Convergent and discriminant validity of the CVLT (dementia version). California Verbal Learning Test.

This study investigated the convergent and discriminant validity of the 9-item "dementia version'' of the California Verbal Learning Test (CVLT-9) in a sample of 130 geriatric patients evaluated for memory complaints. Moderate correlations were observed between the CVLT-9 sum of words recalled for trials 1-5 (Trial 1-5 Recall) and Long-Delay Free Recall (LDFR) measures and the immediate and delayed Logical Memory (LM I and LM II) and Visual Reproduction (VR I and VR II) subtests from the Wechsler Memory Scale-Revised (WMS-R). However, the CVLT-9 Trial 1-5 Recall and VR I measures demonstrated significant correlations with a number of additional measures of language and visuospatial ability. The CVLT-9 LDFR, and the WMS-R LM I, LM II, and VR II showed less overlap with non-episodic memory functioning. A principal components analysis yielded a three-component solution consisting of a general or "g'' component, a specific memory component, and a mood component. The CVLT-9 Trial 1-5 Recall and VR I loaded on both the "g'' and the memory components, whereas LM I, LM II, and VR II loaded on only the memory component. We conclude that the CVLT-9 Trial 1-5 Recall and VR I demonstrate low discriminant validity, suggesting diminished specificity as memory measures.

Activin A regulation of gonadotropin-releasing hormone synthesis and release in vitro.

Activin is essential for the regulation of normal mammalian reproductive function at both the pituitary and gonadal levels. However, its central actions in the control of the hypothalamic-pituitary-gonadal axis remain largely unexplored. The present study aims to determine whether activin could regulate the reproductive axis at the level of the hypothalamus, through control of the GnRH neuroendocrine system. Using the GnRH-secreting GT1-7 neuronal cell line as a model system, we demonstrate expression of mRNAs encoding activin receptor types I, IB, and II. We examined the effects of activin A on GnRH protein secretion and mRNA levels in GT1-7 cells. Treatment with rh-activin A regulated both GnRH protein secretion and GnRH mRNA expression in the GT1-7 cells in a time-dependent fashion. Using transient transfection assays, we explored a potential transcriptional basis for these changes. Activin A increased reporter gene activity driven by minimal GnRH enhancer and promoter elements, suggesting that activin may regulate GnRH gene expression at the level of transcription. Lastly, activin A treatment of male rat hypothalami, in vitro, increased GnRH protein secretion. Collectively, molecular and physiological evidence support the presence of an activin system which might act at a hypothalamic site to regulate mammalian reproduction via activation of GnRH synthesis and release.

Pax6 is implicated in murine pituitary endocrine function.

Pax6, an evolutionarily conserved transcription factor, is expressed in the murine and zebrafish embryonic pituitary, but its role in pituitary development and endocrine function has not been described. To study the role of Pax6 in vivo, we examined Pax6 mutant mouse (SeyNeu) pituitaries. Mice homozygous for the SeyNeu mutation die at birth; therefore, we examined peptide hormone expression by the differentiated pituitary cell types as well as developmental marker expression in the intermediate and anterior lobes of the embryonic pituitary. GH- and PRL-immunopositive cells appear severely decreased in an outbred ICR background at embryonic d 17.5, although mRNA expression of these peptide hormones is present, as is expression of other pituitary markers. This suggests that pituitary cell types are able to differentiate in mutant embryos. To identify the cellular or physiologic mechanism responsible for less GH- and PRL-immunoreactivity in Pax6 mutant mice, we tested serum levels of GH and PRL. Pax6 homozygous mutant mice have GH serum levels one fifth that of controls at embryonic d 17.5, and one-third that of controls at postnatal d 0. PRL serum levels, which are very low during embryonic and neonatal stages, were below assay detection limits in both the wild-type and mutant groups. Taken together, these data suggest that Pax6 is not essential for pituitary differentiation, but rather functions to establish appropriate neonatal homeostatic levels of GH and PRL, possibly through regulation of translational or secretory mechanisms.

Identification of an inhibin receptor in gonadal tumors from inhibin alpha-subunit knockout mice.

Inhibins and activins are dimeric proteins that are functional antagonists and are structurally related to the transforming growth factor-beta (TGFbeta) family of growth and differentiation factors. Receptors for activin and TGFbeta have been identified as dimers of serine-threonine kinase subunits that regulate cytoplasmic proteins known as Smads. Despite major advances in our understanding of activin and TGFbeta receptors and signaling pathways, little is known about inhibin receptors or the mechanism by which this molecule provides a functionally antagonistic signal to activin. Studies described in this paper indicate that an independent inhibin receptor exists. Numerous tissues were examined for inhibin-specific binding sites, including the developing embryo, in which the spinal ganglion and trigeminal ganglion-bound iodinated inhibin A. Sex cord stromal tumors, derived from male and female inhibin alpha-subunit-deficient mice, were also identified as a source of inhibin receptor. Abundant inhibin and few activin binding sites were identified in tumor tissue sections by in situ ligand binding using iodinated recombinant human inhibin A and 125I-labeled recombinant human inhibin A. Tumor cell binding was specific for each ligand (competed by excess unlabeled homologous ligand and not competed by heterologous ligand). Based on these results and the relative abundance and homogeneity of tumor tissues versus the embryonic ganglion, tumor tissues were homogenized, membrane proteins were purified, and putative inhibin receptors were isolated using an inhibin affinity column. Four proteins were eluted from the column that bind iodinated inhibin but not iodinated activin. These data suggest that inhibin-specific membrane-associated proteins (receptors) exist.

Expression of activin and follistatin in the rat hypothalamus: anatomical association with gonadotropin-releasing hormone neurons and possible role of central activin in the regulation of luteinizing hormone release.

The central role of activin in the regulation of the reproductive axis remains largely unexplored. Evidence suggests that activin may play a role in control-linggonadotropin-releasing hormone (GnRH) release. We assessed potential neuroanatomical associations between activin- and GnRH-neuronal systems via examination of the distribution of activin betaA-subunit and activin binding protein (follistatin) protein and mRNA signals relative to GnRH neurons in the adult rat brain. Activin betaA-subunit-immunostained fibers were distributed throughout the hypothalamus and GnRH-positive perikarya, and fibers were in close association with betaA-subunit-immunoreactive fibers. Follistatin mRNA-expressing cells were also identified throughout the hypothalamus with GnRH fibers often observed juxtaposed to follistatin cell bodies. Colocalization of either the betaA-subunit or follistatin within GnRH neurons was not detected. The functional significance of central activin in the regulation of the reproductive axis was also demonstrated. The intracerebroventricular infusion of rh-activin A significantly increased luteinizing hormone, but not follicule-stimulating hormone, serum levels in adult male rats. Taken together, the present results support an interaction between activin and GnRH neuronal systems in the rat hypothalamus, and suggest activin may act within the brain to regulate the reproductive axis.

Tissue-specific expression of inhibin/activin subunit and follistatin mRNAs in mid- to late-gestational age human fetal testis and epididymis.

Inhibin/activin subunit (alpha, beta A, and beta B) immunoreactive protein localization patterns and cell type specific inhibin alpha-subunit mRNA expression have been examined in early- to midgestational age human fetal testes. The scarcity of available third trimester human fetal tissue has, however prevented a complete examination throughout the gestational period and the cell specific expression of follistatin and beta A- and beta B-subunit mRNAs are currently unknown at any gestational age. In the present study, this gap is filled and report mRNA expression patterns of inhibin/activin subunits in mid- and late-gestational age (21-33 wk) human fetal testes and testicular duct system. We also report the first examination of follistatin mRNA signals in the human fetal gonad is also resent in both tubular and interstitial cells, and beta B-subunit mRNA is expressed in seminiferous tubules, in mid- and late-gestational age human fetal testes. Inhibin/activin beta A-subunit mRNA was detected in the interstitial cells of remarkably well preserved mid (21 and 22 wk) and late (29 wk) gestational age testes, and is the only activin-system factor mRNA also expressed in tissue of the duct system of the testis (smooth muscle cells of the epididymis). Follistatin mRNA signal was equal to background levels in testicular and duct tissues at all ages examined. These cell specific expression patterns suggest prominent and possibly differential roles for the inhibins and activins, unopposed by gonadal follistatin, in the human fetal male reproductive system.

Functional outcome of cognitively impaired hip fracture patients on a geriatric rehabilitation unit.

OBJECTIVES: To compare outcomes (physical functions and discharge destinations) of cognitively impaired and intact older hip fracture patients, and to identify cognitive skills related to functional gains. DESIGN: Prospective longitudinal study of hip fracture patients treated on an acute inpatient rehabilitation service, with evaluation of functional performance and living status determined at admission and discharge. SETTING: A specialized inpatient geriatric rehabilitation program at Wesley Woods Geriatric Hospital, which is affiliated with Emory University School of Medicine. SUBJECTS: Fifty-eight hip fracture patients, 35 with and 23 without cognitive impairment. MEASUREMENTS: Cognitive functioning measured by the Mattis Dementia Rating Scale (MDRS); functional outcome assessed by the Functional Independence Measure (FIM); comparison of pre-fracture with discharge living environments. MAIN RESULTS: Both cognitively impaired and intact hip fracture patients exhibited similar overall FIM motor improvements as well as functional gains in specific FIM areas measuring self-care, sphincter control, and locomotion (e.g., walking). Cognitively intact patients, however, displayed significantly greater gains in mobility (e.g., transfers) at discharge. Cognitively impaired patients who lived in the community were as likely as intact patients to return to the community. Patients who entered the program at a modified dependence level (FIM 3-5) and achieved motor independence at discharge (FIM 6-7) had higher MDRS initiation/ perseveration and memory scores. CONCLUSIONS: Hip fracture patients with cognitive impairments can achieve positive outcomes as defined by functional improvement and discharge destination. Intensive post-fracture rehabilitation in the early phase of recovery may promote functional independence and a return to the community for older patients at risk for nursing home placement. Future research should examine the long-term maintenance of these improvements and explore how rehabilitation interventions can be altered to enhance outcome.

Prosody impairment and associated affective and behavioral disturbances in Alzheimer's disease.

We examined the ability to produce, repeat, and comprehend emotional prosody in 20 patients with Alzheimer's disease (AD) and in 11 elderly normal control subjects. In addition, caregivers of AD patients completed affective and behavioral measures with reference to the patient. Relative to control subjects, comprehension of emotional prosody was marginally impaired in mildly demented AD patients, whereas production, comprehension, and repetition of emotional prosody were significantly impaired in moderately demented AD patients. The moderately demented patients performed significantly poorer than the mildly demented patients on the production and repetition tasks. In contrast, there was no significance difference between the two groups on the prosody comprehension task. Additional analyses revealed an inverse relationship between the ability to correctly produce and repeat emotional prosody and the frequency of agitated behaviors and depressive symptomatology in moderately demented patients. This latter findings suggests that the inability to communicate emotional message is associated with disturbances in mood and behavior in AD patients. Implications for the management of disruptive behavior in agitated and aprosodic AD patients include the development of caregiver sensitivity to unexpressed emotion and caregiver assistance with emotional expression.

Short-form alternatives to the Judgment of Line Orientation Test.

The Judgment of Line Orientation Test (JLO; Benton, Hamsher, Varney, & Spreen, 1983) is frequently used as a motor-free method of evaluating visuospatial processing but can be time-consuming to administer. We investigated the internal consistency, validity, and utility of two parallel JLO short forms in a mixed clinical sample of 386 patients. Mean scores were equivalent, and correlational analyses supported the internal consistency and validity of both short forms. When compared to the standard JLO, the odd- and even-item short forms demonstrated good sensitivity, specificity, overall hit rate and predicted positive and negative accuracy. We conclude that the JLO short forms possess sufficient internal consistency, validity, and utility for serial assessment in research studies. The JLO short forms may potentially be used in clinical screening situations by applying a single cut-off score to differentiate levels of performance. However, more detailed clinical use of these JLO short forms will necessitate collection of normative data in order to generate accurate percentile rankings.

Tissue-specific binding of radiolabeled activin A by activin receptors and follistatin in postimplantation rat and mouse embryos.

Activin affects the growth and differentiation of many cultured cell types, including rat anterior pituitary cells and gonadal and neuronal cell lines. Endogenous activins regulate mesoderm induction, body axis formation, and organogenisis in the developing embryo. The messenger RNAs encoding inhibin/activin subunits, follistatin (an activin-binding protein), and activin type II receptors (ActRII and IIB) are expressed in various cell types and tissues of the embryonic rat and mouse. Follistatin-deficient mice have numerous embryonic defects, including shiny taut skin, allowing relatively easy identification by the later stages of embryogenesis. ActRII-deficient mice, on the other hand, show limited developmental defects, with some (22%) embryonic day 18.5 (E18.5) ActRII-deficient embryos showing various skeletal and facial abnormalities. The present study was undertaken to identify the target tissues for biologically active activin A and assess the significance of its association with ActRII and follistatin in developing rat and mouse embryos. Fresh-frozen, slide-mounted, rat (E13 to E19) and mouse (E18.5) embryo sections were incubated with 125I-labeled recombinant human activin A. Nonspecific binding was evaluated by competition with an excess of cold activin A. As determined by image analysis, the highest levels of activin A binding were observed throughout the brain, spinal cord, and trigeminal and spinal ganglia at all ages. Lower levels of binding were found in the dermis of the skin starting on E15. Follistatin-deficient mice demonstrated similar patterns and levels of activin A binding in the neural tissues compared to wild-type controls, but binding was absent in the skin. In ActRII-deficient mice, activin A binding was completely absent in neural tissues, but was similar to wild-type control levels in the dermal layer of the skin. The data indicate that activin A binds to specific tissues of mouse and rat embryos and that binding is dependent upon the presence of ActRII in the central and peripheral nervous system and on follistatin in the skin.

Distribution of follistatin messenger ribonucleic acid in the rat brain: implications for a role in the regulation of central reproductive functions.

Follistatin (FS), which binds to the inhibin/activin beta A- or beta B-subunit is localized with and modulates the biological actions of activin in many systems. However, in contrast to the wide distribution of the activin beta-subunit proteins and messenger RNAs (mRNA) in the brain, demonstration of FS mRNA signal has been limited to the olfactory tubercle and layer II of the frontal cortex. We have hypothesized a more extensive distribution of central FS gene expression and localization in regions coinciding with inhibin/activin beta-subunits and possible activin-mediated effects. In the present study, we examined the central distribution of FS mRNA expression in the normal adult male rat. With in situ hybridization analysis, using a 33P-labeled RNA probe specific for rat FS, gene expression is shown to be widely distributed throughout the brain. Abundant FS mRNA expression is localized in several areas of the olfactory bulb as well as the frontal cortex, a few thalamic nuclei, and in septal regions. Moderate FS mRNA is observed in the caudate putamen and various hypothalamic areas including the paraventricular, ventromedial, dorsomedial, and arcuate nuclei. Several brain stem regions are also found to express FS mRNA, including the medial vestibular and solitary tract nuclei. Notably, FS mRNA, including the medial vestibular and solitary septal/diagonal band region is localized in patterns that are highly correlative with those of GnRH gene expression and hence may serve to regulate possible activin-mediated effects in these areas. FS mRNA is also expressed in areas associated with the activin-oxytocin pathway (solitary tract nucleus and paraventricular nucleus) and is therefore in a position to modulate the role of activin in the solitary tract nucleus-paraventricular nucleus pathway (afferent system mediating the milk-ejection reflex). The results suggest that FS is centrally localized in sites compatible with a role in the regulation of central reproductive functions.

Behavioral outcome of posterior parietal cortex injury in the monkey.

Recent studies from our laboratory have characterized the response properties of trigeminal nociceptive neurons located in the posterior parietal cortex of awake monkeys, particularly in the rostral portion of the inferior parietal lobule and parietal operculum within the lateral sulcus. The stimulus intensity-response functions of some nociceptive neurons were significantly correlated to the stimulus intensity-escape frequency functions. The present study provides evidence that trauma to the posterior parietal cortex alters pain sensibility to the contralateral face. Although thermal pain tolerance was dramatically altered, the discriminative aspect of thermosensitivity may have remained intact. Our results complement the recent findings of clinical studies concerned with pain and damage to the posterior parietal cortex and of experimental studies concerned with painful stimulation and changes in regional cerebral blood flow. The role of the posterior parietal cortex in nociception and pain is discussed in relation to the first somatosensory area and to unilateral spatial neglect (inattention).

Hybridization histochemical and immunohistochemical localization of inhibin/activin subunits and messenger ribonucleic acids in the rat brain.

Inhibin and activin are best known as gonadal glycoprotein hormones but have a broad anatomical distribution. We previously described the central distribution ofinhibin/activin beta A- and beta B-subunit proteins in some neuronal cell bodies, fibers, and nuclei of the rat brain and reported a possible role for central activin in suckling-induced oxytocin secretion and corticotropin releasing factor release. In the present report, we mapped the detailed immunohistochemical localization of inhibin/activin alpha-, beta A-, and beta B-subunits throughout the rat brain to further clarify their central distribution. In addition, the localization and distribution of their corresponding mRNAs was assessed. The results are summarized as follows: 1) Both beta A- and beta B-subunit immunoreactivity are found in neuronal cell bodies in the nucleus of the solitary tract and the dorsal and ventral medullary reticular nuclei, and in fibers and terminals of known projection sites for these nuclei. 2) beta B-subunit immunoreactivity is localized in a group of perifornical neurons in the hypothalamus. 3) beta A-subunit immunoreactivity is present in discrete populations of neuronal cell nuclei scattered throughout the CNS. 4) mRNAs encoding each of the inhibin/activin subunits are expressed in all major brain regions as determined by S1 nuclease assay and in a variety of specific neuroanatomical sites as shown by in situ hybridization. The results suggest that central inhibin and activin proteins are produced in the brain where they may potentially serve inter- and intracellular functions in multiple systems.

Localization of Pbx1 transcripts in developing rat embryos.

Recently, a new family of homeodomain proteins has emerged, that includes extradenticle, ceh-20, Pbx1, Pbx2 and Pbx3. The Pbx family has been shown to modulate the biological activities of the Hox proteins. We demonstrate here by in situ hybridization that Pbx1 transcripts are present in many embryonic tissues. Highest levels of Pbx1 expression in the developing embryo, from 12 to 20 days post coitum, are found in neuronal tissues, including brain, spinal cord and ganglia. In addition, Pbx1 transcripts are also detectable in the gut, lung, olfactory epithelium and kidney. The expression pattern of Pbx1 overlaps with that of many of the Hox gene products and is consistent with them acting in parallel to regulate common target genes.

Expression of human gonadotropin-releasing hormone receptor gene in the placenta and its functional relationship to human chorionic gonadotropin secretion.

Placental GnRH is one of the potential paracrine regulators of hCG secretion from the trophoblasts during pregnancy. Maternal serum hCG levels exhibit an exponential rise during the first 6 weeks of pregnancy, peak at 9-10 weeks, decline to a nadir at 20 weeks, and remain at low levels during the rest of pregnancy. However, the placental content of GnRH does not parallel the time course of hCG secretion, and GnRH messenger ribonucleic acid (mRNA) levels in the placenta remain unchanged during pregnancy. These data do not conform with a simple paracrine mechanism of GnRH as a regulator of hCG secretion. We, therefore, examined the potential variation in GnRH receptor gene expression in the placenta, which may account for the GnRH-mediated dynamic pattern of hCG secretion during gestation. First, we established a functional relationship of GnRH and hCG secretion. Using a placental explant culture system, a dose-response effect of hCG secretion was observed in the placental explant at 9 weeks when treated with GnRH ranging from 10(-9)-10(-7) mol/L. The effect of GnRH was completely blocked by a GnRH antagonist (Nal-Glu). The relative responsiveness of hCG secretion to GnRH stimulation at 10(-7) mol/L was further evaluated in placental explants at 6, 9, and 40 weeks gestation. Whereas the 9-week placenta showed a maximal response (> 300%) relative to the 6-week placenta, there was no response in term placenta. Again, the effects of GnRH on hCG secretion were blocked by Nal-Glu, supporting a receptor-mediated event. Localization of mRNA encoding human GnRH receptor in human placenta at 6, 9, 12, 20, and 40 weeks gestation was established by in situ hybridization. The mRNA signals were present in both cytotrophoblast and syncytiotrophoblast cell layers. Signal intensities varied with gestational ages and were abundant at 6 weeks, peaked at 9 weeks, declined at 12 and 20 weeks, and were undetectable at term. The present study demonstrates, for the first time, that GnRH receptor mRNA is expressed in both cytotrophoblasts and syncytiotrophoblasts and exhibits changes paralleling the time course of hCG secretion during pregnancy. These data provide a mechanistic understanding that the paracrine/autocrine regulation of hCG secretion by placental GnRH is mediated through an increase followed by a decline in GnRH receptor gene expression from the first trimester to term placenta.

Expression of inhibin/activin system messenger ribonucleic acids and proteins in ovarian follicles from women with polycystic ovarian syndrome.

The role of inhibin, activin, and follistatin in the pathophysiology of polycystic ovary syndrome (PCOS) was investigated by examining the expression of human inhibin/activin subunit, follistatin, and type II activin receptor (ActRII and -IIB) messenger ribonucleic acid (mRNA) signals (via in situ hybridization) and encoded proteins (via immunocytochemistry) in ovarian follicles (n = 42) from 6 women diagnosed with PCOS. The localization patterns in cellular compartments were compared to those in small antral follicles of comparable size (3-7 mm; n = 40) from 17 normal human ovaries. In small antral follicles of both normal and PCOS ovaries, mRNA signals for all three subunits of inhibin and activin (alpha, beta a, and beta b) were expressed in granulosa cells, whereas in the thecal cell layer, only alpha-subunit mRNA was expressed. The relative intensity of the alpha-subunit mRNA signal was distinctly different in granulosa and thecal cells between PCOS and normal follicles; in small antral follicles of normal ovaries, the alpha-subunit mRNA signal was stronger in the granulosa cell layer than in the thecal cells, and the reverse was found in the polycystic follicles. A light follistatin mRNA signal was found in the granulosa cells of normal small antral follicles, but no follistatin mRNA was detected in any cell type of PCOS follicles. ActRII and -IIB mRNAs were not detected in any cell layer in either normal or PCOS follicles. There were no notable differences in the protein localization pattern of the inhibin/activin system between the PCOS and normal ovaries. In both types of follicles, follistatin and alpha-, beta a-, and beta b-subunit cytoplasmic staining were observed in granulosa cells, as were their corresponding messages, with the exception of the undetectable follistatin mRNA signal in the PCOS follicles. In both normal and PCOS follicles, follistatin and beta a-subunit cytoplasmic staining were occasionally found in thecal interna cells, with no corresponding localization of mRNA, and alpha-subunit protein was not detected in thecal cells despite the presence of the alpha-subunit mRNA. ActRII and -IIB protein localizations were not examined due to the lack of available antisera. These results suggest that granulosa cells of small antral follicles are less active in polycystic than in normal ovaries with respect to inhibin alpha-subunit and follistatin mRNA expression. A consequence of these differences could be an increase in the availability of activin, relative to inhibin, in the arrested follicles in PCOS.

Somatosensory, multisensory, and task-related neurons in cortical area 7b (PF) of unanesthetized monkeys.

1. The goal of this study was to quantitatively characterize the response properties of somatosensory and multisensory neurons in cortical area 7b (or PF) of monkeys that were behaviorally trained to perform an appetitive tolerance-escape task. Particular emphasis was given to characterizing nociceptive thermal responses and correlating such responses to thermal pain tolerance as measured by escape frequency. 2. A total of 244 neurons that responded to somatosensory stimulation alone or to both somatosensory and visual stimulation (multisensory) were isolated and studied in the trigeminal region of cortical area 7b. Thirty neurons responded only to visual stimulation. Thermoreceptive neurons formed approximately 13% (31 of 244) of the neurons that had somatosensory response properties. Thermal nociceptive neurons made up approximately 9% (21 of 244) of the neurons that had somatosensory response properties or approximately 68% (21 of 31) of the neurons that had thermoreceptive response properties. Thermal nociceptive neurons responded either exclusively to noxious thermal stimuli (high-threshold thermoreceptive, HTT) or differentially to nonnoxious and noxious thermal stimuli (wide-range thermoreceptive, WRT). Multimodal HTT neurons had nonnociceptive (low-threshold mechanoreceptive, LTM) and/or nociceptive (nociceptive-specific, wide-dynamic-range) mechanical receptive fields, whereas multimodal WRT neurons had only nonnociceptive (LTM) mechanical receptive fields. Thermal nonnociceptive neurons (low-threshold thermoreceptive, LTT) made up approximately 3% (8 of 244) of the neurons that had somatosensory properties or approximately 26% (8 of 31) of the neurons that were thermoreceptive. The background discharge of two thermoreceptive neurons (6%, 2 of 31) was inhibited by innocuous thermal stimulation. 3. Thermal nociceptive neurons (HTT and WRT) were functionally differentiated by statistical analyses into subpopulations that did encode (HTT-EN, WRT-EN) and did not encode (HTT-NE, WRT-NE) the magnitude of noxious thermal stimulus intensities. The mean slopes and median regression coefficients for the stimulus-response (S-R) functions of HTT-EN and WRT-EN neurons, respectively, were significantly greater than those for the S-R functions of HTT-NE and WRT-NE neurons. In contrast to HTT-NE and WRT-NE neurons, HTT-EN and WRT-EN neurons reliably encoded the magnitude of noxious thermal intensity by grading their mean discharge frequency. 4. The S-R functions of HTT-EN and WRT-EN neurons, unlike those of HTT-NE and WRT-NE neurons, closely approximated stimulus intensity-escape frequency functions.(ABSTRACT TRUNCATED AT 400 WORDS)

Expression of the genes encoding the insulin-like growth factors (IGF-I and II), the IGF and insulin receptors, and IGF-binding proteins-1-6 and the localization of their gene products in normal and polycystic ovary syndrome ovaries.

To discern the potential role of the insulin-like growth factors (IGFs) in polycystic ovary syndrome (PCOS), we examined the expression of the genes encoding the IGFs, IGF receptors (IGFr), insulin receptor (Ir), and IGF-binding proteins (IGFBPs-1-6) as well as the localization of the gene products in specific cellular compartments of normal and PCOS human ovaries. Messenger ribonucleic acid (mRNA) was localized by in situ hybridization with specific 35S-labeled human antisense RNA probes, and protein was detected by immunohistochemistry using specific antisera. Thecal cells, but not granulosa cells (GC), of small antral follicles (3-6 mm) from PCOS ovaries expressed both IGF-I and IGF-II transcripts. Abundant IGF-Ir mRNA was found only in GC, IGF-IIr mRNA was found in both granulosa and thecal cells, and Ir mRNA was detected in all cell types, including granulosa, thecal, and stromal cells. Localization of the gene products revealed no IGF-I immunoreactivity; however, immunostaining for each of the other gene products was colocalized with its corresponding mRNA. The cellular distribution of mRNA and protein in PCOS follicles was indistinguishable from that observed in small antral follicles from normal ovaries. In dominant follicles, however, IGF-I mRNA was no longer detectable, but abundant IGF-II mRNA was expressed exclusively in GC. Although IGF-Ir mRNA was expressed in GC, IGF-IIr mRNA was found in both granulosa and thecal cells. In follicles taken from PCOS ovaries, no IGFBP-1 mRNA was detected, IGFBP-2 mRNA was abundant in both granulosa and thecal cells, moderate IGFBP-3 mRNA was found only in thecal cells, IGFBP-4 and -5 mRNAs were present in all cellular compartments, and IGFBP-6 mRNA was not detected. Localization of the gene products by immunostaining revealed that each protein colocalized with its corresponding mRNA. The cellular distribution of IGFBP mRNA and protein in PCOS follicles was also indistinguishable from that in small antral follicles of normal ovaries, but remarkable differences were found in dominant follicles, where abundant IGFBP-1 mRNA was seen exclusively in GC, IGFBP-2 mRNA in thecal cells, and IGFBP-3 mRNA in both granulosa and thecal cells. Moderate expression of the IGFBP-4 and IGFBP-5 genes was seen in all cell types, including stromal cells, but no IGFBP-6 mRNA was detected. Again, each of the gene products colocalized with its corresponding mRNA. We conclude the following.(ABSTRACT TRUNCATED AT 400 WORDS)

Expression of messenger ribonucleic acids encoding the inhibin/activin system during mid- and late-gestation rat embryogenesis.

We recently demonstrated that inhibin/activin alpha-, beta A-, and beta B-subunit messenger RNAs (mRNAs) are localized in a variety of embryonic rat tissues from 12-20 days post coitum (pc) and reported localizations consistent with possible growth effects of activin during rat embryogenesis. In the present study, we examined the tissue-specific distribution of mRNAs encoding all known players of the inhibin/activin system. In situ hybridization with radiolabeled RNA probes specific for mouse activin receptors (ActRII and ActRIIB), rat follistatin, and rat inhibin/activin subunits was used to examine the spatiotemporal expression of these molecules in adjacent sections of postimplantation rat embryos (8-20 days pc) as well as in midgestation placenta and uterine tissues (8-12 days pc). With the exception of the dorsal root ganglion and salivary gland, alpha- and beta B-subunit mRNAs were found exclusively in reproductive tissues (brain, pituitary, and/or gonads). beta A-Subunit mRNA signal was observed in the brain and gonads as well as in a variety of other tissues during embryogenesis. ActRII mRNA was found exclusively in neuronal tissue from 14 days pc until birth. ActRIIB mRNA was also found in brain, spinal cord, and ganglion, but usually appeared earlier in development than the ActRII message. ActRIIB message was also expressed in a number of other tissues, in some cases along with beta A-subunit mRNA. In these tissues, ActRIIB expression was confined to epithelial and endothelial cell types. Follistatin message was observed in all tissues (except the heart and vessels) localizing beta A-subunit and/or ActRIIB but not in the same cell type. Outside the embryo, beta A-subunit mRNA was localized in the decidua capsularis during midgestation, whereas ActRIIB message was found in placenta as early as 9 days pc. Expression of follistatin message was apparent in decidua from 8-11 days pc, then disappeared from this tissue and was abundant in myometrium at 12 days pc. These data suggest that: 1) inhibin and activin regulate aspects of the fetal reproductive system, whereas activin A may regulate the growth and differentiation of many embryonic tissues; 2) ActRII and ActRIIB serve different roles during development of the rat embryo; and 3) follistatin is in a position to modulate the effects of activin during postimplantation rat embryogenesis.

Developmental changes in atrial natriuretic factor content and localization of its messenger ribonucleic acid in ovine fetal heart.

OBJECTIVES: The current study investigated the developmental changes in atrial natriuretic factor peptide content and messenger ribonucleic acid localization in the atria and ventricles of the ovine fetus throughout the second half of gestation. STUDY DESIGN: Ovine fetuses from 67 to 146 days' gestation (term 147 days) and newborn lambs were used for the study. Tissue atrial natriuretic factor contents were determined by radioimmunoassay, and atrial natriuretic factor messenger ribonucleic acid distribution was determined by in situ hybridization. RESULTS: In fetal atria, atrial natriuretic factor peptide levels were much greater than those in the ventricles. The levels in the atria increased with advancing gestation from 70 to 140 days, reflecting an increase in weight of the atrial chambers. A similar trend was not observed in the ventricles. In the atria, atrial natriuretic factor peptide content (per unit protein) reached high levels at 100 to 110 days' gestation; this was associated with an increase in level of atrial natriuretic factor gene expression. In the ventricles, atrial natriuretic factor peptide content and gene expression were very low throughout the second half of gestation, except for a peak in content that occurred at 100 days. Atrial natriuretic factor messenger ribonucleic acid abundance was much greater in the atria than in the ventricles in fetuses from 90 to 130 days' gestation. The distribution of atrial natriuretic factor messenger ribonucleic acid was homogeneous throughout the thickness of the atria and ventricles of the fetal heart. CONCLUSION: During the second half of gestation in the ovine fetus, the expression of atrial natriuretic factor messenger ribonucleic acid in the atria and ventricles paralleled the appearance of the peptide.