Adrenocorticotropin concentration following administration of thyrotropin-releasing hormone in healthy horses and those with pituitary pars intermedia dysfunction and pituitary gland hyperplasia

OBJECTIVE: To compare the effect of thyrotropin-releasing hormone (TRH) administration on endogenous ACTH concentrations in healthy horses and those with pituitary pars inter-media hyperplasia and compare the test with the dexamethasone suppression test (DST). DESIGN: Prospective case series. ANIMALS: 15 horses with clinical signs of pituitary pars intermedia dysfunction (PPID), 4 horses with equivocal signs of PPID, and 29 horses without signs of PPID. PROCEDURES: ACTH concentrations prior to and after administration of TRH were measured 61 times in 48 horses. Results of the DST (cortisol response) were compared with those of the TRH test in 29 horses. Thirty-three horses (24 with no clinical signs of PPID, 5 with clinical signs of PPID, and 4 with equivocal clinical signs of PPID) were euthanized and necropsied and their pituitary glands evaluated. RESULTS: ACTH concentrations increased in all horses, but magnitude and duration of increase were significantly higher in horses with PPID. Endogenous ACTH concentrations were influenced by season. The ACTH baseline concentrations and response to TRH were not correlated with results of the DST. Results of DST were abnormal only in clinically abnormal horses or those with pars intermedia hyperplasia, but were within reference range in 17 of 26 tests in these horses. CONCLUSIONS AND CLINICAL RELEVANCE: The ACTH response to TRH is a useful test for diagnosis of pituitary gland hyperplasia, particularly in horses in which baseline ACTH concentrations are within reference range. The DST was specific but not sensitive and was inconsistent for individuals, and results often did not agree with the TRH test response.     

Central Diabetes Insipidus in a Dog With a Pro-Opiomelanocortin-Producing Pituitary Tumor ot Causing Hyperadrenocorticism

Central diabetes insipidus was diagnosed by vasopressin measurements during hypertonic stimulation in a 9-year-old male giant Schnauzer with polyuria and polydipsia. The impaired release of vasopressin was believed to be caused by a large pituitary tumor, which was visualized by computed tomography. Studies of the function of the anterior lobe and the pars intermedia of the pituitary gland were conducted, and high concentrations of ACTH and α-melanotrophic hormone (α-MSH) were found without concomitant hyperadrenocorticism. Studies of the molecular size of the immunoreactive ACTH in plasma by gel filtration revealed that most of the circulating immunoreactivity was not ACTH but its precursor pro-opiomelanocortin (POMC) and low-molecular-weight POMC-derived peptides. The pituitary tumor of this dog probably originated from melanotrophic cells of the pars intermedia. The sensitivity of the pituitary-adrenocortical system for the suppressive effect of dexamethasone was unaffected.     

Melanotroph pituitary adenoma in a cat with diabetes mellitus

A 13-year-old male, castrated, crossbred cat was referred for insulin-resistant diabetes mellitus. The cat had a ravenous appetite and a dull coat. Basal urinary corticoid/creatinine ratios were normal. In the low-dose dexamethasone suppression test there was no suppression of the (nonelevated) plasma cortisol concentration, whereas the (nonelevated) plasma adrenocorticotropic hormone (ACTH) concentration declined to low values. Basal plasma alpha-melanocyte-stimulating hormone (alpha-MSH) concentrations were highly elevated (> 1,500 ng/liter). Computed tomography revealed a pituitary tumor originating from the pars intermedia (PI). After microsurgical transsphenoidal hypophysectomy, the clinical signs resolved and the cat no longer required insulin administration. Microscopic examination of the surgical specimen revealed a pituitary adenoma originating from the PI with infiltration into the neural lobe. The adenoma immunostained intensely positive for alpha-MSH and only weakly for ACTH. It is concluded that the ACTH-independent cortisol production was probably due to the (weak) glucocorticorticotropic effects of the extremely high plasma concentration of alpha-MSH and related peptides.     

Regulation of alpha-melanocyte-stimulating hormone secretion from the pars intermedia of domestic cats

OBJECTIVE: To identify factors regulating secretion of alpha-melanocyte-stimulating hormone (alpha-MSH) from the pars intermedia (PI) of the pituitary gland of cats. ANIMALS: 28 healthy adult cats. PROCEDURE: Indwelling catheters were placed in 1 jugular vein of each of 7 to 10 cats, depending on treatment group. Sixteen hours later, 3 blood samples were collected for determination of baseline plasma hormone concentrations, and saline solution or a test substance (haloperidol, corticotropin-releasing hormone, bromocriptine, isoproterenol, insulin, or dexamethasone) was administered via the catheter. Subsequent blood samples were collected at regular intervals for up to 240 minutes after injection. Concentrations of ACTH, cortisol, and alpha-MSH were measured in plasma by use of specific radioimmunoassays. Cats were rested for at least 3 weeks between experiments. RESULTS: Administration of haloperidol and isoproterenol resulted in increased, and bromocriptine and insulin in decreased, circulating concentrations of alpha-MSH from baseline. ACTH and plasma cortisol concentrations increased after administration of all test substances except dexamethasone. Dexamethasone injection resulted in decreased plasma concentrations of ACTH and cortisol. CONCLUSIONS: Secretion of alpha-MSH from the PI of cats appears to be inhibited by dopaminergic activity and stimulated by beta-adrenergic influences. Activation of secretion of alpha-MSH from the PI can be dissociated from activation of secretion of other pro-opiomelanocortin-derived peptides, such as ACTH, arising from the pars distalis. Regulation of secretory activity of the PI of cats resembles that of rats.     

The effect of geographic location, breed, and pituitary dysfunction on seasonal adrenocorticotropin and α-melanocyte-stimulating hormone plasma concentrations in horses

Background: Plasma α‐melanocyte‐stimulating hormone (α‐MSH) and adrenocorticotropin (ACTH) concentrations in horses vary with season, confounding diagnostic testing for pituitary pars intermedia dysfunction (PPID). Hypothesis: The goals of this study were to determine whether seasonal variation in plasma α‐MSH and ACTH concentrations in horses is influenced by geographic location, breed, or PPID. Animals: Healthy light breed horses residing in Florida, Massachusetts, and Finland (n = 12 per group); healthy Morgan horses (n = 13); healthy ponies (n = 9) and horses with PPID (n = 8). Methods: Monthly plasma α‐MSH and ACTH concentrations were measured by radioimmunoassay. Nonlinear regression analysis was used to estimate the time of peak hormone concentrations. Mean hormone concentrations in fall and nonfall months were compared. Results: The fall peak plasma α‐MSH concentration occurred earlier in horses residing at more northern locations. Mean seasonal α‐MSH concentrations were similar in all healthy groups at all locations, but in the fall, plasma ACTH concentrations were higher in horses living in more southern locations. Plasma ACTH but not α‐MSH concentrations were higher in Morgan horses compared with light breed horses from the same location. Hormone concentrations of ponies did not differ from those of horses during either season. Concentrations of both hormones were high in the fall compared with the spring in horses with PPID. Conclusions and Clinical Importance: These findings suggest geographic location of residence and breed may affect the onset, amplitude, or both of the seasonal peak of pars intermedia (PI) hormones and should be considered when performing diagnostic testing for PPID. Horses with PPID maintain seasonal regulation of PI hormone output.     

Prevalence,risk factors and clinical signs predictive for equine pituitary pars intermedia dysfunction in aged horses

Reasons for performing study: Equine pituitary pars intermedia dysfunction (PPID) is an ageing‐related neurodegenerative disorder. The prevalence and risk factors for PPID using seasonally adjusted basal adrenocorticotropic hormone (ACTH) concentrations in aged horses have not been previously reported. Objectives: To determine the prevalence, risk factors and clinical signs predictive for PPID in a population of horses aged ≥15 years in Queensland, Australia. Methods: Owner‐reported data was obtained using a postal questionnaire distributed to an equestrian group. A subgroup of surveyed owners were visited and a veterinary physical examination performed on all horses aged ≥15 years. Blood samples were analysed for basal plasma alpha melanocyte‐stimulating hormone (α‐MSH) and ACTH concentrations, routine haematology and selected biochemistry. Aged horses with elevations above seasonally adjusted cut‐off values for basal plasma ACTH were considered positive for PPID. Positive horses were compared with their aged counterparts to determine risk factors and clinical signs associated with PPID. Results: Pituitary pars intermedia dysfunction was prevalent in aged horses (21.2%) despite owners infrequently reporting it as a known or diagnosed disease or disorder. Numerous clinical or historical signs were associated with an increased risk of PPID in the univariable model, but only age (odds ratio (OR) 1.18; 95% confidence interval (CI) 1.11–1.25, P<0.001) and owner‐reported history of hirsutism (OR 7.80; 95% CI 3.67–16.57, P<0.001) remained in the final multivariable model. There were no routine haematological or biochemical variables supportive of a diagnosis of PPID. Conclusions and potential relevance: Pituitary pars intermedia dysfunction occurs commonly in aged horses despite under‐recognition by owners. The increased risk of PPID with age supports that this is an ageing associated condition. Aged horses with clinical or historical signs consistent with PPID, especially owner‐reported hirsutism (delayed shedding and/or long hair coat), should be tested and appropriate treatment instituted.     

Cerebral neuroblastoma and pituitary adenocarcinoma in two budgerigars (Melopsittacus undulatus)

Case 1: A tumor mass involving the rostral part of left cerebrum was found in a two-year-old female budgerigar (Melopsittacus undulatus) at necropsy. Histologically, the neoplastic cells were arranged in sheets or cords and occasionally showed nest growth patterns. These uniform tumor cells had a little cytoplasm and ovoid or round basophilic nuclei with clearly distinct cytoplasmic membranes. The tumor cells were strong diffusely immunostained with both neuron-specific enolase (NSE) and neurofilament protein and partially for synaptophysin. They lacked chromogranin A, glial fibrillary acidic protein (GFAP), vimentin, S-100, and cytokeratin antigen expression. Moreover, they had no reaction to antibodies against pituitary hormones, such as adrenocorticotrophic hormone (ACTH), growth hormone, and prolactin. The histological and immunohistochemical examination determined the tumor as neuroblastoma. Case 2: An extremely enlarged pituitary mass was found above the sella turcica of a male budgerigar. It was soft and well delineated from the adjacent structures. On histological examination, this tumor consisted of a sheet of large closely packed polyhedral cells that had scant to a large amount of pale to strongly eosinophilic cytoplasm. The pleomorphic nuclei were apparently variable in shape, from small round hyperchromatic to very large vesicular forms. The cell boundaries were not clearly distinct. The multifocal immunolabelling of neoplastic cells for NSE, synaptophysin, GFAP, and ACTH appeared, whereas a few cells reacted with vimentin and S-100 and stained negative for other markers, which were also utilized for case 1. Histological and immunohistochemical findings led to identification of corticotroph adenocarcinoma in the pituitary gland.     

Circadian and circannual rhythms of cortisol,ACTH, and α-melanocyte-stimulating hormone in healthy horses

Cosinor analysis was used to evaluate whether pituitary and adrenal hormones exhibit circadian rhythmicity in horses. The effect of season and animal age on their respective rhythms was also determined. In addition, the usefulness of evaluating cortisol rhythmicity for the diagnosis of pituitary pars intermedia dysfunction (PPID) was assessed. Serum cortisol concentrations (P < 0.01), but not plasma ACTH or α-melanocyte-stimulating hormone (α-MSH), showed a significant circadian periodicity in horses. An effect of season on hormone concentration was observed with plasma ACTH and α-MSH concentration greater in the fall and cortisol concentration greater in the spring (P < 0.001). Age did not affect cortisol rhythm, but it did blunt the variation in cortisol concentration in horses, similar to what has been previously reported to occur in aged people and dogs. In addition, our results suggest that clinically and diagnostically normal, non–PPID-affected horses commonly have a loss of cortisol diurnal rhythm. Therefore, measurement of circadian rhythm is not an appropriate diagnostic test for PPID.     

Expression of leukemia inhibitory factor and leukemia inhibitory factor receptor in the canine pituitary gland and corticotrope adenomas

Leukemia inhibitory factor (LIF) is a pleiotropic cytokine of the IL-6 family that activates the hypothalamic-pituitary-adrenal axis and promotes corticotrope cell differentiation during development. The aim of this study was to investigate the expression of LIF and its receptor (LIFR) in the canine pituitary gland and in corticotrope adenomas, and to perform a mutation analysis of LIFR. Using immunohistochemistry, immunofluorescence, and quantitative expression analysis, LIF and LIFR expression were studied in pituitary glands of control dogs and in specimens of corticotrope adenoma tissue collected through hypophysectomy in dogs with pituitary-dependent hypercortisolism (PDH, Cushing's disease). Using sequence analysis, cDNA was screened for mutations in the LIFR. In the control pituitary tissues and corticotrope adenomas, there was a low magnitude of LIF expression. The LIFR, however, was highly expressed and co-localized with ACTH_1-24 expression. Cytoplasmatic immunoreactivity of LIFR was preserved in corticotrope adenomas and adjacent nontumorous cells of pars intermedia. No mutation was found on mutation analysis of the complete LIFR cDNA. Surprisingly, nuclear to perinuclear immunoreactivity for LIFR was present in nontumorous pituitary cells of the pars distalis in 10 of 12 tissue specimens from PDH dogs. These data show that LIFR is highly co-expressed with adrenocorticotropic hormone (ACTH) and α-melanocyte-stimulating hormone (α-MSH) in the canine pituitary gland and in corticotrope adenomas. Nuclear immunoreactivity for LIFR in nontumorous cells of the pars distalis may indicate the presence of a corticotrope adenoma.     

The Effect of Season on the Histologic and Histomorphometric Appearance of the Equine Pituitary Gland

The objective of this study was to determine the effect of season on the histologic and histomorphometric appearance of the normal equine pituitary gland. Pituitary glands were collected at necropsy from 121 horses throughout the year. Plasma was also collected from 59 of these horses before euthanasia. Hematoxylin and eosin stained median sagittal sections of each pituitary were evaluated and histologically graded by three pathologists. Histomorphometric analysis was performed on the same slides. Plasma α-melanocyte stimulating hormone was measured by radioimmunoassay in a subset of horses (n = 59). A total of 118 pituitary glands were included in the study after exclusions were made on the basis of the presence of pars intermedia (PI) adenomas (>5 mm). There was a positive correlation between PI hormone concentration (α-melanocyte stimulating hormone) and PI area. Pituitary gland measurements and grades from samples collected in the fall were compared with those collected in the nonfall months using t-test. The PI area, total pituitary area, and PI/total pituitary ratio were significantly greater in the fall compared with nonfall months (P < .0001, P < .01, P < .0001, respectively). Pituitary grades were also higher in the fall compared with nonfall months (P < .001). There was no seasonal difference in pars distalis or pars nervosa area. The results of this study show that the normal equine pituitary shows seasonal changes in appearance and size. These changes must be considered when using postmortem histologic evaluations in the diagnosis of pituitary pars intermedia dysfunction or for validation of antemortem diagnostic tests.     

Systemic and pituitary pars intermedia antioxidant capacity associated with pars intermedia oxidative stress and dysfunction in horses

OBJECTIVE: To determine whether a deficiency in systemic or local (pars intermedia) antioxidant capacity is associated with pituitary pars intermedia oxidative stress and pituitary pars intermedia dysfunction (PPID) in horses. SAMPLE POPULATION: Blood samples from 20 horses with PPID and 20 healthy client-owned horses, archived paraffin-embedded adrenal gland and substantia nigra tissues from 20 horses, and pituitary gland tissue from 16 horses. PROCEDURES: Total glutathione, superoxide dismutase, and glutathione peroxidase activities were determined in RBCs. Accumulation of a systemic marker of oxidative stress (3-nitrotyrosine) was assessed in plasma and formalin-fixed, paraffin-embedded adrenal gland and substantia nigra tissues. Local antioxidants (total and manganese superoxide dismutase, glutathione peroxidase, and total glutathione) were measured in pars intermedia tissues. RESULTS: No significant differences existed in systemic antioxidant enzyme activity or accumulation of 3-nitrotyrosine between horses with PPID and control horses. In pituitary gland tissues, glutathione peroxidase activity was increased in horses with oxidative stress, whereas total glutathione concentration and superoxide dismutase activity remained unchanged. There was an age-associated decrease in manganese superoxide dismutase activity in the pars intermedia. CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence of systemic accumulation of oxidative stress markers or deficiencies in antioxidant capacity in horses with PPID, suggesting that these are unlikely to be major predisposing factors in the development of PPID. Manganese superoxide dismutase activity in the pars intermedia decreased significantly with increasing age. Role of an age-associated decrease in antioxidant capacity for the pars intermedia in the development of PPID in horses warrants further investigation.     

Evaluation of pituitary gland anatomy and histopathologic findings in clinically normal horses and horses and ponies with pituitary pars intermedia adenoma

OBJECTIVE: To determine size and weight of the pituitary gland and associations between pituitary gland size and weight and sex and age in horses without clinical signs associated with pituitary pars intermedia adenoma (PPIA) and horses and ponies with PPIA. ANIMALS: Pituitary glands from 100 horses without clinical signs of PPIA and 19 horses and 17 ponies with PPIA. PROCEDURES: Pituitary glands were weighed, measured, and examined histologically by use of H&E stain. Masson trichrome and periodic acid-Schiff staining were used, when appropriate. Histologic lesions in the pars intermedia, pars distalis, or both were classified as no significant lesions, single or multiple cysts, focal or multifocal hyperplasia, single or multiple microadenomas, and adenoma. Relative pituitary weight (RPW) was calculated as pituitary weight (grams) divided by body weight (grams). RESULTS: There was an age-related increase in the presence of pituitary lesions in the pars distalis and pars intermedia in geldings, mares overall, and non-pregnant mares. Mean (+/-SD) RPW in horses with PPIA was not significantly different from ponies with PPIA (15+/-5.9 x 10(-6) and 16+/-72 x 10(-6), respectively). Maximum pituitary weight in a horse with PPIA was 13.9 g (RPW, 2.9 X 10(-5)). Plasma glucose concentration was positively correlated with RPW in ponies with PPIA. CONCLUSIONS AND CLINICAL RELEVANCE: Pituitary lesions may be a factor in horses with insulin resistance and laminitis before development of clinical signs of PPIA. Ovarian steroids may be involved in the pathogenesis of lesions in the pars intermedia.     

Effects of season and sample handling on measurement of plasma alpha-melanocyte-stimulating hormone concentrations in horses and ponies

OBJECTIVE: To investigate effects of sample handling, storage, and collection time and season on plasma alpha-melanocyte-stimulating hormone (alpha-MSH) concentration in healthy equids. ANIMALS: 11 healthy Standardbreds and 13 healthy semiferal ponies. PROCEDURE: Plasma alpha-MSH concentration was measured by use of radioimmunoassay. Effects of delayed processing were accessed by comparing alpha-MSH concentrations in plasma immediately separated with that of plasma obtained from blood samples that were stored at 4 degrees C for 8 or 48 hours before plasma was separated. Effects of suboptimal handling were accessed by comparing alpha-MSH concentrations in plasma immediately stored at -80 degrees C with plasma that was stored at 25 degrees C for 24 hours, 4 degrees C for 48 hours or 7 days, and -20 degrees C for 30 days prior to freezing at -80 degrees C. Plasma alpha-MSH concentrations were compared among blood samples collected at 8:00 AM, 12 noon, and 4:00 PM. Plasma alpha-MSH concentrations were compared among blood samples collected in January, March, April, June, September, and November from horses and in September and May from ponies. RESULTS: Storage of blood samples at 4 degrees C for 48 hours before plasma was separated and storage of plasma samples at 4 degrees C for 7 days prior to freezing at -80 degrees C resulted in significant decreases in plasma alpha-MSH concentrations. A significantly greater plasma alpha-MSH concentration was found in September in ponies (11-fold) and horses (2-fold), compared with plasma alpha-MSH concentrations in spring. CONCLUSIONS AND CLINICAL RELEVANCE: Handling and storage conditions minimally affected plasma alpha-MSH concentrations. Seasonal variation in plasma alpha-MSH concentrations must be considered when evaluating pituitary pars intermedia dysfunction in equids.     

Correlation of pituitary histomorphometry with adrenocorticotrophic hormone response to domperidone administration in the diagnosis of equine pituitary pars intermedia dysfunction

Functional evaluation of the pars intermedia (PI) is required for the early diagnosis of equine pituitary PI dysfunction (PPID), yet most assays target the hypothalamic-pituitary-adrenal axis, which regulates the pars anterior. In contrast, the PI is regulated by dopaminergic tone from hypothalamic neurons. Loss of dopaminergic inhibition is hypothesized to cause the PI hypertrophy and hyperplasia that result in the clinical manifestations of PPID. Domperidone, a dopamine receptor antagonist, should exacerbate the loss of dopaminergic inhibition in horses with PPID and increase the release of endogenous adrenocorticotrophic hormone (eACTH) by PI melanotrophs. To test this, plasma eACTH concentration was determined in horses with or without clinical signs of PPID at 0, 4, and 8 hours after oral administration of 3.3 mg domperidone/kg. Pituitary glands were evaluated postmortem by histologic grading and morphometry. In the 33 horses, median age, plasma ACTH concentration 8 hours after domperidone, and PI area in median sagittal sections were associated with histologic grade as follows: pituitary grade 1 (normal), n = 3, 7.5 years, 20.0 pg/ml, 0.16 cm(2); grade 2 (focal hypertrophy or hyperplasia), n = 9, 14.5 years, 27.1 pg/ml, 0.27 cm(2); grade 3 (diffuse adenomatous hyperplasia), n = 5, 21.0 years, 64.4 pg/ml, 0.48 cm(2); grade 4 (microadenomas), n = 12, 23.3 years, 128.0 pg/ml, 0.87 cm(2); grade 5 (adenoma), n = 4, 24.9 years, 720.5 pg/ml, 2.1 cm(2). Results suggest that horses with pituitary histologic grade > or =3 respond to domperidone with increased plasma ACTH concentration.     

Seasonal changes in plasma adrenocorticotropic hormone and α-melanocyte-stimulating hormone in response to thyrotropin-releasing hormone in normal, aged horses

Background: Results of diagnostic tests for equine pituitary pars intermedia dysfunction (PPID), including endogenous ACTH concentration and the overnight dexamethasone suppression test (DST), are affected by season. New and potentially more sensitive diagnostic tests for equine PPID, such as thyrotropin‐releasing hormone (TRH)‐stimulated ACTH response, have been developed, but have had limited evaluation of seasonality. Objective: Our purpose was to evaluate seasonal changes in plasma ACTH and alpha‐melanocyte‐stimulating hormone (α‐MSH) responses to TRH administration. Animals: Nine, healthy, aged horses with normal DST results. Methods: Synthetic TRH (1 mg) was administered IV. Plasma ACTH and α‐MSH concentrations were measured at 0, 5, 10, 15, 20, 25, 30, 45, 60, and 180 minutes. Testing was performed in February, July, August, September, October, and November. Mean TRH‐stimulated ACTH and α‐MSH concentrations were compared across months and time by repeated measures analysis of variance. Significance was set at the P < .05 level. Results: Concentrations of ACTH and α‐MSH significantly increased after TRH administration. Endogenous and TRH‐stimulated ACTH and α‐MSH concentrations were significantly different across months with higher concentrations in the summer and fall compared with February. Conclusions and Clinical Importance: Plasma ACTH and α‐MSH responses to TRH administration experience seasonal variation, with TRH‐stimulated ACTH and α‐MSH concentrations increasing from summer through fall. These results support previous evidence of a seasonal influence on the equine pituitary‐adrenal axis. More research is warranted with a larger number of horses to determine if seasonal reference ranges for TRH stimulation testing need to be defined.     

Expression of genes related to corticotropin production and glucocorticoid feedback in corticotroph adenomas of dogs with Cushing's disease

Cushing's disease caused by pituitary corticotroph adenoma is a common endocrine disease in dogs. A characteristic biochemical feature of corticotroph adenomas is their relative resistance to negative feedback by glucocorticoids. In this study, we examined gene expression related to adrenocorticotropic hormone (ACTH) production and secretion, and the negative feedback by glucocorticoids in canine corticotroph adenoma. We used resected corticotroph adenomas from 10 dogs with Cushing's disease. In order to investigate the alteration of gene expression between corticotroph adenoma and normal corticotrophic cells, ACTH-positive cells in the anterior lobe were microdissected using a laser-capture microdissection system, and mRNA levels of proopiomelanocortin (POMC), corticotropin releasing hormone receptor 1 (CRHR1), glucocorticoid receptor (GR), mineralocorticoid receptor (MR), and 11 beta hydroxysteroid dehydrogenase (11HSD) type 1 and type 2 were determined using real-time RT-PCR. POMC, CRHR1, and 11HSD2 mRNA levels in corticotroph adenoma were greater than those in normal corticotrophic cells (POMC, 5.5-fold; CRHR1, 4.9-fold; 11HSD2, 4.2-fold, P<0.01, respectively). MR and 11HSD1 mRNA levels in corticotroph adenoma were lower than those in normal corticotrophic cells (MR, 2.2-fold; 11HSD1, 2.9-fold, P<0.01, respectively). GR mRNA levels did not differ between corticotroph adenoma and normal corticotrophic cells. Our results may help to understand the increased ACTH production and the resistance to negative feedback suppression by glucocorticoids in canine corticotroph adenomas. These changes in gene expression may have a role in the growth of canine corticotroph adenoma, and help elucidate the pathophysiology of dogs with Cushing's disease.     

Plasma pro-opiomelanocortin, pro-adrenocorticotropin hormone, and pituitary adenoma size in dogs with Cushing's disease

It is difficult to predict the size of pituitary corticotroph tumors in dogs with Cushing's disease (pituitary-dependent hyperadrenocorticism [PDH]) without pituitary imaging techniques. The purpose of this study was to examine the relationship between plasma adrenocorticotropin hormone (ACTH) precursor concentration and pituitary size in dogs with Cushing's disease. Plasma concentrations of ACTH precursors (pro-opiomelanocortin [POMC]/pro-ACTH) and pituitary tumor height/brain area were measured in 36 dogs with pituitary corticotroph adenomas of various sizes. There was a correlation between tumor size (measured as the pituitary tumor height/brain area ratio [P/B]) and POMC/pro-ACTH concentration (r = .70; P < .0001). Dogs with P/B > or = 0.40 x 10(-2) mm(-1) had higher concentrations of ACTH precursors than dogs with P/B < 0.40 x 10(-2) mm(-1) (median concentration 85 pmol/L, range 15-1,350 pmol/L, n = 14 versus 15 pmol/L, range 15-108 pmol/L, n = 22; P < .0001). With a threshold of 35 pmol/L of POMC/pro-ACTH concentration, the estimated sensitivity and specificity of the kit were 93% (95% confidence interval [CI], 79-100%) and 86% (95% CI, 73-100%), respectively. We interpret these data as indicating that measurement of POMC and pro-ACTH might be of value in the characterization of tumor size in dogs with Cushing's disease. Low POMC/pro-ACTH concentrations make it unlikely that a large pituitary tumor exists in dogs with PDH.     

Agreement in histologic assessments of the pituitary pars intermedia in aged horses

OBJECTIVE: To evaluate concordance among veterinary pathologists in the assessment of histologic findings in the pars intermedia of pituitary gland sections from aged horses with mild signs suggestive of pituitary pars intermedia dysfunction (PPID). Sample Population-10 pituitary glands from aged horses. PROCEDURE: 7 pathologists were provided with signalment, clinical signs, and a single H&E-stained pituitary gland section from 10 aged horses with mild signs suggestive of PPID. Pathologists described histologic findings for each section and stated whether findings were consistent with PPID. Agreement among pathologists and with antemortem diagnostic test results was calculated. RESULTS: Overall, only fair agreement was found among the pathologists as to which horses had histologic findings consistent with disease (mean +/- SE kappa value, 0.34 +/- 0.069). Interpretation of individual sections varied, with minimal agreement (4 or 5/7 pathologists) for 5 of 10 sections evaluated. Postmortem assessment was in agreement with an antemortem endocrine diagnostic test result 79% of the time. CONCLUSIONS AND CLINICAL RELEVANCE: Validation of antemortem diagnostic testing for PPID in horses often relies on the results of postmortem histologic evaluation. The lack of consensus in histologic interpretation of pituitary glands from aged horses with mild clinical signs in our study indicates that postmortem histologic evaluation of pituitary glands is an inappropriate standard in validation of antemortem diagnostic tests for detection of early PPID. Caution should be used when interpreting diagnostic test results in horses in which early PPID is suspected.     

Problems in the diagnosis of pituitary adenoma (Cushing's syndrome) in horses

Thyrotropin releasing hormone (TRH) stimulation tests were carried out on a number of horses, including one clinically affected horse whose age and clinical signs suggested that a pituitary adenoma was most unlikely. The results of these tests indicated that, according to criteria published overseas, the majority of these horses had pituitary adenomas. The fact that clinically normal or affected horses may have marked increases in cortisol concentrations indicates that the TRH stimulation test is not suitable for the diagnosis of pituitary adenoma. Other tests, including alterations in glucose metabolism and, if available, ACTH measurements, are probably more useful for diagnosis. However, most diagnoses will probably rely upon the characteristic clinical signs.