Class III β-Tubulin isotype (β III) in the adrenal medulla: III. Differential expression of neuronal and glial antigens identifies two distinct populations of neuronal and glial-like (sustentacular) cells in the PC12 rat pheochromocytoma cell line maintained in a gelfoam matrix system

Katsetos, Christos D.; Herman, Mary M.; Balin, Brian J.; Vinores, Stanley A.; Hessler, Richard B.; Arking, E. James; Karkavelas, George; Frankfurter, Anthony

doi:10.1002/(sici)1097-0185(199803)250:3<351::aid-ar10>3.0.co;2-n

Cited by 20 publications

(9 citation statements)

References 76 publications

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“…These observations imply that the class II isotype may play a transient role in later stages of neuronal development, such as those of post-mitotic neurons. Additionally, positive immunoreactivity in the extended processes of NGF-stimulated PC 12 cells also suggests that this isotype is important for neurite growth, similar to the class III isotype shown in the previous reports [Joshi and Cleaveland, 1989;Katsetos et al, 1998b]. Cells of mesoderm-origin, such as fibroblasts, myoblasts, and chondrocytes, showed a strong immunoreactivity in developmental non-neuronal tissues, and show a diminished immunoreactivity by varying degrees in adulthood.…”

Section: Discussionsupporting

confidence: 83%

Distribution of the class II β‐tubulin in developmental and adult rat tissues

Arai¹,

Shibutani²,

Matsuda³

2002

Cell Motil. Cytoskeleton

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During a screen of monoclonal antibodies raised against a cytoskeletal preparation of neonatal rat cerebrum, we have identified a monoclonal antibody, MAb58A, that is specific for the class II beta-tubulin isotype. Immunoscreening of a rat brain cDNA library using MAb58A yielded the cDNA retaining a class II-specific nucleotide sequence. The specificity of MAb58A to the class II beta-tubulin isotype was confirmed by immunoreactivity to synthetic peptides corresponding to isotype-specific sequence of class I, II, III, IVa, or IVb. Further, the results of an immunoassay against a series of overlapping octapeptides derived from a class II-specific region revealed that the antibody epitope was a heptapeptide that consists of Glu-Glu-Glu-Glu-Gly-Glu-Asp (EEEEGED). Immunoblot analysis revealed that the class II isotype represented a significant portion of beta-tubulin present in the adrenal gland, brain, and testis of adult rats. In fetal tissues, this isotype was detected in skeletal muscle, as well as in the brain. Immunohistochemically, MAb58A reacted predominantly with components of the developing rat nervous system, such as migrating neuroblasts, peripheral nerves and ganglion cells, and sensory organs. MAb58A-immunoreactivity was also found in developing skeletal and smooth muscle cells, chondrocytes, and vascular endothelia. In adults, MAb58A-immunoreactivity was remarkably diminished, but persisted in peripheral nerves and ganglion cells, chondrocytes, and capillary components. Together, our results demonstrate that MAb58A is specific for the class II beta-tubulin isotype, which may retain an embryonic nature in both neuronal and non-neuronal tissues.

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Section: Discussionsupporting

confidence: 83%

Distribution of the class II β‐tubulin in developmental and adult rat tissues

Arai¹,

Shibutani²,

Matsuda³

2002

Cell Motil. Cytoskeleton

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“…The glia‐like properties of the nestin–GFP positive stem cells and their neural differentiation potential might also explain the frequent observation of pheochromocytomas with sustentacular properties . This dual differentiation was also observed in the rat pheochromocytoma cell line PC12 although it is supposed to be monoclonal , suggesting that PC12 cells might be derived from a multipotent adrenomedullary precursor cell with properties characterized here.…”

Section: Discussionsupporting

confidence: 63%

Multipotent Glia-Like Stem Cells Mediate Stress Adaptation

et al. 2015

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The neural crest-derived adrenal medulla is closely related to the sympathetic nervous system; however, unlike neural tissue, it is characterized by high plasticity which suggests the involvement of stem cells. Here, we show that a defined pool of glia-like nestin-expressing progenitor cells in the adult adrenal medulla contributes to this plasticity. These glia-like cells have features of adrenomedullary sustentacular cells, are multipotent, and are able to differentiate into chromaffin cells and neurons. The adrenal is central to the body's response to stress making its proper adaptation critical to maintaining homeostasis. Our results from stress experiments in vivo show the activation and differentiation of these progenitors into new chromaffin cells. In summary, we demonstrate the involvement of a new glia-like multipotent stem cell population in adrenal tissue adaptation. Our data also suggest the contribution of stem and progenitor cells in the adaptation of neuroendocrine tissue function in general.

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“…At days 0, 1, 3, 5, 7, and 10, cells were lysed and the expression of cdc2 and cdk2 was evaluated by immunoblotting analysis using specific antibodies. Changes in MAP2 expression were also examined to confirm the neuronal differentiation of PC12 cells induced by NGF (27,34). As shown in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Both of these agents have been previously demonstrated to suppress cdc2/cdk2 activity (23, 24, 35). In addition, changes in MAP2 expression were also examined to confirm the neuronal differentiation of PC12 cells by NGF (27,34). A control group, treated with 0.1% dimethyl sulfoxide alone did not show significant neurite outgrowth and expression of MAP2 was much lower than in cells treated with NGF (50 ng/ml) for 3 days (Fig.…”

Section: Expression Of Endogenous Cdc2 and Cdk2 During Differentimentioning

confidence: 99%

Simultaneous Suppression of cdc2 and cdk2 Activities Induces Neuronal Differentiation of PC12 Cells

Dobashi

Shoji

Kitagawa

et al. 2000

Journal of Biological Chemistry

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The involvement of cdc2 and cdk2 during neuronal differentiation in rat pheochromocytoma PC12 cells was examined. When PC12 cells were cultured with nerve growth factor (NGF), expression of cdc2 decreased significantly after day 5, while expression of cdk2 decreased gradually after day 7. Cells overexpressing cdc2 or cdk2 were resistant to NGF-induced differentiation and growth suppression, and maintained high cdc2 or cdk2 kinase activity, respectively, during NGF treatment. In contrast, the NGF-treated parental cells showed a marked decline in these kinase activities after day 3. When PC12 cells were treated with specific inhibitors of cdc2/cdk2 (butyrolactone-I, olomoucin), they showed marked neurite extension and up-regulation of microtubule-associated protein 2 expression. In addition, treatment with mixtures of antisense oligonucleotides for cdc2 and cdk2 resulted in down-regulation of both cdc2 and cdk2 kinase activities as well as significant neurite outgrowth and up-regulation of microtubule-associated protein 2 expression. However, neurite outgrowth was not observed in cells treated with either single antisense oligonucleotide, or antisense cdc2 + cdk4 or cdk2 + cdk4 oligonucleotide mixtures. These results suggest that simultaneous down-regulation of cdc2 and cdk2 activity is sufficient and necessary for neuronal differentiation in PC12 cells.

show abstract

Class III β-Tubulin isotype (β III) in the adrenal medulla: III. Differential expression of neuronal and glial antigens identifies two distinct populations of neuronal and glial-like (sustentacular) cells in the PC12 rat pheochromocytoma cell line maintained in a gelfoam matrix system

Cited by 20 publications

References 76 publications

Distribution of the class II β‐tubulin in developmental and adult rat tissues

Distribution of the class II β‐tubulin in developmental and adult rat tissues

Multipotent Glia-Like Stem Cells Mediate Stress Adaptation

Simultaneous Suppression of cdc2 and cdk2 Activities Induces Neuronal Differentiation of PC12 Cells

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