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S. Korea's Special Forces hold their annual winter training drills - Duration: 1:52.
Winter is a time many Koreans dread, but for some of the most dedicated soldiers of the
nation, this is their moment.
The Special Forces troops are immersed in their regular winter training exercise held
in one of the coldest parts of the country.
Kim Hyun-bin braved the sub-zero climate to witness the process in Pyeongchang.
A platoon of Army Special Forces covertly rappels down from a helicopter into enemy
territory... and infiltrates key facilities.
Moments later a bomb explodes,... destroying the building.
On top of a mountain,...
Special Forces are in full gear, carrying more than 30 kilograms on their backs as they
ski down a steep slope.
Once the enemy is spotted, they switch into battle formation in the blink of an eye and
volley off a wave of bullets to neutralize the targets.
These exercises are part of South Korea's Special Forces winter training drills at Hwangbyeong
mountain in Gangwon-do Province.
"It's below minus 15 degrees Celsius here but the harsh conditions don't discourage
the Special Forces from completing their training."
From hand-to-hand combat to underwater infiltration,... they are all maneuvers that wouldn't look
out of place in an action movie.
"We are learning different skills, and enhancing our teamwork and minds to crush the enemy
at any cost.
We train as realistically as possible so we can become the best Special Forces in the
world."
Each battalion of South Korea's Special Forces will go through ten intense days of winter
training to better fulfill their duty of protecting the nation.
Kim Hyun-bin, Arirang News.
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Новогодний праздник/ Боняша-Юляша - Duration: 21:47.
Polina Vis-Vis
presents
Bonyasha-Yulyasha
New Year`s fiesta
Yulyasha! your grandmother come to us
Grandmother..
Hi
I am glad to see you
My darling
Yulyasha is more
What `s big!
Can you look for Yulyasha?
Our friends called us to the New Year`s fiesta
ok
You don`t worry, relax. You have weared to care about child every day
Now i and Pavlusha are going to the New Year`s fiesta
Are you ready?
Yes
listen to me
i have such order
Declare fiesta today
Fiesta will be fun
All children must be here
Yulyasha, how are you?
You are such big girl now
You are 10 mounth
Our fiesta was opened wonderfully
Lamps are blazed all castle
Guests are on there places
We `ll be jollify together
My favourite daugters and sons
Everybody are beautifull
But Nesmeyana
is sad
Do you want icecream?
No! i don`t
sweets?
No! i don`t
Stop crying
Settle
What is it?
It`s scorelines
Magic scorelines
When we`ll collect 100 smiles
Santa clause come to
I`ll not laugh, i don`t want
How we `ll laugh Nesmeyana
I know how
We`ll prefer one play to you. We must take smiles
Look! Your mother have decorated such beautifull firtree
The firtree is very nice
Numerate, how much smiles are here
100 smiles
Nesmeyana is laughed
Nesmeyana is Snow Maiden now
How long i not laughed
Now i am anouther
Come here
Dance and meet New Year
Don`t you wait for me?
I`ll demage your fiesta
I`ll help you
Can?
Of course. i am magic
I can do magic
1,2,3 firtree blaze
What do you want?
It`s necessary to damage fiesta them
Let`s me sing song about New year
The firtree is so beautiful
Pills, beads..
Frtree-belle, we are liked with you
Hello, children!
We`ll go around the firtree
And sing songs
Sit down, Santa Clause!
Children will tell you verses
Good. Now i `ll give you gifts
Have everybody got a gift?
yes
Good bue
Happy New Year!
I`am tured
It`s time to go home
if you will be liked this video, write likes and subscribe on the chanel
bue
After the cadre
oh, good
oh, ma camera
She droped camera
Good, i catch it
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Pilates+Fitness Diet Tip ♡ Miss Daisy - Duration: 3:36.
Hi everyone! It's time to workout again~~~~
I have shared a lot of different workout exercises and tutorials
For example, weight training, calorie burnings, stretching, etc
and I got a lot of questions and worries from you
that you are afraid of becoming bigger in size because of muscle development
so you might look bigger
because a lot of you did workout to lose weight and look slimmer
That's why today I prepared this video to share
how to do PilaFit!
It's Pilates + Fitness
You can experience Pilates and Fitness at the same time
Today's PilaFit workout is for neck, shoulder and back line
It also works for arm as well
Before furtherado, let's try PilaFit!
First, put your knee down and your elbow
Now pull up one arm and look at the elbow
This workout stimulate your arm and shoulder muscle
so this stretches your arm and shoulder and at the same time this consumes calories
Let's try 15 times for each arm
If you workout too fast or intense, muscles can be exhausted
and sudden workout can make your muscles look more bigger after workout
that's why it's important to stretch before and after workout.
flop your knees and gather your hands on your back to make an arrow shape
Now let's try more advanced workout!
Before, we tried this workout with our elbows down, but now let's try holding hands down.
Try not to keep your elbow straight. This can hurt your albow joint
It's better to have you elbow facing outwards and a slightly bented
This workout gives tension on your back muscle and waist as well.
Today's PilaFit workout is done!
Try this at home with your family!
See you again on my next video! Bye~~
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Rabbit brain is now so popular in China restaurants import heads from France - Duration: 4:04.
Lick, crack and then slurp: How to eat rabbit brains in China... straight from the skull
Chinese diners greedily crack open delicate rabbit skulls and slurp down their contents, tucking into a delicacy so popular in one province that it has to import its supplies from France.
PHOTO: Local delicacy: Spicy rabbit brains are a favourite late night treat in China's Sichuan province, often washed down with beer
Sichuan is renowned for its spicy, peppery local dishes: one of its favourites is bunny brains, often eaten as a late night treat on the streets of its capital, Chengdu.
At the 'Shuangliu Laoma Tutou', a well-known restaurant in the heart of the city, dozens of customers use their gloved hands to prise open the skulls covered in sauce, suck out the brain and nibble on the cheeks amid cries of satisfaction.
'If Sichuanese people don't eat spicy dishes every day, they're unhappy,' said one 20-something woman surnamed Ma, as she dined with friends
'I eat them at least once a week,' she added.
PHOTO: The 'grapple factor': Part of the pleasure of eating rabbits brains is grappling with the food, says food writer Fuchsia Dunlop
Westerners often avoid animal parts - duck beaks, chicken feet, heads and tripe -- that Chinese gourmets treat as delicacies.
But even in China there is little appetite for rabbits' meat, much less their heads, which are overwhelmingly eaten in Sichuan, a remote province long isolated by mountain ranges.
The dish is a speciality of the region - rarely found outside of a few popular restaurants in Beijing and other major cities
'Two out of three rabbit heads consumed in China, are eaten in Sichuan,' said Wang Min, the manager of the Chengdu restaurant, adding that locals were proud of the snack.
'My parents and grandparents ate them. I've been enjoying them since my childhood', she said, adding the tradition goes back several centuries.
PHOTO: It's yummy! A customer had his face wiped after enjoying rabbit heads at a restaurant in Chengdu, capital of Sichuan
'My friends in Guangxi (a southern province) and elsewhere don't understand why we eat them,' she said, adding 'they can't stand the pepper.'
In Wang's restaurant, head chef Yin Dingjun said the rabbit head recipe seemed simple but required a well-established technique.
'You have to drain the rabbits of their blood, then remove the guts' before marinating the head in a broth for several hours,' he said.
'Diners then use their teeth to gnaw at the flesh.'
Rabbits feature in Chinese mythology (a 'jade rabbit' lives on the moon) and are regarded as cute by many young people rather than thought of as a delicacy, although in Sichuanese dialect eating rabbit head is slang for French kissing
Although sucking on a rabbit head might seem odd to some diners even in China, it is par for the course in Sichuanese cuisine, Fuchsia Dunlop, a London-based expert in Chinese gastronomy said.
'There are lots of spicy dishes, such as spicy duck heads, covered in chili and pepper,' she said.
PHOTO: Feeling hungry? A chef handed out a plate of cooked rabbit heads at a restaurant in Chengdu, in south-west China
For people in Sichuan, playing with your food is part of the fun, she said, adding they like 'the grapple factor'.
'Using your fingers and teeth to get a little bit of meat, it's part of the pleasure.'
As night falls in Chendgu, innumerable stalls sell rabbit heads to locals who wash the treat down with beer.
PHOTO: Seriously popular: France exported 166 tonnes of meat and edible offal from rabbits to China in 2014, where rabbit brains are considered a treat. Pictured, a waitress prepared to deliver the order of rabbit heads in a restaurant
'Night markets are part of our culture in Sichuan,' said Rong Lipeng, deputy chairman of Hage, China's leading supplier of rabbit meat and products.
He sells more than eight million rabbit heads each year. But faced with a colossal local demand Chinese farms are struggling to supply enough
As a result, nearly 20 per cent of rabbit heads marketed by the company are imported from Europe, mainly from Italy and France, and mostly frozen.
France exported 166 tonnes of meat and edible offal from rabbits to China in 2014, according to French government figures.
PHOTO: Busy cooking: A group of chefs prepared various dishes for customers at a restaurant in Chengdu, Sichuan Province
Hage, who says the heads are healthy since they contain little flesh, has now agreed a partnership with French firm Hycole to provide breeding rabbits to its farms in China, according to the French company's manager Fabien Coisne.
He hopes that the taste for the treat will one day expand beyond Sichuan, but, he admits, the barrier is high. 'A lot of people outside our province do not dare taste them, because the rabbit heads do look quite terrifying.'
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What Happened When A Girl went to Advocate After Rape ? || جب ایک لڑکی ریپ کے بعد وکیل کے پاس گئی تو - Duration: 2:00.
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LFP 060 Why are Flute Players Such Good Kissers? | Learn Flute Online Video Podcast - Duration: 7:26.
Why are flute players such good kissers?
Stick around and find out here on this podcast episode number sixty!
I'm Rebecca Fuller, the expert and instructor at Learn Flute Online- stay around for this
one.
Hello, and welcome to this episode.
It's a gorgeous day here at Learn Flute Online!
Literally, I'm looking out the window, and it's making me want to climb that mountain.
I actually love every single season - well, Spring not so much here because nothing really
wakes up for a while, but when it does, it's awesome!
I thought I'd give you a little more insight to what I like to do outside of playing and
teaching the flute because I don't really ever get the opportunity to dive outside of
this circle- but one thing I've noticed is that flute players tend to run in the same
types of circles…
I guess I shouldn't generalize, but maybe you'll be interested in knowing some more
about me.
By now I think that you know that I live in the mountain areas in the United States in
the state of Utah.
If you know anything about Utah, you may have seen pictures of orange, sandy deserts with
very large arches and rock formations including slot canyons and incredibly, large lakes.
Well, that's southern Utah, and Iive on the opposite end where it's mountains, lakes,
hiking and snow skiing.
I really love to eat from my orchard and garden, and we have quite a few horses running around
here also.
I hope that gives you a bit of a visual because it's great scenery for fostering great music.
You're probably wondering when I'm going to stop talking about the weather and get
to the subject I have all picked out for today.
Haha, I'm not really sure what made me think of this, but as I was planning out the next
couple of subjects for these audios I thought about how fluent my sons are at learning other
languages.
I have one son who speaks not only two different African languages, but he is also fluent in
Arabic - as well as English.
He recently told me that his next language was going to be French.
And, I told him that with all of his flute training, he would be able to pick up on it
well.
It takes a specially trained 'ear' to notice those little nuances in different languages,
and a musician is definitely cut out for this type of learning.
As I was sending him a message about -that his lips and tongue had been trained all of
his life for learning different languages.
Such a fun subject to talk about!
Also, I have heard it talked about in musician circles where everyone is debating and wondering
which instrument fosters the best kissers.
Haha well it's actually a hilarious subject to me, because first of all, who started this
questioning anyway?
Haha, it just makes me giggle.
But, I think hands down it has to be the flute players who end up on the top of the list
the most often!
We spend every moment from day one of learning how to play the flute in trying to make our
mouths and lips move in certain ways, and after many months and years of trying and
learning - we get really, really good at moving the really small muscles in and surrounding
our lips.
I heard somewhere that it takes around one hundred and twelve muscles of the lips and
face to make a good kiss.
I wonder how many that means we use when we are jumping octaves or playing with real dynamics
on our flute?
I'm guessing it's even more than that!
Haha, well in short, we flutists get really, really good at learning to use the muscles
in our faces - especially the lips and the region around.
The higher level of a flute player, and the longer we've played - the better!
What do you think about this?
Big lips, medium lips, thin lips, it doesn't really matter.
We can all learn how to play the flute well.
I have coached all shapes and sizes, so it's true.
It's all possible, and I'm excited for you to be learning.
I think I'll keep this particular audio a bit shorter today because I want you to
complete an assignment right now- yep an assignment right here in this audio.
That will actually really help you with your flute playing.
I want you to go to your room or bathroom.
Close the door.
And get your face really close to the mirror.
Now, check out your lips.
Really look at them.
You might think I've gone a little cuckoo right now, but hang with me here, this is
important.
We have to be able to understand which directions our lips can move naturally in order to understand
some of the directions I give you in the lesson module videos.
If I talk to you about (for example) stretching your lips back in a closed-mouth smile and
then bringing them forward (while still keeping them closed) into a ducky face pucker - can
you do that?
How full can you make your lips by pushing them forward and out away from your teeth?
How tight and closed can you get them?
Trying that will make them look super thin.
What we need to do is experiment with these different positions, not that we need to use
some of these positions while we play but we need to be able to move in different directions
depending on what we need while playing.
The areas that we talk about a lot while in flute education is the embouchure and the
aperture.
We've already had full episodes on these subjects, and we definitely explore these
to the point where it becomes natural vocabulary in the lessons I teach, but .. just knowing
a bit more and exploring our face and all the muscles we have can really take us to
a whole new level of understanding.
I hope you agree, and I hope that you have a good time checking out your own face in
the mirror this week!
Haha- I hope to see you very soon in a lesson at learn Flute online dot com.
Again, this is audio podcast number sixty - which we titled 'why are flute players
such good kissers'?
A fun little subject to explore - as usual, keep the conversation going by commenting
in the appropriate places here.
It'll be fun!
Bye now!!
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Meizu MX5e |Thes Unboxing (Greek+Eng Subs) - Duration: 7:18.
Hello and welcome I am John for whoever doesn't know me
Don't forget to like, share and subscribe
Today we are going to show you a Mazda MX5...erhmm I mean a Meizu MX5e
Which is sponsored from our friends at store4u.gr
You can find information about them and their phones down below
and how to buy this phone
Phone, phone, phone
Let's check what's inside the box
Meizu MX5e
This phone is from Meizu
Here we can see a really nice box
MX5 not the Miata
Inside we find the phone
The charger
The charger which is bigger than another one we saw with a different Meizu
This charger is 2A
The key, "flyme" brand for removing SIM cards
A guide in Chinese
A white Micro USB cable
And hands-free headphones
The box contains hands-free
These have plus and minus symbols engraved which means...
yes these are buttons
Hands-free with buttons, which are included in the package
Meizu MX5e
The phone has a 5 inch Full HD screen
It has the helios x10 CPU
don't be too surprised, it has 8 cores
It is dual SIM, dual standby and has gorilla glass 3
4G,
it comes in with Android 5.1 Lollipop
8 cores at 2gHz, 3GB RAM, 16GB or 32GB Onboard Storage,
this one has 32GB with 3GB of RAM
It has a 16MP rear camera
and 5MP front camera
The back of the phone as you can see is made out of metal
Really nice to the touch, I feel like Meizu's have good build quality
On the back we can see the dual LED flash,
the 16MP camera, the Meizu logo,
"Designed by Meizu made in China"
Very nice to the touch
On the top we see the slot for the hands-free
On the side we have the SIM tray
where you can either put 2 SIM cards or 1 SIM and a Micro SD Card
On the other side there's the power and volume buttons
On the bottom we have the Micro USB slot,
the speaker and microphone
The same with the previous device, there's a link in the description below to Meizu U20
On the front there's the speaker,
camera and a light sensor
And like the Meizu u20 there aren't any buttons next to the home button
It's comparable to iPhone's, having only the home button at the bottom
Let's see what this phone has to offer
I didn't know about Meizu's, Im suprised by their screens
Incredible phones, I wasn't expecting this
If you see here on the side, the phone has great quality
I wasn't expecting such good build quality from Meizu
because I've never seen them in person, only in photos
The phone is incredible
That's why it's very important to go to a store and see the phones up close
The camera is great
Really good camera, screen and good build quality
MX5e
A really nice phone I've never touched a Meizu before
It was sponsored by store4u.gr
I got it in my hands, I wasn't expecting such good quality, I liked it very much
The cameras this Meizu have are incredible
You can find it at store4u.gr for 229 euro
With tempered glass and a case You can also see it up close at the store,
it's a really nice phone
Don't forget to Subscribe, Like and Share
Spread the word to your friends and we will see each other in the next video
See you
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Adam Hogan-Cann: eNMDARs mediate astrocyte-induced cortical vasodilation - Duration: 16:01.
Adam Hogan-Cann
and the title of his talk is
endothelial NMDA receptors mediate astrocyte-induced cortical vasodilation
Alright, good afternoon everyone
My name is Adam Hogan-Cann
I'm a PhD student from the University of Manitoba, in Chris Anderson's lab
And I'm really excited to be here today
to have the opportunity to speak with you about some of the work we've been doing recently
We've been looking at the role of endothelial NMDA receptors in astrocyte mediated cortical vasodilation
An important regulatory mechanism that controls brain blood flow is functional hyperemia
Just briefly, this is characterized by rapid increase in blood flow in response to elevated neural activity
And a key driving force of functional hyperemia is neurovascular coupling
And this is really characterized by a complex communication
between cells of the neurovascular unit in response to increased synaptic activity
And we know that astrocytes are at a merge as key mediators and important regulators of neurovascular coupling
and its been shown that they can respond to synaptic activity
and they relay these signals between active neurons within the local vasculature
Now interestingly, the cerebrovascular endothelium also seems to be critically important
for whole brain conducted vasodilation for functional hyperemia
But what really needs to be determined is how these vasodilatory signals from the brain
are reaching the cerebrovascular endothelium to initiate these vascular responses
So our lab hypothesizes, the main objective for this study was to determine
whether or not vasodilatory astrocytic signals
can directly communicate with the cerebrovascular endothelium to initiate local vasodilation
And we hypothesize that this is dependent on endothelial nitric oxide synthase
and activation of endothelial NMDA receptors
So there are 3 key peices of evidence to support this hypothesis
Firstly, work that was done previously in our lab where we took isolated brain arteries that were denervated
we showed that these arteries are capable of dilating in response to NMDA receptor co-agonist
specifically glutamate and D-Serine
Importantly, this dilation was dependent upon intact endothelium and could be blocked by eNOS inhibition
As well, this dilation was sensitive to NMDA receptor antagonism
We had consistent results even in a brain slice model
where we've shown that independent and individual activation of a single astrocyte
results in local vasodilation of a penetrating arteriole
And that this is also dependent upon NMDA receptor co-agonist, glutamate and D-serine. and required again eNOS activity
And as well, our group and others have demonstrated that brain endothelial cells
express the critical NMDA receptor subunit, GluN1
which suggests that NMDA receptors are present in these cell types
So collectively, these peices of data suggest that
NMDA receptors expressed on the brain endothelium may be capable of mediating local vasodilation
So while we're hypothesizing that endothelial NMDA receptors are important mediators of hyperemia
what really needs to be first looked at is the expression profile in this cell type
because this hasn't been fully studied in the past
Using whole cell lysates from primary brain endothelial cells cultured from mice
we co-employed a co-immunoprecipitation approach
in which we targeted the critical GluN1 NMDA receptor subunit
And using this immunoprecipitation approach we identified GluN2C co-immunoreactivity
which suggests that GluN1 and GluN2C subunits are directly associated
And this also apparent in B, using the reciprocal approach
in which the GluN2C subunit is low
So these suggest that brain endothelial cells express NMDA receptors
composed of both GluN1 and GluN2C subunits
So if we're hypothesizing that these receptors are important mediators of hyperemia in the brain
then we'd expect these are located on the abluminal or brain facing side of the cerebrovasculature
In this orientation they are configured so that they are capable of recieving vasodilatory signals from the brain
To investigate the subcellular distribution of endothelial NMDA receptors in situ
we employed immuno electron microscopy again targeting the critcal GluN1 subunit
So we took transverse sections of mouse brain
and this allowed us to have a cross section of penetrating arterioles and cortical vessels
And as you can see in figure C
there was a cluster labeling of GluN1 immuno gold beads
localized primarily along the abluminal or brain facing endothelial membrane
This is highlighted by the purple arrows clustered along the yellow trace which is the abluminal membrane
In fact, just over 80% of the total endothelial GluN1 membrane labeling was localized along the abluminal membrane
And this was consistent across all vessels that were sampled within the cortex
So this is an important piece of evidence that demonstrates that
endothelial NMDA receptors have membrane polarity
and show they are in a configuration suitable for recieving dilatory signals from the brian
And that this is an important piece of evidence to support our hypothesis that they are involved in brain hyperemia
We wanted then to look at whether NMDA receptor subunit expression
is correlated with functional NMDA receptor activity
So to do this we looked at nitric oxide production in brain endothelial cells
in response to NMDA receptor activation
Primary brain endothelial cells were loaded with the fluorescent nitric oxide inibitor, DAF-FM
And that shown here in response to glutamate in combination with D-serine
but not on its own significantly increased nitric oxide production
We directly assessed the involvement of the NMDA receptor in this process
This was done by pre-treating endothelial cells with NMDA receptor antagonist
So see in A, pre-treatment with either AP5 or DCKA
which are blocking the glutamate or D-serine binding sites, respectively
completely eliminated the observed production in nitric oxide
We next wanted to assess the involvement of calcium in this process
so this is done by pre-treating endothelial cells with either MK-801
which blocks the NMDA receptor channel pore
or by pre-treating with BAPTA/AM, which is an intracellular calcium chelator
or finally by removing calcium from the buffer
So shown in B, MK-801 blocks calcium through the channel pore
and this is associated with a significant decrease in nitric oxide production
in response to glutamate and D-serine
So in C, we found that chelation of our intracellular calcium and removal of calcium from the buffer
also lowered the observed nitric oxide production
Thus these results from B and C, are consistent with an effect dependent upon an influx of calcium
And lastly, in figure D we demonstrated that this production of nitric oxide is dependent upon eNOS activity
And this was done by pre-treating endoethelial cells with an eNOS specific inhinitor, LNIO
So jst to summarize this slide, it's a little busy, we showed that
glutamate and D-serine co-treatments increased nitric oxide in brain endothelial cells
and this nitric oxide response is dependent upon activation of NMDA receptors, calcium flux, and eNOS activity
So to better understand the role of endothelial NMDA receptor in brain hyperaemia
we developed an endothelial specific loss of function mouse model and this is developed using the cre lox system
in which mice with lox D sites found in critical regions of the GluN1 genes, green one on the left
is crossed with a mouse that expresses the bacterial enzyme cre recombinase
under control of the endothelial specific promotor TIE-2, shown on right
And the resulting offspring have GluN1 levels reduced specifically within the endothelial cells
while all other cell types which don't express cre maintain normal levels of GluN1
And we validated this mouse model using immuno electron microscopy
You can see in figure A, where we looked at GluN1 immuno gold labeling in the brain endothelium
there was about a 50% reduction in immuno gold beads in the cre positive mice relative to cre negative litre mate controls
And importantly, the synaptic levels of GluN1 were unchanged in these mice
And these were in agreement with in vitro data
so brain endothelial cells cultured from cre positive animals
also displayed about a 50% reduction in GluN1 immunoreactivity relative to cre negative cells
So thus, our animal model demonstrates endothelial specific knock down of GluN1
and we hypothesize that the mouse has reduced levels of endothelial GluN1 expression
but they might also have impaired endothelial NMDA receptor signaling
So speficially we are interested in addressing the role of endothelial NMDA receptors
in nitric oxide production and vasodilation in situ
So this is done using acute cortical mouse slice and we loaded these slices with a
fluorescent nitric oxide indicator DAF-FM, which is shown in red
The slices were also loaded with a caged calcium compound
in combination with a calcium fluorescent indicator, Rhod-2, which shown in green is
And this is used to label astrocyte calcium
So using two photon microscopy, we were able to image penetrating corticol arterioles
that have direct ensheathment of perivascular astrocytes
And then by using targeted two photon flash photolysis
we can photoactivate calcium signaling within a single astrocyte
and then monitor production of endothelial nitric oxide
and changes in vascular lumen diameter downstream of astrocyte calcium signaling
So you can see in the video, in response to two photon activation of astrocyte calcium
there's a dramatic increase in astrocyte calcium fluorescence
and this is followed by slower more gradual build up of the red nitric oxide DAF-FM signaling
So firstly, when we looked at the 5 minute accumulation of nitric oxide production in response to two photo activation
NMDA receptor antagonism either by AP5 or DCKA shown in B, significantly reduced the production of nitric oxide
Similarly, endothelial specific GluN1 knock down showed in the white bar in B
also significantly reduced the production of nitric oxide in repsonse to astrocyte calcium signaling
Now we know that nitric oxide contributes to vasodilation
and it does so in part by modulating the metabolism of arachadonic acid
And specifically, it's been shown that nitric oxide can inhibit the production of the smooth muscle vasoconstrictor 20-HETE
and thus it favors production of these more vasodilatory pathways, such as prostaglandins
So we wanted to address whether, NMDA receptors are also involved in modulating the arachadonic acid metabolism
So we did this and this is represented by figure C and D
by using an acute cortical slice model and bath applied glutamate and D-serine
and then analyzed tissue levels of PGE2 and 20-HETE 5 minutes after using the ELISA
And in panel D, you can see that cre negative slices in black have an unchanged tissue level of 20-HETE
However, cre positive slices that have endothelial specific GluN1 knock down have significantly increased levels of 20-HETE
In contrast, in C, the black bars represent cre negative slices
and have a dramatic increase in tissue levels of the vasodilator PGE2, in response to glutamate and D-serine co-treatments
whereas this effect is lost in cre positive slices
So effectively this slide deomonstrates that in response to endothelial NMDA receptor suppression
there's reduced nitric oxide production and this is associated with altered arachadonic acid metaboilsm
favoring the production of the vasoconstrictor 20-HETE
And lastly, the big question of our study was to
determine whether endothelial NMDA receptors influence vascular lumen diameter
So shown in panel A by the green line, we looked at the cumulative vasodilatory response
after two photon activation of astrocytes
You can see in green, that the cre positive slices with again endothelial sepcific knock down of GluN1
had a significantly reduced vasodilatory response relative to cre negative slices
This suggests that arterioles with endothelial specific GluN1 knock down
have reduced vascular reactivity in response to direct astrocyte activation
So in summary, our lab has evidence to support a novel mechanism of neurovascular coupling
and this links astrocyte signaling, endothelial NMDA receptor activation, and eNOS activity
And this helps to provide some information
how these vasodilatory signals from the brain are coming to reach the cerebrovascular endothelium
And therefore may contribute or play an important role in some of the endothelial dependent mechanisms
that require important vasodilation needed for whole brain hyperemia
And we confirm the functionality of this pathway with our novel loss of function mouse model
where we associated the reduction of endothelial NMDA receptors
with reduced nitric oxide production and altered arachadonic acid metabolism
I'd like to finish by thanking my Advisor, Chris Anderson and also colleagues in the lab
As well as our collaborators at the University of British Columbia and University of Manitoba
And to all our various funding sources. Thank you for your attention.
We have time for a couple of questions
So, just speaking of the NMDA receptors here
Do the AMPA receptors lead the voltage block there or how does it
Is this a metabotropic type of NMDA receptor?
That's not entirely clear at this point
Endothelial cells do express AMPA receptors but because of the activation of the GluN2C subunit
that likely suggests they have reduced sensitivity to magnesium blockade
and therefore aren't necessarily requiring that AMPA depolarization to alleviate that magnesium blockade
Thanks
Good question
You showed that glutamate levels are decreased in synapses
Did you ever look at neural activity, check to see if there's differences in network activity?
Nothing that we've looked at at this point
Our plan is to do in vivo to address the role in our mouse model but it's something we haven't done yet
Well thank you very much to all our speakers for wonderful talks
I'm sure if you have any more questions that you can try to find the speakers
Thank you all for your attention
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