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Adrenergic,¹ Cholinergic, and Inactive Human Neuroblastoma Cell Lines with the Action-Potential Na⁺ Ionophore^,2

Department of Biological Chemistry and Laboratory of Nuclear Medicine and Radiation Biology [G. J. W., J. U., H. R. H.], and the Department of Pediatrics [R. C. S.], UCLA School of Medicine, Los Angeles, California 90024

Cultured human neuroblastoma cell lines were assayed for biochemical characteristics of neuronal function. Cell lines studied included LA-N-1, LA-N-2, IMR-32, SK-N-SH, and SK-N-MC. Veratridine-dependent uptake of ²²Na⁺ implied the presence of the action potential Na⁺ ionophore in LA-N-1, LA-N-2, IMR-32, and SK-N-SH. The time course of ²²Na⁺ uptake and inhibition of uptake by tetrodotoxin supported this. SK-N-MC had no veratridine-dependent ²²Na⁺ uptake. Tyrosine hydroxylase (EC 1.14.10.), glutamic acid decarboxylase (EC 4.1.1.15), and acetylcholine contents in neuroblastoma cells were compared to those in brain. LA-N-1 and IMR-32 contained 15 and 5 times as much tyrosine hydroxylase, respectively, whereas LA-N-2, SK-N-SH, and SK-N-MC contained only 0.5 to 5% of that in brain. Acetylcholine was present in LA-N-2 in 15- to 20-fold greater quantities than in brain; other lines had only 10 to 50% of that in brain. None of the cell lines contained glutamic acid decarboxylase. Thus, continuously propogated human neuroblastoma cell lines may have the action potential Na⁺ ionophore and may be adrenergic (LA-N-1 and IMR-32), cholinergic (LA-N-2), or inactive (SK-N-SH and SK-N-MC). This is the first demonstration of the action potential Na⁺ ionophore and of acetylcholine production in human neuroblastoma cell lines.

¹ In this investigation, adrenergic and cholinergic cells were defined as those that had tyrosine hydroxylase and acetylcholine, respectively. Cells defined as "inactive" lacked these markers; this is an operational definition since such cells may have other adrenergic or cholinergic markers such as dopamine ß hydroxylase or cholineacetyl transferase (4) and thus may not truly be inactive.

² This study was supported by the University of California Cancer Research Coordinating Committee, the California Institute for Cancer Research, and National Science Foundation Grant BMS 75-10093.

³ Recipient of Contract E (O4-1) GEN-12 from the Energy Research and Development Administration.

⁴ Recipient of National Cancer Institute Research Career Development Award 1 KO4 CA 00069. To whom requests for reprints should be addressed, at the Department of Pediatrics, UCLA School of Medicine, Los Angeles, Calif. 90024.

Received 12/ 8/76. Accepted 1/24/77.

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