Fragile-X, Astrocytes and BMC Image of the Month

Dr. Laurie Doering and his team at McMaster University are discovering root causes of Fragile X Syndrome. A disease manifested by cognitive impairment, attention deficit and autistic behaviours.

I wanted to share highlights and links to a recent publication as it contains interesting conclusions and some of the best multiple label staining of combined embryonic rat and mouse neurons-astrocytes cultures I have seen. No wonder that this is a highly accessed Biomed Central Article and includes the image of the month. The featured  image references use of our MAP-2 antibody.

Shelley Jacobs , Meera Nathwani and Laurie C Doering. Fragile X astrocytes induce developmental delays in dendrite maturation and synaptic protein expression. BMC Neuroscience 2010, 11:132doi:10.1186/1471-2202-11-132.

Conclusions: These experiments are the first to establish a role for astrocytes in the delayed growth characteristics and abnormal morphological features in dendrites and synapses that characterize the Fragile X syndrome.

Image: Co-culture of embryonic mouse hippocampal neurons and astrocytes. Primary embryonic hippocampal neurons at 7 days in vitro, were stained with Microtubule Associated Protein-2 (MAP, green) to enable the visualization of the dendritic arbors. These neurons were cultured on top of a monolayer of primary cortical astrocytes, stained with an antibody directed against Glial Fibrillary Acidic Protein (GFAP, red). The cell nuclei were visualized by staining with 4',6-diamidino-2-phenylindole (DAPI, blue).

Related Links:
Neuronal-Glial Markers-Astrocytes, Glia, Microglia, Olidogodendrocytes, Progenitors and Schwann Cell Markers
Neurofilament or NF Antibodies
Stem Cell Research Antibodies
Stem Cell Research Reagents

Primary Neurons and Astrocytes-Primary human, rat and mouse neurons and astrocytes.

Staining Neuron-Glial Cultures-Related Markers

I have been receiving a growing number of requests for best techniques related to staining cultures of primary neurons and glia. I wanted to share this short, step by step protocol.

These requests are often catalyzed by a search of our growing Neuron/Glial Markers catalog. The objective being to find the right markers for a particular assay. I wanted to share examples of the potency of several Neurofilament or NF markers for labeling neurons:

1. Neurofilament NF-L-Mouse Monoclonal Antibody (Clone: DA2) and Neurofilament alpha-internexin/NF66-Whole Serum-Rabbit Antibody

Images: Cells grown from adult rat brainLarge cell in middle is stained with mouse monoclonal to NF-L clone DA2 (green). Another type of neuronal lineage cell was stained with rabbit polyclonal to alpha-internexin (red). These cells were mitotic but had several characteristics of neurons. Rat spinal cord homogenate showing the major intermediate filament proteins of the nervous system (lane 1). The remaining lanes show blots of this material stainted with various antibodies including NF-L. Protocols on data-sheet.

2. Neurofilament NF-H, phosphylated-Mouse Monoclonal and Neurofilament NF-L-Purified Chicken Polyclonal.

Image: View of mixed neuron/glial cultures stained with chicken polyclonal NF-L (red) and phosphorylated NF-H The NF-L protein is assembled into neurofilaments which are found throughout the axons, dendrites and perikarya of these cells. In contrast the phosphorylated NF-H has a much rmore restricted expression pattern, being found only in developed axonal neurofilaments. Since both proteins are found in neurofilaments, the red and green patterns overlap, so that neurofilaments containing NF-L and phosphorylated NF-H appear yellowish. In contrast neurofilaments containing only NF-L appear red. Protocol on datasheet.

Neurofilament Markers