Neuro-biological modeling of the brain function (II) — Cloning of glutamate decarboxylase cDNA by PCR-based eficient cloning method —

s153

QUANTITATION

265 TETSURO

HORIKOSHI

Dept. of Biological

OF GENE EXPRESSION

and MANABU

Science

IN THE RAT BRAIN BY USING SIMPLE PCR METHOD

SAKAKIBARA

and Technology.

Tokai Cniv , Numazu,

School of High-Technology,

Shizuoka

410-0321,

JAPAN. Several quantitative special apparatuses

methods

or extra DNA fragments

method based on PCR

dehydrogenase

(GAPDH),

PCR products

linear ranges of amplification

internal

standard.

(GABA

0 I

were visualized

ii I ), or phospholipase

The reproducibility

of the method

NEURO-BIOLOGICAL

266

-

FREQUENCY

FORMAL

C subunit

by ethidium

B4

staining

3-phosphate

(PLC B 4)

in expression

with the data of ltr .3//r/ hybridization

the

volume of cDNA and

using GAPDH expression

was less than 50% difference

After

In each sample.

From the relation between

were calculated

into cDNA using

for glyceraldehyde

using primers

and quantitated

of each cDNA of interest were established

levels of GABA [I I and PLC [j 4 were consistent

expression

most of them require

from a part of the rat brain, then reverse-transcribed

of PCR product in the linear range, relative gene expression

amount

by using PCR. however.

In the present study, we tested a simple quantitative

cDNA was serially diluted and PCR-amplified

GABA receptor subunit

electrophoresis,

have been developed

such as a competitor

Total RNA was extracted

Obtained

random hexamers

agarose-gel

for gene expression

level as an

levels

Relative

reported by others

MODELING OF THE BRAIN FUNCTION (I)

RANGE ASSIGNMENT

CODING AND INFORMATION

NEURONS WITH TEMPORAL

PROCESSING

USING

SUMMATION -

HIROKI NAKAE Advanced

Research

Laboratory,

Research and Development

Center, Toshiba Corporation,

Komukai Toshiba-cho,

Saiwai-ku,

Kawasaki 210-8582 The brain works on the basis of the information images

expressed

through

with trains of action potentials

the regulation

frequency

of muscle

in the labyrinth

excitatory

namely the frequency

and inhibitory

and acted like a high-pass

filter. When they formed synapses

was realized and could sort out a specific frequency neurotransmitters

coded with trains of action potentials,

summation

of a band-pass

it was suggested

filter

that we can

if we can faithfully regulate the

and receptors of neurons to be excitatory or inhibitory. NEURO-BIOLOGICAL MODELING CLONING OF GLUTAMATE CLONING METHOD -

KAZUNORI Advanced

information

of neurons. We

of temporal

on the same target neuron, the function

which

system in order

that can raise responses

In conclusion,

sound and the behavior

however,

an experimental

They showed a character

range of action potentials.

-

267

of dynamics.

It receives

It is still unclear,

In this study, we designed

ranges of the action potentials

neurons with parameters

design a model neural network processing

of action potentials.

and retina. The brain controls

also with trains of action potentials.

range of the trains is assigned to specific information.

to look for units of information, modeled

contraction

coded by the frequency

translated

MIYAZAKI,

Research

MITSUKO

Laboratory,

OF THE BRAIN FUNCTION (II) DECARBOXYLASE cDNA BY

ISHIHARA-SUGANO,

Research and Development

PCR-BASED

EFICIENT

HIROKI NAKAE Center, Toshiba Corporation,

Komukai Toshiba-cho,

Saiwai-ku,

Kawasaki 210-8582 In order to understand during the information information specific

to be processed

information.

the plasticity

processing

with neural networks of excitatory

realizing

the brain function,

For such an investigating,

in the brain. That is, a frequency

Because we showed by computer

be sorted out with a combination rule can be processed

of a neural network

should be clarified.

simulation

modifications

we have defined

range of action potentials

that a train of action potentials

and inhibitory neurons, it is likely that the information

with culture neurons whose neurotransmitter

is regulated to be excitatory

of the neurons

a coding

rule of the

is assigned

in a frequency

coded according

or inhibitory

to

range can to the

by transfecting

cDNA of the enzyme(s) related to the neurotransmitter systhesis. For the source of the cDNA, we have previously constructed a rat brain cDNA library in A Zap express vector (Stratagene). Here we report that the development of an efficient cloning method by using PCR to detect cDNAs containing is applicable characteristics

to cloning

of the key enzyme,

in glutamatergic

and GABAergic

namely neurons.

glutamate

whole coding region, then demonstrated decarboxylase,

determining,

the excitatory

that the method or inhibitory