SEPT5 Human

What is SEPT5 and what are its primary cellular functions?

SEPT5 (Septin-5), also known as CDCREL, CDCREL-1, CDCREL1, H5, and PNUTL1 , belongs to the evolutionarily conserved septin family of GTP-binding proteins. These proteins form heteromeric complexes that assemble into higher-order structures such as filaments and rings.

Methodologically, studying SEPT5 function requires:

• Live-cell imaging with fluorescently tagged SEPT5

• Immunoprecipitation to identify binding partners

• Knockout/knockdown studies to observe phenotypic changes

• Structural studies using X-ray crystallography or cryo-EM

SEPT5 participates in several critical cellular processes including cytoskeletal organization, membrane dynamics, vesicle trafficking, and exocytosis, particularly in neuronal cells.

What techniques are most effective for detecting SEPT5 in human tissue samples?

Researchers have multiple options for SEPT5 detection, each with specific advantages:

ELISA-based detection:

• Sandwich ELISA provides a quantitative method for SEPT5 detection in samples

• Process involves:

  • Pre-coating microplates with SEPT5-specific antibody

  • Addition of samples where SEPT5 binds to immobilized antibody

  • Application of biotin-conjugated SEPT5-specific antibody

  • Addition of Streptavidin-HRP, substrate solution, and measurement of color development

Western blotting:

• Allows detection of SEPT5 protein and assessment of post-translational modifications

• Requires optimization of antibody dilutions, blocking conditions, and sample preparation methods

qPCR for mRNA quantification:

• Enables detection of SEPT5 transcripts

• Requires careful primer design and validation

• Should include appropriate housekeeping genes for normalization

How is SEPT5 expression distributed across human tissues?

While the search results don't provide comprehensive tissue distribution data, methodologically, researchers should approach this question through:

• RT-qPCR to quantify mRNA levels across tissues

• Western blotting to detect protein expression

• Immunohistochemistry to visualize cellular localization

• Single-cell RNA sequencing to identify cell-type specific expression

When designing studies to analyze SEPT5 expression, researchers should consider:

• Appropriate reference genes for normalization

• Antibody specificity for protein detection

• Tissue preparation methods to preserve epitope integrity

• Age and sex variables that may influence expression levels

How should researchers design experiments to study SEPT5 function in cellular models?

When designing experiments to study SEPT5 function, researchers should consider:

Experimental design principles:

• Include appropriate controls (positive, negative, vehicle)

• Ensure internal validity through rigorous design

• Randomize and blind experiments when possible to reduce bias

• Use sufficient biological and technical replicates to ensure statistical power

Cellular models:

• Neuronal cell lines (such as SHSY-5Y used in Parkinson's disease studies)

• Primary neurons for physiologically relevant conditions

• Patient-derived iPSCs for disease modeling

Manipulation approaches:

• CRISPR/Cas9 gene editing for knockout/knockin studies

• siRNA/shRNA for transient knockdown

• Overexpression systems with tagged SEPT5 constructs

What are the best practices for SEPT5 protein detection and quantification?

The Human Septin-5 (SEPT5) ELISA Kit employs a two-site sandwich ELISA to quantitate SEPT5 in samples . Methodological considerations include:

Sample types:

• Cell culture supernatants

• Biological fluids

• Plasma

• Serum

Key procedural elements:

• Antibody pre-coating for SEPT5 capture

• Sample incubation

• Addition of biotin-conjugated SEPT5-specific antibody

• Application of Streptavidin-HRP

• Substrate addition and color development proportional to SEPT5 concentration

Quality control measures:

• Standard curves with known SEPT5 concentrations

• Multiple technical replicates

• Inclusion of appropriate controls

• Validation across different sample types

How can researchers ensure validity when studying SEPT5 in experimental models?

To ensure experimental validity when studying SEPT5:

Internal validity considerations:

• Control for confounding variables

• Establish clear temporal relationships between intervention and outcome

• Utilize proper control conditions

• Apply rigorous experimental design principles

External validity considerations:

• Select models with translational relevance

• Validate findings across multiple experimental systems

• Consider species differences when using animal models

Statistical approaches:

• Determine appropriate sample sizes through power analysis

• Select suitable statistical tests based on data distribution

• Control for multiple comparisons

• Report effect sizes alongside p-values

What evidence links SEPT5 to Parkinson's disease pathophysiology?

Research has established several connections between SEPT5 and Parkinson's disease:

Gene expression evidence:

• Significant differences in SEPT5 gene expression have been observed in the substantia nigra between PD models and controls

• There is a significant alteration (P < 0.05) in SEPT5 expression in the substantia nigra of rotenone-treated rats

Relationship with other PD-associated genes:

• Inhibition of miR-185 gene expression led to increased LRRK2 expression in SHSY-5Y cells

• LRRK2 inhibition decreased rotenone-induced cellular toxicity

Experimental models demonstrating this relationship:

• In vivo model: rotenone-treated male Wistar rats

• In vitro model: SHSY-5Y cell line exposed to rotenone

How does the relationship between miR-185 and SEPT5 impact neurological function?

Studies have identified important regulatory relationships:

Key findings:

• miR-185 expression is significantly decreased in the substantia nigra (SN) and striatum (ST) of rotenone-treated groups compared to healthy controls (P < 0.05)

• This altered expression pattern occurs in brain regions critical for dopaminergic function

• The results suggest a protective role of miR-185 in preventing the development of PD

Experimental evidence:

• In vivo studies utilizing rotenone-treated rats demonstrated altered miR-185 and SEPT5 expression

• In vitro studies showed that inhibition of miR-185 led to increased LRRK2 expression

Methodological approaches:

• SYBR Green Real-Time PCR Method for measuring expression levels

• Analysis of multiple genes (miR-185, SEPT5, LRRK2, PARK2) in dopaminergic areas

• Cell viability assays following gene inhibition

What are the potential therapeutic implications of targeting SEPT5 in neurological disorders?

Based on emerging evidence linking SEPT5 to neurological conditions:

Potential therapeutic strategies:

• Modulation of SEPT5 expression levels

• Targeting the miR-185 regulatory pathway

• LRRK2 inhibition, which showed decreased cellular toxicity effects of rotenone on SHSY-5Y cells

Methodological considerations for therapeutic development:

• Target validation using multiple model systems

• Identification of druggable sites or interactions

• Development of screening assays for small molecule discovery

• Delivery systems for CNS targeting

How do genetic factors influence SEPT5 expression and function in human disease?

Genetic variants affecting SEPT5 may contribute to disease susceptibility:

Chromosomal location considerations:

• SEPT5 is located on human chromosomal location 22q11.2

• This region may be involved in the development of Parkinson's disease

Methodological approaches to study genetic influences:

• Gene expression analysis in affected tissues (substantia nigra, striatum)

• In vivo and in vitro disease models to assess expression changes

• Genomic association studies in patient populations

Experimental techniques:

• SYBR Green Real-Time PCR for expression analysis

• Gene inhibition studies to assess functional consequences

• Cell viability assessments following genetic manipulation

What is the relationship between SEPT5 and other Parkinson's disease-associated genes?

Research suggests complex interactions between SEPT5 and other PD-related genes:

Key gene interactions:

• miR-185 and SEPT5 both show altered expression in PD models

• Inhibition of miR-185 affects LRRK2 expression

• LRRK2 inhibition influences cellular response to neurotoxins

Experimental evidence:

• Expression analysis of multiple genes (SEPT5, LRRK2, PARK2) in the same experimental models

• Functional studies demonstrating the effect of inhibiting one gene on the expression of others

How can SEPT5 detection methods be optimized for clinical biomarker applications?

For optimizing SEPT5 detection in clinical applications:

Current detection technologies:

• Sandwich ELISA methodology allows quantitative measurement of SEPT5

• The assay employs antibodies specific for SEPT5, with biotin-conjugated detection antibodies and Streptavidin-HRP for visualization

Methodological considerations for clinical applications:

• Sample type selection (serum, plasma, CSF)

• Standardization of collection and processing protocols

• Establishment of reference ranges

• Validation across diverse patient populations

Quality control measures:

• Multiple replicates to ensure reproducibility

• Inclusion of standard curves with known concentrations

• Assay validation across different clinical settings

• Cross-platform validation