Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

Users Online : 105238

AbstractMaterial and MethodsResultsDiscussionConclusionReferencesDOI and Others
Article in PDF How to Cite Citation Manager Readers' Comments (0) Audio Visual Article Statistics Link to PUBMED Print this Article Send to a Friend
Advertisers Access Statistics Resources

Dr Mohan Z Mani

"Thank you very much for having published my article in record time.I would like to compliment you and your entire staff for your promptness, courtesy, and willingness to be customer friendly, which is quite unusual.I was given your reference by a colleague in pathology,and was able to directly phone your editorial office for clarifications.I would particularly like to thank the publication managers and the Assistant Editor who were following up my article. I would also like to thank you for adjusting the money I paid initially into payment for my modified article,and refunding the balance.
I wish all success to your journal and look forward to sending you any suitable similar article in future"



Dr Mohan Z Mani,
Professor & Head,
Department of Dermatolgy,
Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



Dr Kalyani R
Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
As an experienced dentist and an academician, I proudly recommend this journal to the dental fraternity as a good quality open access platform for rapid communication of their cutting-edge research progress and discovery.
I wish JCDR a great success and I hope that journal will soar higher with the passing time."



Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




Dr. Arunava Biswas

"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."



Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2022 | Month : December | Volume : 16 | Issue : 12 | Page : BC10 - BC14 Full Version

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


Published: December 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/59645.17235
Shanmugasundaram Tamilarasan, Uthirakumar Devaraj, Balamurugan Elumalai

1. Research Scholar, Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India. 2. Research Scholar, Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India. 3. Associate Professor, Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India.

Correspondence Address :
Balamurugan Elumalai,
Associate Professor, Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India.
E-mail: balamurugan_au@yahoo.co.in

Abstract

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.

Keywords

Glycogen synthase kinase-3, Glycogen synthase, Overfeed

Glycogen is a multibranched polysaccharide of glucose it has a role in the storage and redistribution of lipids, proteins, and carbohydrates (1). Liver is an essential organ that helps the body to adjust dietary changes in both eating and fasting periods (2),(3). Liver glycogen is needed for blood glucose regulation. Insulin stimulates glycogen formation in hepatocytes, which has a substantial direct effect on them (4),(5). Glycogen synthase, which catalyses the addition of glucose to the glycogen chain, and glycogen phosphorylase, which catalyses the breakdown of glycogen to release glucose-1-phosphate, are the two essential enzymes that control the glycogen metabolism that occurs in the liver (6). The primary enzyme in glycogen production, Glycogen Synthase (GS), is triggered by the allosteric stimulator Glucose-6-Phosphate (G6P), as well as by dephosphorylation after insulin inactivates GS kinase-3. Given that glycogen synthesis is crucial for maintaining glucose homeostasis, decreased glycogenic activity is predicted to encourage the accumulation of intracellular G6P levels (7). The fasting/feeding response to glycogen accumulation helps for the better understanding of complications related to high glycogen storage in the liver (8). Nutritional status has an impact on a variety of different pathways, such as those involved in bile acid metabolism, iron metabolism, immunological responses, circadian rhythms, and stress responses. Dietary regulation of hepatic transcription is essential for physiology, and metabolic disorders are characterised by disruptions of these pathways. Future research is anticipated to uncover additional links and deepen our understanding of this crucial physiologic response in progeny with the continuous development of novel techniques and genetic models (9).

The zebrafish has become a popular model organism for developmental biology, neurology, and molecular genetics (10). Currently, zebrafish are being suggested as a viable model organism for research on nutrition and development (11),(12),(13). Utilised to study human metabolic illnesses, with a focus on diabetes and obesity. Zebrafish’s lipid metabolism networks are highly similar to those of humans, according to a comparative transcriptome analysis of visceral adipose tissue from zebrafish, mice, rats, and humans (14). Overfeeding caused significant hepatic steatosis, implying that a high-calorie diet increased fat accumulation in the liver (5). This model could help researchers to better understand the relationship between obesity and hepatic injury, moreover it provides a viable option for developing useful therapeutic products in the field of biomedical research. The zebrafish share similar characteristic features to human condition like overweight/obese pathologies (15),(16). This study also may helps to understand the impact of overweight on cognitive impairments and that adult neurogenesis is involved in memory (17),(18),(19). In this study, authors have analysed the changes in glycogen levels of food-deprived and overfeeding zebrafish and their F1 generation. The physical parameters and glycogen accumulation in liver of both conditions were observed in parent and F1 generation.

Material and Methods

The present in-vivo study was conducted in Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India, experiment’s was carried out over four weeks.

Fish Maintenance

Adult zebrafish (three months) were obtained from a finites aquarium in Chidambaram, Tamil Nadu, India. The fishes were kept seven days for acclimatisation under laboratory condition. The water condition was monitored frequently during study period. The experiments were carried out in Department of Biochemistry and Biotechnology, Annamalai University, Chidambaram, Tamil Nadu, India.

The fishes were maintained under 14-hour light: 10-hour dark cycle at 28°C with water quality monitored using a well-proven method with around 10 fish per 4-L tank (20). Zebrafish were divided into five dietary groups (Table/Fig 1). In each experimental design, 50 fishes were used (n=10) which so in total for this experiment, 200 fishes were used as mentioned, and were refeed three times a day with feed containing moisture, crude protein, crude fat, crude fibre, carbohydrate, and ash. For this study, the experimental conditions were maintained for four weeks and the condition has given for F1 generation zebrafish.

Measurement of Physical Parameters

The weight and length of zebrafish were measured frequently during the study period. The length and weight of each zebrafish body were assessed to calculate BMI (Table/Fig 2). The BMI was calculated by measuring body weight (g) divided by the square of the body length (cm2) (21). (Table/Fig 2) shows the length and weight measurement of zebrafish used in this study.


Breeding and Larval Maintenance

The breeding and larval maintenance of zebrafish is represented in (Table/Fig 3)a,b. The fishes were taken from experimental groups and kept in the breeding tank. The eggs were collected after a certain period with a dropper. The embryos were rinsed and examined under a microscope. Fertilised eggs were incubated for 72 hours at 28.5 °C until the F1 larvae hatched and after 5 dpf (day’s post fertilisation) feeding starts to grow. After three months, embryos were developed into sexually mature adults used for the experimental study (22).

Artemia Cultivation for Live Feed

The artemia nauplii (brine shrimp) was used as a feed for Larvae after fertilisation (dpf) (5 days) (23). Artemia cysts were purchased from a local pet store and cultivated in salt water with a motor for aeration and focus light for temperature maintenance. After 18-36 hours, the cysts were hatched and grew as adult reddish brown artemia which was given as feed to F1 generation.

Histology and Tissue Sectioning

The fish liver was fixed using 10% formalin solution and embedded in blocks. Then transversely sections were cut and stained with Haemotoxylin and Eosin (H&E). The glycogen vacuolation detection and tissue grading were performed on the parent and F1 offspring. To determine the relative amount of glycogen in hepatocyte cytoplasm, two consecutive sections of whole fish were stained for the Periodic Acid–Schiff (PAS). In the glycogen determination, the PAS-stained slide served as a negative control (24).

Statistical Analysis

In terms of statistical analysis, the mean±SEM was utilised to represent all of the data to compare various groups; a one-way Analysis of Variance (ANOVA) was used, followed by a Duncan’s Multiple Range Test (DMRT) test for control, fasting and refeed group comparisons. A p-value <0.05 was considered statistically significant in all groups.

Results

Effects of Intermittent Fasting and Feed on the Physical Measurement of the Zebrafish

The physiological changes of each experimental group of fishes were analysed to differentiate the control, intermittent fasting, and feeding groups. Body weight was found to be significantly higher in the overfeed and refeed fish groups compared to the control and fasting groups. The initial mean body weight of the zebrafish was roughly found to be 0.40-0.55 g. The BMI of the overfeed group was 0.60±0.05 (BMI 0.053±0.05) and the control group was 0.55±0.04 (BMI=0.042±0.04), respectively. Similarly, the BMI values of 0.54±0.04 (BMI=0.039±0.04), 0.52±0.04 (BMI=0.034±0.03), and 0.50±0.03 (BMI=0.032±0.03) were observed in food-deprived groups. The results show loss of weight and BMI in 12 hours, 24 hours, and 48 hours of intermittent fasting. The physical parameters were seen to be changed in zebrafish. This same parent group, process of feeding three fasting experimental groups increased body weight and BMI shown in (Table/Fig 4).

Physical Parameter Comparison of Parent and their F1 Generation

Physical parameter was measured in F1 generation of each group. It was observed that when compared to parent group, the body weight of the F1 generation groups was slightly changed, in the control group (F1 0.53±0.04, BMI=0.041±0.04) and in overfed group (0.57±0.04 (BMI=0.052±0.05). It was also noticed there was a decrease in body weight and BMI level of three fasting groups 12 hours (0.52±0.04, BMI=0.037±0.04) 24 hours (0.48±0.04, BMI=0.033±0.03), and 48 hours (0.46±0.04, BMI=0.030±0.04) (Table/Fig 4). Conversely, the refeed body weight and BMI of the control (0.55±0.05 BMI=0.047±0.05), Overfed group (0.63±0.05 BMI=0.066±0.07), 12 hours of fasting (0.58±0.05, BMI=0.045±0.05), 24 hours (0.60±0.05, BMI=0.046±0.05) and 48 hours (0.61±0.05, BMI=0.039±0.04) of F1 generation increased, respectively (Table/Fig 4).

Liver Histology and PAS Staining

The histology results show amount of glycogen accumulation in liver of experimental group fish. There was decreased amount of hepatic glycogen vacuolation observed in fasting groups and in their F1 generation (Table/Fig 5)a-j. Conversely, the refeed increased level of glycogen deposition observed in control, overfeed group (Table/Fig 6)a-e and also in their F1 generation (Table/Fig 6)f-j. The tissues were stained with PAS and the decreases in glycogen vacuolation were observed. Despite a difference in PAS staining intensity between the low glycogen exposure and intermittent fasting in the glycogen levels were dramatically reduced in the group exposed to intermittent intermittent fasting and its offspring.

Discussion

Given its functions in controlling whole-body energy metabolism, digesting dietary nutrients, and preserving blood glucose levels, the liver serves as a focal point for the coordination of fasting-feeding transitions (4). The liver’s metabolism of glycogen plays a role in controlling blood glucose levels. It involves the conflicting reactions of the enzymes Glycogen Synthase (GYS2) and Phosphorylase (PYGL). When an animal switches between a fed and fasted condition, the liver is the principal organ responsible for maintaining metabolic equilibrium. Fasting causes the liver to produce glucose and ketone bodies, deplete its supply of glycogen, and accumulate triacylglycerol (25),(26). Glycogen breakdown or gluconeogenesis from glycerol, amino acids, or Tricarboxylic Acid (TCA) cycle intermediates are the two mechanisms by which hepatic glucose is produced. A previous study found that feed-restricted male mice perform better physically, have improved insulin sensitivity, and had lower levels of cholesterol and lipoprotein (27),(28),(29). Present study was parent of fasting group BMI level reduced compared than feeding group. However, during the refeeding period earlier research, fish fed a high-carbohydrates diet had much greater plasma glucose and glycogen levels than fish fed a low-carbohydrate diet. In comparison to zebrafish that were fed normally, the overfed animals had higher BMIs, hypertriglyceridaemia, and hepatosteatosis. Zebrafish males and females responded to overnutrition in a similar manner (30). Evidences from the past studies mostly reported intermittent fasting has been studied in animal models and human research to give an outlook on the health benefits (31). Authors explicitly demonstrated that a four weeks considerably reduced BMI measurements in both parent and F1 generation whereas fed alone resulted in significantly larger rises than fasting group participants. It is crucial to understand the immunophysiological reactions triggered by overfeeding-induced obesity.

When calorie intake is reduced, the liver’s storage of glycogen during feeding conditions serves as a type of glucose reserve that can be used. Glycogen storage higher in overfed and feeding groups to be continuously increased their generation activation of glycogen synthase, a fundamental enzyme of glycogenesis (glycogen synthesis), whereas glycogen phosphorylase, a key enzyme of glycogenolysis, is repressed (glycogen breakdown) (32). The histology of liver stained using PAS staining showed depletion in glycogen reserves during intermittent fasting utilising the glycogen reserves for survival (24). However, the glycogen reserves reversed back upon refeeding the parent group subjected to intermittent fasting. More glycogen accumulation observed in overfeed group and it followed in their F1 generation (33). Conversely the fasting group exhibit less amount of accumulation of glycogen in both parent and F1 generation. Under fasting conditions, dephosphorylated and active GSK-3 phosphorylate and inactivate glycogen synthase, leading to the inhibition of hepatic glycogen synthesis (34). The glycogen synthase that appears to be most prevalent is the gene product (Gsy2). Both the phosphorylated (less active) and the dephosphorylated (active) versions of the Gsy1 and Gsy2 isoforms exist, and both have activities that are regulated allosterically. A glycogen synthase kinase enzyme is inhibited by G6P, which also acts as a direct activator and regulates the phosphorylation state of this enzyme (35).

In response to food intake, increased insulin signalling activates the cell’s Akt, which then phosphorylates and inactivates GSK-3, activating glycogen synthase. Additionally, this enzyme is allosterically activated by higher G6P concentrations, which increases its catalytic activity in feeding situations (36),(37). According to some research, GSK-3 may control insulin resistance development and regulate glucose metabolism without affecting glycogen production (35). Also GSK-3 has been linked to the development of neurodegenerative illnesses like Alzheimer disease (38). Additionally connect human psychiatric disorders like schizophrenia and bipolar disorder is GSK-3 deregulation. The modulation of synaptic plasticity and memory is facilitated by GSK-3. Investigation into how GSK-3 affects the onset of obesity and its associated metabolic problems, such as insulin resistance, in adipose tissue is ongoing (39),(40).

This study observed the BMI alterations as well as increased glycogen storage in the liver. Evidence from the previous study created a consistent overfeeding zebrafish model that led to various metabolic problems, including increased body weight, BMI, blood glucose levels, and liver steatosis. Previous researches discussed above on behavioural changes in zebrafish under control, fasting, refeed, and overfeed conditions showed, zebrafish that had been overfeed and refeed had lipid accumulation and glycogen storage in their liver tissue (41),(42),(43),(44),(45). This study was an experimental proof that intermittent fasting can cause glycogen utilisation via glycogen synthase enzyme and regulate glycogen synthase kinase-3 enzyme outreached to obesity, diabetes and neurological disease related conditions. In the current study, it can be said that intermittent fasting had most benefits when compared to the BMI, fasting glucose. Insulin levels gradually decreased over time, implying that long-term CR (caloric restriction) and physical exercise have great benefits on the health of obese patients. This study shows a direct connection between human overeating behaviour and changes in liver glycogen, which may increase your risk of obesity-related health problems.

Limitation(s)

In the future, authors will test obese patients for the GSK-3 enzyme to determine the gene expression level and to find ways to prevent genetic disease which is at the moment, a limitation of this study.

Conclusion

The findings of study have shown that the zebrafish is an ideal model to study intermittent fasting. The result confers the similar physiological and histological responses to the dietary interventions throughout the generation. The present study illustrates intermittent fasting repercussions of dietary modifications that contained the most positive reactions from the parent and its F1 generation. Thus intermittent fasting was necessary to keep GS and GSK-3 under control. The risk of obesity, diabetes, neurodegenerative and cardiovascular diseases were reduced during frequent fasting conditions. The overfeed and refeeded group and its descendent generation continued to increase body parameters composition and glycogen vacuoles in liver histology tests. According to findings, controlled diet may help to prevention of genetic problems dietary related disorders.

Author’s contribution: The BE conceptualise the work and wrote final version of the manuscript. ST did all the experimental work and wrote the draft version of the manuscript. UD helped in the experimental work.

References

1.
Sadava DE, Hillis DM, Heller HC. Life: The science of biology. Macmillan; 2009.
2.
Roach PJ. Glycogen and its metabolism. Curr Mol Med. 2002;2(2):101-20. [crossref] [PubMed]
3.
Bouskila M, Hunter RW, Ibrahim AF, Delattre L, Peggie M, Van Diepen JA, et al. Allosteric regulation of glycogen synthase controls glycogen synthesis in muscle. Cell Metabolism. 2010;12(5):456-66. [crossref] [PubMed]
4.
Yoon JC, Puigserver P, Chen G, Donovan J, Wu Z, Rhee J, et al. Control of hepatic gluconeogenesis through the transcriptional coactivator PGC-1. Nature. 2001;413(6852):131-38. [crossref] [PubMed]
5.
Kersten S, Seydoux J, Peters JM, Gonzalez FJ, Desvergne B, Wahli W. Peroxisome proliferator–activated receptor α mediates the adaptive response to fasting. J Clin Inves. 1999;103(11):1489-98. [crossref] [PubMed]
6.
Adeva-Andany MM, González-Lucán M, Donapetry-García C, Fernández-Fernández C, Ameneiros-Rodríguez E. Glycogen metabolism in humans. BBA Clin. 2016;5:85-100. [crossref] [PubMed]
7.
Shulman GI. Cellular mechanisms of insulin resistance. J Clin Invest. 2000;106(2):171-76. [crossref] [PubMed]
8.
Gonzalez JT, Fuchs CJ, Betts JA, Van Loon LJ. Liver glycogen metabolism during and after prolonged endurance-type exercise. Am J Physiology-Endocrinology and Metabolism. 2016;311(3):E543-53. [crossref] [PubMed]
9.
Bideyan L, Nagari R, Tontonoz P. Hepatic transcriptional responses to fasting and feeding. Genes & Development. 2021;35(9-10):635-57. [crossref] [PubMed]
10.
Dooley K, Zon LI. Zebrafish: A model system for the study of human disease. Curr Opinion Gen Dev. 2000;10(3):252-56. [crossref] [PubMed]
11.
Aleström P, Holter JL, Nourizadeh-Lillabadi R. Zebrafish in functional genomics and aquatic biomedicine. Trends in Biotechnology. 2006;24(1):15-21. [crossref] [PubMed]
12.
Dahm R, Geisler R. Learning from small fry: The zebrafish as a genetic model organism for aquaculture fish species. Marine Biotechnology. 2006;8(4):329-45. [crossref] [PubMed]
13.
De-Santis C, Jerry DR. Candidate growth genes in finfish—Where should we be looking? Aquaculture. 2007;272(1-4):22-38. [crossref]
14.
Wright D, Nakamichi R, Krause J, Butlin RK. QTL analysis of behavioral and morphological differentiation between wild and laboratory zebrafish (Danio rerio). Behaviour Genetics. 2006;36(2):271-84. [crossref] [PubMed]
15.
Vargas R, Vásquez IC. Effects of overfeeding and high-fat diet on cardiosomatic parameters and cardiac structures in young and adult zebrafish. Fish Physiology Biochem. 2017;43(6):1761-73. [crossref] [PubMed]
16.
World Health Organisation. Obesity: Preventing and managing the global epidemic. (2019).
17.
Cifre M, Palou A, Oliver P. Cognitive impairment in metabolically-obese, normal-weight rats: Identification of early biomarkers in peripheral blood mononuclear cells. Molecular Neurodegeneration. 2018;13(1):01-04. [crossref] [PubMed]
18.
Parimisetty A, Dorsemans AC, Awada R, Ravanan P, Diotel N, Lefebvre d’Hellencourt C. Secret talk between adipose tissue and central nervous system via secreted factors-An emerging frontier in the neurodegenerative research. J neuroinflammation. 2016;13(1):01-03. [crossref] [PubMed]
19.
Alam MJ, Kitamura T, Saitoh Y, Ohkawa N, Kondo T, Inokuchi K, et al. Adult neurogenesis conserves hippocampal memory capacity. J Neurosci. 2018;38(31):6854-63. [crossref] [PubMed]
20.
Westerfield M. The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish Danio (“Brachydanio Rerio”). University of Oregon; 2007.
21.
Wang J, Li Y, Lai K, Zhong Q, Demin KA, Kalueff AV, et al. High-glucose/high-cholesterol diet in zebrafish evokes diabetic and affective pathogenesis: The role of peripheral and central inflammation, microglia and apoptosis. Progress in Neuro-Psychopharmacology and Biological Psychiatry. 2020;96:109752. [crossref] [PubMed]
22.
Avdesh A, Chen M, Martin-Iverson MT, Mondal A, Ong D, Rainey-Smith S, et al. Regular care and maintenance of a zebrafish (Danio rerio) laboratory: An introduction. JoVE (Journal of Visualized Experiments). 2012(69):e4196. [crossref] [PubMed]
23.
Veeramani T, Santhanam P, Manickam N, Rajthilak C. Introduction to Artemia Culture. InBasic and Applied Zooplankton Biology 2019 (pp. 209-224). Springer, Singapore. [crossref]
24.
Bui-Nguyen TM, Baer CE, Lewis JA, Yang D, Lein PJ, Jackson DA, et al. Dichlorvos exposure results in large scale disruption of energy metabolism in the liver of the zebrafish, Danio rerio. BMC Genomics. 2015;16(1):01-08. [crossref] [PubMed]
25.
Renquist BJ, Murphy JG, Larson EA, Olsen D, Klein RF, Ellacott KL, et al. Melanocortin-3 receptor regulates the normal fasting response. Proceedings of the National Academy of Sciences. 2012;109(23):E1489-98. [crossref] [PubMed]
26.
Moore MC, Coate KC, Winnick JJ, An Z, Cherrington AD. Regulation of hepatic glucose uptake and storage in vivo. Advances in Nutrition. 2012;3(3):286-94. [crossref] [PubMed]
27.
Ahmed T, Das SK, Golden JK, Saltzman E, Roberts SB, Meydani SN. Calorie restriction enhances T-cell–mediated immune response in adult overweight men and women. J Gerontol A Biol Sci Med Sci. 2009;64(11):1107-13. [crossref] [PubMed]
28.
Liang X, Wang J, Gong G, Xue M, Dong Y, Wu X, et al. Gluconeogenesis during starvation and refeeding phase is affected by previous dietary carbohydrates levels and a glucose stimuli during early life in Siberian sturgeon (Acipenser baerii). Animal Nutrition. 2017;3(3):284-94. [crossref] [PubMed]
29.
Mattson MP, Wan R. Beneficial effects of intermittent fasting and caloric restriction on the cardiovascular and cerebrovascular systems. The Journal of Nutritional Biochemistry. 2005;16(3):129-37. [crossref] [PubMed]
30.
Van Overveld T, Matthysen E. Personality predicts spatial responses to food manipulations in free-ranging great tits (Parus major). Biology Letters. 2010;6(2):187-90. [crossref] [PubMed]
31.
Mizock BA. Alterations in fuel metabolism in critical illness: Hyperglycaemia. Best Practice & Research Clinical Endocrinology & Metabolism. 2001;15(4):533-51. [crossref] [PubMed]
32.
Han HS, Kang G, Kim JS, Choi BH, Koo SH. Regulation of glucose metabolism from a liver-centric perspective. Experimental & Molecular Medicine. 2016;48(3):e218. [crossref] [PubMed]
33.
Van de Werve G, Jeanrenaud B. The onset of liver glycogen synthesis in fasted-refed lean and genetically obese (fa/fa) rats. Diabetologia. 1987;30(3):169-74. [crossref] [PubMed]
34.
Hooper C, Killick R, Lovestone S. The GSK3 hypothesis of Alzheimer’s disease. Journal of Neurochemistry. 2008;104(6):1433-39. [crossref] [PubMed]
35.
Ros S, García-Rocha M, Domínguez J, Ferrer JC, Guinovart JJ. Control of liver glycogen synthase activity and intracellular distribution by phosphorylation. J Bio Chem. 2009;284(10):6370-78. [crossref] [PubMed]
36.
Eldar-Finkelman H, Schreyer SA, Shinohara MM, LeBoeuf RC, Krebs EG. Increased glycogen synthase kinase-3 activity in diabetes-and obesity-prone C57BL/6J mice. Diabetes. 1999;48(8):1662-66. [crossref] [PubMed]
37.
Grimes CA, Jope RS. The multifaceted roles of glycogen synthase kinase 3 β in cellular signaling. Progress in Neurobiology. 2001;65(4):391-26. [crossref] [PubMed]
38.
Hooper C, Markevich V, Plattner F, Killick R, Schofield E, Engel T, et al. Glycogen synthase kinase-3 inhibition is integral to long-term potentiation. European journal of Neuroscience. 2007;25(1):81-86. [crossref] [PubMed]
39.
Peineau S, Taghibiglou C, Bradley C, Wong TP, Liu L, Lu J, et al. LTP inhibits LTD in the hippocampus via regulation of GSK3β. Neuron. 2007;53(5):703-17. [crossref] [PubMed]
40.
Caballero B, Trugo LC, Finglas PM. Encyclopedia of food sciences and nutrition. Academic; 2003.
41.
Oka T, Nishimura Y, Zang L, Hirano M, Shimada Y, Wang Z, et al. Diet-induced obesity in zebrafish shares common pathophysiological pathways with mammalian obesity. BMC Physiology. 2010;10(1):1-3. [crossref] [PubMed]
42.
Zang L, Maddison LA, Chen W. Zebrafish as a model for obesity and diabetes. Frontiers in Cell and Developmental Biology. 2018;6:91. [crossref] [PubMed]
43.
Montalbano G, Mania M, Guerrera MC, Laurà R, Abbate F, Levanti M, et al. Effects of a flavonoid-rich extract from Citrus sinensis juice on a diet-induced obese zebrafish. Int J Mol Sci. 2019;20(20):5116. [crossref] [PubMed]
44.
Montalbano G, Mania M, Abbate F, Navarra M, Guerrera MC, Laura R, et al. Melatonin treatment suppresses appetite genes and improves adipose tissue plasticity in diet-induced obese zebrafish. Endocrine. 2018;62(2):381-93. [crossref] [PubMed]
45.
Montalbano G, Mania M, Guerrera MC, Abbate F, Laurà R, Navarra M, et al. Morphological differences in adipose tissue and changes in BDNF/Trkb expression in brain and gut of a diet induced obese zebrafish model. Annals of Anatomy-Anatomischer Anzeiger. 2016;204:36-44. [crossref] [PubMed]

DOI and Others

DOI: 10.7860/JCDR/2022/59645.17235

Date of Submission: Aug 13, 2022
Date of Peer Review: Oct 15, 2022
Date of Acceptance: Nov 04, 2022
Date of Publishing: Dec 01, 2022

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? No
• Was informed consent obtained from the subjects involved in the study? No
• For any images presented appropriate consent has been obtained from the subjects. No

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Aug 26, 2022
• Manual Googling: Sep 29, 2022
• iThenticate Software: Nov 03, 2022 (7%)

ETYMOLOGY: Author Origin

JCDR is now Monthly and more widely Indexed .
  • Emerging Sources Citation Index (Web of Science, thomsonreuters)
  • Index Copernicus ICV 2017: 134.54
  • Academic Search Complete Database
  • Directory of Open Access Journals (DOAJ)
  • Embase
  • EBSCOhost
  • Google Scholar
  • HINARI Access to Research in Health Programme
  • Indian Science Abstracts (ISA)
  • Journal seek Database
  • Google
  • Popline (reproductive health literature)
  • www.omnimedicalsearch.com