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Neural Responses to Injury: Prevention, Protection and Repair; Volume 7: Role Growth Factors and Cell Signaling in the Response of Brain and Retina to Injury.

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Annual rept. 20 Sep 95-19 Sep 96,

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The experimental animals used during this period for the project, Neural Responses to Injury Prevention, Protection, and Repair, Subproject Role of Growth Factors and Cell Signaling in the Response of Brain and Retina to Injury, are as follows Species RatAlbino Wistar, Number Allowed 78, Number Used, 78, LSU IACUC 1032. The objective of this study is to assay for changes in expression of genes involved in neural growth and differentiation as a flinction of wound healing. We have used the Chalifour procedure 1 to assay for changes in panels of brain cortex RNAs. Materials and Methods Rat Brain Cryogenic Injury Winstar rats weighing 250-275 g were ether anesthetized and a 9 mm diameter probe cooled in liquid nitrogen was placed on the right parietal region of the rat skull for 1 min. The animals were then euthanized and the brains dissected at the specified times. Analysis of Gene Expression Pafterns Double-stranded radiolabeled cDNAs were synthesized from rat cortex RNAs isolated at various time points following brain injury. Panels of nitrocellulose filter-fixed cDNA clones were then screened according to the method of Chalifour et al. 1. Modifications included the use of 50 g of RNA, 2000u reverse transcriptase, 120 Ci 32P-dCTP, and 2u of klenow per sample. Nitrocellulose filters were hybridized to 106 cpmml of brain cDNA in 10 ml of hybridization solution. RNA Collection and Northern Blots RNAs from rat brain were collected by the method of Chomczynski and Sacchi 2. Northern blots were performed using standard techniques.

Subject Categories:

  • Animal Husbandry and Veterinary Medicine
  • Anatomy and Physiology
  • Medicine and Medical Research

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