SMITHSONIAN SC!  CE INFORMATION EXCHA       U.S. DEPARTCIEHT OF        PROJECT NUMBER
          NOT use this space   HEALTH EDUCATI OR AND WELFARE
            PfiBLl;CH;;AT&SERVlCE
                    INTRAYURAL,RESEARC~~ moma   201 HL 00009-02 LBG

PERIOD COVERED

July 1, 1975 through June 30, 1976
1lTLE OF PROJECT (80 characters or less)

Zell Recognition and Synapse Formation

`IAMES, LABORATORY AN0 INSTITUTE AFFILIATIONS, AND TITLES OF PRINCIPAL INVESTIGATORS ARD ALL OTHER
PROFESSIONAL PERSONNEL ENGAGED ON THE PROJECT
?I:    Marshall Nirenberg      Chief, Lab. of Biochemical
                             Genetics                  LBG NHLI
ITHER:  Donald Puro            Research Associate           LBG NHLI
    Hiroyuki Sugiyama      Visiting Associate            LBG NHLI
    Phillip Nelson        Chief, Behavioral Biology
                            Branch                   BB NICHD
     Clifford Christian      Special Fellow           BB NICHD

COOPERATING UNITS (if any)

3ehavioral Biology Branch, NICHD

LAB/BRANCH
Laboratory of Biochemical Genetics
SECT I ON                -

. .
>ect-ron on Molecular Bioloav
INSTITUTE AND LOCATION
IHLI, NIH, Bethesda, Maryland 20014
TOTAL MANY EARS:         PROES I ONAL:           OTHER:
    2.2               2                  .2
SUMMARY OF WORK (200 words or less - underline keywords)

  Long rangeyobjectives are to define reactions which are required for synapse
formation and for transmission of information from cell to cell.

`HS-6040
:12-75 )


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Project Description:

   Major Findings: Since normal neurons do not divide, clonal lines of neu-
roblastoma cells and somatic cell hybrids derived from neuroblastoma cells
were generated and characterized with respect to receptors, neurotransmitters,
action potential ionophores, and other properties which are required for syn-
aptic communication.  Fusion of clonal neuroblastoma cells with glioma
cells yielded clonal hybrid cell lines which synthesize, store and excrete
acetylcholine; properties which are'not expressed by the parental cell lines.
During the past year cells from one hybrid line were found to form synapses
with cultured striated muscle cells.  Synapses between hybrid cells and
muscle cells closely resemble the synapses between normal motor neurons and
striated muscle before they are fully.developed.  Under appropriate condi-
tions, hybrid cells established synaptic connections with almost every muscle
cell tested; thus, synapses are synthesized in abundance.  Marked differences
were observed in the efficiency of transmission across different synapses.
Axonal activities which were found to+be regulated include choline acetyl-
transferase, acetylcholinesterase, Na action potential ionophore specific
'activities, and the rate of choline transport into cells.

   Eight species of receptors have'been found thus far with the hybrid cell
line which forms synapses.  Receptor mediated shifts in CAMP levels, cGMP
levels and membrane potentials have been identified and characterized. Thus
the foundation has been laid for studies on the effects of receptor-mediated
reactions on synaptic transmission.  In addition, more than 100 cell lines
which synthesize acetylcholine have been obtained and are being studied to
determine that some cell lines are defective with respect to synapse formation.

   Significance to Biomedical Research: This is the first time that clonal
cells of neural origin have been shown to form synapses.  Thus the set of
genes which are required for synapse formation are expressed in this cell line.
The model system which has been established affords extraordinary opportuni-
ties to define synapse formation reactions and properties and correlate bio-
chemical events with developmental and electrophysiological phenomena.

   Proposed Course: Current studies focus on determining the reactions
which are required for synapse formation and factors that regulate these
'reactions.

'Publications:

;l. Greene, Lloyd A., Shain, William, Chalazonitis, Alcmene, Breakefield, Xandra,
Manna, John, Coon, Hayden G. and Nirenberg, Marshall: Neuronal properties of
hybrid neuroblastoma x sympathetic ganglion cells.  Proc. Natl. Acad. Sci. USA
72: 4923-4927, 1975.
-

2. Chalazonitis, A., Greene, L.A. and Shain W.: Excitability and chemosensiti-
vity properties of a somatic cell hybrid between mouse neuroblastoma and
-sympathetic ganglion cells.  Exp. Cell Res. 96: 225-238,1975.
                                                   -

2


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3. Nelson, Phillip, Christian, Clifford and Nirenberg, Marshall: Synapse
formation between clonal neuroblastoma x glioma hybrid cells and striated
muscle cells.  Proc. Natl. Acad. Sci. USA 73: 123-127, 1976.
                                               -

4. Nirenberg, Marshall W.: Coding of neural information by neuroblastoma
cells.  In: Talwar, G. P. (Ed,): Regulation of Growth and Differentiated
Function in Eukaryote Cells. New York, Raven Press, pp. 537-539, 1975

5. Thompson, Edward J., Griffith, James M., Glazer-Schoenberg, Deverera and
Nirenberg, Marshall W.: An improved method for extracellular recording of
action potentials from single cultured neuroblastoma cells.  Med. Biol. Eng.
13: 104-106, 1975.
-

6. Hamprecht, Bernd, Kemper, Wayne and Amano, Takehiko: Electrical response
of glioma cells to acetylcholine.  Brain Res..lOl: 129-135, 1976.