In a human body, the sciatic nerve is considered to be the largest single nerve in the body (Wolf et al., 2014). This is because it runs from the lower spine on each side via the buttocks and back of the thighs. In the event there is an injury of the sciatic nerve, a person may experience a painful throbbing of the limbs as well as the lower back CITATION NIH16 \l 1033 (NIH, 2016). In the study of the treatment of the sciatic nerve, lab experiments using mice have been done in the past to determine the best remedy for treating such hurts. This dissertation is a discussion of the advantages as well as the disadvantages of using GAP-43 as a neuroregeneration marker examination tool in a rat sciatic nerve injury examination.
Keywords Used to Search the Journal in NCBI or PubMed and Inclusion and Exclusion Criteria
This dissertation was developed on a research that was published on the NCBI website. The research was published on September 10, 2016, and it focused on the GAP-43, a growth associated protein used in the treatment of a sciatic nerve injury in a Norway rat CITATION NCB16 \l 1033 (NCBI). The keywords used to attain the access the publication included GAP-43, neuroregeneration marker and sciatic nerve injury. The materials for inclusion in the research were all journals summarizing the effects of GAP-43 on a sciatic rat. In addition, scientific websites with relevant information related to the subject matter will also be considered. The materials for exclusion in the research include all other publications with the exception of peer-reviewed scientific journals and publications from the PubMed or NCBI website.
GAP-43 As Neuroregenaration Examination Marker
The term neuroregeneration characterizes the repair as well as regrowth of cells, cell products and nervous tissues (Coletti et al., 2013). In most cases, such mechanisms are inclusive of the generation of glia, new neurons, synapses, axons as well as and myelin (Sakry, Karram & Trotter, 2011). Neuroregeneration is different from the central nervous system (CNS) and the peripheral nervous system (PNS) (Huebner & Strittmatter, 2009). This is in terms of functional mechanisms and also in terms of speed and extent of development.
If an axon is destroyed, the tissue distal segment goes through a Wallerian degeneration (Rotshenker, 2011). As a result, it loses its myelin sheath. In addition, the proximal segment may ether die through the process of apoptosis or it can undergo through chromatolytic reaction (Al-Abdulla & Martin, 1988). The chromatolytic reaction is considered an attempt of the proximal segment to repair the destroyed axons (Siebert, Eade & Osterhout, 2015). In the CNS, synaptic stripping can take place if the glial foot overruns the dead synapse.
The abbreviation GAP-43 is used to denote the growth associated protein 43, which is a protein situated in human beings and it is encoded by the GAP-43 gene (Uittenbogaard, Martinka & Chiaramello, 2003). GAP-43 in the past has also been termed as a growth or a protein. This is because it is manifested in very elevated levels during the development of the neuro growth cones in humans. This is also, during the axonal regeneration. Nevertheless, it can also be used as a neuro regeneration marker when investigating the inflammation of the sciatic nerve in rats (Miao et al. 2013).
The GAP-43 protein is classified to be a very imperative component of both presynaptic terminal and axon, and its non-mutation can lead to the death of rat in a few days after its birth (Meskini et al., 2007). This is essentially due to pathfinding defects of the axon. GAP-43 is considered to be a cytoplasmic protein of the nervous tissue, which becomes stuck to the membrane through the process of dual palmitoylation sequence (Gorgels et al., 1989). This occurs during the cysteines sequences number 3 and 4.
The sequence of neuroregeneration marker targets the lipid rafts (Wang & Schey, 2015). It is a primary protein kinase C substrate (PKC), and it is believed to play a leading role in the formation of neurite, plasticity as well as regeneration (Dumont et al., 2012). The function of GAP-43 in the development of the CNS is not limited to just the effects of axons. It is also considered to be a component of centrosome and the differentiating neurons, which do not show the GAP-43. In addition, it shows mislocalization of both the mitotic spindles as well as centrosome, and this is specifically in the divisions of neurogenic cells (Neumuller & Knoblich, 2009). Consequently, in the cellular cerebrum, there is a failure in the neuronal precursor pool to expand optimally. As a result, the cerebrum becomes essentially small.
Steps for Using GAP-43 in Rats Examination
When using the GAP-43 protein for a rat examination, the rat must first be intracardially perfused with approximately four percent paraformaldehyde (Brenneman et al., 2007). Then, a DRG tissue must be post-fixed overnight using the same fixative. It should then be cryoprotected in approximately twenty percent of sucrose and later frozen inside an OCT. Later, the IHC free floating is carried out on the 20um sections of the rat tissue, which have been fixed, iced and then cut.
The primary antibody to be used is then incubated at a room temperature for 1/300 over a night while the secondary antibody is incubated for two hours at room temperature at 1/1000 (Simmons et al., 2008). In this case, the GAP-43 protein is used as the antibody. The results of this examination can be well observed within two weeks following the sciatic nerve injury in a rat. The staining of GAP-43 proteins in the rat body tissues around the injured region of the rats body can be observed. In most cases, the staining is usually localized in the cytoplasm sections of several of lesioned neurons and also in most axons.
Advantages of GAP-43 When Employed as a Neuroregenaration Marker
There are various merits that can be associated with using GAP-43 protein compound as a neuroregenerational marker. This is during the process of correcting a rat sciatic nerve injury. This is because the GAP-43 plays a fundamental role in the growth as well as the development of the mammalian Central Nervous System (CNS) (Moore, Apara & Goldberg, 2011). First, GAP-43 is essential for the maintenance of both the structure as well as the dynamics of the axonal fibers (Holahan et al., 2007). It is also essential for the correction of the rats synaptic terminals at optimal conditions as well as in the cases of lesion-triggered axonal sprouting.
In addition, the GAP-43 proteins are attributed to ideal nerve growth, since they belong to the neuromodulin family that also possess a 1 IQ domain (Liu, Fisher & Storm, 1994). It is also associate to be a primary constituent of the motile, which is also referred to as the growth cones. The motile is responsible for the formation of the tips of the cells elongating axons (Dent, Gupton & Gertler, 2011). All these characteristics of the GAP-43 make it an ideal choice if used as a marker during a rat sciatic injury investigation. In addition, GAP-43 is also essential for this examination because of its phosphorylation properties through a compound formed by the protein kinase C.
In this case, phosphorylation is precisely matched with particular types of synaptic plasticity. In addition, GAP-43 facilitates cellular localization in the rats cell membranes. It also supports cellular projection that helps in the growth of the cone membranes. In addition, the GAP-43 creates a cell junction through the synapse process. Ultimately the protein creates a cytoplasmic surface made of growth cones as well as synaptic plasma membranes.
The GAP-43 protein also facilitates clear visibility of the healing progress during a sciatic nerve examination in rats. This is because it offers colored results of the dorsal root ganglion tissues, in a rat that is either subjected to a spinal nerve ligation or a spinal cord tissue lysate. In addition, the GAP-43 protein is an efficient indicator as it has been validated as a marker by other tests with excellent results. Some of the tests where the GAP-43 has been used include the ICC/IF, WB IHC-Fr as well as the IHC (PFA fixed).
Tabulation Summary of the Advantages of Using Gap-43 as a Neuroregenerational Marker
Gap-43 protein also aids in grown and development. It plays a fundamental role in the growth as well as the development of the mammalian Central Nervous System (CNS). It is also essential for the maintenance of both the structure as well as the dynamics of the axonal fibers. It is also essential for the correction of the rats synaptic terminals at optimal conditions as well as in the cases of lesion-triggered axonal sprouting.
Supports the ideal growth of nerves. This is because it belongs to the neuromodulin family that also possess a 1 IQ domain. It is also associate to be a primary constituent of the motile, which is also referred to as the growth cones. The motile is responsible for the formation of the tips of the cells elongating axons.
It has phosphorylation properties. This is through a compound formed by the protein kinase C. In this case, phosphorylation is precisely matched with particular types of synaptic plasticity.
Facilitates cellular localization. This is achieved through the rats cell membranes.
Supports cellular projection This helps in the growth of the cone membranes.
Creates a cell junction. This is achieved through the synapse process.
Gap-43 protein creates a cytoplasmic surface. The surface is made of growth cones as well as synaptic plasma membranes.
Facilitates clear visibility of the healing progress during a sciatic nerve examination in rats. This is because it offers colored results of the dorsal root ganglion tissues, in a rat that is either subjected to a spinal nerve ligation or a spinal cord tissue lysate.
It has been validated as a marker by other tests with excellent results. It has been validated by tests such as the ICC/IF, WB IHC-Fr as well as the IHC (PFA fixed).
Disadvantages of GAP-43 When Employed as a Neuroregenaration Marker
The GAP-43 compound has haploinsufficiency characteristics, which is manifested for the cortical phenotypes. Haploinsufficiency is a condition where diploid organisms possess one copy of a functional gene only. The copy is not capable of producing enough gene product for creating a wild type condition in the rat. In such a case, it could make the rat remain in a diseased state. This means that if the correct procedures are not employed when using GAP-43 as a marker for neuroregenaration, it may render the rat used as the specimen to remain in an anomalous condition.
In addition, studies have portrayed that the GAP-43 gene can be extremely lethal if administered on a knockout mouse line. A knockout mouse is a term used to refer to a genetically modified mouse, whose existing gene has been disrupted or entirely replaced with an artificial strand of DNA. The lethality of GAP-43 is manifested just a few days after the mouse line is born. This is because GAP-43 takes an imperative role in the creation of the mammalian CNS. In such a case, the telencephalic commissures may fail to develop, and as a result, the thalamocortical afferents could become mistargeted. This is more so in the rats somatosensory predominant barrel of the cortex.
In addition, GAP-43 is not suitable for use in the IHC-P. The IHC-P the abbreviation for Immunohistochemistry, which is a procedure employed to demonstrate the presence as well as the location of proteins in a body tissue (Ivell, Teerds & Hoffman, 2014). The method has less sensitivity quantitatively when compared to other immunoassays like the ELISA or the Western...
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