Guanethidine-treated Rats Research Articles

Corticotropin-releasing hormone and the sympathoadrenal system are major mediators in the effects of peripherally administered exendin-4 on the hypothalamic-pituitary-adrenal axis of male rats.

Glucagon-like peptide-1 (GLP-1) and the GLP-1 receptor agonist, exendin-4 (Ex-4), potently stimulate hypothalamic-pituitary-adrenal (HPA) axis activity after either central or peripheral administration. Because several GLP-1 derivative drugs, including synthetic Ex-4, are currently in use to treat patients with type II diabetes mellitus, the characterization of Ex-4 effects on the HPA axis is highly relevant. Herein, the roles of CRH and AVP on these effects were investigated by administering the antagonists astressin and d(CH2)5Tyr(Me)AVP, respectively. The role of the sympathoadrenal system (SAS) was explored in bilateral adrenal enucleated and guanethidine-treated rats, whereas primary pituitary cell cultures were used to study direct effects on the corticotropes. Astressin completely abrogated (P < .05) the effects of Ex-4 central administration on ACTH secretion but only slightly reduced (by 35%) the ACTH response to Ex-4 peripheral administration. Moreover, astressin significantly (P < .05) decreased the corticosterone response to centrally but not peripherally administered Ex-4, suggesting different mechanisms depending on the route of administration. Pretreatment with d(CH2)5Tyr(Me)AVP failed to diminish either the ACTH or corticosterone response to Ex-4 and no direct effect of Ex-4 or GLP-1 was observed on pituitary cell cultures. In contrast, a significant (P < .05) reduction in the corticosterone response elicited by Ex-4 peripheral administration was observed in enucleated and guanethidine-treated rats, indicating a role of the SAS in the glucocorticoid stimulatory effects of Ex-4. Our data demonstrate that the effects of Ex-4 on the HPA axis are partially mediated by CRH and the sympathoadrenal system, and stress the relevance of Ex-4 as a corticosterone secretagogue.

Protein metabolism in guanethidine-treated rats.

The aim of the present study was to evaluate the effect of short-term adrenergic blockade on the rate of whole-body protein turnover and leucine oxidation, and on protein synthesis in specific tissues in male rats. Adrenergic blockade was induced by guanethidine (100 mg/kg body weight subcutaneously). The control group was treated with saline. On the second day, the parameters of whole-body protein and leucine metabolism were evaluated using a primed constant intravenous infusion of L-[1-(14)C]leucine. Protein synthesis in tissues was determined on the basis of L-[1-(14)C]leucine incorporation. Guanethidine treatment caused a decrease in norepinephrine in skeletal muscle. Whole-body leucine oxidation and leucine oxidized fraction were higher in guanethidine-treated rats. There was an insignificant effect of guanethidine on whole-body proteolysis, protein synthesis and leucine clearance. However, protein balance was negative due to the larger difference between protein synthesis and proteolysis in guanethidine-treated animals compared to controls. In guanethidine-treated rats, protein synthesis was higher in the gastrocnemius muscle and in the kidneys and lower in liver and spleen. Changes in the small intestine and colon were insignificant. In addition, a marked decrease in concentration of several amino acids has been observed in the liver, the kidneys and the spleen. It is concluded that adrenergic blockade induced by guanethidine is associated with significant changes in protein metabolism, leucine oxidation and amino acid concentrations in several tissues. The most important consequences of treatment are considered to be a negative effect on protein balance, increased protein turnover in skeletal muscle and kidneys and decreased protein synthesis in the liver and spleen. These changes may also be induced by administration of other sympathetic blocking agents, e.g. in treatment of hypertension.

Evidence of angiotensin II involvement in prolactin secretion in response to hemorrhage in adrenodemedullated and guanethidine-treated rats.

The present experiments were designed to investigate the influence of the renin--angiotensin system (RAS) on prolactin secretion in response to hemorrhage (1.2 ml/100 g body weight (bw)/2 min). Male Wistar rats (250-300 g) were divided into the following experimental groups. (i) Sham-operated animals submitted to intravenous administration of [Sar(1),Thr(8)]-angiotensin II (sarthran), an angiotensin II antagonist (750 ng/100 g bw as a bolus plus an infusion of 25 ng/100 g bw/min over 30 min), which did not alter the prolactin secretion in response to hemorrhage. (ii) Animals submitted to adrenodemedullation which by itself increased the prolactin secretion in response to hemorrhage by 274% (P<0.01). However, sarthran infusion into adrenodemedullated rats completely blocked this increased prolactin secretion in response to hemorrhage (P<0.01). (iii) Intact animals submitted to blockade of sympathetic noradrenergic pathways by pretreatment with guanethidine (10 mg/100 g bw), which also increased the prolactin secretion in response to hemorrhage by 55% (P<0.01). This increased prolactin secretion in response to hemorrhage observed in guanethidine-treated rats was completely blocked by sarthran preinfusion (P<0.01). (iv) Adrenodemedullated animals pretreated with guanethidine, which abolished the prolactin secretion induced by hemorrhage. Our data suggest a role for circulating catecholamines in the prolactin secretion response to stress. In addition, the experiments reported here demonstrate that RAS has a stimulatory effect on prolactin secretion in circumstances in which sympathetic activity or adrenomedullary secretion is suppressed. These are the first data demonstrating that a physiological prolactin secretion response to stress depends on the RAS.

Regulation of adrenal angiotensin receptor subtypes: a possible mechanism for sympathectomy-induced adrenal hypertrophy.

Previous studies indicate that the adrenal gland plays a compensatory role in the maintenance of blood pressure in chemically sympathectomized rats. However, the mechanisms responsible for compensatory adrenal responses are poorly understood. This study examined the regulation of adrenal growth and type 1 A, 1 B, and type 2 angiotensin II (Ang II) receptor (AT1A, AT1B and AT2) expression in the adrenal gland induced by sympathectomy. Five-week-old male Sprague-Dawley rats were treated with either guanethidine (50 mg/kg per day, intraperitoneally) or vehicle for 5 weeks. Norepinephrine and epinephrine levels in the atrium of the heart were measured by high-pressure liquid chromatography. Plasma renin activity was determined by radioimmunoassay. Adrenal AT1 and AT2 receptor density was determined by radioligand binding assay. Adrenal AT1A, AT1B and AT2 mRNA levels were determined by Northern blot analysis. Norepinephrine and epinephrine levels in the atrium of the heart were decreased 86% (P < 0.0001) and 58% (P < 0.05) by guanethidine treatment, respectively. Plasma renin activity was decreased 71% (P< 0.001) in guanethidine-treated rats compared with vehicle. In contrast, the ratio of adrenal to body weight was increased 38% in guanethidine-treated rats compared with vehicle (P< 0.001). Adrenal AT1 and AT2 receptor density was increased by guanethidine treatment (P< 0.05). Adrenal mRNA levels for AT2 (P< 0.001) and AT1A (P< 0.01), but not AT1B (P>0.05), were increased in guanethidine-treated rats compared with vehicle (P< 0.01). There were positive correlations between adrenal weight and AT2 (r = 0.9, P< 0.001) and AT1A (r = 0.6, P< 0.05) but not AT1B (r = - 0.01, P > 0.05) expression. Impairment of the sympathetic nervous system with guanethidine withdraws the normal stimulation of this system on the circulating renin-angiotensin system, but upregulates the expression of adrenal Ang II receptors. Increased expression of adrenal AT2 and AT1A receptors may play an important role in adaptive adrenal hypertrophy and hormonal responses to sympathectomy.

Fertility of rat epididymal sperm after chemically and surgically induced sympathectomy.

Guanethidine, a chemical that selectively blocks sympathetic noradrenergic neurons, was used to investigate the role of sympathetic innervation in the fertility of rat epididymal sperm, using both natural mating and in utero insemination protocols. This animal model correlates, at least in part, with spinal cord injury (SCI) in men. Adult male rats were treated daily by i.p. injections, for 21 or 42 days, with 0 or 6.25 mg/kg guanethidine. To compare the effects of guanethidine-induced sympathectomy with those following surgically induced sympathectomy, the inferior mesenteric ganglion and the proximal hypogastric nerves were removed in another group of rats. Both chemically and surgically induced sympathectomy increased the weight of the epididymis and seminal vesicles/coagulating glands as well as the number and the transit time of cauda epididymal sperm. Neither serum testosterone levels nor LH was affected by treatment with guanethidine. Using natural mating, no litters were produced by guanethidine-treated rats. Chemically denervated rats failed to produce copulatory plugs or ejaculate into the uterus. However, distal cauda epididymal sperm from chemically or surgically denervated rats displayed normal fertilization ability (80%) using in utero inseminations. In addition, the sperm of denervated rats did not show abnormal sperm chromatin structure using an assay that detects DNA damage. We conclude that sympathectomy delays the transit of sperm through the cauda epididymidis and produces ejaculatory dysfunction but does not compromise sperm quality in the distal cauda epididymidis. Moreover, these data provide compelling evidence that there is no association between the prolonged transit time of sperm within the epididymis, i.e., pre-ejaculatory sperm aging, and the fertility of those sperm, which has important implications for artificial insemination using sperm from men with SCI.

Brain ischemia and gastric mucosal damage in spontaneously hypertensive rats: the role of arterial vagal adrenoceptors.

Brain ischemia is often accompanied by acute gastric lesions. To clarify the underlying mechanism, the influence of acute ischemic insult to the brain on gastric hemodynamics and mucosal integrity was examined in spontaneously hypertensive rats. One hour after brain ischemia, gastric mucosal blood flow decreased to 71% of the preischemic levels in the control rats but was preserved significantly better, at 94 and 108%, in the prazosin-treated and guanethidine-treated rats, respectively. Vagotomy almost abolished the decrease in gastric mucosal blood flow during cerebral ischemia. Intragastric 0.6 N hydrochloric acid administered just after reperfusion induced more severe hemorrhagic ulcers in the control than in the prazosin-treated and vagotomized groups. These results suggest that noradrenergic neurons acting through alpha1-adrenoceptors contributes to the decrease in gastric mucosal blood flow, and the subsequent disturbed integrity of the gastric mucosa, through the vagal adrenergic pathway during brain ischemia in spontaneously hypertensive rats.

Effect of [1-Sar,8-Thr]-angiotensin II on the hyperglycemic response to hemorrhage in adrenodemedullated and guanethidine-treated rats

The present experiments were designed to further investigate the action of an angiotensin II antagonist on the hyperglycemic response to hemorrhage (1.2 ml/100 g b.wt./2 min). The animals were divided into 3 experimental groups: (1) sham-operated animals submitted to intravenous administration of [1-Sar,8-Thr]-angiotensin II (sarthran), an antagonist of angiotensin II (750 ng/100 g b.wt. as a bolus plus an infusion of 25 ng/100 g b.wt./min over 30 min), which greatly attenuated (51.8% lower than controls; P < 0.01) the hyperglycemic response to hemorrhage; (2) animals submitted to adrenodemedullation which decreased the hyperglycemic response to hemorrhage by 64% ( P < 0.01). However, sarthran infusion into adrenodemedullated rats caused a 38.5% further decrease in hyperglycemic response to hemorrhage ( P < 0.01); and (3) intact animals submitted to blockade of sympathetic noradrenergic pathways by treatment with guanethidine (10 mg/100 g b.wt.), which greatly decreased the baseline value of plasma glucose (64.1 ± 3.5 mg% vs. 125.3 ± 4.5 mg%, P < 0.01), and reduced the hyperglycemic response to hemorrhage by 34% ( P < 0.01). Sarthran infusion into guanethidine-treated rats caused a further 34% decrease in hyperglycemic response to hemorrhage ( P < 0.01). These data indicate that angiotensin II has a direct hyperglycemic effect in addition to its action on sympathetic nervous system activation and adrenomedullary secretion.

Augmented Flow-Induced Endothelin Release from the Rat Mesenteric Arterial Bed after Long-Term Sympathectomy

The effect of long-term guanethidine sympathectomy and short-term 6-hydroxydopamine (6-OHDA) sympathectomy on the release of endothelial vasoactive substances by high flow from the rat mesenteric arterial bed was examined. Preparations were perfused at a constant flow rate of 5 ml/min with Krebs' solution, and at 25 ml/min for 2 min. During increased flow there was a release of adenosine 5′-triphosphate (ATP), endothelin, vasopressin and substance P into the perfusate. In preparations from guanethidine-treated rats the flow-evoked release of endothelin (median: 23.55 pmol/min) was significantly greater than that in the controls (median: 0.37 pmol/min) (P < 0.05). Flow-evoked release of ATP, vasopressin and substance P was similar between the groups. After short-term sympathectomy with 6-OHDA an increase in flow was accompanied by the release of ATP, endothelin, vasopressin and substance P which was not significantly different between control and 6-OHDA treated preparations. These results show that after long-term sympathectomy there is an increase in the flow-evoked release of endothelin from the rat mesenteric arterial bed which does not occur after short-term sympathectomy. It is suggested that sympathetic nerves may have long-term trophic effects on the content of endothelial vasoactive substances which may be expressed as a selective alteration in their patterns of release after stimuli such as increased flow.

Guanethidine sympathectomy increases substance P concentration in the superior sympathetic ganglion of adult rats

Adult rats received intraperitoneal injections of guanethidine or saline for 5 weeks. Six to 8 weeks following completion of treatment, concentrations of substance P and neuropeptide Y (NPY) were measured by radioimmunoassay in the superior cervical ganglion (SCG) and thoracic spinal cord. The SCG was also immunostained for NPY and substance P. No differences were observed in thoracic spinal cord content of either NPY or substance P. We observed depletion of NPY immunoreactive neurons and NPY levels in the SCG, and pharmacologic evidence of postganglionic denervation in guanethidine-treated rats. In guanethidine-treated rats, there was a marked increase of substance P levels in the SCG, where substance P was localized in fibers, but not cell bodies. Thus, sprouting of substance P-containing sensory fibers in the sympathetic ganglia occurs following postganglionic sympathectomy in adult rats.

Effects of neonatal sympathectomy with 6-hydroxydopamine or guanethidine on survival of neurons in the intermediolateral cell column of rat spinal cord

The effects of removing target cells on survival of, and inputs to, sympathetic preganglionic neurons were studied in rats that were sympathectomized with 6-hydroxydopamine (6-OHDA) or guanethidine sulfate. Separate groups of neonatal and 1-week male rats were given injections of 6-OHDA for 10 days and of guanethidine for 3 weeks (5 days/week), respectively. Histofluorescence results suggest that catecholaminergic neurons in most ganglia are destroyed with treatment except for adrenal medulla, which is unaffected [14], and the pelvic ganglion where only partial destruction occurs. Cells in the intermediolateral cell column from representative spinal cord segments of treated and control adult rats were counted. In 6-OHDA-treated rats, cells decreased in number in all segments compared to controls. In guanethidine-treated rats, cells were also decreased in number; in some segments the decrease was significantly greater than with 6-OHDA. Sympathectomy had no effect on neurons in the intermediate gray of L5 or in the ventral horn of T3. The results of this study demonstrate that peripheral sympathectomy causes loss of sympathetic preganglionic neurons and that guanethidine is slightly more effective than 6-OHDA.

The effect of guanethidine treatment of testicular blood flow and testosterone production in rats

Testicular blood flow was measured by means of Xenon-133 clearance in control rats and rats treated with guanethidine for 3 weeks. Plasma and testis testosterone concentrations were also measured, and the effect of hCG-treatment was examined. No difference in testicular blood flow between the control group and the guanethidine-treated group was found. However, in guanethidine-treated rats, plasma and testis testosterone concentrations after hCG-treatment were significantly decreased. The results may indicate that adrenergic nerves are involved in the regulation of Leydig cell function.

Use of enhanced silver staining combined with electron microscopical immunolabelling to demonstrate the colocalization of neuropeptide Y and vasoactive intestinal polypeptide in cerebrovascular nerves

The combination of immunolabelling at the electron microscope level and enhanced silver staining has been used to demonstrate the colocalization of neuropeptide Y and vasoactive intestinal polypeptide in perivascular nerves supplying cerebral arteries of the rat. This has been shown in control tissue, but it is easier to demonstrate after long-term sympathectomy since that leads to an enhancement of neuropeptide Y in vasoactive intestinal polypeptide-containing parasympathetic nerves supplying these vessels. Immunolabelling of the antigens for these peptides was performed sequentially with the biotin-streptavidin-diaminobenzidine method, and the end product to the first antiserum was gold-silver intensified before the visualization of the second antigen. Using this technique, it was shown that all the neuropeptide Y immunoreactivity present in the rat cerebral vessels after long-term sympathectomy with guanethidine was localized in vasoactive intestinal polypeptide-containing nerves. Furthermore, an immunohistochemical analysis of the parasympathetic pterygopalatine ganglia in guanethidine-treated rats showed an increase in the percentage of neurons displaying neuropeptide Y immunoreactivity. In order to clarify if the pterygopalatine ganglion was the origin of those neuropeptide Y/vasoactive intestinal polypeptide-immunoreactive cerebrovascular nerves, which had increased in number after sympathectomy, a fluorescent neuronal tracer (Fast Blue) was applied to the right middle cerebral artery of rats which had undergone guanethidine treatment for six weeks. Immunohistochemical analysis of the ipsilateral ganglion 72 h after application of the tracer revealed the presence of immunoreactivity to both these peptides in retrogradely labelled neurons. It is concluded that neuropeptide Y and vasoactive intestinal polypeptide are colocalized in perivascular parasympathetic nerves supplying the middle cerebral artery of the rat, which have their origin in the pterygopalatine ganglion. Furthermore, long-term sympathectomy with guanethidine leads to an increase in the expression of neuropeptide Y in these vasoactive intestinal polypeptide-immunoreactive neurons.

Baroreceptor mechanisms in rat

Baroreflex control of cardiovascular parameters was studied in control, atropine- and guanethidine-treated rats. Baroreceptor activity was tested by the relationship between the increase in blood pressure produced by a phenylephrine administration (bolus ov infusion) and the induced bradycardia. No differences were observed in basal arterial blood pressure and heart rate between treated- and control rats. Baroreceptor sensitivity was lower in atropine- or guanethidine-treated rats than in control animals. Baroreceptor activity has two components: a first, rapid, predominantly parasympathetic and a second, slower, that is mediated by both parasympathetic and sympathetic efferent pathways.

Effects of Adrenalectomy and Chronic Guanethidine Treatment on Tissue Adrenaline Concentrations in Swimming-Exposed Rats

Adrenaline is known to increase in extra-adrenal peripheral organs as well as urine and plasma in the rat after swimming stress. In order to clarify the physiological significance of adrenaline in stress responses, the origin and location of adrenaline in the peripheral organs were examined by measuring adrenaline in the heart, spleen, submaxillary gland and some other organs after adrenalectomy or guanethidine treatment. After bilateral adrenalectomy, adrenaline in the peripheral organs mostly disappeared. Swimming stress caused no increase in the adrenaline content of the organs in the adrenalectomized rats, while a marked increase was observed in the sham-operated animals. In guanethidine-treated rats, the adrenaline content of the peripheral organs was extremely low as compared to that in the saline-treated animals, and the swimming-induced increase in adrenaline level in the organs was greatly suppressed. These observations suggest that the organ adrenaline, which exists in sympathetic nerve endings, is mostly derived from the adrenal gland. It is considered that adrenaline would be involved in sympathetic neurotransmission, especially under stressful conditions.

Effects of adrenalectomy and chronic guanethidine treatment on tissue adrenaline concentrations in swimming-exposed rats.

Adrenaline is known to increase in extra-adrenal peripheral organs as well as urine and plasma in the rat after swimming stress. In order to clarify the physiological significance of adrenaline in stress responses, the origin and location of adrenaline in the peripheral organs were examined by measuring adrenaline in the heart, spleen, submaxillary gland and some other organs after adrenalectomy or guanethidine treatment. After bilateral adrenalectomy, adrenaline in the peripheral organs mostly disappeared. Swimming stress caused no increase in the adrenaline content of the organs in the adrenalectomized rats, while a marked increase was observed in the sham-operated animals. In guanethidine-treated rats, the adrenaline content of the peripheral organs was extremely low as compared to that in the saline-treated animals, and the swimming-induced increase in adrenaline level in the organs was greatly suppressed. These observations suggest that the organ adrenaline, which exists in sympathetic nerve endings, is mostly derived from the adrenal gland. It is considered that adrenaline would be involved in sympathetic neurotransmission, especially under stressful conditions.

Factors contributing to the maintenance of arterial blood pressure in adult rats treated neonatally with guanethidine.

Adult rats treated neonatally with guanethidine had normal arterial blood pressures, but these were more dependent on the renin-angiotensin system than in control animals. Antagonism of V1-receptors (with d(CH2)5DAVP) enhanced the depressor effects of captopril in guanethidine-treated animals, but blood pressure was unaffected by this manoeuvre in control animals. Although there is evidence that sympathetic efferent vasomotor function is abolished following the schedule of neonatal guanethidine treatment used, pentolinium had a hypotensive effect in the presence of d(CH2)5DAVP and captopril, indicating that the adrenal medulla could have been contributing to the maintenance of blood pressure. This is consistent with the finding of marked supersensitivity to exogenous adrenaline in guanethidine-treated rats.

The influence of neonatal treatment with guanethidine on the development of isolation-induced hypertension in adult rats.

Rats housed individually in glass metabolism cages develop hypertension. Since previous experiments have provided some evidence for the involvement of the sympathetic nervous system in the maintenance of the hypertension, the present work was designed to explore the possible involvement of the sympathetic nervous system in the genesis of isolation-induced hypertension. Male and female Wistar rats were treated neonatally with guanethidine, with a protocol designed to produce an extensive peripheral sympathectomy; control rats received saline. The effects of isolation on systolic blood pressure and fluid and electrolyte balances were studied when the rats were mature. Guanethidine-treated rats did not develop hypertension in response to isolation whereas control rats did. There were no significant differences between the fluid and electrolyte balances of the guanethidine-treated rats compared with controls throughout the period of isolation. It is concluded that a fully functional sympathetic nervous system is required for the development of isolation-induced hypertension, but its involvement is not through a modulation of renal function.

Impairment of trophic response of brown fat to cold in guanethidine-treated rats.

The aim of the present study was to investigate the role of the sympathetic nervous system in the trophic response of brown adipose tissue (BAT) to chronic cold exposure. The hyperplasia and the development of the mitochondria characterizing this response are usually considered as mainly controlled by the sympathetic activity in BAT, but this has never been clearly demonstrated. In the present work rats were sympathectomized by chronic administration of guanethidine and then exposed to cold during two weeks. The treatment induced a strong reduction of the noradrenaline content of BAT. The trophic response of the tissue to cold was largely impaired: no increase of the tissue weight, weak increase in the DNA, protein, and phospholipid content of the tissue. The development of the mitochondria was almost abolished. The increase in the proportion of the 32,000-dalton protein, a protein which regulates heat production by BAT, was suppressed and the GDP-binding to mitochondria, which is an index of BAT thermogenic capacity, was not increased as in normal cold-adapted rats. The ultrastructure of the tissue remained the same as in warm-adapted animals. It is concluded that the sympathetic nervous system plays an essential role in the control of the trophic response of BAT to the cold.

Crypt cell renewal and chemical sympathectomy in the pre-closure ileum of the neonatal rat.

The present study was designed to analyze the effect of guanethidine-induced sympathectomy on ileal crypt cell proliferation before neonatal closure to macromolecular absorption (pre-closure ileum). The present experiments demonstrated an alteration of the crypt cell generation cycle (T&lt;sub&gt;C&lt;/sub&gt;) in the pre-closure ileum (15 days after birth) with guanethidine treatment. Percent labeled mitoses measurements for 15-day-old rats indicated a lengthening of T&lt;sub&gt;C&lt;/sub&gt; from 18.1 to 23.0 h with the elongation primarily due to alteration of T&lt;sub&gt;G1&lt;/sub&gt; (pre-DNA-synthetic gap period) and T&lt;sub&gt;S&lt;/sub&gt; (DNA-synthetic period). Mitotic and labeling indices and the amplitude of the circadian rhythm in mitotic activity were significantly depressed in guanethidine-treated rats.

Alteration of neonatal rat parotid gland acinar cell proliferation by guandethidine-induced sympathectomy.

Newborn rats were injected with guanethidine-sulfate (20 micrograms/g body weight) every 48 hr from 12 hr after birth until day 14 (eight injections per animal). The guanethidine treatment resulted in an 86% absolute reduction in cell number in the superior cervical ganglia of 15 day old rats. The cells which remained after guanethidine treatment showed destruction of mitochondria and an extensive decrease in endoplasmic reticulum. Chemical sympathectomy with guanethidine induced a 3.1 hr lengthening of the acinar cell generation cycle time (17.4 hr to 20.5 hr), resulting from a longer G1 period (6.9 hr in the control group as compared to 10.5 hr in the guanethidine-treated group), as well as a cecrease in the mean percentage of [3H]thymidine-labeled acinar cells (22.3 +/- 0.5% to 19.3 +/- 0.5%) and mean acinar cell mitotic index (2.6 +/- 0.2% to 2.1 +/- 0.1%). A circadian rhythm was found to exist in parotid gland acinar cell mitotic activity of 15 day old rats and the amplitude of the rhythm was reduced from 26.5% to 14.9% in guanethidine-treated rats. This study indicates that the diminution of sympathetic influence on the developing parotid gland results in a slight, but significant alteration in acnar cell proliferation.