Differential startle amplitude and corticosterone response in rats.

Different rat strains exhibit large differences in hypothalamic pituitary-adrenal activity that have been used to determine the role of the neuroendocrine system in susceptibility to autoimmune disease. To further characterize potential behavioral correlates of these differences, the amplitude of the acoustic (ASR) and tactile (TSR) startle response and the corticosterone response to acoustic startle stimuli were compared between two histocompatible strains, Lewis (LEW/N) and Fischer (F344/N) rats, as well as outbred Harlan Sprague-Dawley (SD) rats. Startle stimuli elicited larger ASR and TSR in LEW/N rats than in F344/N rats, with SD rats exhibiting an intermediate response. The ASR habituated at a similar rate in LEW/N and F344 rats, while the ASR did not habituate in SD rats. After handling and placement in the startle chambers, the three strains did not differ in control levels of corticosterone. In contrast, exposure to acoustic startle stimuli increased corticosterone 5-fold in F344/N rats and 2-fold in SD rats, but had no effect on corticosterone in LEW/N rats. These findings suggested an inverse relationship between the amplitude of the ASR and hypothalamic-pituitary-adrenal activation across strains. This relationship was further supported by a high negative correlation between corticosterone level and ASR amplitude within the F344/N group.     

Changes in parvalbumin immunoreactivity with aging in the central auditory system of the rat

Changes in the levels of calcium binding proteins are known to occur in different parts of the brain during aging. In our study we attempted to define the effect that aging has on the parvalbumin-expressing system of neurons in the higher parts of the central auditory system. Age-related changes in parvalbumin immunoreactivity were investigated in the inferior colliculus (IC), medial geniculate body (MGB) and auditory cortex (AC) in two rat strains, normally aging Long-Evans (LE) and fast aging Fischer 344 (F344). The results demonstrate that the changes in PV-immunoreactivity are strain-dependent with an increase in the number of PV-immunoreactive (PV-ir) neurons occurring in the inferior colliculus of old LE rats and a pronounced decline in the number of PV-ir neurons appearing in the auditory cortex of aged F344 animals. In some parts of the AC of old F344 animals no PV-ir neurons were present at all. The number of PV-ir neurons in the MGB in all examined animals was very low independent of the strain and age. The loss of PV-ir neurons in the auditory cortex of Fischer 344 rats with aging may contribute to the substantial deterioration of hearing function in this strain.     

Collagen changes in the cochlea of aged Fischer 344 rats

Hearing function in the Fischer 344 (F344) albino inbred strain of rats deteriorates with aging faster than in other strains, in spite of the small hair cell loss in old F344 animals [Popelar, J., Groh, D., Pelanova, J., Canlon, B., Syka, J., 2005. Age-related changes in cochlear and brainstem auditory function. Neurobiol. Aging, in press.]. This study was aimed at elucidating the structural changes in the inner ear of this rat strain during aging. Cochlear histopathology was examined in 20-24-month-old F344 rats and compared with that of young F344 rats (4 months) and of old rats of the Long-Evans (LE) strain. Hematoxylin/eosin staining in aged F344 rats showed degenerative changes in the organ of Corti, consisting of a damaged layer of marginal cells, reduced vascularization of the stria vascularis and a distorted tectorial membrane detached from the organ of Corti. Age-related changes in collagen distribution were observed with Masson's trichrome staining in the spiral ligament of old F344 rats. The results of immunohistochemical staining for type II collagen revealed a marked decrease in collagen fibers in the area connecting the spiral ligament and stria vascularis and a decrease in area IV fibrocytes in old F344 but not in LE rats. These findings may contribute to an explanation of the substantial hearing loss found in old F344 rats.     

Glial glucocorticoid receptors in aged Fisher 344 (F344) and F344/Brown Norway rats

Glucocorticoid receptors (GR) regulate glial function, and changes in astrocyte gene expression are implicated in age-related pathology. We evaluated changes in astroglial GR expression in two strains of rats – Fisher 344 (F344; 4, 12 and 24 months) and F344/Brown Norway strain (F344/BN; 4, 12 and 30 months). In both strains basal levels of corticosterone were higher in the oldest groups of rats. Age-related increases in GR (+) astrocytes but not the percent of astrocytes expressing GR were observed in the hippocampus CA1 region in F344 rats. Age-related decreases in CA1 GR (+) astrocytes and the percentage of GR (+) astrocytes were observed in the F344/BN strain only. Similar strain-specific changes were observed in the dentate gyrus. In the hypothalamic paraventricular nucleus: (1) F344 rats exhibited significant decreases in the overall number of glial profiles with age, (2) F344/BN rats exhibited decreases in the numbers of GR (+) astrocytes with aging and (3) the proportion of GR (+) astrocytes decreased in older F344/BN, but not F344 rats. Overall, the data demonstrate age- and strain-related alterations in GR astrocytic expression that may explain unique phenotypic differences in brain function observed in both strains.     

Neurohypophyseal aging: differential changes in oxytocin and vasopressin release, studied in Fischer 344 and Sprague-Dawley rats

We had previously shown that the hypothalamo-neurohypophyseal vasopressin secreting system is suppressed in aged rats. In the present study, using aged (26 months) male Fischer 344 (F344) rats, we showed that in contrast to vasopressin, oxytocin plasma concentration and hypothalamic content were unaltered in comparison with young (2-3 months) rats; however, based on data from our past and current studies, the neurohypophyseal concentrations of both hormones were found to be decreased in aged rats. We also compared the effect of aging on the oxytocin and vasopressin in secretory functions. Superfusion technique was employed to examine oxytocin and vasopressin release from isolated neural lobes of young (2-3 months) and old (26 months) male F344 and young (2-3 months), middle-aged (12 months) and old (30 months) Sprague-Dawley (SD) rats. Aging affected basal release of oxytocin and vasopressin in a differential manner. Expressed per gland, basal release of oxytocin increased in aged rats of both strains; whereas vasopressin release decreased in SD, and did not change in F344, old rats. The vasopressin responses to electrical stimulation, 56 mM K+ and initial traumatic release were decreased in aged rats; whereas oxytocin responses were either unaltered or decreased much less. All age-related changes were more pronounced in SD than in F344 rats. Thus, while the aging process is associated with a significant impairment in the vasopressin secretory function, the oxytocin secretory function is much less affected by that process. Significant strain differences were observed in the effects of aging on oxytocin and vasopressin release.     

Genetic diversity contributes to abnormalities in pain behaviors between young and old rats

Aging has profound yet unpredictable effects on pain perception and incidence of anxiety disorders. However, the mechanisms underlying age-related pathologies are confounded by contradictory observations in rodent models. Therefore, the goal of our study was to test the hypothesis that genetic variability contributes to age-related pain behaviors and susceptibility to anxiety. To address this hypothesis, we examined pain and anxiety-like behavior in young or old Brown Norway (BN), Fisher 344, and BN/F344 (F1), three rat strains used in studies to evaluate the effect of aging. Mechanosensitive thresholds were assessed using the Von Frey assay, and visceral pain sensitivity was measured via the visceromotor response to colorectal distension. Anxiety-like behavior and exploration was quantified in the elevated plus maze. In the BN strain, old rats exhibited increased mechanosensitive thresholds compared to young rats; however, age did not affect visceral sensitivity in this strain. In F344-BN rats, the number of abdominal contractions induced by the highest colonic distension pressure was significantly lower in old rats. However, following colonic sensitization, a difference was no longer apparent. In the F-344 strain, visceral hypersensitivity following afferent sensitization was evident in young rats at all distension pressures but was not observed in older animals at 20 mmHg. Aging significantly reduced maze exploration across all strains. Our data demonstrate that age- and strain-related alterations exist in pain behavior and highlight the effects of aging on exploratory behavior. These findings suggest that strain differences contribute to the controversial data on the effects of aging on pain perception.     

Aging cochleas in the F344 rat: morphological and functional changes

The Fischer 344 rat strain has been frequently used as an animal model of rapid aging. The present study was aimed at evaluating the incidence of apoptotic cells in the inner ear of 20-24-month-old F344 rats and to correlate it with cochlear function using otoacoustic emissions. Staining with cresyl violet and the enzymatic labeling (terminal deoxynucleotidyl transferase, TdT) of fragmented DNA revealed large numbers of apoptotic cells in the marginal and basal layers of the stria vascularis and in adjacent cells of the spiral ligament. The amplitudes of distortion products otoacoustic emissions (DPOAEs), which reflect functional state of the outer hair cells, were significantly reduced or totally absent in these animals. In contrast to old F344 rats, no marked DPOAE amplitude reduction and smaller numbers of apoptotic cells were found in young 4-month-old F344 rats or in aged 24-28-month-old Long Evans rats. The accumulation of apoptotic cells, mainly in the basal layer of the stria vascularis and in adjacent cells of the spiral ligament, leads to a detachment of the stria vascularis from the spiral ligament and results in the impairment of outer hair cell function. This specific type of strial deterioration suggests that aged F344 rats can serve as an animal model of strial presbycusis.     

Aging reduces hypoxia-induced microvascular growth in the rodent hippocampus

The effect of aging on microvascular density and plasticity in the rodent hippocampus, a brain region critically important for learning and memory, was investigated in F344xBN rats. Capillary density and angiogenesis were measured in three regions of the hippocampus in young and old rats and in old rats administered growth hormone, a treatment that improves cognitive function in older animals. Animals were housed under control conditions or in hypoxic conditions (11% ambient oxygen levels) to stimulate vascular growth. Our results indicate that aging is not associated with a reduction in hippocampal capillary density. However, aged animals demonstrate a significant impairment in hypoxia-induced capillary angiogenesis compared to young animals. Growth hormone treatment to aged animals for 6 weeks did not alter hippocampal capillary density and did not ameliorate the age-related deficit in angiogenesis. We conclude that aging significantly reduces hippocampal microvascular plasticity, which is not improved with growth hormone therapy.     

Establishment of an Aging Farm of F344/N Rats and C57BL/6 Mice at the National Institute for Longevity Sciences (NILS)

In 1996 the National Institute for Longevity Sciences (NILS) started an aging/aged farm (Aging Farm), an animal farm for producing aging/aged laboratory rodents on inbred strains, F344/N rats and C57BL/6 mice at its Experimental Animal Facility Wing, based on plans prepared by the Laboratory Animal Research Facilities (LARF). The NILS Aging Farm, being well established, began internal supply of aging/aged laboratory rodents in 1999 to promote both aging and longevity science. This report describes development of the NILS Aging Farm under NILS Aging Farm Guide and the effectiveness of the Guide.     

Age-related differences in the adaptive potential of type I skeletal muscle fibers

This study was designed to investigate whether the relationship of fiber size and force is maintained with aging and inactivity. We hypothesized that fiber size and fiber force-generating capacity would decrease in parallel, thus resulting in no change in specific force with either age or inactivity. Thirty male Fischer 344/Brown Norway F1 hybrid rats, 6-23-month old (young adult), 24-33-month old (middle-aged) and 34-40 month old (old) were hindlimb unweighted for 14 days. Single permeabilized type I fibers from the soleus and gastrocnemius muscles were evaluated for size and contractile function. The diameter of the fibers from the soleus muscle declined with age and unweighting. In contrast, the fibers from the gastrocnemius showed no age-related atrophy. In both the soleus and gastrocnemius, there was a significant decrease in the force generation with age and unweighting. When comparing size and peak absolute force in the type I fibers from control young and middle-aged animals a positive relationship was observed whereas no significant relationship between size and peak absolute force was observed in the old animals. Following unweighting, fibers from young and middle aged rats showed a significant relationship between force and size. Fibers from old animals did not exhibit a relationship between size and force following unweighting. These results suggest aged skeletal muscle has an attenuated ability to adapt to inactivity by altering its size in response to inactivity.     

Enhanced prepulse inhibition following adolescent ethanol exposure in Sprague-Dawley rats

OBJECTIVES: Recent studies have demonstrated that ethanol exposure differentially affects adolescents and adults. The current studies were designed to compare the effects of 2-week exposure to ethanol during adolescence or adulthood on the acoustic startle response (ASR) and prepulse inhibition (PPI) METHODS: Male Sprague-Dawley rats were exposed to ethanol vapor 12 hr/d (on from 6 pm to 6 am) for 14 days during adolescence or adulthood. Six days after the cessation of ethanol vapor exposure, the ASR and PPI were assessed. RESULTS: During ethanol treatment, overall blood alcohol levels averaged 230 to 250 mg/dl in the adolescent and adult treatment groups. Assessment of the ASR revealed that latency to startle was more rapid in adolescents than in adults, but ASR latency was not altered by ethanol exposure. In addition, ASR magnitude was lower in adolescents and was decreased in ethanol-exposed rats on startle trials. Ethanol exposure significantly enhanced PPI, but only after adolescent exposure CONCLUSIONS: These data further demonstrate a differential sensitivity of adolescents and adults to the effects of ethanol exposure. Specifically, a 2-week period of ethanol exposure during adolescence selectively enhanced PPI, a neurobehavioral index of sensorimotor gating. However, ASR magnitude was decreased by ethanol exposure regardless of age. On the basis of previous studies, the effects of ethanol exposure on PPI data could indicate that adolescent rats exposed to ethanol are more likely to exhibit behavioral inflexibility and that ethanol exposure acts as a more potent physical stressor in adolescent rats.     

Prevention of cardiovascular and renal pathology of aging by the advanced glycation inhibitor aminoguanidine.

Human aging is impacted severely by cardiovascular disease and significantly but less overtly by renal dysfunction. Advanced glycation endproducts (AGEs) have been linked to tissue damage in diabetes and aging, and the AGE inhibitor aminoguanidine (AG) has been shown to inhibit renal and vascular pathology in diabetic animals. In the present study, the effects of AG on aging-related renal and vascular changes and AGE accumulation were studied in nondiabetic female Sprague-Dawley (S-D) and Fischer 344 (F344) rats treated with AG (0.1% in drinking water) for 18 mo. Significant increases in the AGE content in aged cardiac (P < 0.05), aortic (P < 0.005), and renal (P < 0.05) tissues were prevented by AG treatment (P < 0.05 for each tissue). A marked age-linked vasodilatory impairment in response to acetylcholine and nitroglycerine was prevented by AG treatment (P < 0.005), as was an age-related cardiac hypertrophy evident in both strains (P < 0.05). While creatinine clearance was unaffected by aging in these studies, the AGE/ creatinine clearance ratio declined 3-fold in old rats vs. young rats (S-D, P < 0.05; F344, P < 0.01), while it declined significantly less in AG-treated old rats (P < 0.05). In S-D but not in F344 rats, a significant (P < 0.05) age-linked 24% nephron loss was completely prevented by AG treatment, and glomerular sclerosis was markedly suppressed (P < 0.01). Age-related albuminuria and proteinuria were markedly inhibited by AG in both strains (S-D, P < 0.01; F344, P < 0.01). These data suggest that early interference with AGE accumulation by AG treatment may impart significant protection against the progressive cardiovascular and renal decline afflicting the last decades of life.     

Age-related changes in auditory temporal processing in the rat

Presbycusis, as the deterioration of hearing ability occurring with aging, can be manifested not only in a shift of hearing thresholds, but also in a deterioration of the temporal processing of acoustical signals, which may in elderly people result in degraded speech comprehension. In this study we assessed the age-related changes in the temporal processing of acoustical signals in the auditory system of pigmented rats (Long Evans strain). The temporal resolution was investigated in young adult (3-4 months) and old (30-34 months) rats by behavioral and electrophysiological methods: the rats' ability to detect and discriminate gaps in a continuous noise was examined behaviorally, and the amplitude-rate function was assessed for the middle latency response (MLR) to clicks. A worsening of the temporal resolution with aging was observed in the results of all tests. The values of the gap detection threshold (GDT) and the gap duration difference limen (GDDL) in old rats increased about two-fold in comparison with young adult rats. The MLR to a click train in old rats exhibited a significantly faster reduction in amplitude with an increasing stimulation rate in comparison with young adult rats. None of the age-related changes in the parameters characterizing temporal resolution (GDT, GDDL and MLR to a click train) correlated with the degree of the age-related hearing loss. However, the age-related changes in MLR amplitude-rate function correlated with the age-related changes in GDDL, but not with the changes in GDT. The behavioral and electrophysiological data clearly show that aging in rats is accompanied with a pronounced deficit in the temporal processing of acoustical signals that is associated with the deteriorated function of the central auditory system.     

Adolescent HIV-1 Transgenic Rats: Evidence for Dopaminergic Alterations in Behavior and Neurochemistry Revealed by Methamphetamine Challenge

Since the introduction of combination antiretroviral therapy (cART) in the mid-90s, the most severe forms of HIV-1-associated neurocognitive disorders (HAND) have diminished. However, milder forms of HAND remain prevalent. Basic and clinical studies implicate alterations in the dopaminergic (DAergic) system in HIV-1 infection. We used the Fischer 344 HIV-1 transgenic (HIV-1 Tg) rat, which expresses 7 of the 9 HIV-1 genes, to examine potential DAergic alterations. Animals were studied beginning at 35 days of age to assess early-onset DAergic alterations, well before any documented neurological symptoms or clinical signs of "wasting". At 48 hr intervals, animals were administered a single dose of methamphetamine (METH) (0, 0.5, 1, 2.5 and 5 mg/kg/ml s.c.) and tested for the auditory startle response (ASR) and prepulse inhibition (PPI), using an auditory prepulse [85dB(A) broad-band noise stimulus] and an auditory startle stimulus [100 dB(A) broad-band noise stimulus] in a sound-attenuating chamber with a continuous 70dB(A) white noise background. The protocol used a 5-min acclimation period, 6 startle trials, and 36 PPI trials [ISIs of 0, 8, 40, 80, 120, and 4000 ms, 6-trial blocks, Latin square design]. As the dose of METH increased, PPI of the startle response decreased. The HIV-1 Tg rats displayed a greater dose-dependency to the METH-induced disruption of PPI compared to non-transgenic controls. Western blot analysis of midbrain extracts revealed lower tyrosine hydroxylase (TH) protein levels and higher monoamine oxidase A (MAO-A) protein levels in HIV-1 Tg rats treated with METH compared to non-transgenic controls. Early-detected cognitive alterations in the preattentive process of sensorimotor gating may have significant predictive utility regarding the progression of DAergic alterations in HIV-1 infection.     

Stability of neuroendocrine and behavioral responsiveness in aging Fischer 344/Brown-Norway hybrid rats

Aging in rodents and primates is accompanied by changes in hypothalamic-pituitary-adrenal (HPA) activity. We examined behavioral and neuroendocrine responses in 3, 15-, and 30-month-old F344/Brown-Norway rats. Basal corticosterone and ACTH levels did not differ with age, although ACTH responses, but not corticosterone responses to restraint stress, were significantly lower in the 30-month-old group relative to 3- and 15-month-old rats. Induction of c-fos mRNA in the paraventricular nucleus from restraint was not affected by age. Furthermore, there was an enhanced sensitivity to dexamethasone suppression in aged animals as evidenced by lesser ACTH and corticosterone release after dexamethasone administration. Evaluation of emotional behaviors in the forced swim test revealed no differences between the age groups. With fear conditioning, aged rats had decreased freeze times relative to middle-aged or young rats. Regression analysis revealed no significant correlations between the behavioral and HPA axis data in any group. Overall, the data suggest that an apparent decrease in pituitary drive is compensated for at the level of the adrenal, resulting in stable patterns of glucocorticoid secretion. The lack of a correlation between HPA axis measures and emotional as well as fear conditioning-related behaviors indicates that corticosteroid dysfunction may not predict age-related behavioral deficits in this aging model.     

Sugar-induced hypertension in Fischer 344 and F1-hybrid rats (F344/BN) at different ages

Epidemiological evidence suggests that some nutrients, like sodium and potassium, participate in age-related hypertension. A role for refined carbohydrates (CHO), principally sugar, in blood pressure regulation is not generally recognized. This may be unfortunate, since modern lifestyle is associated with large amounts of dietary refined CHO. We examined the effect of a high sugar diet on systolic blood pressure (SBP) in Fischer 344 rats (F344) and the F1-hybrid of this strain (F344/BN). These genotypes have been used to develop experimental models for studies on different aspects of aging. Age-dependent hypertension has not been reported in either strain. In fact, insensitivity to salt-induced hypertension has been found in F344 rats. Upon arrival at our laboratory, the mean difference in SBP between the youngest (4 months) and oldest (18 months) F344 and F1-hybrid rats was 10 mm Hg, the highest mean SBP was 123 mm Hg. These values remained relatively constant over the next month when both strains consumed a low sugar diet. Differently, a steady increase in SBP occurred in both strains when rats of all ages were fed a diet high in sucrose content, mean SBP increasing to over 170 mm Hg at termination of study. Older rats proved more sensitive initially to sugar-induced SBP elevations. Associated with rising SBP was evidence of Na retention. We conclude that a diet containing excess sugar can create a gradual elevation of SBP into a hypertensive range with aging of F344 and F1-hybrid rats. This contrasts with previous findings.     

Age-associated changes in MAPK activation in fast- and slow-twitch skeletal muscle of the F344/NNiaHSD X Brown Norway/BiNia rat model

We compared the tissue content, basal phosphorylation, and stretch-induced phosphorylation of the mitogen-activated protein kinase (MAPK) members; extracellular-signal-regulated kinases (ERK 1/2), p38, and c-Jun NH2-terminal kinase (JNK) in the fast-twitch extensor digitorium longus (EDL) and slow-twitch soleus of young adult (6 month), aged (30 month), and very aged (36 month) F344/NNiaHSD X Brown Norway/BiNia (F344/NXBN) rats. The expression and basal phosphorylation of the ERK 1/2, p38, and JNK MAPK proteins were regulated differently with aging in the EDL and soleus. Stretch induced significant phosphorylation of each signaling molecule in both muscle types of young adult and aged animals. In the very aged animals, stretch stimulated ERK 1/2 MAPK phosphorylation; however, EDL stretch failed to induce JNK MAPK phosphorylation, while soleus stretch was unable to induce the phosphorylation of p38 MAPK. The results suggest that skeletal muscle mechanotransduction processes are affected in very aged F344/NXBN rats and that aging alters load-induced signaling in fast- and slow-twitch muscle types differently.     

Central corticotropin-releasing hormone activates the sympathetic nervous system and reduces immune function: increased responsivity of the aged rat.

CRH acts within the brain to activate the sympathetic nervous system and reduce cellular immune function. To determine the effects of age on CRH-induced elevations of sympathetic activity and suppression of immunity, we examined the responses of plasma catecholamines, neuropeptide-Y (NPY), corticosterone, and splenic natural killer (NK) activity after microinjection of rat CRH (200 pmol) into the lateral ventricle of aged (24-month-old) Fischer 344 (F344) rats compared to those in young (4-month-old) F344 rats. Basal concentrations of plasma norepinephrine and NPY were higher in the aged than in the young animals. In addition, CRH produced a greater elevation of plasma levels of catecholamines and NPY, which persisted for a longer period of time in the aged rats compared to responses in the young animals. Splenic NK activity showed an age-related decrement at baseline, and CRH induced a further significant (P less than 0.01) reduction of lytic activity in the aged rats, but did not alter cytotoxicity in the young rats. Corticosterone basal levels and responses were similar in the aged and young rats. These results show an age-related increase in autonomic outflow and suppression of NK activity after central CRH administration. In aged animals, the central nervous system may have a role in abnormal regulation of sympathetic activity and suppression of natural cytotoxicity in vivo.     

Decline in hepatic microsomal monooxygenase components in middle-aged Fischer 344 rats

Monooxygenase components and drug metabolism activities were determined in liver microsomes from young-adult (3–5 months) and middle-aged (14 months) male Fischer 344 rats. Several components of the monooxygenase system were decreased in middle-aged rats including total cytochrome P-450, cytchtomre b₅, NADPH-cytochrome, reductase activity and phospholipids. However, the reduction of cytochrome P-450 by NADPH, thought to be rate-limiting for monooxygenase activity, was not decreased. Drug metabolism activities in microsomes from middle-aged rats were increased (nitroanisole O-demethylation), decreased (aniline hydroxylation) or unchanged (benzphetamine N-demethylation). Aging decreased the microsomal content of phospholipids and changed the relative percentages of several microsomal fatty acids. The substrate selectivity of the age-related changes in drug metabolism activities may be related to changes in the fatty acid composition of microsomal phospholipids.     

A group of five parameters as a new biological marker on F344/N rats

The National Institute for Longevity Sciences (NILS) established an aging farm (A/F) for producing aging/aged laboratory rodents at the Experimental Animal Facility Wing under the NILS A/F Guide planned by the Laboratory Animal Research Facilities (LARF). Five parameters, the average life span, the number of days of 75, 50, and 25% survival points, and average of the top 10 longest life span among laboratory strains of rodents at NILS-A/F, were reproducible for F344/N rats specifically by strain and sex under the LARF A/F guide. These five parameters may serve as an effective and practical biological marker, especially in aging science including longevity science, to evaluate characteristics of strains of laboratory rodents. The five parameters can identify clear substrain differences between F344/N and F344/Du and breeder differences between F344/DuCrj and F344/DuCrl.