RESUMO
Recurrent panic attacks (PAs) are a common feature of panic disorder (PD) and post-traumatic stress disorder (PTSD). Several distinct brain regions are involved in the regulation of panic responses, such as perifornical hypothalamus (PeF), periaqueductal gray, amygdala and frontal cortex. We have previously shown that inhibition of GABA synthesis in the PeF produces panic-vulnerable rats. Here, we investigate the mechanisms by which a panic-vulnerable state could lead to persistent fear. We first show that optogenetic activation of glutamatergic terminals from the PeF to the basolateral amygdala (BLA) enhanced the acquisition, delayed the extinction and induced the persistence of fear responses 3 weeks later, confirming a functional PeF-amygdala pathway involved in fear learning. Similar to optogenetic activation of PeF, panic-prone rats also exhibited delayed extinction. Next, we demonstrate that panic-prone rats had altered inhibitory and enhanced excitatory synaptic transmission of the principal neurons, and reduced protein levels of metabotropic glutamate type 2 receptor (mGluR2) in the BLA. Application of an mGluR2-positive allosteric modulator (PAM) reduced glutamate neurotransmission in the BLA slices from panic-prone rats. Treating panic-prone rats with mGluR2 PAM blocked sodium lactate (NaLac)-induced panic responses and normalized fear extinction deficits. Finally, in a subset of patients with comorbid PD, treatment with mGluR2 PAM resulted in complete remission of panic symptoms. These data demonstrate that a panic-prone state leads to specific reduction in mGluR2 function within the amygdala network and facilitates fear, and mGluR2 PAMs could be a targeted treatment for panic symptoms in PD and PTSD patients.
Assuntos
Tonsila do Cerebelo/metabolismo , Medo/fisiologia , Pânico/fisiologia , Animais , Complexo Nuclear Basolateral da Amígdala/metabolismo , Encéfalo/metabolismo , Extinção Psicológica/fisiologia , Lobo Frontal/metabolismo , Ácido Glutâmico/metabolismo , Inibição Psicológica , Masculino , Optogenética/métodos , Ratos , Ratos Sprague-Dawley , Receptores de Glutamato Metabotrópico/metabolismo , Transmissão Sináptica/fisiologiaRESUMO
The objective of this study was to evaluate the effect of ferrous lactate infusion on postruminal P absorption in lactating dairy cows. Four ruminally cannulated lactating cows were used in a 4×4 Latin square design with 14 d per period. Cows were fed a basal diet containing 0.39% P, providing 100% of the calculated P requirement. On d 8 to 14 of each period, each cow was infused with 0, 200, 500, or 1,250mg of Fe/d in the form of ferrous lactate solution (ferrous lactate in 1L of double-distilled water) into the abomasum. Infusate was formulated to approximate 0, 2, 5, or 12.5mg of Fe/L in drinking water with 100L of water intake/d. Total fecal collection was conducted in the last 4 d of each period to measure nutrient digestion and excretion. Dry matter intake, milk yield, and milk composition were not affected by treatment. Digestibility of DM, NDF, and nitrogen decreased linearly with increasing ferrous lactate infusion. Infusion of ferrous lactate did not affect intake and digestibility of total P, inorganic P, or phytate P. In lactating cows, P absorption was not negatively influenced by abomasally infused ferrous lactate up to 1,250mg of Fe/d.
Assuntos
Abomaso/efeitos dos fármacos , Bovinos/metabolismo , Compostos Ferrosos/administração & dosagem , Lactatos/administração & dosagem , Lactação/fisiologia , Fósforo na Dieta/farmacocinética , Absorção/efeitos dos fármacos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Digestão/efeitos dos fármacos , Feminino , Ferro da Dieta/efeitos adversos , Ferro da Dieta/farmacocinética , Leite/química , Fósforo na Dieta/administração & dosagem , Rúmen/metabolismoRESUMO
The composition of water given to dairy cattle is often ignored, yet water is a very important nutrient and plays a major role in milk synthesis. The objective of this study was to study effects of elevated levels of iron in bovine drinking water on milk quality. Ferrous lactate treatments corresponding to 0, 2, 5, and 12.5mg/kg drinking water concentrations were delivered through the abomasum at 10 L/d to 4 lactating dairy cows over 4 periods (1 wk infusion/period) in a Latin square design. On d 6 of infusion, milk was collected, processed (homogenized, pasteurized), and analyzed. Mineral content (Fe, Cu, P, Ca) was measured by inductively coupled plasma mass spectrometry. Oxidative stability of whole processed milk was measured by the thiobarbituric acid reactive substances (TBARS) assay for malondialdehyde (MDA) and sensory analysis (triangle test) within 72 h of processing and after 7d of storage (4°C). Significant sensory differences between processed milks from cows receiving iron and the control infusion were observed. No differences in TBARS (1.46±0.04 mg of MDA/kg) or mineral content (0.22±0.01 mg/kg Fe) were observed. A 2-way interaction (iron treatment by cow) for Ca, Cu, and Fe concentrations was seen. While iron added directly to milk causes changes in oxidation of milk, high levels of iron given to cattle have subtle effects that initially may not be obvious.
Assuntos
Água Potável/química , Ferro/análise , Leite/química , Leite/normas , Animais , Cálcio/análise , Bovinos , Cobre/análise , Relação Dose-Resposta a Droga , Feminino , Compostos Ferrosos/farmacologia , Lactatos/farmacologia , Malondialdeído/análise , Espectrometria de Massas , Fósforo/análise , Substâncias Reativas com Ácido TiobarbitúricoRESUMO
In skeletal muscle, insulin activates glycogen synthase by reducing phosphorylation at both NH2- and COOH-terminal sites of the enzyme and by elevating the levels of glucose-6-phosphate, an allosteric activator of glycogen synthase. To study the mechanism of regulation of glycogen synthase by insulin and glucose-6-phosphate, we generated stable Rat-1 fibroblast clones expressing rabbit muscle glycogen synthase with Ser-->Ala substitutions at key phosphorylation sites. We found that 1) elimination of the phosphorylation of either NH2- or COOH-terminal sites did not abolish insulin stimulation of glycogen synthase; 2) mutations at both Ser-7 and Ser-640 were necessary to bypass insulin activation; 3) mutation at Ser-7, coupled with the disruption of the motif for recognition by glycogen synthase kinase-3 (GSK-3), did not eliminate the insulin effect; and 4) mutation of either Ser-7 or Ser-640 increased the sensitivity of glycogen synthase to glucose 6-phosphate >10-fold. We conclude that Ser-7 and Ser-640 are both involved in mediating the response of glycogen synthase to insulin and activation by glucose 6-phosphate. In Rat-1 fibroblasts, GSK-3 action is not essential for glycogen synthase activation by insulin, and GSK-3-independent mechanisms also operate.
Assuntos
Glucose-6-Fosfato/farmacologia , Glicogênio Sintase/química , Glicogênio Sintase/metabolismo , Insulina/farmacologia , Alanina , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Sítios de Ligação , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Linhagem Celular , Ativação Enzimática , Glicogênio Sintase/efeitos dos fármacos , Quinase 3 da Glicogênio Sintase , Quinases da Glicogênio Sintase , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fosforilação , Mutação Puntual , Coelhos , Ratos , Proteínas Recombinantes/química , Proteínas Recombinantes/efeitos dos fármacos , Proteínas Recombinantes/metabolismo , Serina , TransfecçãoRESUMO
The basolateral amygdala (BL) is a putative site for regulating anxiety, where inhibition and excitation respectively lead to decreases and increases in anxiety-like behaviors. The BL contains local networks of GABAergic interneurons that are subdivided into classes based on neurochemical content, and are hypothesized to regulate unique functional responses of local glutamatergic projection neurons. Recently it was demonstrated that lesioning a portion of the BL interneuronal population, those interneurons that express neurokinin1 receptors (NK(1r)), resulted in anxiety-like behavior. In the current study, these NK(1r) expressing cells of the BL are further phenotypically characterized, demonstrating approximately 80% co-expression with GABA thus confirming them as GABAergic interneurons. These NK(1r) interneurons also colocalize with two distinct populations of BL interneurons as defined by the neuropeptide content. Of the NK(1r) positive cells, 41.8% are also positive for neuropeptide Y (NPY) and 39.7% of the NK(1r) positive cells are also positive for cholecystokinin (CCK). In addition to enhancing the phenotypic characterization, the extent to which the NK(1r) cells of amygdala nuclei contribute to anxiety-like responses was also investigated. Lesioning the NK(1r) expressing interneurons, with a stable form of substance P (SSP; the natural ligand for NK(1r)) coupled to the targeted toxin saporin (SAP), in the anterior and posterior divisions of the BL was correlated to increased anxiety-like behaviors compared to baseline and control treated rats. Furthermore the phenotypic and regional selectivity of the lesions was also confirmed.