Induction of tyrosine hydroxylase and neuropeptide Y by carbachol: modulation with age.

With aging, circulating catecholamines are elevated in both humans and animals. This may be related to the increased basal levels of dopamine beta-hydroxylase (DbetaH) and tyrosine hydroxylase (TH) mRNA levels and TH enzyme activity in the adrenal medulla of senescent compared with younger animals. Cold exposure induces TH and DbetaH mRNA, and the cholinergic pathway is believed to be involved in the cold-stimulated increase in TH expression in the adrenal medulla. However, TH gene expression in the senescent rat is resistant to stimulation by cold exposure, suggesting that the cholinergic pathway may be impaired with age in the adrenal medulla. To investigate this possibility, we administered carbachol (0.5 mg/kg i.p., every 12 hours for 3 consecutive days), a mixed nicotinic-muscarinic agonist, to young (4-month-old) and senescent (24-month-old) male F-344 rats. We examined the induction of TH mRNA, TH immunoreactivity, and TH enzyme activity in the adrenal medulla in young and old rats. In addition DbetaH and NPY mRNA levels were determined in the adrenal medulla with or without carbachol administration. Basal levels of TH mRNA, TH immunoreactivity, and TH activity as well as DbetaH and neuropeptide Y (NPY) mRNA were 1.5- to 4-fold greater in the adrenal medullae of old rats compared with young rats. Carbachol administration increased TH mRNA, TH immunoreactivity, and TH activity as well as DbetaH and NPY mRNA to the same or a greater extent in the senescent compared with the young rats. The present study indicates that the cholinergic induction of TH or DbetaH are not impaired with age, and that senescent rats retain the capacity to respond to carbachol stimulation. The present findings cannot explain why the adrenal medullae from senescent rats are resistant to the cold-induced elevation of TH mRNA and TH activity observed in young rats.

Exercise training reverses the age-related decline in tyrosine hydroxylase expression in rat hypothalamus.

Tyrosine hydroxylase (TH) is the rate-limiting enzymatic step in the catecholamine biosynthesis pathway. Some studies have demonstrated that aging is associated with a decrease in TH activity and TH mRNA in rat hypothalamus. We previously demonstrated that exercise training can decrease TH gene expression in the adrenal medulla of young but not senescent rats. This study was designed to examine the effects of endurance training on the TH expression in hypothalamus with aging. To this end, we assessed TH mRNA, TH immunoreactivity, and TH activity with or without exercise training. Young and old F-344 female rats were trained by treadmill running for 8 weeks. All parameters examined were significantly lower in hypothalamus of old (25-month) compared with young (5-month) control animals (p < .05). Exercise training significantly elevated TH mRNA (n = 5-7 in each group), TH immunoreactivity (n = 5-8 in each group), and TH activity (n = 12-13 young groups and n = 6 old groups) in the hypothalamus of old animals (p < .05), but there was no significant change in any of these parameters in young animals following training. These data indicate that endurance training can reverse the age-related decline in catecholamine biosynthesis in the hypothalamus.

AP-1 transcription factor binding activity in rat adrenal medulla and hypothalamus with age and cold exposure.

The AP-1 regulatory element has been implicated in the cold-induced expression of tyrosine hydroxylase in the adrenal medulla. Since in this tissue, the cold-induced increase in tyrosine hydroxylase expression is impaired with age and in other tissues, there is some evidence that AP-1 transcription factor binding is diminished with age, we examined the cold-stimulated AP-1 transcription factor binding to an oligonucleotide with the consensus sequence of the AP-1 response element in nuclear extracts from adrenal medulla and hypothalamus of young and senescent rats. AP-1 transcription factor binding activity diminished by 38% with age in unstimulated adrenal medulla. Following cold stimulation, the AP-1 binding activity increased by 21-25% in the adrenal medulla of both young and senescent rats. However, the level of AP-1 binding in cold-stimulated senescent rats was still less than in cold-stimulated younger rats. There were no changes in AP-3 binding activity with either age or cold exposure in the adrenal medulla. Similarly, in the hypothalamus, there was a 25% decrease with age and a 25% increase following cold stimulation in the level of AP-1 binding. There was a 62% age-related increase in AP-3 binding activity but no change with cold exposure. These data indicate that there is reduced AP-1 binding activity in senescent control rats. Moreover, the demonstration that cold stimulus evokes similar increases in AP-1 binding activity in both young and old rats suggests that the stimulation pathway that increases AP-1 transcription factor is maintained in the senescent animal.

Induction of tyrosine hydroxylase by forskolin: modulation with age.

With aging, circulating catecholamines are elevated in both humans and animals. This may be related to the increased basal levels of tyrosine hydroxylase messenger RNA (mRNA) levels and tyrosine hydroxylase enzyme activity in the adrenal medulla of senescent compared with younger animals. In addition, tyrosine hydroxylase gene expression in the senescent rat is resistant to further stimulation by cold exposure as compared with younger animals. Collectively, these observations suggest either that tyrosine hydroxylase expression is already maximally stimulated in senescent rats or that tyrosine hydroxylase gene induction pathways are impaired with senescence. To help distinguish between these possibilities, we examined the induction of tyrosine hydroxylase mRNA, tyrosine hydroxylase immunoreactivity and tyrosine hydroxylase enzyme activity in the adrenal medulla following forskolin administration to young and old F-344 rats. Forskolin at doses of 1.8 and 3.5 mg/kg increased tyrosine hydroxylase mRNA levels 2.5-fold in adrenal medulla from young rats but did not increase either tyrosine hydroxylase immunoreactivity or tyrosine hydroxylase enzyme activity 5 h after administration. Prolonged treatment with forskolin (3 doses, 12 h apart) increased tyrosine hydroxylase mRNA levels and tyrosine hydroxylase immunoreactivity and tyrosine hydroxylase enzyme activity. In senescent rats, the baseline level of tyrosine hydroxylase mRNA was more than 2-fold higher compared with young rats. A single injection of the lower dose of forskolin increased tyrosine hydroxylase mRNA levels by the same increment in senescent as compared with young rats. These data indicate that the tyrosine hydroxylase gene in the adrenal medulla from senescent rats is still capable of further stimulation.

Tyrosine hydroxylase expression in rat adrenal medulla: influence of age and cold.

Chronic and cold exposure is associated with an increase in adrenal medullary tyrosine hydroxylase (TH) activity and expression that may be important for the regulatory response to cold. Senescent rats do not maintain their body temperature as well as young rats. We investigated the ability of the catecholaminergic system of older rats to respond to cold stimulus. TH activity, TH immunoreactivity, and TH mRNA were assessed in adrenal medullae of male F-344 rats of 3 and 24 months of age following 48 h of mild (8 degrees C) cold exposure. In control rats, basal levels of TH activity were increased by 2.9-fold, TH immunoreactivity by 1.3-fold, and TH mRNA by 2.3-fold with age. In the young rats there were increases after a 48-h cold exposure in TH activity, TH immunoreactivity, and TH mRNA per pair of adrenal medullae. In contrast, in senescent rats there were no significant changes in these parameters following cold exposure. These data suggest that the induction of TH activity is impaired in senescent rats following cold exposure and that there is a loss of plasticity with respect to the TH gene expression.

Myosin heavy chain composition in young and old rat skeletal muscle: effects of endurance exercise.

The objective of this study was to determine the effects of age and exercise on the myosin heavy chain (MHC) composition of skeletal muscle. Young (3 mo) and old (22 mo) female specific pathogen-free barrier-reared Fischer 344 rats were randomly assigned to young untrained or young trained and old untrained or old trained groups, respectively. Young trained and old trained animals performed endurance exercise training on a motorized treadmill for 8 wk. Succinate dehydrogenase activity and MHC isoforms were measured in the plantaris (Plan), lateral and medial gastrocnemius (Gast), and soleus (Sol) muscles. In sedentary animals, aging resulted in a decrease (P < 0.05) in type IIb MHC and an increase (P < 0.05) in type IIa MHC in both the Gast and Plan muscles. Also, aging resulted in a small but significant increase (approximately 4%; P < 0.05) in type I MHC in the Sol. Exercise training resulted in significant (P < 0.05) increases in Gast, Plan, and Sol succinate dehydrogenase activity in both young and old animals. Furthermore, exercise training resulted in a decrease (P < 0.05) in the percentage of type IIb MHC and an increase (P < 0.05) in the percentage of type IIa MHC in the Plan in both young and old animals. These data suggest that there is an age-related shift in locomotor muscle MHC isoforms from a faster to a slower isoform.