Salsolinol is a putative endogenous neuro-intermediate lobe prolactin-releasing factor.

The isolation and identification of a prolactin-releasing factor (PRF) from the neuro-intermediate lobe of the pituitary gland has been pursued for over a decade. Using high-pressure liquid chromatography with electrochemical detection (HPLC-ECD) and gas chromatography/mass spectrometry (GC/MS) (R)-salsolinol (SAL) (a dopamine-related stereo-specific tetrahydroisoquinoline) was found to be present in neuro-intermediate lobe as well as median eminence extracts of male, intact-, and ovariectomized female rats. Moreover, analysis of SAL concentrations in neuro-intermediate lobe revealed parallel increases with plasma prolactin in lactating rats exposed to a brief (10 min) suckling stimulus following 4-h separation. SAL appears to be a selective and potent stimulator of prolactin secretion in vivo and it was without effect on the secretion of other pituitary hormones. We have also found that SAL can elevate prolactin release, although to a lesser extent, in pituitary cell cultures as well as in hypophysectomized rats bearing anterior lobe transplants under the kidney capsule. Lack of interference of SAL with [3H]-spiperone binding to AP homogenates indicates that SAL does not act at the dopamine D2 receptor. Moreover, [3H]-SAL binds specifically to homogenate of AL as well as neuro-intermediate lobe obtained from lactating rats. Taken together, our data clearly suggest that SAL is synthesized in situ and this compound can play a role in the regulation of pituitary prolactin secretion.

Decreased expression of fos-related antigens (FRAs) in the hypothalamic dopaminergic neurons after immunoneutralization of endogenous prolactin.

In our previous studies we found that administration of exogenous prolactin increased dopamine turnover in the terminal areas of the hypothalamic dopaminergic neurons controlling prolactin secretion from pituitary lactotrophs. In this study we investigated the effect of immunoneutralization of endogenous prolactin on the expression of FRAs in the tuberoinfundibular dopaminergic (TIDA), tuberohypophysial dopaminergic (THDA), and periventricular hypothalamic dopaminergic (PHDA) subpopulations of the hypothalamic dopaminergic neurons. Female rats were ovariectomized on d 0 of the experiment. At 1000 h of d 10, all animals were injected with 20 microg of 17-beta-estradiol sc to induce a proestrous-like surge of prolactin at 1700 h the next day. At 1000 h on d 11, half of the animals were injected with 200 microL of rabbit anti-rat prolactin antiserum ip, while the controls received normal rabbit serum. Groups of animals were sacrificed for immunocytochemistry in 2 h intervals between 1300 and 2100 h. Double-label immunocytochemistry for FRAs and tyrosine hydroxylase (TH) was performed and the results are presented as percentage of TH-immunoreactive neurons expressing FRAs. In the control animals, expression of FRAs decreased at 1500 h, gradually increased by 1900 h, but was lower than the basal levels by 2100 h. Expression of FRAs was significantly lower at 1900 h in the PHDA, THDA and TIDA neurons of prolactin antiserum treated rats than in the controls. These results indicate that elimination of endogenous prolactin from the circulation lowers the activity and/or prevents the reactivation of neuroendocrine dopaminergic neurons at the beginning of the dark phase.

Ovarian steroids influence the activity of neuroendocrine dopaminergic neurons.

The secretion of prolactin (PRL) from the anterior lobe (AL) of the pituitary gland is tonically inhibited by dopamine (DA) of hypothalamic origin. While ovarian steroids play a role in the regulation of the secretion of PRL, their effect on all three populations of hypothalamic neuroendocrine dopaminergic neurons is not fully understood. In this study we describe the effects of ovarian steroids on regulation of the release of DA from tuberoinfundibular dopaminergic (TIDA), tuberohypophyseal dopaminergic (THDA) and periventricular-hypophyseal dopaminergic (PHDA) neurons. Adult female rats were bilaterally ovariectomized (OVX) and, 10 days following ovariectomy (day 0), injected with corn oil (vehicle), estrogen, or estrogen plus progesterone (day 1). Animals were sacrificed every 2 h from 09.00 to 21.00 h by rapid decapitation. Trunk blood was collected and the concentration of PRL in serum was determined by radioimmunoassay. The median eminence (ME) and the AL, intermediate (IL) and neural (NL) lobes of the pituitary gland were dissected and the concentration of DA and DOPAC in each was measured by HPLC-EC. OVX rats presented small but significant increases in the secretion of PRL at 15.00 and 17.00 h. Replacement of estrogen or estrogen plus progesterone increased the basal concentration of PRL. Moreover, injection of estrogen only, or estrogen plus progesterone increased the concentration of PRL in serum at 15.00 h through 19.00 h, respectively, followed by a decrease to baseline thereafter. The turnover of DA in the ME and NL of OVX rats increased at 13.00 and returned to low levels. Turnover of DA in the IL of OVX rats increased in the morning by 11.00 h and remained elevated before decreasing by 17.00 h. The turnover of DA in the ME, IL and NL of OVX rats increased by 19.00 h. Injection of estrogen advanced the increase of TIDA activity by 2 h in the ME compared to OVX rats. Moreover, administration of estrogen suppressed the activity of THDA and PHDA neurons in the afternoon compared to OVX rats. In estrogen plus progesterone-treated rats, the activity of hypothalamic neuroendocrine dopaminergic neurons terminating in the ME, IL, and NL was inhibited prior to the increase in the secretion of PRL. The concentration of DA in the AL diminished prior to the estrogen-induced increase of PRL. Administration of progesterone, in concert with estrogen, delayed the increase of PRL in serum and the decrease of DA in the AL, compared to estrogen-treated rats, by 4 h. These data suggest a major role for ovarian steroids in controlling increases in the secretion of PRL by not only stimulating PRL release from lactotrophs, but also by inhibiting the activity of all three populations of hypothalamic neuroendocrine DAergic neurons.

Immunoneutralization of prolactin prevents stimulatory feedback of prolactin on hypothalamic neuroendocrine dopaminergic neurons.

We have found that exogenous prolactin (PRL) stimulates all three populations of hypothalamic neuroendocrine dopaminergic neurons. In this study, we investigated the effects of immunoneutralization of endogenous PRL on the activity of these neurons. Injection of 17beta-estradiol (E2) (20 microg subcutaneously) 10 d after ovariectomy induced a proestrus-like increase in PRL in peripheral plasma the following afternoon. At 1000 h the day after E2 injection, rats received either rabbit antirat PRL antiserum (PRL-AS) (200 microL) or normal rabbit serum (NRS, 200 microL, controls) intraperitoneally. Groups of rats were then decapitated every 2 h from 1100 h to 2100 h. Trunk blood was collected and serum extracted with protein A to remove the PRL-AS/PRL complex, and the remaining free PRL was measured by radioimmunoassay. Sites of neuroendocrine dopaminergic nerve terminals, the median eminence (ME), and intermediate and neural lobes of the pituitary gland were excised and stored for determination of dopamine (DA) and 3,4-dihydroxyphenyl acetic acid (DOPAC) concentrations by high-performance liquid chromatography electrochemical detection (EC). In addition, the anterior lobe of the pituitary gland, the locus of DA action, was collected. The concentration of PRL in NRS-treated animals increased by 1500 h, peaked by 1700 h, and returned to low levels by 2100 h. PRL-AS prevented the increase in PRL secretion in response to E2. The turnover of DA (DOPAC:DA ratio; an index of dopaminergic neuronal activity) in the ME of NRS-treated animals increased at 1500 h and rapidly returned to basal levels. Treatment with PRL-AS prevented the increase in DA turnover in the ME. DA turnover in the intermediate lobe increased coincident with the peak of PRL in serum of NRS-treated rats. PRL-AS administration prevented increased DA turnover in the intermediate lobe. The turnover of DA in the neural lobe increased by 1300 h and decreased steadily through 2100 h. However, administration of PRL-AS minimally suppressed the turnover of DA in the neural lobe. Moreover, administration of PRL-AS attenuated the rise of DA in the anterior lobe associated with the waning phase of the E2-induced PRL surge. These results clearly indicate that endogenous PRL regulates its own secretion by activating hypothalamic neuroendocrine dopaminergic neurons.

Dopamine transporters participate in the physiological regulation of prolactin.

Three populations of hypothalamic neuroendocrine dopaminergic (NEDA) neurons, arising from the arcuate and periventricular nuclei of the hypothalamus release dopamine (DA) that acts at the pituitary gland to regulate the secretion of PRL. It is generally accepted that NEDA neurons lack functional DA transporters (DATs), which are responsible for uptake of DA from the synaptic cleft into the presynaptic axon terminal. This study localized DATs to the hypothalamo-pituitary axis and evaluated the effect of DAT blockade on the hypothalamo-pituitary regulation of PRL. After 7 days of treatment with cocaine (a nonspecific amine transporter blocker) or mazindol (a specific DAT blocker), the relative abundance of PRL messenger RNA (mRNA) in the anterior lobe (AL) of OVX rats was significantly decreased, whereas the relative abundance of tyrosine hydroxylase mRNA in the hypothalamus was significantly increased. The effect of cocaine or mazindol administration on DA turnover and serum PRL concentration was examined in estradiol (E2)-treated OVX rats. E2 administration (i.v.) resulted in a significant increase in serum PRL within 4 h; however, cocaine or mazindol administration abolished the E2-induced increase of PRL. Cocaine or mazindol significantly increased the concentration of DA at the site of the axon terminals within the median eminence (ME), intermediate lobe (IL) and neural lobe (NL), indicating blockade of uptake. Because formation of DOPAC requires uptake of DA, concentrations of DOPAC in the ME, IL and NL decreased following treatment with either cocaine or mazindol. These data, together with the presence of immunopositive DAT in the ME, pituitary stalk, IL, and NL, suggest that a functional DAT system is present within all three populations of NEDA neurons. Moreover, similarity between the effects of cocaine and mazindol treatment indicate that blockade of the DAT, but not other amine transporters, is responsible for suppression of PRL gene expression and secretion. Blockade of DATs prevent uptake of DA into NEDA neurons and consequently increases the amount of DA that diffuses into the portal vasculature and reaches the AL. These data provide evidence that DATs play a physiological role in the regulation of DA release from and TH expression in NEDA neurons and consequently PRL secretion and PRL gene expression and further support our previous observation that the regulation of PRL secretion involves all three populations of NEDA neurons.

Prolactin activates all three populations of hypothalamic neuroendocrine dopaminergic neurons in ovariectomized rats.

Prior studies suggest that prolactin (PRL) stimulates release of dopamine (DA) from tuberoinfundibular dopaminergic (TIDA) neurons. In the present study, the time course over which PRL exerts its effects on all three populations of neuroendocrine dopaminergic (DAergic) neuron populations [TIDA, tuberohypophyseal (THDA) and periventricular-hypophyseal (PHDA)] was determined. Ten days following ovariectomy (OVX), groups of female rats were injected either with 15 microg of ovine PRL (oPRL) or saline at 0900 h. Rats were decapitated every 30 min from 0830 h-1100 h and hourly from 1200 h-1500 h. Trunk blood was assayed for rat PRL (rPRL) and oPRL using species-specific radioimmunoassays (RIAs). The concentration of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME), as well as the anterior (AL), intermediate (IL) and neural (NL) lobes of the pituitary gland were determined by HPLC-EC. The concentration of rPRL in oPRL-treated animals, compared to saline-treated animals, was diminished by 1000 h and again between 1200 h-1500 h. DOPAC/DA ratio, an indicator of dopaminergic neuronal activity, increased spontaneously in the ME, IL, and NL during the afternoon in OVX rats. In animals injected with oPRL at 0900 h, the DOPAC/DA ratio increased in the ME, IL and NL within 1 h. Moreover, a secondary increase in the DOPAC/DA ratio in the IL and NL occurred during the afternoon in oPRL-treated rats. However, the second increase of DA turnover present in the ME of control animals never occurred in oPRL-treated animals. Furthermore, there were two increases in the concentration of DA in the AL: the first coincided with the increased turnover of DA in all three terminal areas and the second with increased DA turnover in the IL and NL. These data suggest that all three populations of hypothalamic neuroendocrine DAergic neurons are activated by PRL and that PHDA/THDA neurons have a second 'delayed' activation.

Increased NPY activity in the PVN contributes to food-restriction induced reductions in blood pressure in aortic coarctation hypertensive rats.

We hypothesized that hypothalamic NPYergic mechanisms mediate the blood pressure lowering effect of caloric restriction in hypertensive rats. Aortic coarctation-induced (AC) hypertensive rats (n=25) were assigned to either an ad libitum fed control group (AL) or food restricted group (FR; 60% of AL consumption) for 3 weeks. Rats were instrumented chronically with vascular catheters and bilateral guide cannulae directed at the paraventricular hypothalamic nuclei (PVN). Blood pressure (BP) and heart rate (HR) responses to bilateral PVN microinjection of saline (200 nl) or the putative NPY receptor antagonists [D-Trp32]NPY(1-36) (3.3 micrograms/200 nl) and [D-Tyr27,36 Thr32]NPY(27-36) (D-NPY(27-36); 3.3 micrograms/200 nl) were determined. The FR rats were then refed and cardiovascular responses to PVN injections of NPY receptor antagonists were again determined. FR rats had significantly reduced resting BP (159+/-4 vs. 129+/-4 mmHg) and HR (360+/-11 vs. 326+/-9 bpm) compared to AL controls. Refeeding restored BP and HR of FR rats to levels similar to AL (BP=153+/-4 mmHg, HR=359+/-11 bpm). PVN administration of [D-Trp32]NPY produced foraging behavior and concurrent increases in BP and HR in FR, AL and Re-fed rats. The behavioral activation suggests that [D-Trp32]NPY(1-36) produced activation of NPY receptors. In contrast, D-NPY (27-36) did not produce any behavioral response or affect BP or HR in AL or Re-fed rats. In FR rats, D-NPY (27-36) produced significant increases in BP (peak=15+/-3 mmHg) which partially reversed the effect of FR on BP. Thus, in FR rats with reduced BP, PVN administration of an NPY receptor antagonist increases BP. NPY blockade in the PVN accounted for about 50% of the BP effect of food restriction, thus other mechanisms are likely to be involved. These findings are consistent with the hypothesis that NPYergic mechanisms may contribute to the reduction of BP produced by food restriction.

The effect of neurointermediate lobe denervation on hypothalamic neuroendocrine dopaminergic neurons.

The contribution of tuberohypophyseal and periventricular-hypophyseal dopaminergic neurons to the regulation of the secretion of prolactin (PRL) has yet to be clarified. In this study, we used pituitary stalk compression to disrupt hypothalamic neural input to the neurointermediate lobe (NIL). Neurointermediate lobe denervation (NIL-D) selectively disrupts the axons of tuberohypophyseal and periventricular-hypophyseal dopaminergic neurons, while leaving tuberoinfundibular dopaminergic neurons and the vascular supply of the pituitary gland intact. NIL-D was performed in ovariectomized (OVX) rats. The concentration of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME) and various regions of the pituitary gland of OVX and OVX+NIL-D rats were measured by HPLC-EC. The concentration of PRL, alpha-melanocyte stimulating hormone (alpha-MSH), and luteinizing hormone (LH) in serum were determined by radioimmunoassay. Successful NIL-D was confirmed by increased water intake. One week after NIL-D, serum PRL and alpha-MSH were elevated, but there was no change in the concentration of LH in serum. The concentration of DA was increased in the median eminence (ME), decreased in the outer zone of the anterior lobe (AL-OZ), as well as the intermediate (IL) and neural lobes (NL), and remained unchanged in the inner zone of the anterior lobe (AL-IZ). The concentration of DOPAC was increased in the ME and NL, decreased in the IL, and remained unchanged in both the AL-IZ and AL-OZ. These data confirm that pituitary stalk compression denervates the NIL. Moreover, decreases in the concentration of DA in the IL and AL-OZ, coupled with elevation of serum PRL and alpha-MSH indicate that DA from the NIL contributes to the increased inhibition of the secretion of PRL and alpha-MSH in OVX rats.

Characterization of the dopaminergic input to the pituitary gland throughout the estrous cycle of the rat.

Changes in the concentrations of dopamine (DA) and its major metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), were characterized in the pituitary gland throughout the 4-day estrous cycle of the rat. Female rats were sacrificed at 2- to 3-hour intervals throughout each day of the 4-day estrous cycle. Pituitary glands were removed, and the concentrations of DA and DOPAC were determined by high-performance liquid chromatography coupled to electrochemical detection. The concentration of prolactin (PRL) in serum from these same animals was determined by radioimmunoassay. The concentration of DA in the anterior lobe was constant throughout most of the 4-day estrous cycle. Prior to initiation of the proestrous surge of PRL, there were significant (p < 0.05) decreases in the concentrations of both DA and DOPAC in the anterior lobe which returned to elevated baseline levels just prior to the termination of the proestrous surge of PRL. The concentrations of DA and DOPAC in the intermediate lobe exhibited a daily rhythm. However, in the intermediate as well as in the anterior lobe, there were significant (p < 0.001) decreases in the concentrations of both DA and DOPAC, coincident with the initiation of the proestrous surge of PRL. Similarly, coincident with the peak of the proestrous surge of PRL, there were significant (p < 0.001) increases in the concentrations of DA and DOPAC in the intermediate lobe, followed by a return to basal levels and resumption of the daily rhythm. The pattern of the concentrations of DA and DOPAC in the neural lobe was also daily in nature, with peaks occurring between 13.00 and 15.00 h each day of the 4-day estrous cycle. These data, taken together: (1) confirm that a decrease of the concentrations of DA and DOPAC occurs in the anterior lobe prior to the proestrous surge of PRL; (2) reveal that DA is released in a daily pattern at intermediate and neural lobes, and (3) suggest an apparent role for DA released to the intermediate lobe in the regulation of the proestrous surge of PRL.

Food restriction attenuates the blood pressure response to paraventricular hypothalamic nuclei lesions in aortic coarctation hypertension.

Chronic food restriction reduces blood pressure (BP) and sympathetic support of BP in aortic coarctation hypertension. The purpose of this study was to test the hypothesis that chronic food restriction would reduce sympathetic support of BP mediated by the paraventricular hypothalamic nuclei (PVN). Hypertension was induced in male Sprague-Dawley rats (n=40) by suprarenal aortic coarctation. Rats were assigned to either an ad libitum fed (AL) group or a food restricted (FR) group that received 60% of the food consumed by AL for 3 weeks. One week prior to data collection, catheters were implanted in the left carotid artery and right jugular vein. BP was measured for 2 days prior to, and 7 days after rats in AL and FR groups received either bilateral electrolytic lesions of the PVN (PVNx) or sham lesions (SHAM). Prior to either PVNx or SHAM, FR rats had significantly lower BP (AL=152+/-5; FR=113+/-2 mmHg), less of a depressor response to ganglionic blockade (AL=-58+/-4; FR=-35+/-2 mmHg), and lower plasma norepinephrine levels (AL=758+/-71; FR=380+/-23 pg/ml) compared to AL. PVNx reduced BP in both AL and FR rats (AL-PVNx=105+/-6 mmHg, FR-PVNx=101+/-3 mmHg). PVNx also lowered the depressor response to ganglionic blockade (AL-PVNx=-28+/-5 mmHg, FR-PVNx=-29+/-4 mmHg) and plasma norepinephrine levels (AL-PVNx=372+/-74 pg/ml, FR-PVNx=248+/-31 pg/ml). FR decreased the magnitude of the reductions in resting BP and in sympathetic activity in response to PVNx. These results indicate that intact PVN are required for maintenance of aortic coarctation hypertension, and implicate the PVN as a site involved in BP reductions produced by chronic food restriction.

Changes in the local metabolism of dopamine in the anterior and neural lobes but not in the intermediate lobe of the pituitary gland during nursing.

Dopamine (DA), produced by tubero-infundibular dopaminergic (TIDA) neurons of the arcuate nucleus (ARN) is the established inhibitor of the secretion of prolactin (PRL). Changes in dopaminergic (DAergic) neuronal activity in the median eminence-long portal vessels (ME-LPV) and/or the concentration of DA in the anterior lobe (AL) are inversely related to the secretion of PRL. However, conflicting reports concerning DAergic neuronal activity during the suckling-induced release of PRL persist. In addition to TIDA neurons, PeVN-hypophysial DAergic (PHDA) and tubero-hypophysial DAergic (THDA) neurons which, respectively, innervate the intermediate lobe (IL) and the IL/neural lobe (NL) also have a significant role. We measured the concentrations of DA and its main metabolite, 3, 4-dihydroxyphenylacetic acid (DOPAC), in the median eminence and the three pituitary lobes of lactating mothers. Concentrations of DA and DOPAC from tissues and the concentration of PRL in plasma were measured by HPLC-EC and RIA, respectively. There were no changes in the concentration of DA and DOPAC of the IL due to the suckling stimulus. In the NL, a decrease in the concentration of DOPAC was detected due to the suckling stimulus. In addition, there were no changes of DA or DOPAC in the outer zone of the AL (AL-OZ) due to suckling. However, a decrease in the concentrations of DA and DOPAC was detected in the inner zone of the AL (AL-IZ). These data suggest lactotrophs from the AL-IZ are responsible for the changes in the concentration of plasma PRL in response to the suckling stimulus. In addition TIDA and THDA neurons, but not PHDA neurons, regulate the control of the secretion of PRL in response to suckling.

Abdominal distension in Kaufman-McKusick syndrome.

A female infant with Kaufman-McKusick syndrome redeveloped respiratory distress and abdominal distention at 5 weeks of age. Ultrasonography demonstrated recurrence of peritoneal cysts and hydrometrocolpos. It is postulated that refluxing vaginal secretions may contribute to the abdominal distention seen in many infants with Kaufman-McKusick syndrome.

Carcinoma of the prostate.

Prostatic carcinoma is the third most common cause of death from cancer among males. Selection of appropriate therapy and evaluation of results is often difficult, since patients present at different stages of the disease. Methods of staging, diagnosis, treatment of localized tumor, radiation, surgery and treatment of metastases are described.

Renal function in continent patients after surgical closure of bladder exstrophy.

We assessed the upper urinary tracts and renal function in 22 children who had achieved continence after staged correction of bladder exstrophy. All patients had been followed for an average of 8 years after completion of the reconstructive operation. In 15 patients the upper urinary tract was normal and only 1 of the remaining 7 required surgical correction (transureteroureterostomy) of the residual anomaly. Renal function was normal in all children. We attribute these satisfactory results to careful selection of patients, improved surgical techniques and good postoperative care.

The first 10 years of the dialysis-transplantation program at The Hospital for Sick Children, Toronto. 1: Predialysis and dialysis.

Renal dialysis and transplantation have been used for many years for adults with kidney failure but only recently for children. In May 1967 a renal-dialysis-transplantation program was established at The Hospital for Sick Children, Toronto for patients aged 6 to 18 years living within 240 km of Toronto. In 1973, children aged 1 to 5 years began to be accepted into the program, and by August 1977, 90 children (mean age 11 years) from all parts of Canada had been admitted to the program. The creation of vascular access in very small patients is difficult; the most successful types of access have been central shunts (established above the knee or the elbow) and bovine grafts. Specially made dialysis equipment is necessary for young patients. Young children should only be accepted in a dialysis-transplantation program that has a medical staff expert in meeting the specific needs of such children.

The first 10 years of the dialysis-transplantation program at The Hospital for Sick Children, Toronto. 2. Transplantation and complications of chronic renal failure.

Between January 1969 and August 1977, 78 children received 100 kidney transplants (94 from cadavers and 6 from living donors) at The Hospital for Sick Children, Toronto. Since 1971 the average wait for a first cadaveric transplant has been less than 5 months. Preferably the kidney is placed in a location that has not previously undergone an operation, usually the iliac fossa on the side opposite that from which the donor kidney was taken. Immunosuppressive therapy begins with prednisone (or methylprednisolone), 3 mg/kg body weight per day; the dose is gradually decreased until a maintenance dose of 10 to 20 mg every 48 hours is reached 3 to 6 months postoperatively. Azathioprine, 2 to 3 mg/kg body weight, is also given each day. Early recognition or prevention of renal osteodystrophy, the toxic effects of steroids, psychosocial problems, growth retardation and hypertension minimize their effects on these patients.

Renal transplantation in patients with bilateral Wilm's tumor.

Review of cases and of published reports of patients with bilateral Wilms' tumor revealed a significantly higher mortality rate in those who had received a renal transplant than in those who had not. The increased mortality is attributed to overwhelming sepsis resulting from chemotherapy, radiotherapy and immunosuppression. Growth of the tumor did not appear to be accelerated by immunosuppression and transplantation.