Robert J. Contreras

The central projections of the trigeminal, facial, glossopharyngeal and vagus nerves: an autoradiographic study in the rat☆

The central distributions of primary afferent axons in the facial, trigeminal (mandibular branch), glossopharyngeal, and vagal nerves of the rat have been re-examined using the autoradiographic tracing technique after injections of [3H]proline or [3H]leucine into their peripheral ganglia. Within the nucleus of the solitary tract (NST), the labeled terminals from VII, V, IX and X form a continuous distribution that spans the length of this nucleus. Sensory axons in VII terminate mainly within the lateral division of the rostral NST, although some of the terminals extend further caudally within the nucleus. Immediately caudal to the rostral NST, the distribution continues with major contributions from V and IX. Both are confined mainly to the lateral division of the NST, although some of the fibers in IX terminate within the medial division. Injections into the inferior ganglion of X confirm the extensive distribution of vagal axons as they ramify significantly within the lateral division, and virtually monopolize the medial division of the NST. Thus, the major zone of convergency for these 4 cranial nerves is the lateral division of the nucleus from the level of the entering fascicles of IX caudally to the level of the area postrema. Furthermore, only X has a crossed projection as vagal axons invade the commissural nucleus and the medial division of the contralateral NST. Vagal fibers also enter the area postrema bilaterally. Finally, some afferent fibers from VII, IX and X descend in the dorsal part of the spinal trigeminal tract and terminate within the marginal subdivision of the spinal trigeminal nucleus pars caudalis, as well as the dorsal horn of the cervical spinal cord.

Weight cycling in female rats increases dietary fat selection and adiposity

The effect of repeated food restriction-refeeding (weight cycling) on macronutrient selection and adiposity was investigated in female Sprague-Dawley rats. Rats were maintained on ad lib macronutrient self-selection and were put on one of two types of restriction. One group was reduced to 75% of their body weight on restricted amounts of chow and a second group was given ad lib chow during the concurrent period and were voluntarily hypophagic. During refeeding on macronutrient self-selection, animals previously restricted selected a higher percentage of dietary fat, had larger adipose depots and plasma insulin values, and had lower heart weights both expressed in grams and as a percentage of body weight than non-restricted groups. This suggests that both severe and moderate periods of restriction may have negative health consequences.

Sex differences in behavioral taste responses to and ingestion of sucrose and NaCl solutions by rats

Sex differences in the ingestion of food and concentrated NaCl solutions by rats have been investigated for more than a quarter of a century, though the underlying mechanism(s) and the role of reproductive hormones remain the subject of debate. We hypothesized that sex differences in the ingestion of sucrose and NaCl solutions are attributable, in part, to sex differences in taste responses/taste perception. We employed short-access, 10-s tests along with 18-h, two-bottle preference tests to examine sex differences in sensitivity to and ingestion of sucrose and NaCl solutions. To evaluate the role of estrogen, we ovariectomized (OVX) female rats and then used an estrogen-replacement schedule that mimics the pattern of fluctuation of estrogen levels in intact female rats. We observed striking sex differences in the rate of licking NaCl mixed in a dilute sucrose solution. Compared to males, OVX rats with or without estrogen licked at higher rates to more concentrated NaCl solutions, suggesting that female rats are less sensitive to concentrated NaCl solutions. Although less pronounced, we also observed sex differences in the rate of licking to sucrose, particularly at lower concentrations. Compared to males, OVX rats with or without estrogen licked less, suggesting that female rats are less sensitive to lower concentrations of sucrose. Estrogen appeared to play, at most, a small role in mediating taste responses to specific concentrations of sucrose in these testing procedures. Nonetheless, sex differences in taste responses were clear, and it seems likely that such differences underlie, in part, observed differences in ingestion.

High dietary NaCl early in development enhances mean arterial pressure of adult rats

We investigated the long-term influence of early dietary NaCl on adult mean arterial pressure (MAP) and heart rate (HR), assessed continuously for 7 weeks when fed water and chow containing 1% NaCl (weeks 1, 4, 7), 0% NaCl (weeks 2-3), and 3% NaCl (weeks 5-6) while on a 12:12 light/dark cycle. Subjects were offspring of female Sprague-Dawley rats fed solid chow consisting of either 0.1% (basal), 1% (intermediate), or 3% (high) NaCl throughout pregnancy and lactation. After weaning on postnatal day (PD) 21, offspring were fed the same NaCl diet of their mother until PD 30, at which time all offspring were given Purina 5001 diet (1% NaCl) as their solid chow. On PD 60, 22 adult male offspring (eight basals, six intermediates, eight highs) were implanted with an aortic electronic sensor for transmitting blood-pressure signals by telemetry. MAP and HR varied significantly across the 7-week testing period in association with dietary NaCl levels. The three perinatal salt groups had similar HR levels on normal 1% NaCl chow. However, HR increased in all groups during 0% NaCl and decreased in all groups during 3% NaCl. In contrast, the average MAP levels were significantly greater in the highs compared to both the intermediate and basal groups during both dark and light periods. MAP increased in high and basal groups during both 0% and 3%; however, the increase was more pronounced in the highs. In conclusion, exposure to high levels of dietary NaCl early in development led to a persistent increase in MAP in adulthood.

Sodium intake linked to amiloride-sensitive gustatory transduction in C57BL/6J and 129/J mice

Mice of 129/J and C57BL/6J inbred strains received two-bottle, 48-h preference tests of NaCl solutions vs. distilled water. 129/J mice exhibited a greater preference for 0.08 M NaCl than did C57BL/6J mice. To determine if this strain difference was mediated by taste, the integrated neural responses of the chorda tympani nerve to a concentration range of NaCl and KCl were examined. Gustatory neural responses to NaCl were similar for 129/J and C57BL/6J mice. However, lingual application of 0.5 mM amiloride hydrochloride significantly suppressed chorda tympani responses to a range of NaCl concentrations in C57BL/6J mice but did not do so consistently in 129/J mice. Amiloride failed to significantly suppress responses to a range of KCl concentrations in both mouse strains. The results suggest that for 129/J mice, sodium reception and transduction are primarily amiloride insensitive, whereas for C57BL/6J mice, both amiloride-sensitive and amiloride-insensitive components are present. The strain difference in NaCl intake may be mediated, in part, through gustatory mechanisms, with reduced preference for NaCl influenced by amiloride-sensitive sodium transduction mechanisms.

Estrogen influences stimulated water intake by ovariectomized female rats

To further elucidate the influence of estrogen on water consumption, we examined water intake by adult female rats stimulated by water deprivation, injection of hypertonic saline or injection of isoproterenol (ISOP), a beta-adrenergic agonist that activates the renin-angiotensin system (RAS). Rats were ovariectomized (OVX) then injected with estradiol benzoate (EB; 10 microg/0.1 ml oil) or the oil vehicle (OIL; 0.1 ml) for 2 consecutive days. Twenty-four hours after the second injection, rats were deprived of food and water. On the following day, rats were given water and intake was measured after 2 h. EB significantly decreased water intake compared with that by OIL-treated rats following water deprivation. Two additional groups of adult female rats were OVX and treated with EB or OIL. Forty-eight hours after EB or OIL treatment, rats were injected with hypertonic saline (1 ml of 2 M NaCl) or ISOP (30 microg/kg in 0.15 M saline) and water intake was measured after 2 h. EB significantly attenuated water intake following ISOP but not after hypertonic saline. Finally, we examined plasma sodium concentration (pNa) after hypertonic saline and plasma renin activity (PRA) after ISOP in EB- and OIL-treated rats and found no differences in pNa or PRA. These results suggest that the stimuli for water intake after hypertonic saline and ISOP were comparable in EB- and OIL-treated rats. Taken together, these results raise the possibility that EB attenuation of stimulated water intake is specific to water intake elicited by activation of the RAS.

Chorda tympani nerve transection alters linoleic acid taste discrimination by male and female rats

Taste is intimately associated with food choice, yet little is known about the role of taste in preferences for dietary fat, a major component of many foods. We measured the taste threshold for linoleic acid (LA), an essential free fatty acid found in dietary fat, before and after bilateral transections of the chorda tympani nerve (CTX) in adult male and female rats. We conditioned a taste aversion to 88 microM LA and assessed the generalization of the aversion to lower LA concentrations to determine LA discrimination thresholds. We discovered that female rats had a lower LA discrimination threshold (approximately 2.75 microM LA) than did male rats (approximately 11 microM LA). In another set of animals, we performed CTX and found that CTX elevated LA threshold to the same level (approximately 22 microM LA) in male and female rats. Finally, we evaluated licking responses to 11, 22, 44 and 88 microM LA mixed in sucrose by male rats and ovariectomized (OVX) female rats treated with estradiol benzoate or oil vehicle. All rats increased licking to increasing LA concentrations, but OVX rats responded to a lower LA concentration (22 microM) than did males (44 microM) in 10-s trials. However, estradiol did not affect this outcome. Collectively, these experiments show that male and female rats use taste to discriminate LA and that the chorda tympani nerve, which innervates taste buds on the anterior tongue, plays a role in this discrimination. Furthermore, sex differences in fat preferences may depend on differences in fatty acid taste thresholds as well as on the taste stimuli with which fat is combined.

Gustatory adaptation as an explanation for dietary-induced sodium appetite

To test the hypothesis that a Na deprived rat takes longer to adapt to a salt stimulus than a normal rat the temporal characteristics of drinking 0.4 M NaCl and distilled water were investigated. Analysis showed that Na Deprived rats took less time between drinking episodes (interdrink intervals) and drank consecutively for longer periods of time (drinking time) than normal controls. These results were attributed primarily to taste factors because postingestional and thirst influences were at a minimum. Research was also directed at determining the urinary and blood chemical changes associated with dietary sodium deprivation. The levels of sodium in plasma were unchanged because of reduced sodium excretion but the levels of potassium were significantly increased after 20 days of deprivation. Thus, sodium appetite might be important to combat against hyperkaelemia (high plasma potassium) although the appetite develops long before an increase in potassium is detected.

Area postrema lesions disrupt food intake induced by cerebroventricular infusions of 5-thioglucose in the rat

The present experiment assesses the contribution of the area postrema (AP) to glucoprivic feeding in the rat. Lateral ventricular infusions of 5-thioglucose increased pelleted food intake in sham animals but not in animals with AP lesions. Drinking in response to infusion of angiotensin II into the lateral ventricle was not reduced by AP lesions. The data suggest that the AP is part of the brainstem circuitry underlying glucoprivic feeding.

Temperature and amiloride alter taste nerve responses to Na+, K+, and NH4+ salts in rats

The effects of adaptation/stimulus temperature (25 degrees C vs. 35 degrees C) on taste nerve responses to salt stimulation and amiloride suppression were assessed in rats. We measured the integrated responses of the chorda tympani nerve to 500 mM concentrations of NaCl, Na2SO4, sodium acetate (NaAc), KCl, K2SO4, potassium acetate (KAc), NH4Cl, (NH4)2SO4, and ammonium acetate (NH4Ac) mixed with or without 100 microM amiloride hydrochloride at 35 degrees C. Taste nerve responses to all Na+ and NH4+ salts, but not K/ salts, were significantly smaller at 25 degrees C than at 35 degrees C. Amiloride significantly suppressed taste nerve responses to all salts (Na+ salts > K+ salts > NH4+ salts); amiloride suppression of Na+ and NH4+ salts was significantly greater at 25 degrees C than at 35 degrees C. Benzamil-HCl, a more potent Na+ channel blocker compared to amiloride, strongly suppressed taste nerve responses to NaCl and KCl, but not to NH4Cl. Amiloride and benzamil suppression of NaCl responses were similar; however, amiloride suppressed KCl responses more than did benzamil. The results suggest that: (1) amiloride-sensitive Na+ channels are involved to varying degrees in the transduction of sodium and potassium salt taste, and (2) amiloride may inhibit membrane proteins other than passive Na+ channels during stimulation with potassium and ammonium salts.