Thank you Countess Bettina and Wolfgang Schurer and the other members of the Lindau foundation for inviting me here.
I had a great time last year and this year looks like it’s the same way, so I’m very happy to be here.
I want to start off by explaining the title.
As you must have an idea now from several of the talks about what translational research is,
it’s usually defined as taking the discoveries from the laboratory
and applying them either for the cure or treatment of human diseases.
And unfortunately in my opinion this has become almost the sole rationalisation sometimes of how things should be funded.
To give you one example, when the Obama administration gave a very nice amount of money to NIH,
the NIH first took the money from the stimulus package and had a series of challenge grants.
And the challenge grants were 100 different challenges that people could apply for money and they were going to be funded.
Of those 98 of the 100 challenges were translational, 2 were not.
I have nothing at all against translational research, as I get older I actually think it’s extremely important
but I think this proportion of 98 to 2 is completely wrong.
And to me one of the very best definitions, and so I’ve entitled this 'non translational' and I’ll think you’ll see why,
it doesn’t mean there’s not implications of the work but that’s not the direction the work has gone.
To me one of the best definitions or descriptions of what I would call non translational research
and the importance of basic research as Bob pointed out was actually done in part in testimony by Robert Wilson,
a different physicist, Robert Wilson to the one that’s there,
this is the physicist, sculpture, architect who designed the Fermi Labs.
And in 1969 he was asked by congress to come and testify to, because congress was going to fund this lab
and they wanted some justification.
And senator John Pastore from Rhode Island tried to sort of ease Wilson into the discussion by asking him questions like
"How will the discoveries made at the Fermi Lab help the national defence?"
And Wilson said "They won’t."
And he said "Well no, isn’t there other things that it will help?", he said "No, none of these things."
And finally after being pushed for quite a while he came up with this wonderful statement
that the research has only to do with the respect with which we regard one another,
the dignity of man, our love of culture, it has to do with whether we are good painters, good sculptures, great poets.
I mean all the things we really venerate in our country and are patriotic about.
It has nothing to do directly with defending the country except to make it worth defending.
So with that as the background I want to tell you a little bit of my history
and some of the things that have gone on in my lab and what I’ve been interested in.
And I’m going to start with what has motivated me for much of my life.
Now actually only since I’m a post doc have I been really interested in the particular problem that I’m going to mention
but for means of demonstration I’m going to use a younger self of mine here.
And what I’m going to try to demonstrate on this picture is, what I’ve been fascinated about now for over 30 years
and that is the mechanical senses.
Biologists have a very good idea about the chemical bases, the molecular bases of vision
and the molecular bases of chemical senses,
whether they’re smell or taste or neurotransmitter or function of hormones within the body.
But we have a whole series of senses, hearing, our sense of balance and of vestibular system, tendon organs,
stretch of our muscles and our senses of touch and which there are actually 5 different senses of touch,
harsh painful touch, gentle touch, texture and to be able to detect through the skin either low or high frequency vibrations.
And all of these mechanical senses really work because cells are pushed around, there’s some physical manipulation of the cells.
They also have one other thing in common, we are clueless as to how they work at a molecular level,
so again there’s a lot of work to be done for this.
And this is what has preoccupied me and I also work on the nematode caenorhabditis elegans
and this is a diagram of the cells that sense touch.
To give you an idea of what a touch to the animal looks like, when you touch the animal it moves backwards.
Animals that when we touch them don’t move backwards, could be touch insensitive animals
and this is how we’ve actually isolated quite a number of mutant strains.
Now of course we have to be a little more careful because animals that are dead also don’t move when you touch them
but we do have ways of differentiating this.
And the end result has been that we’ve been able to find quite a number of genes that encode components of this complex
that are actually needed for touch sensitivity.
Now when we started isolating, identifying the genes and characterizing their products,
one of the very first questions we wanted to answer was what cell makes the protein, what cell activates the gene.
And at the time which was about 1989 there were a number of methods of being able to localise gene activity,
you could look for the protein by antibody, one could use a reporter like beta galactosidase
and let it do its enzymatic reaction and produce a product.
One could use in situ hybridization actually, look where the RNA was made from the activated gene.
And in all of these cases what we needed to do was fix the tissue and then permeabilise it,
so we were looking at dead tissue and we were looking only at a snap shot in time.
And once we did the experiment we had to repeat the whole thing all over again.
And went through the whole procedure.
Now one of the things about C-elegans that for the 10 years before 1989 that I used to always tell people
and you can sort of get a hint from that, of that middle picture there, is that the animal is transparent,
we can put it on a microscope stage and really look right through it and see, using various means the cells within the animal.
So I’ve been saying it’s a transparent animal, it’s a transparent animal, I sort of got that part down.
And then we were doing all of this work trying to look at where genes were expressed.
And it’s at that point that inspiration struck.
Now to me one of the nicest examples of inspiration and more of this should be done in bars, is this, Cambridge 1953,
shortly before discovering the structure of DNA, Watson and Crick, depressed by their lack of progress visit the local pub,
"I’ll have a double, Felix."
For me the inspiration was a talk by Paul Brehm in which he talked about the work of Osamu Shimomura
who will talk tomorrow or Thursday, his work in discovering first aequorin as a light producing molecule, bioluminescent molecule.
And then discovering this very interesting protein, green florescent protein GFP, that produced green light in the system.
And Paul Brehm in his talk said "All you have to do is shine blue light or ultraviolet light on this protein
and you’re able to see green."
And for the first time in a long time I did not fall asleep in the seminar, I actually listened and I said to myself,
I work on a transparent animal, this would be terrific.
The next day I got in touch, I thought this would be a great thing but what I actually learned about it
was that GFP is a rather unusual protein, it doesn’t need any other cofactor and that was good
because then we didn’t have to add anything, all we needed to add to the organism was the DNA,
it would make the protein and we would be set.
Unfortunately as what had been known is that there’s the peptide backbone,
at this position GFP when it was matured formed this 5 membered ring.
And no one at the time knew how this was going to be made and the best guess was
that it was going to require at least 1 enzyme, maybe 2 or 3 or 4 converting enzymes to make the final florescent product.
And if that were the case then it wouldn’t be a very good, useful tool.
I was able to find that the person who was getting, making the cDNA was Douglas Prasher in Massachusetts
and Douglas and I got together and talked for quite a long time on the telephone and decided to collaborate.
We had a couple of years of being out of touch with each other, I thought he hadn’t actually finished the cloning,
he thought I had dropped out of science, but eventually we got back together and discovered that we were both still interested
and the catalyst was that I had a new student in the lab Ghia Euskirchen who had come in to work
and she had just gotten her masters in engineering, chemical engineering in our engineering school at Columbia
which was just the building next door to mine.
And she had worked on florescence.
And so it was really quite wonderful, we called up Douglas, renewed the collaboration
and then we got the clone from him and one month after she started graduate school she had green florescent E-coli.
And it was really wonderful, we were very excited, but you can’t see in this because, right under the words florescent,
right here, use scope in engineering, our microscope was no good, we couldn’t actually see the florescence in our microscope
but fortunately she had worked in that other lab in engineering
and knew where there was a good microscope to look at the florescence.
We then continued to do the experiment but without an appropriate microscope, that was a little hard,
so I called up all the representatives of the microscope companies and told them that I was interested in buying a scope
and could they bring by a demo and could they leave it for a couple of months
and so we finished all the experiments on demo microscopes.
I’ve recently done something similar, I don’t know if I should say this too much, in any case we were very excited about this.
This led to us also showing that all we really needed was the DNA encoding for the coding sequence of the protein,
that was it, no other converting enzymes were needed, only GFP.
And so we eventually wrote up this work and sent it to Science and I’m going to say a little bit about publishing
and how hard it is sometimes, even when you’re really excited about the work.
I sent the paper to them to be reviewed, it was entitled "Green florescent protein, a new marker for gene expression."
And the editors wouldn’t even send it out for review, they said "We’re sorry, the title is no good".
And I said “what?” and they said “you have the word ‘new’, and you know this is Science magazine, everything is new in Science.
So you have to change the title.”
I was a little miffed, I changed the title to this long one:
"The Aequorea Victoria green florescent protein needs no exogenously added component
to produce a florescent product in prokaryotic and eukaryotic cells", that’s actually the whole paper.
And they didn’t balk at that, the reviewers were very nice, they accepted it and then they came back
and said "You know the title, it’s a little long", so finally became "Green florescence as a marker for gene expression".
The next problem was this cover, which I’ve sort of enhanced the colour of here, and that was actually the problem,
the art editor called me up and said "Well, we really like the picture that you have for the cover",
I did too because it was showing a growing nerve cell, this is the growth cone in the living animal.
And they said "We really like it but you know there’s one colour we really don’t like to have on the cover
and that’s green, so can we change the colour", and I said "No, that’s no good".
And the final problem with publishing is that we had already given samples to many people to work on
and we wanted to cite their unpublished results and many people just said oh yeah sure that’s fine, go ahead.
One person was a problem, hopefully this will come up here, and this is a letter from Tulle Hazelrigg
in which she makes the demand that I make coffee every Saturday morning for the next 2 months,
I prepare a special French dinner and I empty the garbage nightly for the next month, this is my wife.
(laughter).
What she did was actually though the next really important experiment with regard to GFP
because she is the person that made the first protein fusion of GFP and she used it to study a drosophila protein
that she was looking at, that was made in these nerve cells and then transported
and deposited into the developing oocyte in drosophila, comes to lie along here
and as this demonstrated also in the posterior part of the oocyte and she was able to watch this happen in the living tissue.
And so this was a very important thing.
She claims I haven’t paid this up, the latest thing of this, I’ve shown this slide quite a number of times,
several months ago she went to the local drug store to pick up some prescription and the pharmacist said to her:
"Wait a second aren’t you the person who’s married to that guy who won the Nobel Prize?", and she said "yes",
and he said "I really liked your letter".
(laughter).
In any case the advantages of GFP are that it’s inheritable, one can give the DNA to an organism,
it will be passed on to subsequent organisms and you can do repeated observations.
It’s a relatively non invasive technique, we just shine blue light and that activates the protein
and looks for the florescent light, so we don’t really interfere with life when we do this.
It’s small monomeric and this really means it can diffuse throughout a cell to give you the entire shape of the cell
and I’ll show you that in a second.
And of course it can be used in living organisms,
so really for the first time we have a dynamic view of cellular events rather than a static view in the tissues.
So some examples, here are a bunch of animals, worms, drosophila, that's canola plant, mice, zebra fish, this is Alba,
the GFP bunny that the artist Eduardo Kac commissioned so that he could bring it around to art installations
and talk about the interaction of science and arts.
These are some cells in culture.
This is drosophila, Arabidopsis, this is a Purkinje cell
and you see how all the branches of the cell are lit up by the GFP that’s in it.
I want to show you a couple of other examples.
I also want to point out that GFP itself, the protein is probably one of the most beautiful proteins in the world
as far as I’m concerned and I had nothing to do with it, I should explain the colour code.
People whose names that are in red are from my lab, people in blue they are people that I’ve collaborated with,
people whose names are in black as you see here did something I wish I had done.
So what this is is a lantern and the florescent centre is right here in the middle of it,
physically it’s a beautiful, beautiful structure.
In any case let me give you some background to the 2 movies I’m going to show.
This is one of those standard descriptions of cell division, I want to just point out 2 aspects of cell division
that you can look at, one is the nuclear envelop which of course has to break down for the chromosomes to separate,
so there’s no nucleus during this time and then the nucleus reforms.
And the second is the fact that these microtubules, these lines that you see here
and these lines make what's called the mitotic spindle.
And so one can watch them.
So I’m going to show you 2 movies, the first movie uses GFP attached to a protein in this spindle,
so you're going to see the spindle forming and this is in the developing embryo of drosophila.
Now in that case there are no cellular divisions, it’s just nuclei that continue to divide.
The second movie I’m going to show you is where GFP has been attached, not to any component of the cell
but to a small peptide that will cause it to be inserted into the nucleus.
So whenever there are nuclei around it has some place to go, when the nuclei are not there it will go away
or it will go all over the place.
So first here on the left cell division, both of these are taken from the American society for cell biology website,
you can see them on there.
And what I hope you can see in this picture is not only how these wonderful spindles are formed
but the fact that everything is being done synchronously.
And being able to see this synchrony is one of the things that we want to be able to do in living organisms.
And while that’s going this is, and this is false coded so the more concentration of the GFP,
the redder it will look, so it’s not different colour florescence coding.
So now all the nuclei are formed, now they break down but now they’re breaking down in a wave,
they’re not synchronise, so these 2 different aspects of nuclear division are being done or controlled in different ways.
And we can watch them by watching what happens with the GFP signals.
Of course there has been a lot of improvements of GFP and Roger has already told you about some of the work
that he’s done in making other colours and that’s of course led to things like Josh Sanes and Jeff Lichtman’s use of these
in labelling neurons, so what they call brainbow.
Roger has also talked about the idea of making various molecular monitors for calcium imaging
and I’m going to go sort of quickly through this, for peptide cleavage, for protein, protein interactions,
basically using this force to resonance energy transfer, FRET as a way of monitoring the state of molecules within a living cell.
So it’s really expanded much more than I ever imagined when we started doing our work on GFP.
GFP has been transformative in my lab, actually I’m going to try an experiment I have not ever tried before here,
how many people consider themselves biologists, if you are just raise your hand, ok so sort of look around,
how many of you have, keep your hands up please, how many of you,
keep your hand up if you’ve worked on GFP in one way or another, so almost everybody has their hand still up,
so it’s been remarkable, about how many people have actually, how this has been used really all over.
In my lab we’ve used it to examine gene expression, to look at the localisation of proteins, to find mutants
because if we can label the cells we can mutate the animals and look for animals that had abnormal cells,
nerve cells that don’t grow appropriately, nerve cells that don’t have the right connections and so on.
We can also use it to label cells so we can isolate them and in one of my favourite set of experiments
we’ve used it to actually be able to identify the first eukaryotic transducer of touch.
So we had identified a number of components here that are part of this channel complex, including these 2 proteins,
mec-4 and mec-10, which we thought were going to be the basis of touch in the worms.
And we needed to record from them molecular physiologically and although we can look in the animal
and we can see every cell, these are really tiny cells.
So if you stick an electrode into the animal, it explodes and that’s not really good for the experiment but it also,
even if you open it up, you don’t know what cell is what unless you label it with GFP and then you can see where it is.
So we’ve been able to do that, I should explain the double colour code here,
Miriam started as a post doc and then ended this project as a collaborator.
And we were able to show by the electrical recordings that these components were important
and it worked and I’m actually not going to talk about here,
that we were able to actually demonstrate genetically that this was the transduction molecule.
So it’s work that I’m very proud about.
Since I only have a couple more minutes and you’re probably all dying of thirst,
I’m going to skip talking about what we’re doing now, I’ll be glad to talk to people in the discussion part
about what that is and really just go to my last slide.
I’m not going to talk about any of this work but I am going to say something about this and GFP
and that is that even before these various lessons that I want to go into, is that technology,
and you’ve seen this also in other talks, Theodor Hänsch’s talk exemplified this also, Roger’s talk exemplified it,
technology really drives a lot of science and having new ways of looking at things really allows you to make amazing findings.
And we’ve been very fortunate to be able to continue to make new tools and to use new tools.
But what I really want to talk about here are a couple of these lessons that I’ve learned.
And the first is that scientific success comes in, by many different ways.
If you look at the 3 of us that have, that shared the GFP prize, we’re 3 very different scientists.
Osamu Shimomura is an incredibly dedicated, hard working, passionate about his work, biochemist
who really has stuck with the problem and really works it until he gets it finished.
Roger was a person who was recognised even in high school as being a really brilliant scientific mind,
winner of the Westinghouse Award.
My path is a little bit stranger and I won’t really go into it but it’s sort of an embarrassment to me
that I very rarely if ever will talk about my chemistry grades after I’ve gotten this chemistry prize
but be that as it may, there is no route to this, you just have to enjoy what you’re doing and work it
and there is no right or wrong way of doing this, just being committed I think is the most important.
The second thing is scientific progress is cumulative.
I was really delighted to share this prize with the other 2 and in fact I could name a whole series of other people
that I would love to share this prize with, because it’s really been hundreds and thousands actually
of people that have modified GFP and made it the molecule that it is and increased its usefulness.
So we’ve all added on to one another’s work and that’s really been the wonderful aspect of it.
Third to me the students and post docs are the real innovators in the research.
When we started letting people know that we had gotten GFP to work, I sort of went through all the great finds
and I get people calling me up and they would say tell me about this GFP, my graduate student told me about it
and I want to find out if we really want this in my lab or my post doc told me about it.
So the people that were disseminating information and were getting excited about it were the students and post docs.
And they still are the most important part of my lab.
Finally I want to say 2 things, one that basic research is essential, it’s the engine that drives innovation
and leads to the insights, that will lead to translational research and to advances in agriculture and industry in general.
But basic research is essential, Bob also said this earlier.
And finally we should be studying all of life, not just the model organisms, not just the flies and the yeast
and the mice and the zebra fish, we should be studying everything.
Look at what we almost missed by someone not studying the jellyfish
and looking at a problem that’s a very fundamental problem, how is light produced by an organism.
That's not a translational project, but to look at a non traditional animal,
ask a very interesting and fundamental question, there are lots more of these questions to be asked and lots more to learn.
Thank you very much.
(applause).