Intermittent Fasting Modulates the Glycogen Level in Zebrafish (Danio rerio) and their Next Generation

Introduction: Zebrafish (Danio rerio) has become the best model organism to study the evolutionary biological process and human developmental studies. The liver glycogen plays a vital role in maintaining cellular metabolism, accumulation of glycogen in liver affects the enzymes related to glycogen metabolism. Aim: Impact of intermittent fasting, refeed and overfeeding in glycogen homeostasis on Zebrafish (Danio rerio) and their F1 generation. Materials and Methods: The present in-vivo study demonstrates the effect of intermittent fasting on glycogen storage in zebrafish and their F1 generation. The study was conducted at Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India. The duration of study was carried out for one month (December, 2021) for both parental and their F1 generation (April, 2022) groups. The F1 generation fishes involved after its matured (three months). The zebrafish (AB strain) were randomised and split into five experimental groups such as control, overfed, 12 hours, 24 hours, and 48 hours intermittent fasting. The F1 generation from each group was treated as same as parenting groups. The physiological and histological changes were observed in the study group. Significant results were evaluated as p<0.05 values turkey’s method was used. Results: The fasting and overfeeding significantly affects the physiological condition like body weight, length and Body Mass Index (BMI). The parental control and their F1 have a BMI of 0.042±0.04 g/cm² and 0.041±0.04 g/cm². The maximum fasting treated groups (48 hours) of both parent and their F1 generation shows reduced BMI such as 0.032±0.03 g/cm² and 0.030±0.04 g/ cm². The over feed group shows a BMI of 0.053±0.05 g/cm² and 0.052±0.05 g/cm². The result demonstrates that the food-deprived groups and their F1 generation showed less glycogen storage in histological observation. The refeed and overfed groups and their F1 generation exhibit more glycogen accumulation in the liver. The result confers normal regulation of glycogen synthase and glycogen synthase kinase 3 in normally in control and fasting groups as well as in their F1 generation. Conversely, the overfeeding and refeed groups show modulated glycogen activity in both parent and their F1 generation. Conclusion: Glycogen accumulation leads too many diseases and it also affects the generations. The frequent fasting may help to minimise glycogen accumulation and BMI level reduces the complications of disorders related to glycogen homeostasis

Glutamate Elicits Therapeutic Responses in Light-Induced Sleep-Deprived Zebrafish, Danio rerio

Sleep deprivation disrupts most neurotransmitters, which can lead to adverse behavioural changes and other psychiatric illnesses. Many neurotransmitter systems, including dopamine (DA), serotonin (5-HT), norepinephrine (N.E.) and GABA, have been implicated in the pathophysiology of mood disorders. The precise significance of sleep deprivation (S.D.) changes in the neurotransmitter levels and the mechanism underlying behavioural alterations is unknown. According to research, sleep deprivation (S.D.) has a major effect on an individual’s quality of life and ability to perform essential physiological functions. As a result, we wanted to confirm the levels of neurotransmitters and behavioural modifications in zebrafish after 24, 48, and 72 hours of sleep deprivation and glutamate treatment on the sleep-deprived groups. The T-maze test was used to assess learning and memory alterations in zebrafish. We used the Novel Tank Test (NTT) and Light and Dark Test (LDT) to examine the anxiety-like behaviour. The spectrofluorimetric method was used to determine the quantities of DA, 5-HT, N.E. and GABA. From this study, it is evident that 72h sleep-deprived fish had a loss of learning and memory via T-maze test and also the anxiety levels were very high in the sleep-deprived group than the other groups. The groups that received glutamate after sleep deprivation showed betterment in the behavioural response. Also, the levels of neurotransmitters were increased in the glutamate treated groups than the sleep-deprived groups. Our findings indicate that sleep loss dramatically impairs behavioural responses and disrupts most neurotransmitter concentrations. When sleep-deprived fish were given glutamate, their behaviour and neurotransmitter levels were nearly identical to those of the control group. This study will have a greater impact on sleep deprivation therapy and pave the way for using the neurotransmitters as external therapeutic agents in treating sleep deprivation and other behavioural changes related to sleep deprivation.It has been suggested that zebrafish is an excellent testing subject for loss of sleep on cognition and that it may also be an efficient model for unravelling the pathways that underpin learning and memory formation.</jats:p

Intermittent Fasting Modulates the Glycogen Level in Zebrafish (Danio rerio) and their Next Generation

Introduction: Zebrafish (Danio rerio) has become the best model organism to study the evolutionary biological process and human developmental studies. The liver glycogen plays a vital role in maintaining cellular metabolism, accumulation of glycogen in liver affects the enzymes related to glycogen metabolism. Aim: Impact of intermittent fasting, refeed and overfeeding in glycogen homeostasis on Zebrafish (Danio rerio) and their F1 generation. Materials and Methods: The present in-vivo study demonstrates the effect of intermittent fasting on glycogen storage in zebrafish and their F1 generation. The study was conducted at Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India. The duration of study was carried out for one month (December, 2021) for both parental and their F1 generation (April, 2022) groups. The F1 generation fishes involved after its matured (three months). The zebrafish (AB strain) were randomised and split into five experimental groups such as control, overfed, 12 hours, 24 hours, and 48 hours intermittent fasting. The F1 generation from each group was treated as same as parenting groups. The physiological and histological changes were observed in the study group. Significant results were evaluated as p&lt;0.05 values turkey’s method was used. Results: The fasting and overfeeding significantly affects the physiological condition like body weight, length and Body Mass Index (BMI). The parental control and their F1 have a BMI of 0.042±0.04 g/ cm² and 0.041±0.04 g/cm². The maximum fasting treated groups (48 hours) of both parent and their F1 generation shows reduced BMI such as 0.032±0.03 g/cm² and 0.030±0.04 g/cm². The over feed group shows a BMI of 0.053±0.05 g/cm² and 0.052±0.05 g/ cm². The result demonstrates that the food-deprived groups and their F1 generation showed less glycogen storage in histological observation. The reefed and overfed groups and their F1 generation exhibit more glycogen accumulation in the liver. The result confers normal regulation of glycogen synthase and glycogen synthase kinase 3 in normally in control and fasting groups as well as in their F1 generation. Conversely, the overfeeding and refeed groups show modulated glycogen activity in both parent and their F1 generation. Conclusion: Glycogen accumulation leads too many diseases and it also affects the generations. The frequent fasting may help to minimise glycogen accumulation and BMI level reduces the complications of disorders related to glycogen homeostasis.</jats:p

A Study on the Alleviating Effects of N-phthaloyl-γ-Aminobutyric Acid (P-GABA) on the Behavioral, Histopathological and Biochemical Adverse Effects of Sleep Deprivation in Zebrafish (Danio rerio)

Background: Sleep is defined as a reversible behavioural state of perceptual disconnection from insensitivity to the environment that facilitates the interaction of physiological and behavioural processes. Sleep Deprivation (S.D.) is defined as a decrease in sleeping duration below the recommended minimum, which has been linked to learning and memory problems.&#x0D; Aim: The primary objective of this work was to determine the effect of P-GABA on metabolic parameters, behavioural changes, whole-body cortisol, and brain histology in light-induced sleep-deprived zebrafish, as well as the optimal dose of P-GABA neutralizing undesirable effects.&#x0D; Methodology: The present study was conducted for ten days, consisting of three days in a row of sleep deprivation and seven days of treatment with P-GABA. The current investigation used six fishes in a group (n=6).&#x0D; Group 1: Control ; Group 2: 24h Total SD ; Group 3: 48h Total SD ; Group 4: 72h Total SD ; Group 5: 24h Total SD + P-GABA (100 mg/L) ; Group 6: 48h Total SD + P-GABA (100 mg/L) ;Group 7: 72h Total SD + P-GABA (100 mg/L)&#x0D; Results: The current study provides scientific data demonstrating the positive effects of P-GABA in treating sleep deprivation and associated cognitive impairment. To test if P-GABA treatment can alleviate the cognitive and memory impairment caused by S.D., we established non-toxic concentrations and treated the zebrafish with a safe dose of 100mg/L. The use of P-GABA increased cognitive performance in the T-maze, demonstrating that it has a favourable effect in a sleep-deprivation condition. The SD group exhibited neutrophil infiltration, and this S.D fish treated with P-GABA at a concentration of 100 mg/L demonstrated a moderate reduction in neuronal cell degeneration compared to controls. The levels of biochemical parameters during sleep deprivation and treatment phase with P-GABA were checked. It was evident from the results that the SOD, CAT and GPX levels in the S.D groups were drastically decreased, whereas treatment with P-GABA could show a significant increase in the levels of biochemical parameters.&#x0D; In contrast to the control group, zebrafish subjected to sleep deprivation showed enhanced AChE activity in the brain. The results of the P-GABA indicated an anti-AChE profile, which corresponds to improved memory parameters in zebrafish, as observed in the NTT and T-maze tests. When comparing the sleep-deprived fish to the control group, the MDA level, which indicates lipid peroxidation, was higher. Treatment with P-GABA considerably reduced the amount of MDA produced compared to the amount produced in sleep-deprived fish. The cortisol levels gradually increased in the single row 24h, 48h, and 72h sleep deprived groups. There was a gradual decrease in cortisol levels in the groups that received P-GABA treatment. The levels of neurotransmitters were seen to be decreased in the sleep-deprived groups when compared with the control. Upon treatment with P-GABA, the neurotransmitters were restored to near normal.&#x0D; Conclusion:  This study showed that P-GABA counteracts cognitive performance decrease and anxiety increase resulting from sleep deprivation through a mechanism implying mitigation of brain oxidative stress and regulation of AChE activity.</jats:p