Image Analysis of Neurofilament Immunoreactivity in Human Spiral Ganglion

Abstract

Computer-assisted image analysis was used to study the immunoreactivity to NF-L, NF-M and NF-H in human spiral ganglion cells. The concentrations, represented by the relative mean grey of NF-L, NF-M and NF-H, were calculated and compared. The cell-area mean-grey correlations for NF-L, NF-M and NF-H were analysed and calculated, showing that NF-M is more specific to the larger cells (type 1?) and NF-H is more specific for the smaller cells (type 2?), while NF-L is non-specific for cell size. These findings confirm several previous assumptions by providing a quantitative basis. We conclude that image analysis is a useful – even essential – tool for the analysis of immunostained temporal bone sections.

Age-Related Changes of Myelin Basic Protein in Mouse and Human Auditory Nerve

“…When tested by immunoblotting on pig neurofilament polypeptides, the antibody reacts with an epitope in the tail domain of neurofilament 200, also referred to as the H-subunit, which is present on both the phosphorylated and non-phosphorylated forms of this polypeptide. The staining patterns with neurofilament 200 seen here in the mouse and human auditory nerve were similar to those in previous descriptions [87], [107]. The neuronal class III β-tubulin (TUJ1) rabbit monoclonal antibody was raised against microtubules derived from rat brain and is highly reactive to mammalian neuron-specific class III β-tubulin, but not to glial β-tubulin (manufacturer's technical information).…”

“…Immunohistochemical techniques have been used successfully to define the expression of several proteins in paraformaldehyde-fixed cryostat sections of human ears. These include basement membrane proteins [82], peripherin [83], synaptophysin [84], connexin [85], neurofilament [86], [87], aquaporins [88], a subclass of the ATP-binding cassette transporter proteins [89], tyrosine kinase receptor B and BDNF [90]. Using paraffin-embedded sections, Robertson et al [91] were the first to successfully demonstrate aggregation of cochlin expression in a human ear obtained from a DFNA9 donor.…”

Estimation of the number of nerve fibers in the human vestibular endorgans using unbiased stereology and immunohistochemistry

“…The identification of nerve fibers in vestibulo-cochlear endorgans can be made with the use of specific antibodies against neurofilaments and immunohistochemistry (Dau and Wenthold, 1985;Hafidi and Romand, 1989;Hsu et al, 1997;Nishizaki and Anniko, 1995). Neurofilament proteins are intermediate filaments that are specific to neurons, dendrites and axons.…”