Puzzle of Endothelium by Tom Slattery You look at junk fragments collected in our genome over eons of evolution and

wonder how all of that irrelevant garbage got there. But then you look at the finished product that was crafted by natural selection, and it is an efficient organism. And we humans have become the efficient organisms at the top of the heap, our efficient bodies and our powerful dominating brains. I daily wonder about what we are and how we got here because my apartment sits at the edge of a high cliff of sedimentary layers, most of which were deposited in the geological Devonian epoch beginning 360 million years ago. I daily look out across the canyon of the Rocky River Valley nature preserve and see the opposite cliff of layers of sedimentary sandstones and shales. And I cannot help being reminded that one of those layers was deposited at the exact time when a miserable tetrapod crawled up out of the amniotic fluid of a Devonian sea and awkwardly struggled onto the land. With its minimal sentience it would hardly have known that it began a whole new phase in natural history, a long line of animals living and crawling and walking around on the land that would eventually be capable of leaving the planet and reaching for the stars. And as luck, fate, or natural selection had it, the animal had four limbs. Think what would have happened if instead of two front limbs and two hind limbs our ancestral creature had, as do insects, six limbs. Or what if it had eight limbs, as do spiders and octopuses. Or what, even, if it had had a theoretical hundred limbs, as do centipedes. And then what might we all look like today. Consider buying a hundred shoes each time you went to the shoe store, some of which would wear out sooner, others later. But all joking aside, natural selection settled for an efficient four limbs, a minimum necessary to allow heads, mouths, and a pair of stereoscopic eyes to be held above ground level and front appendages free to grab and grasp. Natural selection has in general honed efficient animals. And that tendency toward efficiency in natural selection is why I am slightly puzzled by endothelium. What, you may ask, is endothelium? Endothelium is a single layer of living cells that line our blood vessels. But it is more than a mere lining. Taken together these cells make up a body organ that works in sync with the endocrine system to control blood pressure. And it is no small organ. Endothelium makes up about three percent of the mass of our bodies. Maintaining and nourishing it thus costs our bodies in energy, material, and the complex digestive processes that manufacture body nutrients. What is puzzling is that not all animals have endothelial cells. Some get by quite nicely without having to bear the biological costs of having endothelium.

Endothelial cells line blood vessels, but they only line blood vessels in vertebrates. Vertebrates -- animals like us mammals as well as reptiles, birds, and amphibians with internal skeletons -- have endothelial cells. Invertebrates -- animals like insects, octopuses, squids, spiders, earthworms, and jellyfish without internal skeletons -- do not have endothelial cells or in a few cases have small patchworks of incomplete and leaky endothelial systems. Why would endothelial cells have to line the blood vessels in vertebrates and not have to line the blood vessels in invertebrates? What justifies the additional cost to vertebrates? Well, these are good questions. Among the things that endothelial cells do is to reduce some friction between flowing blood and blood vessel walls. This allows a faster blood flow and that, in turn, allows for larger size of animal bodies. But an invertebrate like a giant squid, for instance, is bigger than most vertebrates on the planet. Size in itself means little in survivability that would justify the extra biological cost of having endothelium. The division between vertebrate animals that have endothelium and invertebrate animals that do not have endothelium goes back a half billion years. And to put it in context, it goes back to way before the above first land-crawling tetrapod was born. Somewhere around a half billion years ago marks the time when endothelial cells came into being. A close surviving -- well, rather than surviving we should say thriving -- relative of that unknown first animal with endothelium is the present-day hagfish, an eel-looking slimy creature living in cold ocean water in all of the oceans. Hagfish, need I say more, have endothelium. The fact of hagfish having endothelium raises questions even while it implies answers to other questions. The questions it answers have to do with blood clotting, calcium, and vessel walls. Calcium ions are one of the ingredients needed for blood clotting, and bare blood vessel walls without endothelium would offer opportunities for clotting mechanisms. Vessels might soon be clogged. An example of this in us in our time is arterial blockages due to plaque. An injury to our human endothelium from, say, a herpes virus puts blood and blood-carried nutrients in contact with a bare section of arterial wall. This soon leads to plaque buildup and eventually to blockage of an artery. And this, in turn, may lead to a deadly heart attack. So what does this have to do with our friend the hagfish? It has to do with hard mineralized bones. Hard calcium phosphate bones have a number of functions. They offer strength and support for vertebrates. In geology the calcium phosphate bone mineral is a rock called apatite. It is literally rock hard and hard to break.

But bones are also a reservoir for calcium and phosphorous. Bodies of creatures with calcium phosphate bones use the calcium and phosphorous from these bones to keep blood acidity levels constant and to provide calcium to repair wounds. How do they do it? Inside mineralized bones like ours are living cells called osteoblasts and osteoclasts. The osteoblasts make new bone by taking calcium and phosphorous out of the blood and spitting out calcium phosphate into the bone matrix to make new hard mineralized bone mineral. The osteoclasts perform the opposite. When the body needs calcium or phosphorous for, say, maintaining a constant level of blood acidity, the osteoclasts chew up some bone and spit out calcium and/or phosphorous for the body's needs. When in old age the osteoblasts no longer make new bone at the same rate that the osteoclasts continue to supply bone calcium and phosphorous to the body, bones get porous and weak. These weak porous bones present a disease called osteoporosis. But there's a catch to this speculation about hard, mineralized calcium phosphate bones and endothelium. Endothelium did not evolve to protect against calcium-caused blood clotting inside blood vessels. The hagfish tells us this. Hagfish and sharks both have endothelium, but they do not have mineralized calcium phosphate bones. They are vertebrates, but they have hard cartilage for bones. So their endothelium may not be present to prevent calcium ions from bumping up against bare blood vessel walls and causing blood clots inside blood vessels. You might point out that hagfish and sharks are cold-blooded animals. But many animals like reptiles and amphibians have mineralized calcium phosphate bones and are also cold blooded. And these all have endothelium. The hagfish is probably the closest living species to the time of the great divide a half billion years ago between vertebrates that have endothelium and invertebrates that do not. It is a singularly ugly-looking creature without eyes as we know them. It merely has two light-sensitive indentations on its head where eyes would normally be. Probably it can only make out light and shadow. And this may hint at something. The hagfish would appear to be more primitive than sharks. The hagfish's primitive eye, optic nerve-to-brain system would seem lower on the evolutionary line than sharks with their fully developed eyes and optic nerve-to-brain systems. There must have been some evolutionary going forward and then backtracking. But because scientists cannot pin it down, no one knows how many times endothelium went out of style and how many times it came back again. By coincidence or not, the timeframe may put the missing link creature somewhere in the Ordovician Mass Extinction when a huge variety of ancient animals died off and never

came back. For some reason our small blue planet suddenly cooled at the time of the Ordovician Mass Extinction. This brought on several ice ages of heroic proportion that dominated the climate and dramatically changed sea temperature, sea oxygen content, sea acidity, and sea salinity, fluctuating back and forth over millions of years. Surviving animals probably would have had some edge that gave them survivability during climatechange fluctuations. During this great planetary trauma, endothelium would appear to have had some yet unknown usefulness to justify its biological cost and yet not enough usefulness to make it absolutely necessary to species survival. The missing-link creature of the future evolutionary line that includes us would probably be more primitive than either the hagfish or the shark. Sharks, with bones made of hard cartilage, would seem to show that intelligent and capable animals need endothelium for some reason yet unknown to us. But clearly sharks do not need endothelium as blood-brain-barrier filtering system. What is this blood-brain barrier? In all animals with higher functioning brain elements, vertebrates and invertebrates, there is a barrier between the variety of chemicals and molecule sizes carried in the blood stream and the brain and its pure brain fluid. This barrier is effectively a filter that allows oxygen and energy chemicals in and burnedup waste chemicals and carbon dioxide out. It is known as the blood-brain barrier. Neurons need this purity to function and without this blood-brain barrier advanced brains could not function. While sharks have endothelium, for some unknown reason their blood-brain barriers came to be made up of glial cells. In the hagfish, on the other hand, endothelial cells function, as in us humans, to maintain the blood-brain barrier. One is led to believe that there were evolutionary steps prior to both the hagfish and the shark. These then branched out into both blood-brain barriers using endothelium in the hagfish and bloodbrain barriers using glial cells in the shark. And this suggests that endothelium probably did not evolve to fill a needed blood-brainbarrier function. Endothelium would seem to have evolved to fill some other need, maybe a now long gone and no-longer-needed need. Nevertheless, some kind of a blood-brain barrier is vital. Except for the case of sharks, this vital blood-brain-barrier function calls for the continued organism outlay for endothelium at its present level of biological cost. The endothelial blood-brain barrier, as opposed to the glial barrier, apparently had a natural selection advantage for vertebrates. Maybe for some reason it allowed living on land. All land-dwelling vertebrates now utilize endothelium for their blood-brain barrier.

But was this use of endothelium simply some great evolutionary accident that was later utilized for something vital to growth and dominance of us vertebrates? Or way back a half billion years ago was there something critical that endothelium evolved to solve and now no longer is needed for that? Tom Slattery, Rocky River, Ohio May 2012. Coda, May 2013 A curious possible answer may be hinted at in a new paper, "Alzheimer's Disease: The Great Morbidity of the 21st Century," by Charles T. Ambrose, in American Scientist, May-June 2013 (vol 100, num 3). Endothelium cells generate new blood supplies to old and new neurons through processes called neuroangiogenesis. Without neuroangiogenesis new neurons die of suffocation and starvation. That seems to be what is happening when brains with Alzheimer's shrivel up. As a result, endothelium cells permit higher animals to have longer-lived, larger, and routinely refreshed brains. At the top of this chain we "thinking beings" (homo sapiens) have our huge thinking capacities because our brain neurons regularly rejuvenate and capillaries to supply oxygen and nutrients regenerate with them. It's a stab in the dark. But it sounds reasonable. May 8, 2013.