MK-801 Neurotoxicity in Cupric Silver-Stained Sections: Lesion Reconstruction by 3-Dimensional Computer Image Analysis

Abstract

Routine histopathologic evaluation of the brain (paraffin embedding, hematoxylin and eosin staining) makes it difficult for an investigator to identify the overall location and relative extent of lesions as they relate to neural substructures. Moreover, it is very difficult to convey this information to others who are less familiar with neuroanatomy. This study combined a 3-dimensional imaging program with a cupric silver stain for neuronal degeneration in order to determine the location and extent of a focal lesion produced by (dizocilpine maleate), a glutamate receptor antagonist that induces necrosis in a small population of neurons in the cortex of rats. A male Sprague-Dawley rat was treated with a subcutaneous dose of MK-801 (10 mg/kg) and was perfused with fixative through the left ventricle 3 days after treatment, a time point known to reveal maximal neurotoxic effects. The brain was embedded in a gelatin matrix, frozen, and serially sectioned at a thickness of 40 μm. The cupric silver method of de Olmos was used to stain frozen sections at 320-μm intervals. Using a color charged-couple device (CCD) camera and a macro lens, a series of 2-dimensional images, which encompassed the entire rostral to caudal extent of the brain, was captured. A computer program was written to define internal and external boundaries in these 2-dimensional images. Then, 3-dimensional reconstructions were generated on a Silicon Graphics workstation using IRIS "Explorer." The quality of the 3-dimensional reconstructions allowed for easy identification of various neural substructures while clearly revealing the exact location and extent of the resulting necrotic neurons that were positively identified by the cupric silver stain. This 3-dimensional lesion reconstruction method provides a powerful tool for conveying spatial information about the nature of neurotoxic lesions in the brain. In addition, it may be used to investigate further dose-response relationships and the effects of other neurotoxicants.

A single application of MK801 causes symptoms of acute psychosis, deficits in spatial memory, and impairment of synaptic plasticity in rats

“…Systemic application results in impairment of learning and memory in rodents (Ahlander et al, 1999;Fix et al, 2000) as well as in dose-related motor dysfunctions like ataxia and hyperactivity. Antagonists of the An acute schizophreniform psychotic episode has no effect on locomotion, rearing, grooming or defecation Acute application of MK801 (5 mg/kg, n 5 8) 4 weeks prior to the commencement of the 10 day radial maze study had no effect on locomotion (A), grooming (B) or rearing (C) compared with vehicle injected controls (n 5 8).…”

Acute and long‐term effects of MK‐801 on direct cortical input evoked homosynaptic and heterosynaptic plasticity in the CA1 region of the female rat

“…In contrast, MK‐801‐induced neurodegeneration of hippocampal CA1 cells and EC cells have been reported in animals treated with high concentrations of intraperitoneal MK‐801 (10 mg/kg). In most of these studies, neurodegeneration was successfully illustrated by using the amino–cupric–silver method to assess the degree of neuronal cell death (Fix et al ., 2000; Bueno et al ., 2003a).…”

Registering Histological and MR Images of Prostate for Image-Based Cancer Detection

“…The scale-space representation of this blob f can be thus obtained as: (6) At any scale s, the normalized Laplacian of L(x, y, z; s) reaches its maximum at the location (x 0 , y 0 , z 0 ), i.e., (7) The maximum of Q(s) across scales is calculated by differentiating Q(s) with respect to s, (8) As Equation (8) After finding the local maxima in normalized Laplacian maps across different scales, the centers of blob-like structures are detected and used as candidates for internal landmarks. The expected internal landmarks are further determined as follows.…”