Chronic Organic Magnesium Supplementation Enhances Tissue-Specific Bioavailability and Functional Capacity in Rats: A Focus on Brain, Muscle, and Vascular Health methods
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Materials and Methods
Thirty-eight adult outbred Sprague Dawley rats (Dokuz Eylul University School of Medicine, Experimental Animal Laboratory, Izmir, Turkey) were used in this study. All rats were housed in individual cages with free access to water and standard laboratory chow (Optima rodent 20, Bolu, Turkey). They were kept in a 12-h light/12-h dark cycle at constant room temperature (22 ± 1 °C) and humidity (60%). Dokuz Eylul University School of Medicine, Animal Care Committee, with 36/2018 protocol number, approved all experimental procedures.
Materials and Methods
Mg-citrate ( n = 10): 35.4 mg/kg/day.
Materials and Methods
Mg-glycinate ( n = 10): 35.4 mg/kg/day.
Materials and Methods
Mg-malate ( n = 10): 35.4 mg/kg/day.
Morris Water Maze Test
Spatial learning and memory performance were tested in the Morris water maze. The Morris water tank is a black circular pool with a diameter of 140 cm and a depth of 75 cm, made of non-reflective material. The tank is filled with 22 °C water up to 50-cm level, and a cylindrical hidden platform with a diameter of 10 cm is placed hidden below the water level. Various colored and illuminated objects were placed around the environment where the pool was located to be used as clues. The Morris water tank was divided into four equal quadrants, southeast (SE), southwest (SW), northeast (NE), and northwest (NW), by two lines that intersect each other right in the middle of the tank, forming a cross shape. The points where the plus shape cut the tank's walls were determined as north, south, east, and west. The platform was placed in the southwest quadrant of the tank, 1-2 cm b...
Tissue Extraction and Homogenization of Tissues
Rats were gone under CO 2 via cardiac puncture to achieve exsanguination before tissue harvesting. After exsanguination, all tissues (brain and regions such as hippocampus, prefrontal cortex, amygdala, uterus, aort, gastrocnemius muscle, and soleus muscle) were dissected. After the animals were euthanized via CO₂ inhalation followed by cardiac puncture to ensure exsanguination, dissection was initiated immediately to prevent post-mortem degradation. All procedures were performed under sterile conditions using autoclaved surgical instruments. First, the thoracic cavity was opened to extract the heart and aorta, followed by carefully separating the descending thoracic aorta using micro-scissors and forceps. Subsequently, the abdominal cavity was exposed to isolate skeletal muscles: gastrocnemius and soleus muscles were bilaterally excised. For brain extraction, a midline scalp inc...
Behavioral and Emotional Consequences: Anxiolytic Potential of Mg
The reduced thigmotaxis duration in Mg-treated groups during the open-field test, alongside increased central area activity in the Mg-glycinate group, demonstrates that chronic Mg supplementation modulates anxiety-related behavioral responses. These findings align with prior evidence indicating Mg's ability to attenuate stress responses via suppression of the hypothalamic-pituitary-adrenal (HPA) axis [,,, ]. However, the absence of significant differences in the elevated plus maze test implies that Mg's anxiolytic effects may be context- or stressor-dependent, highlighting the importance of methodological variables in interpreting anxiety-related outcomes.
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Thirty-eight adult outbred Sprague Dawley rats (Dokuz Eylul University School of Medicine, Experimental Animal Laboratory, Izmir, Turkey) were used in this study. All rats were housed in individual cages with free access to water and standard laboratory chow (Optima rodent 20, Bolu, Turkey). They were kept in a 12-h light/12-h dark cycle at constant room temperature (22 ± 1 °C) and humidity (60%). Dokuz Eylul University School of Medicine, Animal Care Committee, with 36/2018 protocol number, approved all experimental procedures.
Mg-citrate ( n = 10): 35.4 mg/kg/day.
Mg-glycinate ( n = 10): 35.4 mg/kg/day.
Mg-malate ( n = 10): 35.4 mg/kg/day.
Spatial learning and memory performance were tested in the Morris water maze. The Morris water tank is a black circular pool with a diameter of 140 cm and a depth of 75 cm, made of non-reflective material. The tank is filled with 22 °C water up to 50-cm level, and a cylindrical hidden platform with a diameter of 10 cm is placed hidden below the water level. Various colored and illuminated objects were placed around the environment where the pool was located to be used as clues. The Morris water tank was divided into four equal quadrants, southeast (SE), southwest (SW), northeast (NE), and northwest (NW), by two lines that intersect each other right in the middle of the tank, forming a cross shape. The points where the plus shape cut the tank's walls were determined as north, south, east, and west. The platform was placed in the southwest quadrant of the tank, 1-2 cm below the water surface. Its position was not changed during the learning trials. The subjects were placed into the water from a different point each day, facing the tank's walls. After placing the animal into the pool, the subject was expected to find the platform within 60 s. If the subject c...
Rats were gone under CO 2 via cardiac puncture to achieve exsanguination before tissue harvesting. After exsanguination, all tissues (brain and regions such as hippocampus, prefrontal cortex, amygdala, uterus, aort, gastrocnemius muscle, and soleus muscle) were dissected. After the animals were euthanized via CO₂ inhalation followed by cardiac puncture to ensure exsanguination, dissection was initiated immediately to prevent post-mortem degradation. All procedures were performed under sterile conditions using autoclaved surgical instruments. First, the thoracic cavity was opened to extract the heart and aorta, followed by carefully separating the descending thoracic aorta using micro-scissors and forceps. Subsequently, the abdominal cavity was exposed to isolate skeletal muscles: gastrocnemius and soleus muscles were bilaterally excised. For brain extraction, a midline scalp incision was made, and the skull was opened using rongeurs. The brain was gently removed en bloc and placed on a chilled dissection platform. The whole brain was sectioned coronally to isolate the hippocampus, prefrontal cortex, and amygdala using fine micro-dissecting tools, guided by a rat brain atla...
The reduced thigmotaxis duration in Mg-treated groups during the open-field test, alongside increased central area activity in the Mg-glycinate group, demonstrates that chronic Mg supplementation modulates anxiety-related behavioral responses. These findings align with prior evidence indicating Mg's ability to attenuate stress responses via suppression of the hypothalamic-pituitary-adrenal (HPA) axis [,,, ]. However, the absence of significant differences in the elevated plus maze test implies that Mg's anxiolytic effects may be context- or stressor-dependent, highlighting the importance of methodological variables in interpreting anxiety-related outcomes.
Machine-readable layer
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