akfg1.jpg (41982 bytes)   Emerging Area of Aging Research:
  Long-lived Animals with
  "Negligible Senescence"
Research 1-8
Research 9-14
Publications on Long Lived Animals
Principal Investigators


(9 - October 2000) Lysosomal markers of intracellular proteolytic activity.
Ana Maria Cuervo, Albert Einstein College of Medicine, Bronx, NY. In a very extensive pilot study, Ana Maria analyzed levels and activity of several lysosomal components in liver of different aged rockfish. She sought to determine if increased protein turnover might contribute to the preservation of systems for removal of damaged proteins and consequently to better cellular functioning (see Cuervo 2000 for protocol). In addition she compared levels of selected substrate proteins, and ran zymograms to analyze specific protease activities (shown are figures for lysosomal enzymes, proteasome enzymatic activities, zymograms and oxidized proteins). Her age groups of five samples each were young (14-23 years old), middle (27-37 years old), and oldest (43-77 years old).

She did not find significant differences in the activity of proteases between age groups, suggesting that the normal levels of protein degradation found in the oldest animals results directly from the normal functioning of their proteolytic systems. A standardized method to detect the presence of oxidized amino acid residues in proteins also showed no increase with age. She concluded that the groups analyzed here did not exhibit the dramatic decline in protein degradation with age described in other species such as rodents.

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(10 - September 2000) Tocopherol derivatives. Robert A. Floyd, Free Radical Biology and Aging Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK. Bob analyzed tocopherol derivatives in rockfish brain and heart up to 83 years old (Hensley et al 2000 for protocol). He found unexplained differences in two comparisons, albeit with significant scatter in the data points (shown are tocopherol comparisons). In gamma over alpha tocopherol and 5-nitro gamma over total gamma measurements, brain tissue showed a positive correlation with age or no trend, respectively, whereas heart tissue showed an inverse relationship with both.

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(11 - September 1999) Telomerase expression. Guido Krupp, Institute for Hematopathology, Christian-Albrechts University, Kiel, Germany. Guido and Wolfram Klapper analyzed telomerase expression in brain, heart and liver tissues from rockfish up to 93 years old. According to the telomere hypothesis, DNA replication leads to telomere shortening, resulting in a cellular mitotic clock (Klapper, Krupp et al 1998a, 1998b). Telomerase resets it by telomere synthesis. Since most rockfish grow throughout their life, they must perform continuous cell proliferation. For maintaining this cell proliferation capacity, telomerase should be active in cells of all somatic tissues, irrespective of fish age. Results of the pilot study confirmed this expectation: in all three tissues, significant telomerase activity was detected (shown are figures depicting the telomerase cycle, and chart comparing expression in rockfish brain, heart and liver tissues). Most importantly, there was no age-dependent change in expression. Unanswered questions of interest are the length of rockfish telomeres, the correlation of telomerase activity with cellular proliferation, and quantification of levels of apoptosis.

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(12 - May 1999) Anion exchange protein in relation to Alzheimer's disease.
Giel Bosman, Department of Biochemistry, Faculty of Medicine, University of Nijmegen, The Netherlands. Giel researched anion exchange (AE) proteins in rockfish brain and heart over 90 years old (Bosman 1997 for protocol). These proteins are known in humans to increase with aging, and especially with degeneration in Alzheimer's disease-affected brain areas. Elucidation of the molecular nature of these changes, and the underlying mechanisms, can lead to insight into the processes that govern aging- and degeneration-associated perturbation of membrane integrity. Immunoblots showed several reactive protein bands that had approximately the same molecular weights as those observed in human brain tissue (shown is immunoblot; the vertical axis is the apparent molecular weight of marker proteins, the horizontal axis is the rockfish tag numbers). In a preliminary analysis, there appeared to be a decrease in total AE protein expression with age, in at least one out of three protein bands in both rockfish brain and heart tissue. A larger sample size will be needed to clarify if this is an age-dependent effect.

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(13 - May 1997) Oxidative damage. David E. Williams, Linus Pauling Institute, Oregon State University, Corvallis, OR. David exposed both Rougheye and Yelloweye rockfish liver samples up to 101 years old to oxidative damage (Kelly et al 1992 for protocol). He found the generation of TBARS (a marker of lipid peroxidation) was dramatically reduced compared to rat or monkey liver microsomes (this experiment is shown below, followed by a figure showing dose-dependent effect of rockfish cytosol protection). Paradoxically, the polyunsaturated fatty acid (PUFA) content (and hence intrinsic oxidizability) of rockfish is relatively high compared to trout, for example, suggesting that rockfish have additional protection from oxidative reactions. Attempts to duplicate this research a year later with samples that had been stored at -80 degrees C. were unsuccessful. Dr. Williams suspects the protection may be thermolabile, possibly a protein or peptide.

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(14 - May 1997) Histology.
Jerry D. Hendricks, Department of Food Science and Technology, Oregon State University, Corvallis, OR. Jerry performed histological examinations of young-, medium- and older-aged rockfish spleen, liver and kidney. He found an increase of melanomacrophage centers in older specimens, although the physiological consequences of these melanomacrophage centers does not seem to affect survival (shown in figure is liver from a 15 year old rockfish in top frame, and an 83 year old rockfish in bottom frame, both 25X). No other cellular indicator existed to differentiate between the cells of young and old rockfish, which ranged over 80 years between youngest and oldest samples.
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