, including enzyme-linked immunosorbent assays (ELISA), provide an useful tool for screening animals for the presence of antibodies (Abs) against a wide range of infectious agents (including viruses that cause respiratory disease in cattle) and are mainly used in veterinary medicine to assist to the control and disease's monitoring. The aim of the present study was developing and validating one indirect enzyme-linked immunosorbent assay (ELISA) based on semi purified bovine parainfluenza virus type 3 (BPIV3).This test would allow to detect and quantify Abs against PI3 in serum sample from cattle and guinea pigs on both purposes diagnostic and typify/specify the quality of vaccines. The diagnostic sensitivity and specificity from the assay was 88% and 100% for bovine samples, using a threshold of corrected optical density, ODc =0.300, and 91% and 100% for guinea pig samples with a ODc =0.250.The intermediate precision expressed as the assays positive control coefficient of variation (CV) was 20% for bovines and 8.5% for guinea pigs. Both techniques reproducibility obtained in interlaboratory assays was CV=17% for bovines and 15% for guinea pigs, which found the requirements of OIE (CV<30%). The efficacy of biological medicinal products, such as vaccines, relies an optimal model testing quality control. The validated ELISAs represents an important tool for testing vaccine quality, and quantifying and controling BPIV3 infections on cattle.
Background: There is increasing evidence that environmental factors, particularly stressful events experienced early in life, increase the risk of developing a psychiatric illness and/or a behavioural disorder. The aim of this study was to evaluate the effects of acute and chronic maternal separation (AMS and CMS) plus cold stress on the expression patterns of Glutamate Transporters (TGlus) in the developing and young adult Central Nervous System (CNS). As regulation of Glutamate (Glu) extracellular levels is of key importance, sodium-dependent Glu uptake using synaptosome-enriched fractions isolated from Frontal Cortex (FC) and Hippocampus (Hic) was also studied.
Serological assays, including enzyme-linked immunosorbent assays (ELISA), provide an useful tool for screening animals for the presence of antibodies (Abs) against a wide range of infectious agents (including viruses that cause respiratory disease in cattle) and are mainly used in veterinary medicine to assist to the control and disease's monitoring. The aim of the present study was developing and validating one indirect enzyme-linked immunosorbent assay (ELISA) based on semi purified bovine parainfluenza virus type 3 (BPIV3).This test would allow to detect and quantify Abs against PI3 in serum sample from cattle and guinea pigs on both purposes diagnostic and typify/specify the quality of vaccines. The diagnostic sensitivity and specificity from the assay was 88% and 100% for bovine samples, using a threshold of corrected optical density, ODc =0.300, and 91% and 100% for guinea pig samples with a ODc =0.250.The intermediate precision expressed as the assays positive control coefficient of variation (CV) was 20% for bovines and 8.5% for guinea pigs. Both techniques reproducibility obtained in interlaboratory assays was CV=17% for bovines and 15% for guinea pigs, which found the requirements of OIE (CV<30%). The efficacy of biological medicinal products, such as vaccines, relies an optimal model testing quality control. The validated ELISAs represents an important tool for testing vaccine quality, and quantifying and controling BPIV3 infections on cattle.
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