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Biomedical subjects

J F Wilber

Publications and source records attributed to J F Wilber.

At least 37 records · Page 2Linked to original sources
PubMed1993

Identification of thyrotropin-releasing hormone receptor messenger RNA in the rat central nervous system and eye.

TRH exerts a wide variety of neuropharmacological actions by interacting with specific receptors in the central nervous system (CNS). Specific binding sites for TRH have been identified also in the mammalian retina. However, whether TRH receptors (TRH-R) in the brain and retina are identical in structure with those in the anterior pituitary gland is presently unknown. In this study, TRH-R gene expression was examined by Northern blot analysis in the CNS and eye using a cloned rat pituitary TRH-R cDNA. Northern analysis demonstrated a specific hybridization band of approximately 3.8 kb in hypothalamus, cerebrum, cerebellum, brain stem, spinal cord, and eye, indistinguishable from that characterized in pituitary gland. These data strongly support the hypothesis that a TRH receptor similar or identical to that cloned from the pituitary occurs in the retina and throughout the CNS.

Animals↗
PubMed1992

Preprothyrotropin-releasing hormone mRNA and TRH are present in the rat heart.

PreproTRH mRNA has been identified in rat cardiac tissues by Northern analyses and RNase protection assays with a specific rat 32P-TRH cRNA probe. Densitometric analyses revealed that atrial ppTRH mRNA concentrations were approximately five-fold greater than those of the ventricles. TRH concentrations (RIA), by contrast, were two-fold higher in ventricles. These data suggest that TRH and TRH mRNA are present in the rat heart, but their concentrations are dissociated, possibly because of differential post-transcriptional or post-translational processing. TRH is postulated to play an autocrine or paracrine role in cardiac physiology.

Animals↗
PubMed1992

Influences of hypothyroidism on TRH concentrations and preproTRH mRNA levels in rat hypothalamus: a simple and reliable method to detect preproTRH mRNA level.

To gain further insight into the regulation of hypothalamic TRH by thyroid hormones, we measured TRH concentration in specific hypothalamic nuclei and preproTRH mRNA levels in the anterior hypothalamus. Adult male rats were decapitated 1, 7, 14 days after thyroidectomy. Micropunches by the method of Palkovitz from seven hypothalamic nuclei and median eminence were used for measurement of TRH by radioimmunoassay. As compared with normal levels, TRH concentration significantly decreased in the median eminence and five hypothalamic nuclei including paraventricular nucleus (PVN), posterior nucleus, anterior nucleus, arcuate nucleus, and ventromedial nucleus pars lateralis, by 7 days after thyroidectomy. No significant changes were observed in dorsomedial nucleus or ventromedial nucleus pars medialis until 14 days after thyroidectomy. A rapid and simple method to detect specific mRNA for preproTRH was developed using the polymerase chain reaction and a single anterior hypothalamic section. PreproTRH mRNA levels in the anterior hypothalamus increased approximately twice 7 days after thyroidectomy. These data indicate that thyroidectomy caused a marked increase in preproTRH mRNA levels of the anterior hypothalamus, while it significantly reduced TRH concentrations not only in PVN and median eminence but also in other specific hypothalamic nuclei, suggesting that these nuclei might be involved in the thyrotropin regulation in the hypothalamus.

Animals↗
PubMed1991

Assignment of human preprothyrotropin-releasing hormone (TRH) gene to chromosome 3.

The human gene encoding preproTRH (thyrotropin-releasing hormone) was assigned to chromosome 3, using human-Chinese hamster ovary somatic cell hybrids, analyzed by Southern hybridizations. Hybridization was carried out with a 32P-labeled human preproTRH cDNA labeled by the method of random priming. Hybridization of the cDNA probe to a human specific 4.8-kb DNA fragment of EcoRI-digested WBC DNA was used to localize the human preproTRH gene. No hybridization, by contrast, was seen with human preproTRH cDNA probe and hamster DNA after EcoRI treatment. Results from 29 somatic cell hybrids corroborated unequivocally that the human preproTRH gene can be assigned to human chromosome 3.

Blotting, Southern↗
PubMed1990

Reciprocal regulation of preprothyrotropin-releasing hormone (TRH) mRNA in the rat anterior hypothalamus by thyroid hormone: dissociation from TRH concentrations during hypothyroidism.

TRH mRNA has not been quantified concomitantly with TRH itself to examine graded effects of thyroid hormones (TH) upon TRH gene transcriptional regulation and post-transcriptional expression. To examine such TH effects, rats were rendered thyrotoxic with L-T3 (50 micrograms per 100g body weight) or hypothyroid by total thyroidectomy. After decapitation, frozen coronal brain sections were prepared in a matrix for hypothalamic micropunches. PreProTRH mRNA was quantified in punch pools by slot-blot hybridization and densitometry, using a 32P 1.2Kb rat riboprobe (gift of Drs. S. Lee and R. Goodman). T3 toxicosis resulted in parallel reductions in PreProTRH mRNA (-45%) and TRH concentrations determined by RIA (-43%), 1.1 versus 1.9 ng/mg protein, p less than 0.01. Conversely, elevations in PreProTRH mRNA were stimulated by hypothyroidism on Day 14 (+32%). However, TRH concentrations were reduced paradoxically from 2.2 +/- 0.05 ng/mg protein to 0.68 +/- 0.03, p less than 0.01. No changes, in contrast, were identified in whole hypothalamic extracts in either PreProTRH mRNA or TRH after T3 treatment. It is concluded that TH do exert inhibitory effects upon PreProTRH mRNA transcription, and in the specific hypothalamic nucleus concerned with thyroid regulation (PVN). The failure of TRH concentrations to rise concordantly with activated TRH gene transcription in hypothyroidism suggests that TH may exert inhibitory effects upon PreProTRH mRNA translation, in addition to inhibition of TRH gene transcription and TRH secretion.

Animals↗
PubMed1990

Acute alterations of cyclo(His-Pro) levels after oral ingestion of glucose.

We analyzed the response of plasma cyclo(His-Pro) (CHP) (N = 14), insulin (N = 8), and glucose (N = 8) to oral ingestion of 75 grams of glucose in normal volunteers. Mean Fasting CHP levels prior to glucose were 614 +/- 112 pg/ml (+/-SE). After glucose ingestion there occurred an acute rise of plasma CHP within 15 minutes (12/14) followed by a fall in plasma concentrations to below baseline values (11 of 14). The mean of highest levels within the first 15 minutes after glucose ingestion was 1035 +/- 300 pg/ml (+/-SE), significantly above baseline (p = 0.002). The mean of lowest values below baseline was 490 +/- 94 pg/ml (+/-SE), p less than 0.001. We conclude that levels of CHP are acutely and reversibly altered by the ingestion of glucose. The possible significance of these perturbations in CHP concentrations is discussed.

Blood Glucose↗
PubMed1990

Change in circulating cyclo(His-Pro) concentrations in rats after ingestion of oral glucose compared to intravenous glucose and controls.

Changes in circulating cyclo(His-Pro) (CHP) levels after ingestion of 0.5 g/kg of glucose (oral) were measured in 32 Sprague Dawley rats at varying time points between 0 (before glucose) and 60 minutes. Each rat provided one time point. CHP levels displayed a biphasic response rising from 933 +/- 56 pg/ml to 1083 +/- 287 pg/ml (p greater than 0.05), followed by a fall to 543 +/- 46 pg/ml (p = 0.002) and then a recovery to near baseline values. To further investigate this response, a separate group of rats were sampled sequentially after 3 g/kg oral glucose (n = 7), 0.6 g/kg I.V. glucose (n = 6) or controls given saline only (n = 14). CHP levels were significantly elevated in the oral glucose group at 5 (p = 0.048) and 12 (p = 0.02) minutes compared to controls, while the i.v. glucose group was not statistically different from controls. Comparison of the mean of the highest incremental response to baseline values in each group revealed a greater excursion in CHP levels after oral (p = 0.027) glucose than after I.V. (p = 0.08) glucose. These data suggest CHP is acutely and reversibly elevated after glucose ingestion in rats and that the response is greater after oral glucose than after i.v. glucose.

Administration, Oral↗
PubMed1990

Cloning and structure of human genomic DNA and hypothalamic cDNA encoding human prepro thyrotropin-releasing hormone.

The gene encoding human preproTRH was isolated from a human lung fibroblast genomic DNA library with a rat prepro TRH cDNA fragment. The transcriptional unit is 3.3 kilobases in size and contains three exons interrupted by two introns of approximately 1050 and 650 base pairs, respectively. Exon 1 encodes the 5'-untranslated region of the mRNA, exon 2 the putative signal sequence and the initial portion of propeptide, and exon 3 encodes the remainder of the propeptide, which contains six copies of the TRH sequence in contrast to five copies in the rat preproTRH gene. The predicted human preproTRH peptide structure has 242 amino acids compared to 255 amino acids in the rat. Homology with rat preproTRH is 73.3% and 59.5% at the nucleic acid and amino acid levels, respectively. Intron-exon splicing sites and 5' and 3' mRNA borders were confirmed rigorously by sequencing a human preproTRH cDNA using the polymerase chain reaction and human hypothalamic cDNA.

Amino Acid Sequence↗
PubMed1989

Radioimmunoassay of cyclo(His-Pro) in unextracted human plasma: report of a normal range and definition of factors critical for successful assay.

A radioimmunoassay for cyclo(His-Pro) (CHP) in unextracted human plasma that can detect 40 pg has been developed. Elution profile of CHP-like immunoreactivity (CHP-LI) corresponded precisely to those of [3H] cyclo(His-Pro) by both high pressure liquid chromatography and Sephadex G-25 column chromatography. In addition plasma CHP-LI exhibited close immunoidentity with authentic CHP. Charcoal treatment of plasma containing [3H] cyclo(His-Pro) resulted in loss of both CHP-LI and [3H] cyclo(His-Pro) activity. Plasma frozen at draw and assayed at 0, 6 and 24 hours displayed no change in CHP-LI while an aliquot from the same sample maintained at 4 degrees C and assayed at the same time intervals showed a 50% rise (0 hrs-856 +/- 47 pg/ml; 24 hrs-1288 +/- 85 pg/ml) (+/- SE) over 24 hours. In addition, plasma drawn from 14 volunteers and immediately frozen until assay yielded a mean CHP value of 829 +/- 64 pg/ml (+/- SE) while that of 14 volunteers that was maintained at 25 degrees C for 90 minutes was significantly higher at 1085 +/- 34 pg/ml (+/- SE) (p less than 0.03). Taken together, these data suggest that CHP can be easily measured in a direct RIA of human plasma, however, failure to maintain the sample frozen from the time of draw until assay may yield spuriously elevated values.

Adult↗
PubMed1989

Histidyl proline diketopiperazine (Cyclo [His-Pro]) in eating disorders.

The authors explored the role of Cyclo [His-Pro] (CHP) in 50 adolescents who fulfilled DSM III diagnostic criteria for anorexia nervosa and bulimia. CHP, a relatively new neuro modulator which has a role in inducing satiety, was assayed in serum during routine blood work. CHP levels correlated significantly with weight in restrictor (AN-R) (R = -0.449, P less than 0.05) and bulimic anorexics (AN-B) (R = +0.489, P less than 0.01). There was no significant correlation in normal-weight bulimics (NWB) (R = +0.556, P less than 0.01). It did not correlate with weight in a depressed control group. Longitudinal data on six patients showed a similar relationship between weight, purging and CHP binge/purge activity and/or weight changes of around 2 kilograms correlated with average CHP changes of 42%. Clinical material suggests changes in satiety during CHP changes consistent with a satiety disturbance model of these disorders.

Adolescent↗
PubMed1989

TRH analog administration increases endogenous TRH levels in the central nervous system.

Experiments were carried out to increase the endogenous levels of TRH in brain and spinal cord. After 7 days of continuous oral ingestion of TRH analogs (D-pGlu-Leu-ProNH2, pGlu-Leu-Pro, pGlu-Pro-ProNH2, or pGlu-Leu-pyrrolidide), spinal cord and brain levels of TRH were elevated, whereas the content of TRH metabolite, cyclo(His-Pro), was decreased. In addition, the TRH analogs inhibited in vitro metabolism of [3H-Pro]-TRH by brain and spinal cord extracts. Our data show that certain TRH analogs can elevate tissue levels of TRH by inhibiting its metabolism. Furthermore, because these analogs do not affect interaction between TRH and its receptor, the elevated TRH can effectively enhance TRH-related bioactivity.

Animals↗
PubMed1989

Exogenous triiodothyronine lowers thyrotropin-releasing hormone concentrations in the specific hypothalamic nucleus (paraventricular) involved in thyrotropin regulation and also in posterior nucleus.

Recent evidence indicates that thyroid hormones can regulate thyrotropin secretion in vivo in part by inhibiting thyrotropin-releasing hormone (TRH) secretion itself. Therefore, to explore whether triiodothyronine (T3) interacts with the specific hypothalamic area involved in thyrotropin (TSH) secretory regulation, the paraventricular nucleus (PVN), Palkovitz micropunches from eight nuclear regions were obtained from 1,000-microns frozen coronal brain slices for immunoassay determinations of TRH. Rats were treated either with parenteral L-T3 for 6 days to induce experimental thyrotoxicosis, or 0.15 M saline. The induction of thyrotoxicosis was confirmed by demonstrating that mean plasma TSH concentrations fell from 108 to less than 10 microU/ml (p less than 0.01). TRH concentrations in the PVN were reduced concomitantly after L-T3 from 1.9 to 1.1 ng/mg protein (p less than 0.05). No reductions in TRH concentrations during T3 treatment occurred in other nuclear groupings except in the posterior hypothalamic nucleus. Total TRH content in the median eminence declined also in T3-treated animals from 1.77 to 1.29 ng, representing a 32% reduction (p less than 0.01). No significant change was seen in the median eminence content of the TRH structurally related dipeptide, cyclo(His-Pro). The data herein indicate that experimental thyrotoxicosis in the rat is associated with a selective reduction in TRH concentrations in the PVN, documenting T3 effects upon hypothalamic TRH metabolism per se.

Animals↗
PubMed1987

Thyrotropin-releasing hormone and cyclo (His-Pro)-like immunoreactivities in the cerebrospinal fluids of 'normal' infants and adults, and patients with various neuropsychiatric and neurologic disorders.

Levels of thyrotropin-releasing hormone (TRH) - and cyclo(His-Pro) (CHP)-like immunoreactivities and the activity of enzyme Pyroglutamate aminopeptidase (PAPase) were measured in cerebrospinal fluid (CSF) of over 100 normal adults (NA) and infants, and adult patients with various neurologic and neuropsychiatric disorders (NNDA). Levels of TRH and CHP in CSF of over 70% of the NA group were below 50 and 500 pg/ml respectively. The TRH- and CHP-like immunoreactivities in the remainder of the 30% of NA specimens exhibiting higher peptide concentrations were enzymatically and chromatographically characterized and were found to behave like authentic peptides. The levels of both of these peptides were significantly elevated in the CSF of most of the NNDA patients. An elevation in the CSF level of CHP was significantly correlated with the level of TRH, but not PAPase. Results from this study suggest that CSF elevation of TRH level may be due to a nonspecific response to stress that may be associated with hospitalization, myelogram procedure, and/or the neurologic and neuropsychiatric diseases for which the patients were admitted.

Adult↗
PubMed1987

Thyrotropin-releasing hormone (TRH) in murine motor neuron disease (the wobbler mouse).

Clinical benefits of thyrotropin-releasing hormone (TRH) were tested in wobbler mice, an animal model of motor neuron disease. After the disease was clinically recognized at 3-4 weeks, the animals were divided into two groups, each group consisting of 5 pairs of wobbler mice and normal littermates. TRH (50 mg/kg) and normal saline (NS) were injected intraperitoneally daily, 6 times per week for 9 weeks, in a double-blind study. Weekly assessments consisted of front paw grip strength, push walking, body weight, and semiquantitative grading. At the end of the trial, the brain and spinal cord were sampled to measure TRH and cyclo (His-Pro) concentrations. Progression of motor neuron disease was evident in wobbler mice, regardless of treatment. Descriptive semiquantitative gradings showed the tendency of improvement in TRH-treated wobbler mice. In saline-injected controls, TRH levels in the cervical spinal cord were significantly increased (P less than 0.01) in wobbler mice compared to littermates. However, with TRH treatment, there was no significant difference in TRH and cyclo (His-Pro) levels in any neural tissue between wobbler and controls. The lack of clinical benefits with TRH in wobbler mice may be due to increased TRH levels found in diseased spinal cord in murine motor neuron disease.

Amyotrophic Lateral Sclerosis↗