Immunohistochemical analysis of spinal cord lesions in amyotrophic lateral sclerosis using microtubule-associated protein 2 (MAP2) antibodies.

Immunohistochemical evaluation of spinal twine lesions in amyotrophic lateral sclerosis utilizing microtubule-associated protein 2 (MAP2) antibodies.
We have now studied microtubule-associated protein 2 (MAP2) expression in anterior horn neurons within the cervical and lumbar spinal cords of 19 instances of adult-onset sporadic amyotrophic lateral sclerosis (ALS) utilizing immunohistochemistry. Specimens from 7 sufferers with out neurological illness served as controls.
MAP2 expression decreased within the anterior grey horn of all ALS instances and within the intermediate grey of a number of ALS instances. Such discount correlated with the diploma of degeneration or neuronal loss in anterior horn cells and with the scientific signs of limb weak spot.
Cytopathologically, the MAP2 immunoreactivity decreased equivalent to the prevalence of particular person indicators of neuronal degeneration, comparable to chromatolytic neurons, shrunken neurons and pigmented neurons. MAP2 expression was comparatively nicely preserved within the specimens during which spheroids are conspicuous. The findings of this examine display MAP2 to be a superb marker for the detection and quantification of anterior horn degeneration in ALS.

Distribution of phosphate-independent MAP2 epitopes revealed with monoclonal antibodies in microwave-denatured human nervous system tissues.

In distinction with outcomes obtained in experimental animals, antibodies to microtubule related protein-2 (MAP2) preferentially label irregular constructions in human nervous system tissue samples, however the regular websites at which MAP2 is expressed usually are not well-defined.
To find out the distribution of MAP2 within the human central and peripheral (PNS) nervous programs, we ready monoclonal antibodies (MAbs) particular to MAP2, and in contrast the localization of this MAP in postmortem bovine and human tissues in addition to in a number of human neural cell strains that categorical both neurofilament (NF) or glial filament proteins.
Eight MAbs particular for phosphate-independent epitopes in bovine and human MAP2 had been obtained, and people who carried out nicely in tissues produced immunoreactivity confined to the somatodendritic area of neurons in bovine and human CNS and PNS tissues. Different neural cells didn’t categorical MAP2 immunoreactivity utilizing these MAbs.
Postmortem delays of lower than 24 h previous to tissue denaturation didn’t have an effect on the distribution of MAP2 immunoreactivity. Nevertheless, microwave denaturation of those tissues preserved MAP2 immunoreactivity higher than fixation with Bouin’s resolution or formalin. Microwave remedy additionally improved the immunoreactivity of a number of MAbs for NF and GF proteins.
Lastly, MAP2 was not detected in human neural cell strains that categorical NF (2) or GF (1) proteins. We conclude that microwave denaturation gives an efficient means to protect the immunoreactivity of regular human neuronal cytoskeletal proteins, and that this methodology of tissue denaturation permits the conventional distribution of MAP2 to be outlined in postmortem samples of human CNS and PNS tissues.

Differential subcellular localization of tubulin and the microtubule-associated protein MAP2 in mind tissue as revealed by immunocytochemistry with monoclonal hybridoma antibodies.

The distribution and subcellular localization of tubulin and MAP2 in mind tissue had been analyzed by immunocytochemistry with monoclonal hybridoma antibodies ready towards Chinese language hamster mind tubulin and MAP2. We examined three anti-tubulin hybridoma antibodies (Tu3B, Tu9B, Tu12) particular for beta-tubulin, and two anti-MAP2 hybridoma antibodies (AP9,AP13).
The specificity of every of the monoclonal antibodies was characterised by staining nitrocellulose electrophoretic blots of SDS-polyacrylamide gels of entire mind or hippocampal extracts. Every hybridoma antibody sure solely its respective antigen in these preparations.
Polyclonal antisera towards tubulin had been additionally examined. Sections reacted with antisera towards tubulin or monoclonal antibodies towards beta-tubulin revealed all kinds of stained mobile compartments. The response product was discovered to brighten dendritic and axonal microtubles in neurons; glial cells had been additionally stained.
MAP2 immunoreactivity was discovered solely in neurons. Within the case of one of many monoclonal antibodies (AP9), staining was preferentially related to dendritic processes. Nevertheless, mild however important staining of axonal processes was seen with AP13. Inside dendrites, MAP2 was discovered related to dendritic microtubules and postsynaptic densities (psd), each in shaft and backbone synapses.
As well as, sturdy immunoreactivity for MAP2 was discovered inside the cytoplasm of dendritic spines. There was little or no immunoreactivity for tubulin within the backbone cytoplasm, though the psd was stained. The localization of MAP2 in dendritic spines and within the psd means that this protein could have a organic function unbiased of its affiliation with microtubules.
The observations on differential staining of the hybridoma antibodies towards MAP2 recommend that there could also be distinct subtypes or states of MAP2 inside neurons.

Microtubule-associated proteins: a monoclonal antibody to MAP2 binds to differentiated neurons.

Hybridomas that secret IgG reacting particularly with the mind microtubule-associated protein MAP2 have been ready with speen cells from BALB/c mice hyperimmunized with excessive molecular weight neurotubule-associated proteins. Immunofluorecence microscopy utilizing twin fluorochrome labeling of tubulin and MAP2 antigens revealed similar patterns of interphase fiber networks in cells from explants of new child mouse mind.
The anti-MAP2 antibody didn’t stain main mouse kidney cells or CHO, 3T3, HeLa, or PtK1 cell strains. Immunoprecipitation and antibody gel staining methods didn’t display any crossreacting antigen in these cells. MAP2 antigen was not seen in affiliation with the mitotic spindle in any of the cells examined.
Radioimmunoassay confirmed species crossreactivity of the anti-MAP2 antibody with mammalian however not avian neural cell extracts. Glial cells and a few neuroblastoma cell strains didn’t seem to comprise MAP2. Nevertheless, within the B104 rat neuroblastoma cell line the MAP2 antigen seemed to be related to the cytoskeleton concomitant with differentiation induced by dibutyryl cyclic AMP.
In disagreement with most beforehand printed stories, our knowledge recommend that MAP2 is discovered solely in differentiated neuronal cells and raises the likelihood that MAP2 is concerned in neuronal differentiation or neuron-specific processes.

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