Applications

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Golgi staining and analysis is a valuable tool in the neurostructural assessment of dendritic parameters for studies related to:

An overview of some of the applications are shown below. For additional information Click here to see abstracts from recent scientific meetings and publications.

  • Assess extent of dendritic atrophy and spine loss
  • Evaluate the effects of cognitive enhancers, nerve growth factors, neurotrophins, environmental enrichment or other factors resulting in possible neuroplasticity which might ameliorate or reverse the neurodegeneration or damage (new dendritic branching, increased spine density, changes in spine configuration)
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    Normal Cerebellar Purkinje Cell


     Purkinje cell showing age-related dendritic atrophy


    Camera lucida drawing of normal granule cell of the rat dentate Gyrus


    Camera lucida drawing of damaged granule cell neuron from neonatal rat infected with Borna disease virus

    In the graph below we see the effect of neonatal infection of rat brain with Borna disease virus on estimated total dendritic length, granule cells of the dentate gyrus, (borna disease virus-infected vs. controls)

    Subtle changes in genotype and genomic expression can affect neuronal morphology, often leading to neuronal atrophy or neurodegeneration. Changes in dendritic neurostructure can alter regional brain circuitry and ultimately influence behavior (learning and/or memory). Below are representative examples of findings from some of our studies in this area.


    Transgenic PDAPP mice overexpress human amyloid precursor protein. This results in dendritic atrophy in hippocampus and dentate gyrus. This figure shows a comparison of camera lucida drawings of normal control granule cells and atrophic granule cells of the dentate gyrus in the transgenic mice.


    PDAPP tg mice develop senile plaques. Golgi studies show that there is abnormal neuritic sprouting within the plaque. This figure shows a camera lucida drawing of such aberrant sprouting within a plaque.

  • Another example of the value of Golgi analysis in mutant mice is seen in evaluation of compound heterozygous, tottering/leaner mice (tg/tgla), which exhibit abnormal phenotypic expression , including ataxia,. These mice were found to have cerebella with Purkinje cells having smaller dendritic arbors and significant spine loss.


    • This figure shows a normal Purkinje cell from the cerebellum of a wild-type control


    This grossly dysmorphic Golgi stained Purkinje cell was representative of those widely found thoughout the cerebellum of the tottering/leaner mouse.

  • An animal model of human lissencephaly was developed in which there was a heterozygous deletion of the Lis1 gene. During brain development these Lis1 deficient mice showed abnormal neuronal migration which resulted in hippocampal disorganization accompanied by dendritic atrophy of CA1 pyramids which had not migrated to their proper location in stratum pyramidale. This resulted in the data shown in the figure below.
     
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  • Sparce fur (spf/Y) mice mice have a ornithine carbamoyltransferase deficiency, an X-linked trait, which leads to toxic hyperammonemia. Our Golgi studies clearly showed that this genetic deficiency resulted in cortical pyramids with smaller dendritic trees and spine loss. These changes are seen in the camera lucida drawings below.


    •  Figure A is a camera lucida drawing of the basilar tree of a normal layer V pyramidal cell.


    Figure B, a layer V cortical pyramid from a sparce fur mouse. The dendritic atrophy and spine loss seen in these animals is apparent.

    Subtle influences of chronic and subchronic exposure to environmental neurotoxins on various regions of the developing, adult, or aging brain can be assessed.

  • The figure below shows the neurotoxic effect of gp120 – a glycoprotein associated with the shell of the human immunodeficiency virus (HIV) – on dendritic morphology in the rat. Note the swollen portions along the dendrite segment and the patchy loss of dendritic spines as early signs of neuronal damage.
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  • Prenatal exposure of PCBs (polychlorinated biphenyls) has also been broadly associated with neurotoxic consequences. From analysis of Golgi impregnated neurons we show in the graph below that PCB exposure in rats resulted in reduced numbers of dendritic spines on hippocampal CA1 pyramids.
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     Applications of Golgi studies include:


    Appearance of damaged pyramidal neuron following middle cerebral artery occlusion. Dendritic atrophy, spine loss, and branch swellings can be seen.

  • Clinical tissue with reasonably short post-mortem interval to fixation (usually less than 8 hours) can be evaluated to assist in the assessment of neurological and genetic disorders.

    • The above figure shows the appearance of a highly dystrophic Golgi-stained Purkinje cell from the brain of a 3 year-old who was infected with HIV. This child, whose mother was HIV-positive, showed significant neurological involvement and delayed developmental milestones prior to death.

    For additional examples of these various applications click here to see abstracts from some recent representative publications and presentations at scientific conferences

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    Ronald F. Mervis, Ph.D.
    Neurostructural Research Laboratories, Inc.
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