Hensen's node in chicken embryos governs movement of neural cells, study shows

For us living creatures with backbones, existence begins as a single fertilized cell that then subdivides and grows into a fetus with many, many cells. But the details of how those cells end up as discrete organs instead of undifferentiated heaps of cells is only now being understood in microscopic detail.

Why, for example, should some of the cells migrate to the region that will become the brain, while others travel netherward to make a spinal cord? Although some details are known about which cells contribute to particular regions of the nervous system and which signals help to establish the organization of the brain, much less is known about factors that guide the development of the spinal cord.

In a new study, researchers from the California Institute of Technology have gained unprecedented information about the molecular signals and cell movements that coordinate to form the spinal cord. The study takes advantage of recently developed bioimaging and cell labeling techniques to follow individual cell movements in a developing chick embryo through a clear "window" cut into a fertilized egg. The results, reported in the June issue of the journal Nature Cell Biology, suggest that a proliferative stem zone at the tail end of the growing embryo contributes descendants to the growing neuraxis.

"The basic idea is that descendants of cells from Hensen's node, the structure that lays down the trunk, are sequentially distributed along the elongating spinal cord" says Luc Mathis, a former researcher in the lab of Caltech biology professor Scott Fraser, and lead author of the paper. "In the past, we did not have the ability to follow individual cells in living vertebrate embryos and could not determine how neural precursor cells could remain within Hensen's node, while some descendants leave it to form the spinal cord. "

In the paper, the researchers explain that neural precursor cells get displaced into the neural axis by the proliferation in Hensen's node. The researchers labeled cells near Hensen's node in 40-hour old chick embryos by using an external electric field to deliver an expression vector encoding green fluorescent protein (GFP) into cells, a process called electroporation. Using state-of-the-art imaging techniques developed by postdoctoral researcher Paul Kulesa, the group recorded the motion of fluorescent cells in ovo using a confocal microscope set up for time-lapse imaging and surrounded by a heated chamber to maintain embryo development.

"As the cells proliferate, some progenitors are displaced from the stem zone to become part of the neural plate and spinal cord," Mathis says. "Our analyses show that the Hensen's node produces daughter cells that are eventually displaced out of the node zone on the basis of their position in relation to other proliferating cells, and not on the basis of asymmetric cell divisions."

The paper also addresses the molecular signaling involved in the spreading of the cells. Previous work has shown that fibroblast growth factor (FGF) is somehow involved in formation of the posterior nervous system. To test the possibility that FGF could act by maintaining the stem zone of cell proliferation, the researchers disrupted FGF signaling within Hensen's node. Indeed, the result was a seriously shortened spinal cord and premature exit of cells from the node, indicating that FGF is required for the proliferation of neural precursor cells in the stem zone that generates the spinal cord.

A structure similar to Hensen's node—called simply a "node"—is found in mammals, and analogous zones are found in other vertebrates as well. The cell behavior and genetic control discovered in the chick might also be responsible for the development of the spinal cord in mammals, including humans.

"This new understanding of the formation of the spinal cord is the result of a fusion between hypotheses that arose during previous studies that I had conducted in France, the great embryological background and imaging facilities provided by Scott Fraser, and the original experimental systems of cell tracking developed by Paul Kulesa" concludes Mathis."

Scott Fraser is the Anna L Rosen Professor of Biology and the director of the Biological Imaging Center of Caltech's Beckman Institute. Luc Mathis is a former researcher at the Biological Imaging Center who is currently at the Pasteur Institute in Paris. Paul Kulesa is a senior research fellow supported by the computational molecular biology progam and associated with the Biological Imaging Center.

Contact: Robert Tindol (626) 395-3631