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Adenovirus-mediated gene transfer in the ovine pituitary gland is associated with hypophysitis.

Gene therapy for pituitary disease requires evaluation for safety as well as efficacy. We have reported results of adenovirus-mediated gene transfer using the sheep as a large animal model that allows longitudinal evaluation of hormone secretion and have confirmed high levels of transgene expression up to 7 days after direct stereotaxic injection into the pituitary gland. Here we report the results of detailed histological examination of the pituitary glands from animals injected with two recombinant adenoviruses expressing the beta-galactosidase marker gene, or with saline vehicle to control for the potential tissue-disruptive effect of the injection volume itself. Pituitaries injected with saline showed no evidence of inflammatory response apart from occasional minor foci of apoptosis. In all other respects they were indistinguishable from normal uninjected control pituitary glands. Glands injected with recombinant adenoviruses containing either the hCMV-beta-gal or the hPRL-beta-gal transgene, on the other hand, displayed variable degrees of inflammatory response, with periglandular fibrosis, lymphocytic infiltrate and venulitis in almost all cases. Focal necrosis and/or apoptosis was noted in six of nine cases. In summary, we have found evidence of severe inflammatory reaction within the first seven days of adenovirus injection, amounting to significant hypophysitis. The histological extent of this reaction has not previously been recognised by studies of the efficacy of gene transfer in rodents, and was underestimated by immunocytochemical studies of hormone and transgene expression. The findings emphasise the need for careful evaluation of the safety of endocrine gene therapy, and for caution with the dose of vector used.

Adenoviridae↗

Dynamic computed tomographic evaluation of the pituitary gland in healthy dogs.

OBJECTIVE: To determine the contrast enhancement pattern of the pituitary gland in healthy dogs via dynamic computed tomography (CT). ANIMALS: 17 dogs. PROCEDURE: With each dog in sternal recumbency, transverse CT scans were made perpendicular to the skull base from the rostral clinoid processes to the dorsum sellae. At the position of the image that contained the largest cross section of the pituitary gland, a series of 9 to 11 scans was made during and after IV injection of contrast medium (dynamic CT scans). The contrast enhancement pattern of the pituitary gland and surrounding arteries was assessed visually and by use of time-density curves. RESULTS: After strong enhancement of the maxillary arteries, the intracavernous parts of the internal carotid arteries, and the communicating arteries of the arterial cerebral circle, there was a strong enhancement of the central part of the pituitary gland followed by enhancement of its peripheral part. On the last images of the dynamic series of the pituitary gland, the central part was hypodense, compared with the peripheral part. Time-density curves confirmed an early, strong enhancement of the central part and a delayed, less strong enhancement of the peripheral part of the gland. CONCLUSIONS AND CLINICAL RELEVANCE: The difference in enhancement between the central and peripheral parts of the pituitary gland was attributable to a difference in vascularization of the neurohypophysis and adenohypophysis, respectively. Distortion or disappearance of the strong central enhancement (pituitary flush) may be used for the detection and localization of pituitary abnormalities in the adenohypophysis.

Animals↗

In vitro induction of luteinizing hormone synthesis by estrogen in organ-cultured pituitary glands of the Japanese eel, Anguilla japonica.

An organ culture method for pituitary glands isolated from immature Japanese eels (Anguilla japonica) was developed. This method could conserve the histological features of the pituitary glands for at least 21 days. The ability to synthesize gonadotropic hormone (GTH) in cultured eel pituitary glands was examined by detecting luteinizing hormone (LH) beta protein immunohistochemically. In a basal medium (Leibovitz L-15), LH beta-immunoreactive cells were very scarce, but after addition of estradiol-17beta (E2) a large number of immunoreactive cells appeared, particularly in the proximal pars distalis. The stimulatory effects of E2 on LH beta synthesis were dose (1-100 ng/ml)- and time (1.5-7 days)-dependent. Thus, in contrast with previous reports of the lack of a direct effect of E2 on GTH synthesis in primary cultured eel pituitary cells, the present results clearly indicate that E2 can stimulate GTH synthesis in immature eel pituitary glands. This organ culture method is useful to examine the actions of steroids and also other endocrine factors on the eel pituitary gland.

Anguilla↗

Evidence of existence of two isozymes of carbonic anhydrase in the anterior pituitary gland of female rats.

Two molecular forms of carbonic anhydrase were identified in the rat anterior pituitary gland. Electrophoretic and immunological studies disclosed that the low activity carbonic anhydrase (type B) was associated with the particulate fraction of the pituitary gland and the high activity carbonic anhydrase (type C) was localized in the soluble fraction. The 800 times g pellet contained 50% of the total activity of a pituitary homogenate prepared in 0.25 M sucrose, while the 105,000 times g supernatant accounted for an additional 30% of the total. The particulate enzyme was rendered soluble by treatment with Trition X-100. The activity of pituitary gland carbonic anhydrase was about 6 times greater in preparations from female rats as compared with males. The activity of carbonic anhydrase in the particulate fraction decreased, and the activity in the soluble fraction increased without a change in total activity following incubation in vitro of pituitary glands. The intracellular adjustment in enzyme activity was accompanied by the secretion of prolactin into the incubation medium by the pituitary gland. The secretion of prolactin and the solubilization of the particulate carbonic anhydrase was inhibited when the pituitary gland was incubated in the presence of 10-5 M dopamine.

Animals↗

Effects of hypophysectomy, pituitary gland homogenates and transplants, and prolactin on photoreceptor destruction.

Prepubertal removal of the pituitary gland, which in young animals influences sexual maturation, reduces significantly the amount of retinal photoreceptor destruction when the rats are exposed to continuous illumination in adulthood. When crude pituitary gland homogenate is administered to adult rats hypophysectomized prior to puberty, photoreceptor destruction is more severe. Transplantation of whole pituitary glands to the kidney capsule of hypophysectomized rats also reduces the effect of pituitary gland removal and results in more extensive damage to receptor cells than found in hypophysectomized, adult animals. Hypophysectomized rats treated with prolactin had more severe retinal damage than untreated, hypophysectomized rats. The injection of pregnant mare serum and human chorionic gonadotropic hormones into hypophysectomized rats was not effective in reversing the protection afforded by hypophysectomy. Results of these studies indicate the hormones of the pituitary gland have a regulatory influence on the severity of light-induced, retinal photoreceptor damage in the rat.

Animals↗

Formation of a direct arterial blood supply to the anterior pituitary gland following complete or partial interruption of the hypophyseal portal vessels.

If regions of the anterior pituitary gland received systemic blood via a direct arterial blood supply these regions would escape hypothalamic regulation and thus be a sequela in endocrine disorders. Since, in the untreated rat, all of the blood supply to the anterior pituitary gland is via the hypophyseal portal vessels, we hypothesized that partial interruption of the portal vessels could provoke the establishment of a direct arterial blood supply (arteriogenesis). We utilized the injection of polystyrene microspheres (15 or 9 micron diameter) into the left ventricle of the heart to test this hypothesis. Microspheres are trapped in the first capillary plexus they reach since they are too large to traverse the capillaries. No microspheres reached the anterior pituitary gland of control rats, a finding consistent with the fact that the anterior pituitary gland receives all of its blood supply via the hypophyseal portal blood vessels. Microspheres were observed in the primary portal capillary plexus in the infundibulum (median eminence), infundibular stalk (pituitary stalk), and infundibular process (pars nervosa), the first capillary plexus which they reached. A lesion of the medial basal hypothalamus (MBH) which destroyed the long portal vessels did not result in arteriogenesis since few, if any, microspheres were observed in the anterior pituitary gland. We confirmed, using vascular casts, that these lesions resulted in the long-term destruction of the primary portal capillaries in the infundibulum and of the long portal vessels. In MBH-lesioned animals it appears that all of the blood supply of the anterior pituitary gland is via short portal vessels arising from the infundibular stem and process.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Abnormal perfusion of the pituitary gland secondary to dural arteriovenous fistulas in the cavernous sinus: dynamic MR findings.

BACKGROUND AND PURPOSE: If venous congestion is the primary cause of pituitary gland enlargement in cases of dural arteriovenous fistulas (AVFs), other abnormal pituitary findings may be detectable on MR images. We sought to investigate the perfusion abnormality of the pituitary gland secondary to dural AVFs in the cavernous sinus and to clarify its clinical importance. METHODS: Nine consecutive patients (all female; age range, 50-77 years) with dural AVFs in the cavernous sinus underwent prospective MR examinations, including dynamic studies, before and after therapy. Their clinical signs and symptoms were recorded. Two radiologists visually evaluated the enhancement patterns of the anterior pituitary gland. Dynamic MR curves were obtained by locating regions of interest at the center and bilateral peripheral areas of the anterior pituitary gland on coronal images. MR images obtained in five healthy individuals served as controls. RESULTS: No patient had symptoms of hypopituitarism or other endocrine abnormalities. Asymmetric pituitary enhancement was found in five patients; the side with the dural AVF was less enhancing. This finding disappeared after therapy. Although asymmetric enhancement was not detected in the remaining four patients, statistical analysis showed significantly delayed enhancement of the pituitary gland in the patients compared with enhancement patterns in control subjects. After treatment, this delay improved significantly. The pituitary gland significantly decreased in size after treatment. CONCLUSION: Perfusion of the pituitary gland is impaired in patients with a dural AVF in the cavernous sinus. This finding is probably due to venous congestion of the pituitary gland caused by high pressure in the cavernous sinus; it is usually not related to pituitary dysfunction.

Adult↗

MR imaging of the pituitary gland in infants and children: changes in size, shape, and MR signal with growth and development.

It has been reported that on MR images of the brain in neonates the entire pituitary gland is hyperintense. This is quite different from the well-established observation that portion is isointense. To better understand the development of the neonatal pituitary gland, we studied the timing and magnitude of changes in the size, shape, and MR signal of the pituitary gland in early childhood and infancy. The study included 76 infants and children ages 3 days to 4 years old. In each MR study, the signal intensity of the pituitary gland on T1-weighted coronal (n = 24) and sagittal (n = 76) images was compared with the signal intensity of the pons by using region-of-interest methods. The size and shape of the pituitary glands were recorded. Statistical comparisons were made for signal intensity and size of the pituitary gland between boys and girls and among age groups. The results showed that the MR signal of the posterior lobe remained hyperintense relative to the pons for all age groups represented in our study (through 4 years). However, the signal intensity of the anterior lobe gradually diminished and by approximately age 2 months was isointense with the pons. Thus, by approximately age 2 months the MR signal intensity of the infant's pituitary gland has changed from being entirely hyperintense (as seen in neonates) to isointense in the anterior lobe and hyperintense in the posterior lobe (as seen in adults). In the first 2 months of life the pituitary gland is bulbous in shape; later it has a flatter upper surface. We found no significant changes in the length or signal intensity of the pituitary gland; no differences among sexes in size, shape, or signal intensity were demonstrated. Findings of a bulbous pituitary gland and high MR signal of the entire pituitary are normal in neonates up to 2 months of age. After that, the gland gradually flattens and the MR signal of the anterior lobe decreases until it has characteristics similar to the adult pituitary. Lack of high signal or discovery of a small pituitary gland in a neonate should alert the radiologist to the possibility of pituitary malformation or dysfunction.

Child, Preschool↗

Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland.

Distribution and development of growth hormone secretagogue receptor (GHS-R) mRNA expression in rat brain and pituitary gland were examined using ribonuclease protection assay. In adult male rats, GHS-R mRNA levels were highest in the pituitary gland, whereas those in the hypothalamus and hippocampus were 57 and 30% of those in the pituitary gland, respectively. Less abundant but detectable levels of GHS-R mRNA were found in the midbrain, pons, and medulla oblongata, but expression was barely detectable in the cerebellum and cerebral cortex. The expression of GHS-R mRNA was detected at late gestation (embryonic day 19) in the pituitary gland, hypothalamus, and brainstem. The mRNA levels increased with age in the pituitary gland, and decreased postnatally in the brainstem, while they remained constant in the hypothalamus during development. In contrast, GHS-R mRNA was not detectable in the hippocampus during the fetal period, but was first detected on postnatal day 7. Expression of GHS-R mRNA was also examined in the spontaneous dwarf rat (SDR), a model for isolated GH deficiency, to examine alterations in GHS-R mRNA expression in a GH-deficient state. GHS-R mRNA levels in the pituitary gland of SDRs were higher than those of control rats, suggesting negative regulation of GHS-R mRNA by GH in this region. GHS-R mRNA levels increased in the hypothalamus of female, but not in male SDRs. In contrast, GHS-R mRNA levels were not affected by GH in the brainstem and hippocampus. These results indicate that region-specific, developmentally regulated expression of GHS-R mRNA may reflect divergent physiological roles of GHS/GHS-R in distinct regions of the central nervous system and the pituitary gland.

Age Factors↗

The expression of pituitary gland genes in laying geese.

The purpose of this study was to detect differential expression of genes in the pituitary gland in laying geese by suppression subtractive hybridization (SSH). Pituitary glands from prelaying and laying geese were dissected for mRNA extraction. The cDNA from pituitary glands of prelaying geese was subtracted from the cDNA from the pituitary glands of laying geese (forward subtraction); the reverse subtraction was also performed. We screened 384 clones with possible differentially expressed gene fragments by differential screening. Sixty-five clones from the differential screening results were subjected to gene sequencing and further analysis. We found that at least 19 genes were highly expressed in the pituitary glands of laying geese compared with prelaying geese. Among these, 6 genes (including 4 novel genes) were confirmed by virtual Northern analysis. We found that prolactin and visinin-like protein were highly expressed in the pituitary glands of laying geese compared with prelaying geese (P < 0.05). Further investigation is needed to demonstrate specific functions of the novel genes discovered in the current study.

Animals↗

Dislocation of the pituitary gland by sphenocavernous meningiomas.

BACKGROUND: Dislocation of the pituitary gland from the sella turcia by tumor has only very rarely been described. To date, only two cases have appeared in the neurosurgical literature. METHODS: After intraoperative identification of pituitary dislocation in five patients with cavernous sinus meningiomas, a review of our most recent 20 patients with cavernous sinus meningiomas was performed. A subgroup of sphenocavernous meningiomas was identified, as were the associations between pituitary dislocation and the patient's endocrinologic status. RESULTS: Of 20 consecutive patients with cavernous sinus meningiomas, seven tumors could be subclassified as sphenocavernous meningiomas due to the significant amount of tumor extension into the sphenoid sinus. At the time of surgery, dislocation of the pituitary gland from the sella into the suprasellar cistern was observed in five of seven of these patients. Four of the five patients with pituitary dislocation had evidence of hypopituitarism, whereas there was no evidence of this condition in the two patients with sphenocavernous meningiomas without pituitary dislocation. CONCLUSIONS: Pituitary dislocation in these patients is likely due to the physical dislocation of the pituitary gland from the sella, through an incompetent diaphragma sella, and by the tumorous extension in the sphenoid sinus. The extension of a cavernous sinus meningioma into the sphenoid sinus, and specifically the sella turcica, may be via the intercavernous venous sinuses. The lack of endocrinologic improvement suggests that the hypopituitarism noted in these patients is likely due to disturbance of the pituitary microcirculation with subsequent pituitary ischemia. The possibility of pituitary dislocation in patients with sphenocavernous meningiomas, especially those with panhypopituitarism, should be considered and searched for at the time of surgery.

Adult↗

Effects of monoaminergic and cholinergic drugs on release of follicle stimulating hormone from co-incubated pituitary glands and hypothalami.

1. Male rat pituitary glands and hypothalami were incubated with agonists and antagonists of various neurotransmitters, and release of follicle stimulating hormone (FSH) measured. 2. Noradrenaline and cholinergic agonists stimulated release of FSH, responses blocked by the alpha antagonist, phentolamine, and by atropine respectively. 3. Dopamine did not affect basal, but inhibited noradrenaline-stimulated, release of FSH. The dopamine antagonist, pimozide, stimulated release of FSH. 4. Neither serotonin nor its antagonist, methysergide, affected release of FSH. 5. None of these drugs acted directly on the pituitary gland, suggesting that they act by altering secretion of the gonadotrophin releasing hormone.

Animals↗

Suckling-induced change in oxytocin but not in alpha-MSH concentrations of the median eminence, the neural-, intermediate- and anterior lobes of the pituitary gland.

Previous reports have implicated that pituitary-derived prolactin (PRL) is secreted from two distinct zones of mammotropes within the anterior lobe (AL). The inner zone (AL-IZ), located adjacent to the neuro-intermediate lobe (NIL), is supposed to be involved in the rapid and massive discharge of PRL from the pituitary gland due to suckling stimulus. Whereas the outer-zone (AL-OZ) gives the basal secretion and it does not play a role in the acute secretory response during nursing. Anatomically, the AL-IZ has an intimate contact with the NIL because the blood passing through the short portal vessels (SPV) bathes it first. Based on this fact it would be hypothesized that locally released and/or produced compounds, like OXY and alpha-MSH, can be delivered to the AL-IZ. In conjunction, OXY and alpha-MSH have already been implicated to play a role in the regulation of PRL release during suckling. Therefore, the purpose of this study was to examine the possible local transportation of these hormones into the median eminence and various regions of the pituitary gland of lactating rats. We have measured the concentrations of OXY and alpha-MSH from tissue samples of nonsuckled (NS) and 10 or 30 min after suckling (S) was initiated using specific RIAs. It has been shown that there are no changes in the concentration of OXY and alpha-MSH in theAL-IZ and AL-OZ due to suckling stimulus. In contrast, our data provide compelling evidence that OXY is transported into the IL, which can be further increased by suckling stimulus. These data suggest that blood transfusing NL passes through the IL before it is drained into the cavernous sinus, which opens the road for OXY into the general circulation. In addition, our data have unequivocally shown a lack of local delivery of either alpha MSH or OXY into the AL that raises serious doubt about their possible role in PRL secretion during suckling stimulus.

Animals↗

Partial transcriptome of the developing pituitary gland.

The anterior lobe of the pituitary gland is composed of five hormone-producing cell types and develops from Rathke's pouch, an invagination of oral ectoderm. In mice, rapid cell proliferation occurs in the pouch from embryonic day 12.5 (e12.5) to e14.5, preceding the appearance of most hormone transcripts. Cell-type-specific commitment probably occurs prior to e14.5, but cell differentiation can be demonstrated only by detection of hormone transcripts. Although several transcription factors critical for pouch expansion are known, few of their target genes have been identified. To identify putative transcription factor target genes and cell-type-specific markers, we used differential display PCR analysis of RNA prepared from e12.5 and e14.5 Rathke's pouches. We present an expression profile of the developing pituitary gland including 83 transcripts, 40% of which are novel. The tissue distribution, cell specificity, and developmental regulation were determined for a subset of the transcripts.

Animals↗

Release and metabolism of [3H]dopamine in the neurointermediate lobe of the rat pituitary gland.

Neurointermediate lobes of the rat pituitary gland were incubated with [3H]dopamine in the presence of desipramine and then superfused with radioactivity-free medium. The outflow of tritium was studied and in most experiments [3H]dopamine and its metabolites were separated by column chromatography. After 60-70 min of superfusion, the spontaneous rate of tritium outflow was 1.2%/min. The spontaneously released radioactivity consisted of 52% O-methylated and deaminated metabolites, 28% 3,4-dihydroxyphenylacetic acid, 18% dopamine and 2% 3-methoxytyramine. In the presence of pargyline (10 microM) the spontaneous rate of total tritium outflow decreased by 46%, that of the O-methylated and deaminated metabolites by 72% and that of 3,4-dihydroxyphenylacetic acid by 79%. The spontaneous rate of outflow of dopamine was unchanged and that of 3-methoxytyramine increased 3-fold. Further addition of nomifensine (10 microM) doubled the rate of outflow of dopamine and 3-methoxytyramine, but had no effect on the other metabolites. Electrical stimulation of the pituitary stalk (0.2 ms, 80 V, 3 Hz, 2 min) caused a tritium release of 8.5% of the tissue tritium. The evoked tritium release was only partially dependent on the extracellular calcium and not affected by tetrodotoxin. In contrast, vasopressin release evoked by stimuli of the same strength was completely calcium-dependent and blocked by tetrodotoxin. After modification of the stimulation conditions (1 ms, 10 V, 10 Hz, 2 min) the evoked tritium release was 4.1% of the tissue tritium. This tritium release was reduced by 73% in the presence of tetrodotoxin. The total evoked tritium release was decreased by 30% in the presence of pargyline and increased by 150% after further addition of nomifensine. Under the latter conditions, tetrodotoxin reduced the evoked tritium release by 67%, but nearly all of the tetrodotoxin-resistant tritium release could be identified as dopamine metabolites. Thus, the electrical stimulation appears to liberate some [3H]dopamine metabolites from an extraneuronal compartment. In conclusion, oxidative deamination and O-methylation are important pathways of the catabolism of dopamine in the neurointermediate lobe of the pituitary gland. After labelling of the transmitter stores with [3H]dopamine, the total tritium release is a poor indicator of [3H]dopamine release from the nerve terminals. Only the isolated [3H]dopamine fraction appears to reflect the release of neuronal [3H]dopamine.

Animals↗