Barry J. Connell

Autonomic and cardiovascular reflex responses to central estrogen injection in ovariectomized female rats

The role of estrogen in central autonomic nuclei was examined in ovariectomized female Sprague-Dawley rats supplemented daily for 7 days with either estrogen (5 microg/kg; sc) or saline (0.9%; sc). Animals were subsequently anaesthetized with sodium thiobutabarbital (Inactin; 100 mg/kg; ip) and instrumented to record blood pressure and heart rate. Efferent vagal parasympathetic (VPNA) and renal sympathetic (RSNA) nerve activities were recorded and used to assess baseline and reflexive changes in autonomic tone. The cardiac baroreflex was evoked using a single bolus injection of phenylephrine (0.1 mg/kg) both before and following either intrathecal injection of estrogen (0.5 microM; 1 microl) or bilateral injection of estrogen (0.5 microM; 100 nl/side) into several central autonomic nuclei. In estrogen-replaced rats, both the baseline and PE-evoked values for mean arterial pressure and RSNA were significantly decreased following injection of estrogen into the nucleus tractus solitarius (NTS), rostral ventrolateral medulla (RVLM), parabrachial nucleus (PBN), central nucleus of the amygdala (CNA) and the intrathecal space. Baseline heart rate and VPNA were significantly decreased following injection of estrogen into NTS, nucleus ambiguous (Amb), PBN and the intrathecal space. PE-evoked changes in heart rate and VPNA were significantly enhanced following injection of estrogen into these same nuclei. Injection of estrogen into the insular cortex (IC) produced significant decreases in baseline and PE-evoked RSNA only. The cardiac baroreflex was significantly enhanced following injection of estrogen into all nuclei and the intrathecal space. In saline-replaced females, injection of estrogen into NTS, RVLM, Amb and the intrathecal space had similar effects on both baseline and PE-evoked parameters although of a reduced magnitude compared to estrogen-replaced rats. However, no significant changes in autonomic tone and baroreflex function were observed following the injection of estrogen into the PBN, CNA or IC of saline-replaced rats. These results demonstrate a role for estrogen in central autonomic nuclei in female rats and suggest a possible alteration of estrogen receptor distribution or efficacy within the central nervous system of estrogen-deficient female rats.

17β-Estradiol modulates baroreflex sensitivity and autonomic tone of female rats

The following experiments examine the role of estrogen as a central modulator of autonomic tone and baroreflex sensitivity in the female rat. Female Sprague-Dawley rats were ovariectomized and then supplemented daily for 7 days with a fixed dose of estrogen (5 microg/kg; sc) to produce a stable level of estrogen similar to that present at proestrous (17 pg/ml). The rats were then anaesthetized with sodium thiobutabarbital (100 mg/kg) and instrumented to record blood pressure, heart rate and both vagal and renal efferent nerve activities. The sensitivity of the cardiac baroreflex was tested using intravenous injection of multiple doses of either phenylephrine hydrochloride or sodium nitroprusside. Estrogen-supplemented female rats exhibited a significantly enhanced BRS as compared to male rats from a previous study (0.78 vs. 0.5). Furthermore, bolus injection of estrogen (1x10(-2) mg/kg; iv) in estrogen-supplemented female rats produced a significant increase in vagal nerve activity and a significant decrease in renal nerve activity which together resulted in a further enhancement of the BRS (0.78 vs. 2.4). Injection of the selective estrogen receptor antagonist, ICI 182,780, into nucleus ambiguus and the intrathecal space of the spinal cord blocked the respective changes in parasympathetic and sympathetic nerve activities indicating that intravenously administered estrogen modulates baseline autonomic tone via the activation of central estrogen receptors.

Centrally mediated effect of 17β-estradiol on parasympathetic tone in male rats

The following experiments were conducted to determine if peripherally administered estrogen has an effect on central autonomic tone and whether this change in tone results in an alteration in cardiovascular reflex control. Male Sprague-Dawley rats were anesthetized with thiobutabarbitol sodium (50 mg/kg) and instrumented to record blood pressure, heart rate, and vagal parasympathetic or renal sympathetic efferent nerve activity. Additional rats were instrumented to test the sensitivity of the cardiac baroreflex using intravenous injections of phenylephrine hydrochloride (0.025, 0.05, 0.1 mg/kg) or sodium nitroprusside (0.0025, 0.005, 0.01 mg/kg) and plotting the cardiovascular responses. Intravenous injection of estrogen (10-4, 10-2, and 10-1 mg/kg) produced a significant increase in vagal efferent activity and in baroreflex sensitivity. The bilateral microinjection of an estrogen receptor antagonist, ICI-182,780 (1 pM, 50 nl/side) into the nucleus ambiguus blocked both the estrogen-induced increase in vagal efferent activity and baroreflex sensitivity. These results demonstrate that in male rats estrogen acts centrally to enhance baroreflex sensitivity by increasing parasympathetic efferent tone.

Acute injection of 17β-estradiol enhances cardiovascular reflexes and autonomic tone in ovariectomized female rats

Abstract Among the many benefits of long-term hormone replacement therapy to postmenopausal women is a significant reduction in risk for and progression of cardiovascular disease. However, long-term estrogen replacement therapy has been associated with several undesirable, and likely dose-dependent, side-effects. There is some evidence to suggest that the dose of estrogen which confers optimal beneficial effects on the cardiovascular system is much lower than that which is currently prescribed for postmenopausal women. The following experiments were conducted to determine the dose–response relationship of acutely administered estrogen on autonomic tone and reflex control of heart rate in ovariectomized Sprague–Dawley female rats. Rats were anaesthetized with sodium thiobutabarbital (100 mg/kg) and instrumented to record blood pressure, heart rate and efferent parasympathetic and sympathetic nerve activities. The sensitivity of the cardiac baroreflex was tested using intravenous injection of either phenylephrine hydrochloride (0.025–0.1 mg/kg) or sodium nitroprusside (0.0025–0.01 mg/kg). Intravenous injection of estrogen produced dose-dependent increases in the magnitude of the baroreflex sensitivity and parasympathetic tone while reducing sympathetic tone with a maximal effect observed at 1×10−3 mg/kg. Prior administration of the selective estrogen receptor antagonist, ICI 182,780 blocked the estrogen-induced changes in baroreflex sensitivity and autonomic tone. These results demonstrate that acutely administered, low-dose estrogen has beneficial effects on autonomic tone and cardiovascular reflexes.

Medullary and intrathecal injections of 17β-estradiol in male rats

The following experiments were designed to investigate the role of estrogen in central autonomic nuclei on autonomic tone and reflex control of heart rate. Male Sprague-Dawley rats were anesthetized with sodium thiobutabarbital (100 mg/kg) and instrumented to record blood pressure and heart rate. Efferent vagal and renal nerve activities were recorded and used to assess changes in parasympathetic and sympathetic tone, respectively. The cardiac baroreflex was evoked using a single bolus injection of phenylephrine (0.1 mg/kg) both before and following either intrathecal injection of estrogen (0.5 microM; 1 microl) to influence sympathetic preganglionic neurons of the intermediolateral cell column or bilateral injection of estrogen (0.5 microM; 100 nl/side) into the nucleus tractus solitarius, rostral ventrolateral medulla or nucleus ambiguus. The cardiac baroreflex was significantly enhanced following both intrathecal and medullary injections of estrogen. Efferent vagal nerve activity was significantly increased following injection of estrogen into the nucleus tractus solitarius, nucleus ambiguus and the intrathecal space. Renal sympathetic nerve activity was significantly depressed following injection of estrogen into the nucleus tractus solitarius, rostral ventrolateral medulla and the intrathecal space. In all cases, simultaneous injection of estrogen with the selective estrogen receptor antagonist, ICI 182,780 (1 pM) blocked all previously observed changes in baroreflex function and autonomic tone. These results demonstrate a role for estrogen in the reflex control of heart rate and as a central modulator of autonomic tone in male rats.

Lipoic acid protects against reperfusion injury in the early stages of cerebral ischemia.

Lipoic acid (LA) is a known antioxidant currently used as a therapy in patients with vascular and metabolic disorders. We tested the hypothesis that lipoic acid is protective against the cell death observed following stroke. Lipoic acid was administered 30minutes prior to, or immediately following removal of sutures used to occlude the middle cerebral artery (MCA) in male Sprague-Dawley rats. Following removal of the sutures, the MCA territory was allowed to undergo 5.5hrs of reperfusion. This ischemia/reperfusion (I/R) resulted in a focal infarct restricted to the prefrontal cortex (24±3mm(3)). Pretreatment with LA 30minutes prior to occlusion resulted in a dose-dependent reduction in infarct volume. This reduction in infarct volume was not observed when the LA was administered immediately prior to reperfusion (30minutes post-occlusion). To investigate a potential hemodynamic mechanism for this LA-induced neuroprotection, blood pressure, heart rate and baroreceptor reflex sensitivity (BRS) were measured. Intravenous administration of LA did not result in any significant changes in any of these parameters compared to saline-treated rats. Similarly, there was no significant contribution of systemic nitric oxide or alteration in cerebral perfusion measured following pretreatment with lipoic acid or during the course of occlusion and reperfusion compared with saline-treated rats. Western blot analysis of tissue from the ischemic cortex showed an increase in protein expression of superoxide dismutase (SOD2), but not SOD1, in LA pretreated rats. This suggests a potential mechanism of action contributing to the LA-induced neuroprotection observed. Furthermore, the data in the present investigation suggest the potential use of LA pretreatment as a neuroprotectant in stroke patients.

Estrogen-induced autonomic effects are mediated by NMDA and GABAA receptors in the parabrachial nucleus

The present study was done to determine if estrogen interacts with excitatory and/or inhibitory amino acid neurotransmitters to alter neuronal excitability within the parabrachial nucleus (PBN) and modulate autonomic tone. First, the role of estrogen in modulating autonomic tone was investigated in male rats anesthetized with Inactin (100 mg/kg). Animals were instrumented to record blood pressure, heart rate, vagal parasympathetic and renal sympathetic nerve activities as well as baroreflex sensitivity. Direct, bilateral injection of 17beta-estradiol (0.5 microM; 200 nl/side) into the PBN resulted in a significant decrease in blood pressure (17+/-4 mmHg), sympathetic tone (20+/-5%) and heart rate (22+/-5 beats/min) while increasing parasympathetic tone (34+/-4%) 30 min post-injection. These estrogen-induced effects were completely blocked by the co-injection of estrogen with the estrogen receptor antagonist, ICI 182,780 (20 microM; 200 nl/side). Co-injection of the NMDA receptor antagonist, (+/-)-3-(2-carboxypiperazine-4-yl) propyl-1-phosphonic acid (CPP; 10 microM; 200 nl/side), with estradiol resulted in complete blockade of the estrogen-induced decrease in heart rate and increase in parasympathetic tone only. Co-injection of estradiol with the GABA(A) receptor antagonist, (+)-bicuculline (0.1 microM; 200 nl/side), resulted in complete blockade of the estrogen-induced decrease in blood pressure and sympathetic nerve activity only. These results suggest that estrogen acts on estrogen receptors on neurons in the PBN to modulate GABA(A)-receptor mediated inhibitory neurotransmission to alter sympathetic tone and blood pressure and on neurons in a separate, parallel pathway to modulate NMDA-receptor mediated neurotransmission to alter parasympathetic tone and heart rate.

Role of 17β-estradiol in the modulation of baroreflex sensitivity in male rats

Female mammals have an enhanced baroreflex sensitivity compared with their male counterparts, leading researchers to speculate that estrogen modulates autonomic tone. Therefore, this study tests the hypothesis that exogenous estrogen can enhance the baroreflex sensitivity of male rats. Male Sprague-Dawley rats anesthetized with thiobutabarbitol sodium (50 mg/kg) were instrumented to measure blood pressure and heart rate and for the intravenous injection of drugs. The baroreflex was tested using intravenous injections of phenylephrine (0.025, 0.05, and 0.1 mg/kg), and the cardiovascular responses were plotted to obtain a measure of the sensitivity of the cardiac baroreflex. Intravenous injection of estrogen produced dose-related increases in the baroreflex sensitivity due to an increase in the magnitude of the reflex bradycardia. In a separate group of animals, stimulation of the vagus nerve for 2 h resulted in a decrease in baroreflex sensitivity. This effect was blocked when estrogen (1 × 10-2 mg/kg) was administered immediately before the end of stimulation. In conclusion, intravenous injection of estrogen in male rats significantly enhanced baroreflex sensitivity and blocked the attenuation in the baroreflex sensitivity observed after vagal stimulation.

ROLE OF OESTROGEN IN THE CENTRAL REGULATION OF AUTONOMIC FUNCTION

1 In recent years, the role of oestrogen in womens health has been a subject of considerable scientific and popular debate. There is unquestionable evidence that oestrogen has both potent and long‐lasting effects on several vital organ systems, including the cardiovascular system, the autonomic nervous system and, most recently, within the central nervous system itself. 2 The research and medical community continues to debate whether the benefits of oestrogen therapy outweigh the risks in the treatment of the symptoms of menopause, the attenuation of the risk for cardiovascular insults, such as stroke and heart disease, and even the retardation of the progression of Alzheimers disease. 3 The recent evidence provided by the Heart and Estrogen/Progestin Replacement Study (HERS) II clinical trial suggesting that long‐term exposure to combined oestrogen and progestin in post‐menopausal women who have previously had a heart attack or stroke (for secondary prevention) may actually increase their risk of a subsequent cardiovascular insult has further fuelled the debate. However, there remain considerable gaps in our knowledge with respect to the actual mechanisms by which oestrogen exerts its various beneficial effects at the cellular level for the primary prevention of cardiovascular disease. This information is essential if we are to harness the positive aspects of oestrogen therapy in such a manner as to avoid or minimize the associated risks of increased oestrogen exposure in women who we know, with some certainty, to be at an increased risk of cancers of the uterus, cervix and breast tissue.

Role of the insular cortex in the modulation of baroreflex sensitivity.

Cervical vagal stimulation for 2 h results in a depressed baroreflex sensitivity produced by an enhanced sympathetic output, as indicated by increased plasma norepinephrine levels. The current study examined the role of the insular cortex in modulating the vagal stimulation-induced changes in baroreflex sensitivity. Male Sprague-Dawley rats were anesthetized with thiobutabarbitol sodium and instrumented for recording blood pressure, heart rate, intravenous drug administration, and vagal afferent nerve stimulation. Stereotaxic microinjections (300 nl) of either 5% lidocaine or 0.9% saline were made bilaterally into the insula. Thirty minutes after 2 h of vagal stimulation, the baroreflex was significantly depressed and plasma norepinephrine levels were significantly elevated in both groups. The baroreflex was also significantly depressed after bilateral lidocaine injections into the insula, independent of vagal stimulation. However, no significant change in plasma norepinephrine was observed, suggesting that an attenuated parasympathetic output contributed to the altered baroreflex. Taken together, the results suggest that the insular cortex modulates the cardiac baroreflex through a modulation of parasympathetic output.