LMNA Rat
Description
Definition and Biochemical Characteristics
LMNA Rat is a recombinant protein produced in Escherichia coli and fused with a His tag for purification . Key features include:
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Structure: Single polypeptide chain of 614 amino acids, corresponding to lamin A/C isoforms .
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Formulation: Supplied in a buffer containing 20 mM Tris-HCl (pH 7.5), 1 mM DTT, 0.3 M NaCl, 1.5 mM EDTA, and 10% glycerol .
| Property | Specification | Source |
|---|---|---|
| Production Organism | E. coli | |
| Post-Translational Modifications | Non-glycosylated | |
| Stability | Store at 4°C (short-term) or -20°C (long-term); avoid freeze-thaw cycles |
Research Applications in Disease Modeling
LMNA Rat is utilized to study laminopathies, including cardiomyopathy, muscular dystrophy, and progeroid syndromes. Key applications include:
Cardiomyopathy and Muscle Dysfunction
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Zebrafish Models: Transgenic zebrafish expressing LMNA mutations (e.g., L35P, R453W) exhibit reduced swim speed, muscle endurance, and fiber size .
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Mouse Models:
Therapeutic Interventions
Associated Diseases and Mutations
LMNA mutations are linked to a spectrum of disorders, including:
Mechanistic Insights
LMNA Rat studies reveal critical pathways disrupted in laminopathies:
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Nuclear Envelope Rupture: Leads to cytoplasmic DNA exposure and macrophage-driven inflammation in cardiomyopathy .
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Cytoskeletal Disruption: Lamin A/C interacts with actin, microtubules, and desmin, impacting mechanotransduction .
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Mitochondrial Dysfunction: SIRT1/Parkin axis regulates mitophagy, mitigating oxidative stress .
Therapeutic Challenges and Future Directions
Product Specs
Prelamin-A/C, Lamin-A/C, 70 kDa lamin, LMNA, LMN1, Renal carcinoma antigen NY-REN-32, Progerin.
METPSQRRAT RSGAQASSTP LSPTRITRLQ EKEDLQELND RLAVYIDRVR SLETENAGLR LRITESEEVV SREVSGIKAA YEAELGDARK TLDSVAKERA RLQLELSKVR EEFKELKARN TKKEGDLLAA QARLKDLEAL LNSKEAALST ALSEKRTLEG ELHDLRGQVA KLEAALGEAK KQLQDEMLRR VDAENRLQTL KEELDFQKNI YSEELRETKR RHETRLVEID NGKQREFESR LADALQELRA QHEDQVEQYK KELEKTYSAK LDNARQSAER NSNLVGAAHE ELQQSRIRID SLSAQLSQLQ KQLAAKEAKL RDLEDSLARE RDTSRRLLAE KEREMAEMRA RMQQQLDEYQ ELLDIKLALD MEIHAYRKLL EGEEERLRLS PSPTSQRSRG RASSHSSQSQ GGGSVTKKRK LESSESRSSF SQHARTSGRV AVEEVDEEGK FVRLRNKSNE DQSMGNWQIR RQNGDDPLMT YRFPPKFTLK AGQVVTIWAS GAGATHSPPT DLVWKAQNTW GCGSSLRTAL INATGEEVAM RKLVRSLTMV EDDEDEDGDD LLHHHHGSHC SSSGDPAEYN LRSRTVLCGT CGQPADKASA SGSGAQVSSQ NCSIM
Q&A
Experimental Design for Studying LMNA Mutations in Rats
Q: How can researchers design experiments to study the effects of LMNA mutations in rats, particularly focusing on cardiac and skeletal muscle laminopathies? A: To study LMNA mutations in rats, researchers can employ genetic engineering techniques to introduce specific mutations into the LMNA gene. This involves using CRISPR-Cas9 or similar methods to create rat models with mutations akin to those found in human laminopathies. Experimental designs should include controls for wild-type LMNA expression and should assess phenotypic changes such as muscle endurance, cardiac function, and histological alterations in muscle tissues. Techniques like immunoelectron microscopy can be used to study lamin organization and nuclear morphology in cardiomyocytes .
Data Analysis and Contradiction Resolution
Q: How can researchers resolve contradictions in data when studying LMNA-related laminopathies, particularly when different studies report varying effects of LMNA mutations on muscle and cardiac tissues? A: Resolving data contradictions involves a thorough review of experimental methodologies, including differences in mutation types, animal models used, and the specific tissues analyzed. Bioinformatic tools can help in comparing gene expression profiles across studies to identify consistent patterns or discrepancies. Additionally, meta-analysis techniques can be applied to pool data from multiple studies, providing a more comprehensive understanding of LMNA mutation effects .
Advanced Research Questions: Mechanisms of LMNA Mutations
Q: What are the underlying mechanisms by which LMNA mutations lead to laminopathies, and how can researchers investigate these mechanisms at the molecular level? A: LMNA mutations disrupt the nuclear lamina structure, affecting nuclear stability and gene expression. Researchers can investigate these mechanisms by using techniques like superresolution microscopy to visualize lamin networks and by analyzing gene expression changes through RNA sequencing. Additionally, studying the interaction between lamins and other nuclear components, such as nuclear pore complexes, can provide insights into how mutations affect cellular function .
Methodological Approaches for Drug Screening
Q: How can researchers use rat models to screen for potential therapeutic agents targeting LMNA-related laminopathies, and what methodologies are most effective? A: Drug screening in rat models involves introducing LMNA mutations and then testing various compounds for their ability to rescue or mitigate the phenotypic effects of these mutations. Techniques like high-throughput screening can be employed to rapidly assess the efficacy of numerous compounds. Additionally, using zebrafish models, as seen with l-carnitine and creatine treatments, can provide preliminary insights before moving to rat models .
Bioinformatic Analysis of LMNA Gene Expression
Q: What bioinformatic tools and databases are available for analyzing LMNA gene expression and its regulation in rat models, and how can these tools aid in understanding laminopathies? A: Researchers can utilize databases like SwissProt and InterPro for protein domain analysis and gene expression databases such as GEO for transcriptomic data. Bioinformatic tools like BLAST and ClustalW can help in comparing LMNA sequences across species and identifying conserved regions. These analyses can provide insights into how LMNA mutations affect gene expression and cellular function .
Comparative Analysis of LMNA Mutations Across Species
Q: How do LMNA mutations manifest differently in rats versus humans, and what are the implications for translational research? A: While LMNA mutations in rats and humans can lead to similar laminopathies, differences in genetic background and physiology may affect the severity and presentation of the disease. Comparative studies should focus on the conservation of lamin A/C functions across species and how specific mutations impact these functions. This can involve comparing gene expression profiles and phenotypic outcomes in both rat models and human patients .
Advanced Techniques for Studying Nuclear Lamina Dynamics
Q: What advanced microscopy techniques can researchers use to study the dynamics of the nuclear lamina in rat cardiomyocytes, particularly in the context of LMNA mutations? A: Techniques such as superresolution microscopy and live-cell imaging can provide detailed insights into the organization and dynamics of the nuclear lamina in cardiomyocytes. These methods allow researchers to visualize how LMNA mutations affect nuclear stability and morphology, which is crucial for understanding the pathogenesis of laminopathies .
Integrating Bioinformatics and Experimental Data
Q: How can researchers integrate bioinformatic analyses with experimental data to better understand the effects of LMNA mutations on rat models? A: Integrating bioinformatic and experimental data involves using computational tools to analyze gene expression changes and predict functional impacts of LMNA mutations. This can be combined with experimental validation using techniques like qPCR and Western blotting to confirm bioinformatic predictions. Databases and pipelines like those mentioned in can facilitate this integration.
Experimental Models for Studying LMNA-Related Cardiomyopathies
Q: What experimental models are most suitable for studying LMNA-related cardiomyopathies in rats, and how can these models be optimized for therapeutic research? A: Mouse models, such as the Lmna N195K mutation, have been extensively used to study cardiomyopathies. For rat models, similar genetic modifications can be applied. Optimizing these models involves ensuring that the mutations accurately replicate human disease phenotypes and using techniques like cardiac-specific gene editing to focus on cardiomyocyte function .
Challenges in Translating LMNA Research to Humans
Q: What are the main challenges in translating research findings from rat models of LMNA mutations to human therapeutic applications, and how can these challenges be addressed? A: Challenges include differences in genetic background, physiology, and disease progression between rats and humans. Addressing these involves using multiple animal models, including larger mammals, and conducting thorough clinical trials to validate therapeutic efficacy. Additionally, understanding the molecular mechanisms underlying LMNA mutations can help in developing targeted therapies applicable across species .
Data Table Example: LMNA Mutations and Their Effects
| Mutation | Phenotypic Effects | Experimental Model |
|---|---|---|
| LMNA(L35P) | Reduced muscle endurance | Zebrafish |
| LMNA(R453W) | Decreased swim speed, improved with creatine | Zebrafish |
| LMNA(N195K) | Cardiac dysfunction, nuclear instability | Mouse |
Product Science Overview
Lamin A/C belongs to the lamin family of proteins, which are intermediate filament proteins. These proteins are highly conserved across species, indicating their fundamental role in cellular biology . Lamin A and Lamin C are produced from the same gene (LMNA) through alternative splicing. They share a common structure, including a central rod domain flanked by a head and a tail domain .
During mitosis, the nuclear lamina disassembles as lamin proteins are phosphorylated. This disassembly is crucial for the breakdown of the nuclear envelope, allowing chromosome segregation . Lamin A/C is also involved in maintaining nuclear stability and regulating gene expression .
Phosphorylation of Lamin A/C at specific sites, such as Ser22, has been identified in vivo in several cell lines . This phosphorylation is part of a typical MAPK/CDK phosphorylation motif, implicating a role in the cell cycle and mitosis . The phosphorylation state of Lamin A/C can influence its function and interactions with other nuclear components.
Recombinant Lamin A/C proteins, such as those derived from rats, are used in various research applications. These recombinant proteins are valuable tools for studying the structure and function of Lamin A/C in different cellular contexts. They are also used in assays to investigate the role of Lamin A/C in diseases and to develop potential therapeutic interventions.
