AUCTORES
Globalize your Research
Research Article
*Corresponding Author: Thiago Bassi, Research and Development at Lungpacer Medical Inc. Vancouver, Canada.
Citation: Bassi T, Rohrs E, Fernandez K, Ornowska R, Reynolds S, An Easier Method to Analyze Stereologically the Pig’s Hippocampus. J Neuroscience and Neurological Surgery. 4(3): Doi:10.31579/2578-8868/059
Copyright: ©2019. Thiago Bassi . This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Received: 17 July 2019 | Accepted: 23 July 2019 | Published: 25 July 2019
Keywords: neuroscience; stereological method; hippocampus; pigs
Most neuro-pathophysiological research involving the hippocampus uses rodents as a nonhuman model, due to the extensive experimental descriptive literature and favorable cost. The findings from the rodent hippocampus may not be generalizable to humans, though, as ontogenetically they have different hippocampal components. While the rodent model has these limitations, the use of nonhuman primates is often not feasible due to economic and ethical considerations. Pigs are a translational alternative due to the anatomic similarity of the hippocampus in humans and pigs. Materials and Methods: Eight pigs’ brains were harvested and then analyzed according to our previously established technique. Five brains were frozen and three were stored in formalin. All eight brains were then sent to an independent histology service, where they were sectioned according to the methodology established by Holm 1994. The slabs were 10 µm with 2.5 cm2 of hippocampus cross-sectional area. Results: The mean total hippocampus volume was 892.84 mm3 ± 198.91 mm3 using Holm’s methodology. The mean number of cells per sample (20X magnification settings) was 9996.75, using automated ImageJ cell counting. Discussion: In this study, the counts of hippocampus cells were divided into two regions of interest: CA1 and CA3. Our results show that the mean number of hippocampus cells observed was 5.75 million and 2.25 million, in the CA1 and CA3 regions respectively. Holm reported 4.12 million cells in the CA1 region and 1.51 million cells in the CA3 region. The results presented here indicate the CA1 and CA3 cell percentages being 23% and 9% respectively, which are similar to the percentages reported by Holm (21% and 12%). Conclusion: These results corroborate previous findings and demonstrate a novel and cost-effective way to study the hippocampus of pigs in translational neurological research.
Most neuro-pathophysiological research involving the hippocampus uses rodents as a nonhuman model, due to the extensive experimental descriptive literature and favorable cost. The findings from the rodent hippocampus may not be generalizable to humans, though, as ontogenetically they have different hippocampal components. Moreover, the hippocampus proportions and architectonics are significantly different between rodents and humans. While the rodent model has these limitations, the use of nonhuman primates is often not feasible due to economic and ethical considerations. Pigs are a translational alternative due to the anatomic similarity of the hippocampus in humans and pigs. In addition, pigs are a well-accepted animal research model and have lower cost when compared to nonhuman primates.
Human-size pigs have been used in research due to their anatomical and physiological similarity to humans. [1,2] The pig’s brain has a gyrencephalus similar to humans, which facilitates surgical procedures and interventions. Pigs are genetically relatively homogeneous, similar to inbred laboratory rats, leading to a minimal interindividual variation.
A publication by Holm in 1994 provided a quantitative description of fundamental structural parameters, regional volumes and neuron numbers in the hippocampus of the domestic pig [3]. The volumetric and numerical data presented by Holm provided a unique opportunity to evaluate structural differences in homologous hippocampus areas between pigs and humans, and obtain a better understanding of the functional consequences of the differences in size. Despite this strong foundation, the pig hippocampus has not been clearly established as the translational model of choice for neurological studies. In 2016, van Dijk published a systematic review comparing the hippocampus in 18 different species, where one pig study was reported [4].
The hippocampus is well recognized as the hub of neuroplasticity and neurogenesis in the central nervous system. Many diseases begin in areas CA1 and CA3 before spreading to the cortex. As an example, Masurkar, 2018 explains in great detail the progression of Alzheimer’s disease, where the CA1 area is the starting point [5]. Recently, there has been an increased interest in derangements in the CA1 and CA3 areas as the basis for important clinical syndromes such as memory impairment and spatial recognition deficit [5]. Historically, many researchers have been using rodents to analyze the hippocampus, but this poses significant translational limitations, as discussed above.
Herein, we report work that can serve as a basis for pig-based neuroscience research. The aim of this study is to extend previous work by Holm with a focus on CA1 and CA3 volume and cell count
Materials and methods:
Animal Care and Ethical approvals were obtained, and post-euthanasia surgical procedures were conducted at UBC Research Centre in Vancouver, Canada. Eight pigs’ brains were harvested and then analyzed according to our previously established technique [6]. Five brains were frozen and three were stored in formalin. All eight brains were then sent to an independent histology service, where they were sectioned according to the methodology established by Holm 1994. The slabs were 10 µm with 2.5 cm2 of hippocampus cross-sectional area. The hippocampus cell calculations were made according to Holm’s formula:
Hippocampus Cells (N)=Cells per slab*2500
According to the previous work by Holm, the formula above was determined using the number of cells per slab multiplied by the size of the hippocampus extension. To replicate this work, the ITCN ImageJ software plugin was used to count the number of cells per slab. Next, the ImageJ ruler was used to measure the area of the hippocampus. Holm’s formula was applied to obtain the final number of hippocampal cells per subject per area. Finally, the CA1 and CA3 percentage were calculated applying the same formula on the respective areas (Figure 1).
Results and Discussion:
The mean total hippocampus volume was 892.84 mm3 ± 198.91 mm3 using Holm’s methodology. The mean number of cells per sample (20X magnification settings) was 9996.75, using automated ImageJ cell counting. The percentages of hippocampal cells in areas CA1 and CA3 were 9% and 23% respectively, after normalizing by weight. Results are listed in table 1.
The cytoarchitecture of the pig hippocampus is similar to humans. Thus, it is crucial to establish a stereological number of hippocampal cells in pigs as a basis for future studies.
It is important to correct the cell counts in proportion to body weight, as we did, and as Holm did.
Our results showed an average number of hippocampus cells of 24.99 million. This is similar to those reported by Holm (21.61 million).
In this study, the counts of hippocampus cells were divided into two regions of interest: CA1 and CA3. Our results show that the mean number of hippocampus cells observed was 5.75 million and 2.25 million, in the CA1 and CA3 regions respectively. These results are comparable to previous studies. Holm reported 4.12 million cells in the CA1 region and 1.51 million cells in the CA3 region. The results presented here indicate the CA1 and CA3 cell percentages being 23% and 9% respectively, which are similar to the percentages reported by Holm (21% and 12%). It is important to highlight that the proportion of cells is similar in the human hippocampus.
This study provides pig hippocampus data to be used in future studies. So far, only one paper was found to address this topic.[4] However, the use of open source software to stereologically study the hippocampus was not mentioned as part of the reported methodology. The use of open source software (in our case, ImageJ) is a novel and cheaper approach to stereologically analyze the brain tissue.
Limitations: On this study we did not analyze pigs' diets,which might change the hippocampus structure and volume [7] being a variable to be considered in future comparative studies.
Conclusion:
These results corroborate previous findings and demonstrate a novel and cost-effective way to study the hippocampus of pigs in translational neurological research.
Acknowledgements
This study was funded by Lungpacer Medical Inc.
Clearly Auctoresonline and particularly Psychology and Mental Health Care Journal is dedicated to improving health care services for individuals and populations. The editorial boards' ability to efficiently recognize and share the global importance of health literacy with a variety of stakeholders. Auctoresonline publishing platform can be used to facilitate of optimal client-based services and should be added to health care professionals' repertoire of evidence-based health care resources.
Journal of Clinical Cardiology and Cardiovascular Intervention The submission and review process was adequate. However I think that the publication total value should have been enlightened in early fases. Thank you for all.
Journal of Women Health Care and Issues By the present mail, I want to say thank to you and tour colleagues for facilitating my published article. Specially thank you for the peer review process, support from the editorial office. I appreciate positively the quality of your journal.
Journal of Clinical Research and Reports I would be very delighted to submit my testimonial regarding the reviewer board and the editorial office. The reviewer board were accurate and helpful regarding any modifications for my manuscript. And the editorial office were very helpful and supportive in contacting and monitoring with any update and offering help. It was my pleasure to contribute with your promising Journal and I am looking forward for more collaboration.
We would like to thank the Journal of Thoracic Disease and Cardiothoracic Surgery because of the services they provided us for our articles. The peer-review process was done in a very excellent time manner, and the opinions of the reviewers helped us to improve our manuscript further. The editorial office had an outstanding correspondence with us and guided us in many ways. During a hard time of the pandemic that is affecting every one of us tremendously, the editorial office helped us make everything easier for publishing scientific work. Hope for a more scientific relationship with your Journal.
The peer-review process which consisted high quality queries on the paper. I did answer six reviewers’ questions and comments before the paper was accepted. The support from the editorial office is excellent.
Journal of Neuroscience and Neurological Surgery. I had the experience of publishing a research article recently. The whole process was simple from submission to publication. The reviewers made specific and valuable recommendations and corrections that improved the quality of my publication. I strongly recommend this Journal.
Dr. Katarzyna Byczkowska My testimonial covering: "The peer review process is quick and effective. The support from the editorial office is very professional and friendly. Quality of the Clinical Cardiology and Cardiovascular Interventions is scientific and publishes ground-breaking research on cardiology that is useful for other professionals in the field.
Thank you most sincerely, with regard to the support you have given in relation to the reviewing process and the processing of my article entitled "Large Cell Neuroendocrine Carcinoma of The Prostate Gland: A Review and Update" for publication in your esteemed Journal, Journal of Cancer Research and Cellular Therapeutics". The editorial team has been very supportive.
Testimony of Journal of Clinical Otorhinolaryngology: work with your Reviews has been a educational and constructive experience. The editorial office were very helpful and supportive. It was a pleasure to contribute to your Journal.
Dr. Bernard Terkimbi Utoo, I am happy to publish my scientific work in Journal of Women Health Care and Issues (JWHCI). The manuscript submission was seamless and peer review process was top notch. I was amazed that 4 reviewers worked on the manuscript which made it a highly technical, standard and excellent quality paper. I appreciate the format and consideration for the APC as well as the speed of publication. It is my pleasure to continue with this scientific relationship with the esteem JWHCI.
This is an acknowledgment for peer reviewers, editorial board of Journal of Clinical Research and Reports. They show a lot of consideration for us as publishers for our research article “Evaluation of the different factors associated with side effects of COVID-19 vaccination on medical students, Mutah university, Al-Karak, Jordan”, in a very professional and easy way. This journal is one of outstanding medical journal.
Dear Hao Jiang, to Journal of Nutrition and Food Processing We greatly appreciate the efficient, professional and rapid processing of our paper by your team. If there is anything else we should do, please do not hesitate to let us know. On behalf of my co-authors, we would like to express our great appreciation to editor and reviewers.
As an author who has recently published in the journal "Brain and Neurological Disorders". I am delighted to provide a testimonial on the peer review process, editorial office support, and the overall quality of the journal. The peer review process at Brain and Neurological Disorders is rigorous and meticulous, ensuring that only high-quality, evidence-based research is published. The reviewers are experts in their fields, and their comments and suggestions were constructive and helped improve the quality of my manuscript. The review process was timely and efficient, with clear communication from the editorial office at each stage. The support from the editorial office was exceptional throughout the entire process. The editorial staff was responsive, professional, and always willing to help. They provided valuable guidance on formatting, structure, and ethical considerations, making the submission process seamless. Moreover, they kept me informed about the status of my manuscript and provided timely updates, which made the process less stressful. The journal Brain and Neurological Disorders is of the highest quality, with a strong focus on publishing cutting-edge research in the field of neurology. The articles published in this journal are well-researched, rigorously peer-reviewed, and written by experts in the field. The journal maintains high standards, ensuring that readers are provided with the most up-to-date and reliable information on brain and neurological disorders. In conclusion, I had a wonderful experience publishing in Brain and Neurological Disorders. The peer review process was thorough, the editorial office provided exceptional support, and the journal's quality is second to none. I would highly recommend this journal to any researcher working in the field of neurology and brain disorders.
Dear Agrippa Hilda, Journal of Neuroscience and Neurological Surgery, Editorial Coordinator, I trust this message finds you well. I want to extend my appreciation for considering my article for publication in your esteemed journal. I am pleased to provide a testimonial regarding the peer review process and the support received from your editorial office. The peer review process for my paper was carried out in a highly professional and thorough manner. The feedback and comments provided by the authors were constructive and very useful in improving the quality of the manuscript. This rigorous assessment process undoubtedly contributes to the high standards maintained by your journal.
International Journal of Clinical Case Reports and Reviews. I strongly recommend to consider submitting your work to this high-quality journal. The support and availability of the Editorial staff is outstanding and the review process was both efficient and rigorous.
Thank you very much for publishing my Research Article titled “Comparing Treatment Outcome Of Allergic Rhinitis Patients After Using Fluticasone Nasal Spray And Nasal Douching" in the Journal of Clinical Otorhinolaryngology. As Medical Professionals we are immensely benefited from study of various informative Articles and Papers published in this high quality Journal. I look forward to enriching my knowledge by regular study of the Journal and contribute my future work in the field of ENT through the Journal for use by the medical fraternity. The support from the Editorial office was excellent and very prompt. I also welcome the comments received from the readers of my Research Article.
Dear Erica Kelsey, Editorial Coordinator of Cancer Research and Cellular Therapeutics Our team is very satisfied with the processing of our paper by your journal. That was fast, efficient, rigorous, but without unnecessary complications. We appreciated the very short time between the submission of the paper and its publication on line on your site.
I am very glad to say that the peer review process is very successful and fast and support from the Editorial Office. Therefore, I would like to continue our scientific relationship for a long time. And I especially thank you for your kindly attention towards my article. Have a good day!
"We recently published an article entitled “Influence of beta-Cyclodextrins upon the Degradation of Carbofuran Derivatives under Alkaline Conditions" in the Journal of “Pesticides and Biofertilizers” to show that the cyclodextrins protect the carbamates increasing their half-life time in the presence of basic conditions This will be very helpful to understand carbofuran behaviour in the analytical, agro-environmental and food areas. We greatly appreciated the interaction with the editor and the editorial team; we were particularly well accompanied during the course of the revision process, since all various steps towards publication were short and without delay".
I would like to express my gratitude towards you process of article review and submission. I found this to be very fair and expedient. Your follow up has been excellent. I have many publications in national and international journal and your process has been one of the best so far. Keep up the great work.
We are grateful for this opportunity to provide a glowing recommendation to the Journal of Psychiatry and Psychotherapy. We found that the editorial team were very supportive, helpful, kept us abreast of timelines and over all very professional in nature. The peer review process was rigorous, efficient and constructive that really enhanced our article submission. The experience with this journal remains one of our best ever and we look forward to providing future submissions in the near future.
I am very pleased to serve as EBM of the journal, I hope many years of my experience in stem cells can help the journal from one way or another. As we know, stem cells hold great potential for regenerative medicine, which are mostly used to promote the repair response of diseased, dysfunctional or injured tissue using stem cells or their derivatives. I think Stem Cell Research and Therapeutics International is a great platform to publish and share the understanding towards the biology and translational or clinical application of stem cells.
I would like to give my testimony in the support I have got by the peer review process and to support the editorial office where they were of asset to support young author like me to be encouraged to publish their work in your respected journal and globalize and share knowledge across the globe. I really give my great gratitude to your journal and the peer review including the editorial office.
I am delighted to publish our manuscript entitled "A Perspective on Cocaine Induced Stroke - Its Mechanisms and Management" in the Journal of Neuroscience and Neurological Surgery. The peer review process, support from the editorial office, and quality of the journal are excellent. The manuscripts published are of high quality and of excellent scientific value. I recommend this journal very much to colleagues.
Dr.Tania Muñoz, My experience as researcher and author of a review article in The Journal Clinical Cardiology and Interventions has been very enriching and stimulating. The editorial team is excellent, performs its work with absolute responsibility and delivery. They are proactive, dynamic and receptive to all proposals. Supporting at all times the vast universe of authors who choose them as an option for publication. The team of review specialists, members of the editorial board, are brilliant professionals, with remarkable performance in medical research and scientific methodology. Together they form a frontline team that consolidates the JCCI as a magnificent option for the publication and review of high-level medical articles and broad collective interest. I am honored to be able to share my review article and open to receive all your comments.
“The peer review process of JPMHC is quick and effective. Authors are benefited by good and professional reviewers with huge experience in the field of psychology and mental health. The support from the editorial office is very professional. People to contact to are friendly and happy to help and assist any query authors might have. Quality of the Journal is scientific and publishes ground-breaking research on mental health that is useful for other professionals in the field”.
Dear editorial department: On behalf of our team, I hereby certify the reliability and superiority of the International Journal of Clinical Case Reports and Reviews in the peer review process, editorial support, and journal quality. Firstly, the peer review process of the International Journal of Clinical Case Reports and Reviews is rigorous, fair, transparent, fast, and of high quality. The editorial department invites experts from relevant fields as anonymous reviewers to review all submitted manuscripts. These experts have rich academic backgrounds and experience, and can accurately evaluate the academic quality, originality, and suitability of manuscripts. The editorial department is committed to ensuring the rigor of the peer review process, while also making every effort to ensure a fast review cycle to meet the needs of authors and the academic community. Secondly, the editorial team of the International Journal of Clinical Case Reports and Reviews is composed of a group of senior scholars and professionals with rich experience and professional knowledge in related fields. The editorial department is committed to assisting authors in improving their manuscripts, ensuring their academic accuracy, clarity, and completeness. Editors actively collaborate with authors, providing useful suggestions and feedback to promote the improvement and development of the manuscript. We believe that the support of the editorial department is one of the key factors in ensuring the quality of the journal. Finally, the International Journal of Clinical Case Reports and Reviews is renowned for its high- quality articles and strict academic standards. The editorial department is committed to publishing innovative and academically valuable research results to promote the development and progress of related fields. The International Journal of Clinical Case Reports and Reviews is reasonably priced and ensures excellent service and quality ratio, allowing authors to obtain high-level academic publishing opportunities in an affordable manner. I hereby solemnly declare that the International Journal of Clinical Case Reports and Reviews has a high level of credibility and superiority in terms of peer review process, editorial support, reasonable fees, and journal quality. Sincerely, Rui Tao.
Clinical Cardiology and Cardiovascular Interventions I testity the covering of the peer review process, support from the editorial office, and quality of the journal.
Clinical Cardiology and Cardiovascular Interventions, we deeply appreciate the interest shown in our work and its publication. It has been a true pleasure to collaborate with you. The peer review process, as well as the support provided by the editorial office, have been exceptional, and the quality of the journal is very high, which was a determining factor in our decision to publish with you.
The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews journal clinically in the future time.
Clinical Cardiology and Cardiovascular Interventions, I would like to express my sincerest gratitude for the trust placed in our team for the publication in your journal. It has been a true pleasure to collaborate with you on this project. I am pleased to inform you that both the peer review process and the attention from the editorial coordination have been excellent. Your team has worked with dedication and professionalism to ensure that your publication meets the highest standards of quality. We are confident that this collaboration will result in mutual success, and we are eager to see the fruits of this shared effort.
Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, I hope this message finds you well. I want to express my utmost gratitude for your excellent work and for the dedication and speed in the publication process of my article titled "Navigating Innovation: Qualitative Insights on Using Technology for Health Education in Acute Coronary Syndrome Patients." I am very satisfied with the peer review process, the support from the editorial office, and the quality of the journal. I hope we can maintain our scientific relationship in the long term.
Dear Monica Gissare, - Editorial Coordinator of Nutrition and Food Processing. ¨My testimony with you is truly professional, with a positive response regarding the follow-up of the article and its review, you took into account my qualities and the importance of the topic¨.
Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, The review process for the article “The Handling of Anti-aggregants and Anticoagulants in the Oncologic Heart Patient Submitted to Surgery” was extremely rigorous and detailed. From the initial submission to the final acceptance, the editorial team at the “Journal of Clinical Cardiology and Cardiovascular Interventions” demonstrated a high level of professionalism and dedication. The reviewers provided constructive and detailed feedback, which was essential for improving the quality of our work. Communication was always clear and efficient, ensuring that all our questions were promptly addressed. The quality of the “Journal of Clinical Cardiology and Cardiovascular Interventions” is undeniable. It is a peer-reviewed, open-access publication dedicated exclusively to disseminating high-quality research in the field of clinical cardiology and cardiovascular interventions. The journal's impact factor is currently under evaluation, and it is indexed in reputable databases, which further reinforces its credibility and relevance in the scientific field. I highly recommend this journal to researchers looking for a reputable platform to publish their studies.
Dear Editorial Coordinator of the Journal of Nutrition and Food Processing! "I would like to thank the Journal of Nutrition and Food Processing for including and publishing my article. The peer review process was very quick, movement and precise. The Editorial Board has done an extremely conscientious job with much help, valuable comments and advices. I find the journal very valuable from a professional point of view, thank you very much for allowing me to be part of it and I would like to participate in the future!”
Dealing with The Journal of Neurology and Neurological Surgery was very smooth and comprehensive. The office staff took time to address my needs and the response from editors and the office was prompt and fair. I certainly hope to publish with this journal again.Their professionalism is apparent and more than satisfactory. Susan Weiner
My Testimonial Covering as fellowing: Lin-Show Chin. The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews.
My experience publishing in Psychology and Mental Health Care was exceptional. The peer review process was rigorous and constructive, with reviewers providing valuable insights that helped enhance the quality of our work. The editorial team was highly supportive and responsive, making the submission process smooth and efficient. The journal's commitment to high standards and academic rigor makes it a respected platform for quality research. I am grateful for the opportunity to publish in such a reputable journal.
My experience publishing in International Journal of Clinical Case Reports and Reviews was exceptional. I Come forth to Provide a Testimonial Covering the Peer Review Process and the editorial office for the Professional and Impartial Evaluation of the Manuscript.
