Research Article
Volume 19 Issue 8 - 2020
Transfection of Human Gingival Fibroblasts with Cementum Protein 1 does not Alter the Ultrastructure of the Interchromatin Granule Clusters in the Cell Nucleus
Edith Florely Del Toro-Rangel1, Reyna Lara-Martínez1, María de Lourdes Segura-Valdez1, Higinio Arzate2, Carlos Esteban Villegas-Mercado3 and Luis F Jiménez-García1*
1Electron Microscopy Laboratory and Cell Nanobiology Laboratory, Department of Cell Biology, Faculty of Sciences, National Autonomous University of Mexico (UNAM), Circuito Exterior, Mexico
2Faculty of Dentistry, Periodontal Biology Department, DEPeI, National Autonomous University of Mexico (UNAM) Mexico City, Mexico
3Unidad de Posgrado, Facultad de Odontología, Universidad Autónoma de Sinaloa, Culiacán, Sinaloa, México
*Corresponding Author: Luis F Jiménez-García, Electron Microscopy Laboratory and Cell Nanobiology Laboratory, Department of Cell Biology, Faculty of Sciences, National Autonomous University of Mexico (UNAM), Circuito Exterior, Mexico.
Received: July 17, 2020; Published: July 29, 2020




Abstract

The cell nucleus contains domains related to gene expression. The nucleolus is involved in pre-rRNA metabolism while interchromatin granules are involved in pre-mRNA metabolism. It was recently shown that no ultrastructural changes are detected in the nucleolar organization, but no data of the effect of transfection in speckles are known. The aim of this work is to know if the interchromatin granule clusters display ultrastructural changes when introducing cement protein 1 (CEMP1) genes into human gingival fibroblasts (HGFs), a cell line of interest since they do not mineralize and upon transfection they perform mineralization, a potential to use in treatments of periodontal disease. Here we explored ultrastructural changes in interchromatin granule clusters after transfection of CEMP1. Non transfected and transfected cells display similar nuclear particles that form clumps in the interchromatin space and contain 10 - 15 nm in diameter individual granules intermingled with fibers, which are composed of ribonucleoproteins. We conclude that they correspond to interchromatin granule clusters. The absence of ultrastructural changes in the morphology of interchromatin granule clusters suggests that after transfection the general ultrastructure of particles related to pre-mRNA metabolism is not altered. These data may support the development of new dental materials.

Keywords: Cementum Protein 1; Human Gingival Fibroblasts; Interchromatin Granule Clusters; Cell Nucleus; Transfection

References

  1. DL Spector. “Macromolecular domains within the cell nucleus". Annual Review in Cell Biology 9 (1993a): 265-315.
  2. DL Spector., et al. “Dynamics of transcription and pre-mRNA splicing within the mammalian cell nucleus”. Cold Spring Harbor Symposium in Quantitative Biology 58 (1993b): 799-805.
  3. T Misteli., et al. “The dynamics of a pre-mRNA splicing factor in living cells”. Nature 387 (1997): 523-527.
  4. T Misteli. “Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function”. Journal of Cell Science 113 (2000): 1841-1849.
  5. M Dundr and T Misteli. “Functional architecture in the cell nucleus”. Biochemistry Society 356 (2001): 297-310.
  6. A Monneron and W Bernhard. “Fine structural organization of the interphase nucleus in some mammalian cells”. Journal of Ultrastructure Research 27 (1969): 266-288.
  7. S Fakan. “Perichromatin fibrils are in situ forms of nascent transcripts”. Trends in Cell Biology 4 (1994): 86-90.
  8. AI Lamond and D Spector. “Nuclear speckles: a model for nuclear organelles”. Nature Reviews in Molecular and Cell Biology 4 (2003): 605-612. 
  9. DL Spector and AI Lamond. “Nuclear speckles”. Cold Spring Harbor Perspective in Biology 3 (2011): a000646.
  10. P Sacco-Bubulya and DL Spector. “Disassembly of interchromatin granule clusters alters the coordination of transcription and pre-mRNA splicing. Journal of Cell Biology 156 (2002): 425-436. 
  11. I Melcak., et al. “Nuclear pre-mRNA Compartmentalization: Trafficking of released transcripts to splicing factor reservoirs”. Molecular Biology of the Cell 11 (2000): 497-510.
  12. T Misteli and DL Spector. “The cellular organization of gene expression”. Current Opinion in Cell Biology 10 (1998): 323-331.
  13. R George-Téllez., et al. “Cellular organization of pre-mRNA splicing factors in several tissues. Changes in the uterus by hormone action”. Biology of the Cell 94 (2002): 99-108.
  14. DJ Abraham., et al. “New developments in fibroblast and myofibroblast biology : Implications for fibrosis and scleroderma”. Current Rheumatology Reports 9 (2007): 136-143.
  15. HY Chang., et al. “Diversity, topographic differentiation and positional memory in human fibroblasts”. Proceedings of the National Academy of Sciences U. S. A. 99 (2002): 12877-12882.
  16. F Guo., et al. “Mechanical tension increases CCN2/CTGF expression and proliferation in gingival fibroblasts via a TGFβ-dependent mechanism”. PloS One 6 (2011): 19756.
  17. X Shi-wen., et al. “Endogenous endothelin-1 signaling contributes to type I collagen and CCN2 overexpression in fibrotic fibroblasts”. Matrix Biology 26 (2007): 625-632. 
  18. MA Alvarez-Pérez., et al. “Molecular cloning, expression and immunolocalization of a novel human cementum-derived protein (CP-23)”. Bone 38 (2006): 409-419. 
  19. B Carmona-Rodríguez., et al. “Human cementum protein 1 induces expression of bone and cementum proteins by human gingival fibroblasts”. Biochemical and Biophysical Research Communications 358 (2007): 763-769. 
  20. J Nuñez., et al. “Human cementoblasts express enamel-associated molecules In vitro and In vivoJournal of Periodontal Research 45 (2010): 809-814. 
  21. AS Narayan and RC Page. “Biochemical characterization of collagens synthesized by fibroblasts derived from normal and diseased human gingiva”. Journal of Biological Chemistry 251 (1976): 5464-5471.
  22. CE Villegas-Mercado., et al. “Cementum protein 1 transfection does not lead to ultrastructural changes in nucleolar organization of human gingival fibroblasts”. Journal of Periodontal Research 53 (2018): 636-642. 
  23. W Bernhard. “A new staining procedure for electron microscopical cytology”. Journal of Ultrastructural Research 27 (1969): 250-265.
  24. H Arzate., et al. “Immunolocalization of a human cementoblastoma-conditioned medium-derived protein”. Journal of Dental Research 81 (2002): 541-546. 
  25. L Hoz., et al. “Cementum protein 1 (CEMP1) differentiation by human periodontal ligament cells under three-dimensional culture conditions”. Cell Biology International 36 (2011): 129-136.
Citation: Luis F Jiménez-García., et al. “Transfection of Human Gingival Fibroblasts with Cementum Protein 1 does not Alter the Ultrastructure of the Interchromatin Granule Clusters in the Cell Nucleus”. EC Dental Science 19.8 (2020): 179-186.

PubMed Indexed Article


EC Pharmacology and Toxicology
LC-UV-MS and MS/MS Characterize Glutathione Reactivity with Different Isomers (2,2' and 2,4' vs. 4,4') of Methylene Diphenyl-Diisocyanate.

PMID: 31143884 [PubMed]

PMCID: PMC6536005


EC Pharmacology and Toxicology
Alzheimer's Pathogenesis, Metal-Mediated Redox Stress, and Potential Nanotheranostics.

PMID: 31565701 [PubMed]

PMCID: PMC6764777


EC Neurology
Differences in Rate of Cognitive Decline and Caregiver Burden between Alzheimer's Disease and Vascular Dementia: a Retrospective Study.

PMID: 27747317 [PubMed]

PMCID: PMC5065347


EC Pharmacology and Toxicology
Will Blockchain Technology Transform Healthcare and Biomedical Sciences?

PMID: 31460519 [PubMed]

PMCID: PMC6711478


EC Pharmacology and Toxicology
Is it a Prime Time for AI-powered Virtual Drug Screening?

PMID: 30215059 [PubMed]

PMCID: PMC6133253


EC Psychology and Psychiatry
Analysis of Evidence for the Combination of Pro-dopamine Regulator (KB220PAM) and Naltrexone to Prevent Opioid Use Disorder Relapse.

PMID: 30417173 [PubMed]

PMCID: PMC6226033


EC Anaesthesia
Arrest Under Anesthesia - What was the Culprit? A Case Report.

PMID: 30264037 [PubMed]

PMCID: PMC6155992


EC Orthopaedics
Distraction Implantation. A New Technique in Total Joint Arthroplasty and Direct Skeletal Attachment.

PMID: 30198026 [PubMed]

PMCID: PMC6124505


EC Pulmonology and Respiratory Medicine
Prevalence and factors associated with self-reported chronic obstructive pulmonary disease among adults aged 40-79: the National Health and Nutrition Examination Survey (NHANES) 2007-2012.

PMID: 30294723 [PubMed]

PMCID: PMC6169793


EC Dental Science
Important Dental Fiber-Reinforced Composite Molding Compound Breakthroughs

PMID: 29285526 [PubMed]

PMCID: PMC5743211


EC Microbiology
Prevalence of Intestinal Parasites Among HIV Infected and HIV Uninfected Patients Treated at the 1o De Maio Health Centre in Maputo, Mozambique

PMID: 29911204 [PubMed]

PMCID: PMC5999047


EC Microbiology
Macrophages and the Viral Dissemination Super Highway

PMID: 26949751 [PubMed]

PMCID: PMC4774560


EC Microbiology
The Microbiome, Antibiotics, and Health of the Pediatric Population.

PMID: 27390782 [PubMed]

PMCID: PMC4933318


EC Microbiology
Reactive Oxygen Species in HIV Infection

PMID: 28580453 [PubMed]

PMCID: PMC5450819


EC Microbiology
A Review of the CD4 T Cell Contribution to Lung Infection, Inflammation and Repair with a Focus on Wheeze and Asthma in the Pediatric Population

PMID: 26280024 [PubMed]

PMCID: PMC4533840


EC Neurology
Identifying Key Symptoms Differentiating Myalgic Encephalomyelitis and Chronic Fatigue Syndrome from Multiple Sclerosis

PMID: 28066845 [PubMed]

PMCID: PMC5214344


EC Pharmacology and Toxicology
Paradigm Shift is the Normal State of Pharmacology

PMID: 28936490 [PubMed]

PMCID: PMC5604476


EC Neurology
Examining those Meeting IOM Criteria Versus IOM Plus Fibromyalgia

PMID: 28713879 [PubMed]

PMCID: PMC5510658


EC Neurology
Unilateral Frontosphenoid Craniosynostosis: Case Report and a Review of the Literature

PMID: 28133641 [PubMed]

PMCID: PMC5267489


EC Ophthalmology
OCT-Angiography for Non-Invasive Monitoring of Neuronal and Vascular Structure in Mouse Retina: Implication for Characterization of Retinal Neurovascular Coupling

PMID: 29333536 [PubMed]

PMCID: PMC5766278


EC Neurology
Longer Duration of Downslope Treadmill Walking Induces Depression of H-Reflexes Measured during Standing and Walking.

PMID: 31032493 [PubMed]

PMCID: PMC6483108


EC Microbiology
Onchocerciasis in Mozambique: An Unknown Condition for Health Professionals.

PMID: 30957099 [PubMed]

PMCID: PMC6448571


EC Nutrition
Food Insecurity among Households with and without Podoconiosis in East and West Gojjam, Ethiopia.

PMID: 30101228 [PubMed]

PMCID: PMC6086333


EC Ophthalmology
REVIEW. +2 to +3 D. Reading Glasses to Prevent Myopia.

PMID: 31080964 [PubMed]

PMCID: PMC6508883


EC Gynaecology
Biomechanical Mapping of the Female Pelvic Floor: Uterine Prolapse Versus Normal Conditions.

PMID: 31093608 [PubMed]

PMCID: PMC6513001


EC Dental Science
Fiber-Reinforced Composites: A Breakthrough in Practical Clinical Applications with Advanced Wear Resistance for Dental Materials.

PMID: 31552397 [PubMed]

PMCID: PMC6758937


EC Microbiology
Neurocysticercosis in Child Bearing Women: An Overlooked Condition in Mozambique and a Potentially Missed Diagnosis in Women Presenting with Eclampsia.

PMID: 31681909 [PubMed]

PMCID: PMC6824723


EC Microbiology
Molecular Detection of Leptospira spp. in Rodents Trapped in the Mozambique Island City, Nampula Province, Mozambique.

PMID: 31681910 [PubMed]

PMCID: PMC6824726


EC Neurology
Endoplasmic Reticulum-Mitochondrial Cross-Talk in Neurodegenerative and Eye Diseases.

PMID: 31528859 [PubMed]

PMCID: PMC6746603


EC Psychology and Psychiatry
Can Chronic Consumption of Caffeine by Increasing D2/D3 Receptors Offer Benefit to Carriers of the DRD2 A1 Allele in Cocaine Abuse?

PMID: 31276119 [PubMed]

PMCID: PMC6604646


EC Anaesthesia
Real Time Locating Systems and sustainability of Perioperative Efficiency of Anesthesiologists.

PMID: 31406965 [PubMed]

PMCID: PMC6690616


EC Pharmacology and Toxicology
A Pilot STEM Curriculum Designed to Teach High School Students Concepts in Biochemical Engineering and Pharmacology.

PMID: 31517314 [PubMed]

PMCID: PMC6741290


EC Pharmacology and Toxicology
Toxic Mechanisms Underlying Motor Activity Changes Induced by a Mixture of Lead, Arsenic and Manganese.

PMID: 31633124 [PubMed]

PMCID: PMC6800226


EC Neurology
Research Volunteers' Attitudes Toward Chronic Fatigue Syndrome and Myalgic Encephalomyelitis.

PMID: 29662969 [PubMed]

PMCID: PMC5898812


EC Pharmacology and Toxicology
Hyperbaric Oxygen Therapy for Alzheimer's Disease.

PMID: 30215058 [PubMed]

PMCID: PMC6133268


News and Events


December Issue Release

We always feel pleasure to share our updates with you all. Here, notifying you that we have successfully released the November issue of respective journals and the latest articles can be viewed on the current issue pages.

Submission Deadline for Upcoming Issue

ECronicon delightfully welcomes all the authors around the globe for effective collaboration with an article submission for the upcoming issue of respective journals. Submissions are accepted on/before December 23, 2022.

Certificate of Publication

ECronicon honors with a "Publication Certificate" to the corresponding author by including the names of co-authors as a token of appreciation for publishing the work with our respective journals.

Best Article of the Issue

Editors of respective journals will always be very much interested in electing one Best Article after each issue release. The authors of the selected article will be honored with a "Best Article of the Issue" certificate.

Certifying for Review

ECronicon certifies the Editors for their first review done towards the assigned article of the respective journals.

Latest Articles

The latest articles will be updated immediately on the articles in press page of the respective journals.