 Alright, so next we're going to have Ning Tian, he's a research professor, he's going to talk about retinal neuron degeneration after optic nerve injury. Okay, so my name is Ning Tian, I was initially trained as a retinal surgeon in China. So I've been spending my last 27 years on the research of the basic science as a type of retina. Recently we started to use more disease-related projects that we might have. So to fit this into the traditional research, I'm going to show you some training results we got for our disrelief research. So our research has been focused on retina gangestyles. For most of you, I guess it's a different part of the science. But I'm still going to go through it quickly. So retina gangestyles are the only cell type. I cannot get the eye to the brain, so it's the only alpha neuron. And retina gangestyles are very widely used in many diseases including, I could call it Omen, it's a very common disease. And they're also like a brain injury, so on some of them the car extended, you can get a head down to the dashboard and the optic nerve can be injured there. And the swollen optic nerve or bone fracturing behind the eye can cause an optic nerve injury. And the big study of the projects that's supported by the baby, because a lot of the soldiers in the battlefield, and they have a blood site in the bottom of the glass to bring, can cause the optic nerve to be affected to me. So the question we're interested in is that, when you look at those, any of the disease, problem, cancer, retina gangestyles, from those gangestyles are actually, there are many types of, some type of retina cells, the response to those pathological retina cells is very different. Some cells are much easier to be injured or damaged or killed, and some cells are much tougher and more resistant to those diseases. So we're trying to figure out what are the reasons for the mechanism which can show the vulnerability of those gangestyles and hopefully to find a way to actually enhance the resistance that retina cells cause different injuries. So that's the purpose. So this is just to show the examples. So when we try to study how gangestyles die, so we don't want to forgot the process of their deaths. So these are the pictures from the live animal performance. When we use the transgenic mouse, we can express the fluorescent proteins in the gangestyles so we can actually watch the cell, the morphology every day for a long time period. So this is an example of when we put those gloomies injected into the eyes of gloomies, the commonly excited neurotransmitter naturally. So in our brain and our retinas, most of the excited cells actually release gloomies as a neurotransmitter. But when they release too much of them, it causes them to become toxicity, so we could decide on toxicity. So those happen if someone has a brain injury. You have a primary cell death and those death cells release a lot of gloomies into the adjacent cells and cause those cells, the secondary deaths of the cells along with the primary injury. So gloomy toxicity is a very, very common cause. So in the hospital, in the emergency room, about 40% of the patients have a brain injury due to the secondary injury, not the primary injury. So gloomy toxicity is a very, very important issue. But in some people even believe that gloomy toxicity is the cause of the, or partially of the aspect of the cause of gangestyles in glaucoma, because sometimes in particularly of the angle of close glaucoma cause immediate IOP increase cause a lot of cell death and cause secondary death by gloomy toxicity. Okay, so when we inject gloomy into the eye, we look at how the cell died. So before the injection into the gangestyles and the half of the dendrites. And then just 10 minutes on that, the dendrites started to become signal. And about three hours on that, you lose maybe 90% of the dendrites and the six hour of that, the cells loose all the dendrites, become a soul. So when we look at the cell death process what we found is, and we used the Capsule 3, it's a cell death monocode when the cell started to synthesize Capsule 3 which means the cells in the process are blind. So, and then we compare the time course of the disappear of the forest in the path of the dendrites we found the gangestyles always lose their dendrites completely before they started to cell death process. So it starts like, loose of the gangestyles the loose of the dendrites is a condition for the cell to die or it's the earlier stage of the cell death. So, and then, but when the cells they lose their dendrites they can stay, they can keep alive for several days in this condition. So, what actually keep them die not die without dendrites or can we actually promote in real dendrites and what that saved them from dying so it's a question we're trying to start like that. So that's one example. So another example is this this is actually often earth crashed in the life and also we identify these cells and then we go back and I use the four Capsule mechanically crashed option earth on the life animal and then we watch the cells how far they will die, how they will die what you can say it's like a 31 hours after never crashed that same gangestyles lose all the dendrites and four days later that cell is gone. So again, so in addition to the boom in toxicity, mechanical injury or option earth even you injure the axon of the dendrites the cell don't lose that axon actually they lose the dendrites first before they die. Okay. And the third example is this this is even much longer this is a light induced cell damage so we look at a bunch of cells we usually relate to a high intensity laser to just scan on this particular cell and then we wait like for 13 days and those cells start to actually lose their dendrites and then they become sick minded and then 14 days later well one days after this they lose their dendrites and another two days after that the cell is dead. So it doesn't matter what kind of injury you use when you injure the dendrites they lose their dendrites first and then they die. Okay. So then we have as I said before so if we look at mouse and even human or rabbit all those in the medians there are multiple type of dendrites cells morphologically and physiologically. Okay. And those cells actually respond so first we want to know we know the cell response to disease can be very different because some cells die sooner than others and then we want to know okay so what are the control markers for the model unit? So that's one that's one goal we want to look at so to characterize again there's a subtype of specific model unit that's one goal. The second good one we want to look at is when we lose dendrites cells in the rat and the way it's not going to affect other cells even they are not directly injured is there any secondary cell death after dendrites are injured? Okay. And then because that's very important because if we want to say to the stem cell transplantation to restore the basin and if you don't have the normal cell morphology or well-being of other cells even you're getting a cell in retina they're not going to function normally because the whole retina has to function as a network not even with your cells because it works it can all function in a network. So that's the second. So the third purpose where we have some candidates have a neuroprotective canopy so we want to try those in Europe and those candidates with a neuroprotective protective capability while they actually protect themselves from death is the model we're using. And the fourth purpose where it means with 23.5 there's any genetic approach we can actually protect yourself by overexpression or non-regularity gene expression to change yourself more or promote and survive these things. So any model we're using is as I said we're using two common models one is the auto-nerve injury by partial auto-nerve and the second is by injected movement into the eye to cause the movement inside the toxicity. And then we use transgenic immune cells which express the fluorescent in this guidance cell so we can actually very precisely monitor the structure change of individual guidance cells. So we choose a guidance cell model and then another animal the mouse model is the apricot cell specifically express GFP so we basically have four animal models which is the alpha guidance cell we use one model specifically label alpha type cells and then those two other guidance cells we call either BDE guidance cell and GFV cell and then there's this apricot cell so we just pick up a four type of cells to test their vulnerability and their response disease or their pathological result. So the the technique we're using is just look at some numbers and then look at some morphology and this is an example of our three dimensional tracing of single guidance cell dynamics so by looking at right now we can actually precisely identify every small branches of dynamics and look at them and this is how this changes or if we change the gene of the morphology. So this is the first result we're looking at just look at the morphability of three types of subtype of guidance cells and then look at if we take it right now we just put it in gloomage on the right there and then watch individually ourselves how fast they're going to lose their guidance what we found is from those three subtype of guidance cells one, the two type of subtype they may actually lose their guidance very quickly but another type which is a BDE cell they actually done this can last much longer than other two types of cells so prove that on the common cause of cell there's movement toxicity and the guidance cell to three types of guidance is a very different model and this is the one point I'm going to touch that later is if we mutate the gene intrinsically express the gene in the guidance cell that changes that cell will appear from there to here like other cells so there's some gene actually controls cell morphability but I will get back to that so the second model we're using is to do optical nerve crash and then look at how cell die and how it's going to be found in the gene cell and the guidance cells so this is the way we do the surgery we just anesthetize the mouse and cut the skin a little bit and then approach to the optical nerve from the back of the eye and put any of the four cells behind there just crush it for 10 seconds and then when we put the die into the guidance cells and then look at how the die has been transported along the optical nerve they can say from the crash side the die can not pass that which confirms the optical nerve is completely crushed at the surgery site so we now the axon are very now then we look at so we use the nutrient die to label all the cells the cell nutrients in the guidance cell later and then count the number of those cells so the die for the die so this is the this is the red nerve the red nerve and this is the red nerve after optical nerve crash you can say the balloon dies much smaller if you're in this because they're cell died so the number of cells is reduced now so when we look at the time course of cell death you can say before the crash and after the crash about the same day's crash we can do something about eating the same cells you can crush heart in that so that's what it's very reliable now when you really know how cell died how fast it died and then we have a very reliable way to do that so now for the dying cells so that's what we know and then we start looking at another model it's looking at amyquid cells so one of the amyquid cells just in your own which is synapses the amyquid cell in Iran is very important for the amyquid cell to have their normal function so the amyquid cell we're looking here is a transgenic amyquid cell which is expressed in the green fluorescent protein in those amyquid cell type and then if we use antibody to against the label that cell that cell release a super-con so we call it a coninergic amyquid cell okay if we use the anti-acidic antibody label that we found it kind of this is easier to say those green cells are all red the red label is anti-acidic antibody and the green is the GFB spread by the cell what that means is not all the we call subversive amyquid cells express the GFB but all the YFB particles are the starvers amyquid cells so we know what cells are looking then we look at the amyquid structure of the cells so after eye injury after amyquid cell injury what we found here is we get a much lighter crash on the cell 10 days later women lose about 28-30% of amyquid cells and then we so in that in this sense we don't lose a lot of amyquid cells we try to say we only took 30% of the amyquid cell test while the amyquid cell be infected immediately what we found here is actually just like seven days and a half of crash even if there is no directly injury on amyquid cells the amyquid cell die after amyquid cells I'm sorry amyquid cell did the amyquid cell didn't die but amyquid cell started lose their amyquid structure they lose their amyquid cell by 30% in seven days after amyquid cell die so even the cell don't die like this the cell don't die the amyquid cell don't die but they lose their structure the amyquid structure very quickly in that way so so that's the conclusion about that then we start to look at this we have those games of death by groom can we protect them from those injuries so what we try is a growth factor on amyquid so we inject those drugs chemical into the eye and then either crash off the nerve or or co-inject those movement into that to cause toxicity and then come the cell members if they still cause a cell death what we found here is the drug we've given cannot completely protect itself from dying but those drugs can't protect it for the gloomy toxicity induced death by reduced by 20 4% so we go from here an innovative fact so but when we look at the optical nerve crash and that same drug has no effect at all so which means even you know those cells die it looks like the same way they die but when you put it on the neuron protected drug there and the drug can protect itself so this is the same slide that I pointed to before so we're trying to look at if there's any genetic way we can actually find out how that the molecules expressed by cell per se control the cell death if we manipulate the activity of the gene expressing can we protect the cells so the first sense to do is to find is there any gene actually can control the cell death which would be we found one gene for that particular cell so this is before the lack of gene expressing and the cells very adjacent to gloomy toxicity but when you look at the lack of which is the triangle red which is here so when we lack of the gene expressing in that particular cell and then we put the gloomy on the red nerve we say that cell is no longer adjacent to the gloomy toxicity so we're working on the process of so this is actually when we lack of the gene expressing which means we lack of gene activity in the cell so the next step will be if we all activate the gene expressing the cell can we protect the cell and then we're working on that and we'll look at the cells again so this is the conclusion so Genghisel Lusthenes before cell death with all kinds of various pathological in cells you can survive when you're testing and the cell has a specific transgenic model which provides an invigorated time for cell death and some of those neuro-protective agents which can protect the cell Genghisel from death for some causes but on other causes so the cell responds to the same agent for Genghisel since very often depends on what's the cost to cause the cell death and then since then we seem to define the gene which could potentially activate so this work is most of the work is by an MPPH student Zebier and then they're basically just kind of home viewing and then there are two other people who are not in the picture thank you so I'm going to ask a question I have to say is there support about model for aging questions are there other thoughts which we don't know actually so which is a very good which is the question we really want now if you don't know that you could support it now because I guess you heard the Brian Johnson talk you know the earlier we take you can at least not the which are for the it could really be a stop in the whole retinal circuit because there's not much so so we probably have to stop how how how how