JPH03163887A - Bioelement - Google Patents

Abstract

PURPOSE:To utilize a bioelement for constructing a bio-computer having an information processing type simulating the gustation and olfaction of an organism by arraying liposomes composed of a living organism substance generating membrane potential fluctuation by the adsorption of one or both of a gustatory substance and an olfactory substance and a living-organism simulated substance onto a substrate in a two-dimensional manner. CONSTITUTION:Liposomes (artificial follicles) 30 formed by using one or both of a living organism substance generating membrane potential fluctuation by the adsorption of one or both of a gustatory substance and an olfactory substance and a living-organism simulated substance are arrayed onto a substrate 41 in a two-dimensional manner. Consequently, the field of the same membrane potential fluctuation as membrane potential fluctuation in the taste cell of a living organism and/or the field of membrane potential fluctuation in the olfactory cell of the living organism is acquired. Accordingly, a bioelement can be utilized for constructing a bio-computer having an information processing type simulating the gustation and olfaction of the living organism.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a bioelement that imitates the chemical information processing function of living organisms.

(Prior Art) A conventional computer is constructed of silicon semiconductor elements, etc.
mann) method (hereinafter referred to as a Neumann computer). However, although this method was able to perform logical operations accurately, it had the disadvantage that it was difficult to process a large number of information in parallel, and it was not good at pattern recognition.

On the other hand, higher organisms easily perform pattern recognition, etc., as is well known. Therefore, if we can clarify the principles on which the pattern recognition and learning/memory functions found in the brain are carried out, and what kinds of elements are used to carry out the functions, then we can imitate these functions. It is hoped that it will become possible to realize computers with various functions that could not be satisfied with the Neumann type computer, such as biocomputers.

For example, among biological functions, chemical senses such as taste and smell, that is, chemical information processing functions, are
Lee J Vol. 02, No. 12. p. 2(
(1987) Maruzen), and is as described below. Figures 5(A) to 5(C) are diagrams for explaining this, in particular, Figure 5(A) is a diagram schematically showing the olfactory cell, and Figure 5(B) is a diagram schematically showing the olfactory cell. The diagram shown, FIG. 5(C), is a diagram showing the relationship between the concentration of a chemical substance (sometimes referred to as a stimulant), cell potential, and nerve impulse.

When a chemical substance is adsorbed to the taste receptor membrane 11 of a taste cell or the olfactory receptor membrane 21 of an olfactory cell, the cell potential changes. This potential change becomes larger in an analog manner as the concentration of the chemical substance becomes higher (see FIG. 5(C)).

In the case of a parasitic cell, when the cell potential changes, a transmitter is released, which acts on the terminal of the parasitic nerve 13 and generates an impulse in the parasitic nerve 13. In addition, Fig. 5 (A)
15 is Cinnabus.

On the other hand, in the case of olfactory cells, when the cell potential changes, an impulse is generated directly from the olfactory nerve 23 via synapses.

These impulses propagate between nerve cells and reach the brain, which is made up of neurons, where information is processed at a high level.

In this way, in the sense of taste, the field of potential changes in olfactory cells is processed, and in the sense of smell, the field of potential changes in olfactory cells is processed in the brain and recognized as a pattern.

As for research that imitates taste, for example, the literature (
MEMBRANE. 12(4), p. 231-
237 (1987), “Taste Response in Synthetic Lipid Membranes”
) was disclosed. According to this document, it is stated that a membrane in which a synthetic lipid dioleyl phosphate is adsorbed onto a Millipore filter responds to bitter substances.

In addition, as for research that imitates the sense of smell, for example, see the literature ■ (Biochemistry),
6, p. 6135 (1987)) or literature ■ (Biochemistry), 26.
p. 614 [1987)). According to literature ■, a suspension containing liposomes formed using azolectin or a suspension containing liposomes formed using phosphatidylcholine, phospatidylethanolamine, etc. It is stated that it responds to substances.

(Problems to be Solved by the Invention) However, the conventional technology that imitates the sense of taste described above is a study that utilizes the small pores of a filter, and the conventional technology that imitates the sense of smell is a study using a suspension. taste of living things,
The technical aspects of building a biocomputer with a form of information processing that mimics the sense of smell were unsatisfactory.

This invention has been made in view of these points, and therefore, the purpose of this invention is to provide a bioelement that can be used to construct a biocomputer that has an information processing form that imitates the sense of taste and smell of living things. It is in.

(Means for Solving the Problems) In order to achieve this object, the biodevice of the present invention provides a liposome formed using one or both of a biological substance and a biosimilar substance, which contains a taste substance and It is characterized by a two-dimensional arrangement of liposomes on a substrate that cause a change in membrane potential upon adsorption of one or both of the olfactory substances.

In carrying out the present invention, it is preferable that the aforementioned liposome is composed of one or more substances selected from the lipids phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, sphygomyelin, and cholesterol.

Alternatively, it is preferable to construct the aforementioned liposome with the synthetic lipid dioleyl phosphate.

Furthermore, it is preferable to encapsulate a membrane potential-sensitive fluorescent dye within the aforementioned liposome.

Furthermore, the aforementioned liposome is preferably immobilized on a substrate using an antigen-antibody reaction.

(Function) According to the biodevice of the present invention, liposomes (artificial vesicles) that cause a change in membrane potential by adsorption of one or both of taste substances and olfactory substances are arranged two-dimensionally on the substrate.
A field of membrane potential change similar to that of a living body's olfactory cells and/or a membrane potential change field of a living body's olfactory cells can be obtained.

Here, liposomes are combined with the lipids phosphatidylcholine, phosphatidylethanolamine, phosphonoatidylserine,
When it is composed of one or more substances selected from sphygomyelin and cholesterol, the olfactory substances muscone, 0ionone, citral, menthone, camphor monoacetate 10-amyl, nonanol, oritanol,
It responds to one or more substances such as hexanol and pentanol and causes a change in membrane potential.

In addition, when liposomes are composed of synthetic lipid dioleyl phosphate, bitter substances such as strychnine, bicric acid, quinine, nicotine, caffeine, and bromine, as well as sweet substances such as sugar syrup, lactose, and glucose, can be added. , nolactose, and galactose, causing a change in membrane potential, and moreover, responding to monovalent ions K+, Na+, and 10, causing a change in membrane potential.

Furthermore, when a membrane potential-sensitive fluorescent dye is encapsulated in a liposome, a field of membrane potential change and a field of fluorescence intensity change corresponding to this field are obtained.

In addition, when immobilizing liposomes on a substrate by an antigen-antibody reaction, the procedure is to first form a film from the antibody on the substrate, and then denature unnecessary parts of this film by irradiating it with an ion beam such as gallium ions. Any two-dimensional pattern can be created using the removed and remaining antibody, and then this antibody C ribosome can be immobilized. Therefore, since the liposome itself is not directly processed by an ion beam or the like, a bioelement with an arbitrary two-dimensional pattern can be obtained without causing any deterioration of the liposome.

(Example) Hereinafter, examples of the biodevice of the present invention will be described in detail with reference to the drawings. Note that each figure used in the explanation schematically shows the dimensions, shapes, and arrangement relationships of each component to the extent that the present invention can be understood.

At the beginning of the Zuyo era and the 2nd 1st and 4th eras, it was formed using biological materials and biological analogues.
A liposome that causes a change in membrane potential upon adsorption of an olfactory substance is prepared as described below.

First, 99% by weight of soybean phospholipid azo-lectin (manufactured by Sigma Co., Ltd., hereinafter sometimes abbreviated as azo-lectin) whose main component is lecithin shown by the following formula (■), and a phospholipid antigen shown by the following formula (■) A mixed lipid containing 1% by weight is prepared. However, the white in the formula (■) represents an alkyl group (C,,
H2. , +1). In addition, the phospholipid antigen represented by formula (1) can be obtained by synthesis.

On the other hand, as a membrane potential-sensitive fluorescent dye, for example, 3,3'-dibrobylthiocyanine iodide represented by the above formula (1) (trade name diS-C3 (5) manufactured by Nippon Kanko Shiki Co., Ltd.)
%, KCj2V93rnM and NaCl 7'm
An aqueous solution containing M at a concentration is prepared. Here, diS-C
The content of 3(5) is set to such a value that the required fluorescence intensity can be obtained. If no fluorescence intensity is obtained, dis-Cs
(5) is not used.

Next, the above mixed lipid is dispersed in this aqueous solution. After applying ultrasonic waves to the obtained lipid suspension, the suspension is centrifuged under the conditions of 100.0009. Liposomes contained in the supernatant after centrifugation are used to construct the biodevice of this invention.

A schematic diagram of the structure of the liposome prepared as described above is shown in the second figure.
Figure ζ is shown. As can be understood from FIG. 2, this lipome 30 has a lipid bilayer composed of azo lectin 31 and contains di as a membrane potential-sensitive fluorescent dye.
S-C3 (5) Contains 33G.

Furthermore, as can be understood from the formula (2), the phospholipid antigen 35 has an aromatic ring in its parent group, so the head polar group is bulky. Therefore, there is a high probability that the phospholipid antigen 35 exists on the side of the lipid bilayer with greater curvature, that is, on the outside of the lipid bilayer, and as shown in FIG. 2, it exists on the outer wall of the lipid bilayer.

Examination of liposomes In order to investigate the properties of the liposomes prepared as described above, a small amount of the liposome suspension (supernatant as described above) was
CuV7mM and NaCu! It is added to an aqueous solution containing a concentration of 93 mM and stirred. Next, nonanol, which is an odorant, is added to the stirred solution, and after 10 seconds, the solution is irradiated with light with a wavelength of 622 nm to excite this solution, and the fluorescence intensity (F) at a wavelength of 670 nm is obtained. Measure.

In addition to the above-mentioned measurement of fluorescence intensity, changes in the membrane potential of liposomes (positive depolarization) are directly measured using a ΔEV microelectrode. Furthermore, nonanol is added one after another, and the fluorescence intensity and membrane potential change are measured each time according to the method described above. Such treatments and measurements are carried out when citral is used as the dioxic substance, and when amyl acetate is used as the dioxygen substance.

The measurement results of the fluorescence intensity ( ) and membrane potential for each concentration of two monomers are plotted on the vertical axis as the rate of change in fluorescence intensity Δ(%).
and membrane potential change ΔE (mV), and the concentration of odorant 12 0 9 (CXM) is plotted on the horizontal axis, as shown in FIG. 3C. Here, ΔF is determined from the following formula (2). However,■
F in the formula. is the fluorescence intensity when no two-year-old substance is added. Further, in FIG. 3, the properties indicated by 1 are those of nonanol, the properties indicated by II are those of citral, and the properties indicated by ■ are those of amyl acetate.

ΔF=Ioo (F Fo )/Fo...
■As is clear from Figure 3, the degree of change in 1, ΔF, and ΔE is almost the same for both, and the liposome prepared as described above converts the concentration change of the 2-year substance into a change in membrane potential. It can be seen that the change in membrane potential can be roughly converted into a change in the fluorescence intensity of the fluorescent dye encapsulated in the liposome.

The properties shown in Figure 3 of liposome suspensions are described in the above-mentioned literature
istry), 26. p. 6135 (1987))
This is almost in agreement with the experimental results shown in .

几・Next, in order to construct the biodevice of the present invention, a processing operation for forming an antibody as a monomolecular film on a substrate will be explained.In this example, the flatness of the substrate surface and the processing operation For simplicity, a case will be described in which a glass substrate is used as the substrate. However, the substrate may be made of any other suitable material such as polystyrene.

First, the substrate surface is hydrophobized by pre-immersing the substrate in octadecyltrichloride silane.

In addition, the antibody γ-globulin obtained against phospholipid antigen 35 (see Figure 2, especially near the aromatic ring of the
A monomolecular film of the antibody is formed on the aqueous solution by spreading it on the subphase aqueous solution in the water tank of the membrane-forming device and compressing the surface of the aqueous solution. Due to the polar distribution of antibody molecules, this monolayer is formed in a certain direction, ie, with the antigen-binding region of the antibody facing down.

Subsequently, this monomolecular film made of the antibody is transferred onto the surface of a substrate whose surface has been made hydrophobic by a horizontal adhesion method, thereby forming 18 films on the substrate. FIG. 4 is a cross-sectional view of the substrate 41 schematically showing the substrate 41 in this state. 43 indicates an LB membrane made of antibodies. The antibody is transferred so that the antigen-binding region faces 1Fr on the side opposite to the substrate 41 side (in the upward direction of the substrate).

In this example, a 1B film forming apparatus is used to form the substrate 4.
However, for simplicity, it is possible to dissolve the antibody in an appropriate solution and directly immerse the hydrophobically treated substrate 41 in this solution. It can be adsorbed.

Next, the substrate 41 on which the LB film of the antibody is formed is immersed in the ribosome suspension (the above-mentioned supernatant) to perform an antigen-antibody reaction, and the liposome is transferred to the substrate 41. Make a two-dimensional array on top. As a result, the biodevice of Example is obtained.

FIGS. 1(A) and (B) are diagrams for explaining this biodevice, and in particular, FIG. 1(A) is a diagram schematically showing a state in which liposomes 30 are fixed to a substrate 41; FIG. 1(B) is a diagram schematically showing a part of the biodevice of the example.

Note that when arranging liposomes in a predetermined two-dimensional pattern, first the unnecessary portions of the LB film 43 made of antibodies on the substrate 41 are denatured by irradiating them with an ion beam such as gallium ions or a scanning electron beam. After removal, the remaining antibodies create a predetermined two-dimensional pattern,
It is then preferable to immobilize the liposome on this antibody. According to this method, since the liposome itself is not processed by an ion beam or an electron beam, deterioration of the liposome can be prevented, which is preferable.

Next, the biodevice of the example prepared as described above was placed in the KC
U was immersed in an aqueous solution containing 7mM and Na (1% 93mM), and in this aqueous solution, nonanol, citral, and n-amyl acetate, which are odorants, were added.
Fluorescence intensity changes and membrane potential changes are measured according to the same procedure as in the case of the liposome suspension described above.

As a result, it was found that even in the state of the biodevice, the fluorescence intensity changes corresponding to changes in the membrane potential of the liposomes, as shown in FIG. 3.

Although the embodiments of the biodevice of the present invention have been described above, the present invention is not limited to the above-mentioned embodiments, and various changes and modifications can be made as described below. .

First, the method for producing a biodevice, the numerical conditions used therein, and the materials used for producing the biodevice described in the above-mentioned examples are preferred examples to facilitate understanding of the present invention.
It is clear that changes in design are possible within the scope of the purpose of this invention.

In addition, in the above-mentioned embodiment, a case was described in which the same type of liposomes were two-dimensionally arranged on the entire surface of the substrate, but for example, the above-mentioned document
). 26. p. 614 [1987)], several types of liposomes having different lipid structures and exhibiting different response characteristics to dioxin substances may be arranged in different regions of the substrate. With such a structure, more sophisticated recognition becomes possible by enabling responses of various dioxin substances and by simultaneously measuring changes in fluorescence intensity in each region. In addition, we prepared multiple types of liposomes containing membrane potential-sensitive fluorescent dyes with different fluorescence wavelengths,
These may be arranged on a substrate.

In addition, the above-mentioned document ■ (MENBRANE), +2
(4). p. 231-237 (1987), by two-dimensionally arranging liposomes using the lipid dioleyl phosphate that responds to various taste substances on a substrate as in the example, it is also possible to construct a bioelement that recognizes taste substances. It is possible.

(Effects of the Invention) As is clear from the above description, the biodevice of the present invention is capable of converting information regarding chemical substances into a field of changes in membrane potential of two-dimensionally arranged liposomes.

Furthermore, by incorporating a membrane potential-sensitive fluorescent dye into liposomes, the field of membrane potential changes in the liposome can be converted into a field of fluorescence intensity changes. Therefore, it can be used in an information processing device, an input device, an output device, etc. of a biocomputer, which has an information processing form that imitates the sense of taste and smell of living things.

[Brief explanation of the drawing]

Figures 1 (A) and (B) are diagrams for explaining the biodevice of the example, Figure 2 is a diagram schematically showing the structure of the liposome of the example, and Figure 3 is a diagram for explaining the biodevice of the example. Figure 4 is a diagram showing the bioresponse characteristics of liposomes and biodevices, and Figure 5 is a diagram for explaining the antibody-attached substrate.
) is a diagram showing olfactory cells, Figure 5 (B) is a diagram showing olfactory cells, and Figure 5 (C) is a diagram showing the relationship between chemical concentration, membrane potential changes, and nerve impulses. be. 30... Liposome, 31... Azolectin 33... Membrane voltage sensitive fluorescent dye 35... Phospholipid antigen 41... Substrate, 43... Antibody. Patent Applicant Oki Electric Industry Co., Ltd. Figure 5 (A) A diagram showing the olfactory cells showing the Oi response characteristics of the ribbon and biodevice of the Example Example. Figure 4 shows the olfactory cells. Figure 5 (B) Diagram showing the relationship between chemical yellow concentration, membrane potential change, and impulse (C)

Claims (5)

[Claims] (1) Liposomes formed using one or both of a biological substance and a biosimilar substance, which cause a change in membrane potential by adsorption of one or both of a taste substance and an olfactory substance, are two-dimensionally arranged on a substrate. A bio-element characterized by: (2) The bioelement according to claim 1, wherein the liposome is composed of one or more substances selected from the lipids phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, sphygomyelin, and cholesterol. . (3) The biodevice according to claim 1, wherein the liposome is composed of a synthetic lipid dioleyl phosphate. (4) Any one of claims 1 to 3, wherein the liposome contains a membrane potential-sensitive fluorescent dye.
The bioelement described in section. (5) The liposome is immobilized on the substrate using an antigen-antibody reaction.
4. The bioelement according to any one of 4.