Overview of Neurobiology Antibody Applications

Neuroscience or neurobiology is the study of the nervous system. This area of biology studies the structure and function of the nervous system at the molecular, developmental, and physiological level. Neurobiology has long been considered one of the most challenging areas of study to fully comprehend. The nervous system is very complex with organs that are hard to reach physically and components such as emotions and consciousness are difficult to measure. However, scientific advances in recent decades have given us better insight to the functions of the nervous system, how these functions are regulated, what causes developmental defects or diseases, and how they can be managed or cured. Research tools, such as antibodies, can help examine these aspects at a molecular level, where proteins can be identified, and their functions can be derived.

Explore Neurobiology Antibodies
 

Antibodies are used in a variety of applications in neurobiology research. Each application aids in the study of the structure and function of specialized cells within the nervous system. These applications include:

• Detection in western blotting
• Visualization in live and fixed cells or tissues using immunohistochemistry, immunocytochemistry, or immunofluorescence
• Protein enrichment/the study of protein interactions using immunoprecipitation
• Quantitate, characterize, identify, and isolate cell types/populations using flow cytometry and FACS

Antibodies are used in neurobiology to study neuronal markers, cellular differentiation, and disease markers associated with the nervous system. Target-specific antibodies can help detect low-abundance proteins in complex cellular systems such as neurons. The following sections discuss in further detail how antibodies are used as neuronal cell markers, in in vitro differentiation, and as biomarkers for neurodegeneration.

• Glial cells or neuroglia are non-neuronal cells that function to hold neurons in place, supply nutrients to neurons, and defend neurons against pathogens.
• Astrocytes, which are a sub-type of glial cells, are star-shaped cells that support the blood-brain barrier, as well as, provide structural and metabolic support to neurons.
• Oligodendrocytes, another type of glial cell, provide insulation to neurons in the form of myelin.
• Neuroectoderm, which consists of cells from the ectoderm, are important in the development of the nervous system.
• Microglia, a type of glial cell, that function as macrophage cells and are the first form of immune defense in the central nervous system.

Each cell type also expresses specific protein marker profiles that can be used to identify and characterize the cells. This allows researchers to use antibodies, that target these specific protein markers, as neuronal cell markers. For example, in the data below, antibodies specific to GFAP (an astrocyte marker) and NeuN (a mature neuron marker) are used in immunofluorescence to identify localization of these two cell markers. GFAP is found in the cytoskeleton and NeuN is found in the nucleus. In addition to immunofluorescence data, there is western blot data demonstrating that these antibodies are specific to their targets.

Further markers of interest against commonly studied neuronal cell types are listed in the table below.

Access to appropriate samples in neuroscience research can be challenging. Samples can include animal models and primary neurons, which can be difficult to collect, preserve, and process for sample preparation. Cell repositories carry few immortalized neuronal cell lines and the ones that are available are transformed cells obtained from resected tumors. One way to overcome this challenge is by generating specialized cell/organoid structures in the lab using stem cells. Learn more about using in vitro differentiation models to study neuronal protein function and the critical role antibodies play in characterizing these populations in the Behind the Bench Blog: DIY Neurons for Antibody Validation.

Stem cells from various sources can be differentiated into several neuronal lineages to study developmental and disease physiology. For example, Neural Stem Cells (NSCs) that are positive for proteins such as SOX2 and Nestin can be obtained from Embryonic Stem Cell (ESC) differentiation. The NSCs can further be differentiated into mature neurons that express proteins including beta-III tubulin and MAP2. Similarly, induced Pluripotent Stem Cells (iPSCs) can be differentiated into specialized cell types like Retinal Ganglion Cells (RGCs), a neuronal cell type located in the retina, that produce Optineurin, a Golgi-associated retinal protein.

Below, antibodies against four important markers used to characterize cells during cellular differentiation experiments are showcased. SOX1 is a transcription factor that promotes neuronal cell date determination and differentiation. SOX2 is a protein that is required for stem cell maintenance in the central nervous system. PAX3 is known to regulate neurogenesis during development of the peripheral nervous system. And finally, TUBB3 is an early marker of neuronal differentiation.

Further markers of interest used to characterize different neuronal lineages are listed in the table below.

The nervous system is vulnerable to several disorders that can be brought about by injury, tumors, infarcts, autoimmune conditions, degeneration, and structural/developmental defects. Infectious diseases, such as encephalitis and meningitis, can also affect the brain.

There is currently an urgent need to understand neurodegenerative diseases such as Parkinson’s disease (PD), Alzheimer’s disease (AD), and Huntington’s disease at a cellular and molecular level. This level of understanding could reveal target proteins impacted by these conditions. However, many neurodegenerative diseases share common pathogenic mechanisms and can make the study of the proteins involved even more challenging.

For example, defects in axonal transport has been implicated in both Parkinson’s and Alzheimer’s disease. The SNARE family of proteins that include SNAP25 (synaptosomal-associated protein, molecular mass of 25 kDa) and VAMP1 (vesicle-associated membrane protein-1) play critical roles in facilitating vesicular transport of neurotransmitters across synapses and are found to be mutated in both neurodegenerative disorders.

Another example, where diseases share a pathogenic mechanism, is Amyotrophic Lateral Sclerosis (ALS) and in Spinal Muscular Atrophy (SMA). Abnormal assembly of neurofilaments is found in both diseases. Neurofilament proteins are found in neuron cytoplasm and are important for signal transmissions across axons. Degeneration of neurons in disease conditions releases neurofilaments into the blood or cerebrospinal fluid where they can be detected to aid diagnosis.

To tease apart common disease mechanisms, antibodies can be used in a variety of applications to provide insights into protein signaling pathways and identifying the difference in the structures and functions between normal and diseased conditions. Given the complexity, low-abundance, and often overlapping nature of proteins involved in neuronal disorders, it is imperative that scientists choose antibodies that are certain to recognize their intended targets. Below are examples of data using antibodies tested for specificity against four proteins important to some neurodegenerative diseases.

Further proteins of interest used as biomarkers of some neurodegenerative diseases are listed in the table below.

The nervous system is a challenging area of biology. Research in the neuroscience field helps us understand the intricacies behind the processes, the developmental cues, and the identification of treatment modalities for different neurological diseases. The scope of neuroscience in recent decades has widened significantly to include cellular and molecular aspects of the nervous system and has led to advancements in research models and tools. Neuroscientists are increasingly curious to find ways to predict, prevent, and treat anomalies. However, a lack of quality research reagents poses challenges. Tools such as specificity-verified antibodies can help overcome these challenges and allow researchers to unveil the curiosities surrounding the nervous system.