Metabolic insights from a GHSR-A203E mutant mouse model.
JavierSeptember 20, 20200 Comments
Ghrelin binding to its receptor, growth hormone secretagogue receptor (GHSR) stimulates GH release, induce eating, and increase blood glucose. This process can also be influenced by the constitutive activity of GHSR (ghrelin-independent), as suggested by the findings of short stature in people with naturally occurring mutations GHSR-A204E and reduces food intake and blood glucose in rats given GHSR inverse agonists both harmful GHSR constitutive activity.
Here, we aim to further define the physiological relevance of constitutive GHSR activity.We GHSR generated mice with the mutation that replaces alanine at position 203 with glutamate (GHSR-A203E), in accordance with the A204E mutation-human GHSR above-referenced, and using them to perform ex vivo neuronal electrophysiology and in vivo assessment of metabolism. Also, we measure the signal in COS-7 and HEK293T cells transfected with wild-type GHSR (GHSR-WT) or construction GHSR-A203E.
RESULTS
In COS-7 cells, GHSR-A203E IP3 accumulation resulting lower base than GHSR-WT; ghrelin induced IP3 accumulation was observed for both construction. In HEK293T cells co-transfected with the voltage-gated CaVVVOur the data indicate that the A203E mutation ablates GHSR constitutive activity and constitutive activity GHSR original contribution to the conductance depolarizing GHSR-express NPY curve.
Although the A203E mutation does not block the ghrelin-generates a signal that is assessed using in vitro and ex vivo models, rat-A203E GHSR acute food shortages typically respond to ghrelin intake given in vivo. Also, GHSR-A203E mutation blunts GH release, and, in aged mice, leading to a decrease in body length and femur length, which is consistent with short stature human carrier GHSR-A204E mutation.
Metabolic insights from a GHSR-A203E mutant mouse model.
Effects of Alcohol and cocaine in Mutant Rat Model predisposition Post-Traumatic Stress Disorder
Comorbidity between substance abuse and post-traumatic stress disorder (PTSD), a stress-related dysregulation of fear responses, very high. While some drugs known to increase fear and anxiety, there are few data regarding the interaction between voluntary consumption of drugs and the fear memory.
Either spontaneous chronic consumption of alcohol or cocaine under 3 weeks of free choice progressive paradigm alcohol (3/6/10%) or cocaine (0.2 / 0.4 / 0.6 mg / ml), assessed in VGV transgenic mice, have full 5-HT2C receptor editing and displaying PTSD-like behaviors. The consequences of the consumption of these drugs on the potential contextual and cued fear conditioning response of VGV mice was assessed. Drug effects on hippocampal neurotrophic factor derived from the brain (BDNF) mRNA expression was measured as previously was found to decrease in VGV mice.
Chronic alcohol consumption was similar in WT and VGV mice. Under conditions of alcohol, the acquisition of fear is no different at the end of the learning session and cue-facilitated fear extinction. Regarding cocaine, in contrast to WT mice, rats VGV not improve their drug consumption along with increased dosage, effects that may be associated with stimulation of discrimination was increased through an increase in 5-HT2C receptor.
Total Protein - Human Adult Normal Tissue: Bladder
Description: Bladder cancer tissue array with normal bladder tissue, including pathology grade, TNM and clinical stage, 48 cases/48 cores, replaced by BL481c
Description: Bladder tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
cDNA - Human Adult Normal Tissue: Heart: Ventricle (left)
Description: Our tissue products are produced by strictly following the IRB ethical standards and procedures and from highest quality tissues. Immediately after collection the tissues are placed in liquid nitrogen and examined by certified pathologists. The thickness of each individual section is ~5um. They are Hematoxylin and Eosin stained and quality tested by immunostaining with anti-beta-actin antibodies. Our tissue products are suitable for various studies on cellular level (RNA localization, Protein expression, etc.) on both normal and pathological cases. It is also an excellent control and educational tool.
Description: Bladder cancer tissue array, set 1, containing 24 cases in 48 cores, with normal paired with tumor tissues from the same patients, with grading and TNM staging data.
Matched Pair (Normal and Carcinoma) Human Bladder Tissue Array (5 slides/pk)
Description: Gall bladder cancer with matched cancer adjacent normal tissue array, including pathology grade, TNM and clinical stage, 20 cases/46 cores (core size 1.5mm),replacing GA461
Cocaine-intake VGV mice do not display contextual generalization fear VGV usually observed in control rats. In addition, BDNF expression was upregulated after alcohol intake or chronic cocaine either. Overall, these results indicate that both chronic alcohol and cocaine consumption may exert some voluntary oral therapeutic effect in models like the mutants PTSD predisposition.
