Computer program sequence for analysis and summary of radioimmunoassay data

A sequence of three programs is described for efficient design and analysis of radio-immunoassays. The first program designs the assay; the second program analyzes all available data according to the organizational base provided by the first program; and the third program summarizes the results of several assays in either tabular or graphical form. The analysis program uses a logit response-log dose transformation to obtain a linear inhibition curve for all preparations assayed at multiple levels. The curves are subjected to a weithted, least squares, regression analysis. All curves are tested for linearity and parallelism with the curve chosen as standard, and a weighted mean potency estimate with error limits is computed for all preparations run at single or multiple dose levels with any degree of replication.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/34088/1/0000369.pd

Molecular determinants of a competent bovine corpus luteum: first vs final wave dominant follicles

Reproductive management in cattle requires the synchrony of follicle development and oestrus before insemination. However, ovulation of follicles that have not undergone normal physiological maturation can lead to suboptimal luteal function. Here, we investigated the expression of a targeted set of 47 genes in (a) a first-wave vs final-wave dominant follicle (DF; the latter destined to ovulate spontaneously) and (b) 6-day-old corpora lutea (CLs) following either spontaneous ovulation or induced ovulation of a first-wave DF to ascertain their functional significance for competent CL development. Both the mass and progesterone-synthesising capacity of a CL formed following induced ovulation of a first-wave DF were impaired. These impaired CLs had reduced expression of steroidogenic enzymes (e.g. STAR and HSD3B1), luteotrophic receptors (LHCGR) and angiogenic regulators (e.g. VEGFA) and increased expression of BMP2 (linked to luteolysis). Relative to final-wave DFs, characteristic features of first-wave DFs included reduced oestradiol concentrations and a reduced oestradiol:progesterone ratio in the face of increased expression of key steroidogenic enzymes (i.e. CYP11A1, HSD3B1 and CYP19A1) in granulosa cells and reduced expression of the HDL receptor SCARB1 in thecal cells. Transcripts for further components of the TGF and IGF systems (e.g. INHA, INHBA, IGF2R and IGFBP2) varied between the first- and final-wave DFs. These results highlight the importance of hormones such as progesterone interacting with local components of both the TGF and IGF systems to affect the maturation of the ovulatory follicle and functional competency of the subsequent CL

Can luteal regression be reversed?

The corpus luteum is an endocrine gland whose limited lifespan is hormonally programmed. This debate article summarizes findings of our research group that challenge the principle that the end of function of the corpus luteum or luteal regression, once triggered, cannot be reversed. Overturning luteal regression by pharmacological manipulations may be of critical significance in designing strategies to improve fertility efficacy

Glucose Metabolism In Vivo in Four Commonly Used Inbred Mouse Strains

OBJECTIVE—To characterize differences in whole-body glucose metabolism between commonly used inbred mouse strains

Leptin Receptor Signaling and Action in the Central Nervous System

The increasing incidence of obesity in developed nations represents an ever‐growing challenge to health care by promoting diabetes and other diseases. The discovery of the hormone, leptin, a decade ago has facilitated the acquisition of new knowledge regarding the regulation of energy balance. A great deal remains to be discovered regarding the molecular and anatomic actions of leptin, however. Here, we discuss the mechanisms by which leptin activates intracellular signals, the roles that these signals play in leptin action in vivo, and sites of leptin action in vivo. Using “reporter” mice, in which LRb‐expressing (long form of the leptin receptor) neurons express the histological marker, β‐galactosidase, coupled with the detection of LRb‐mediated signal transducer and activator of transcription 3 signaling events, we identified LRb expression in neuronal populations both within and outside the hypothalamus. Understanding the regulation and physiological function of these myriad sites of central leptin action will be a crucial next step in the quest to understand mechanisms of leptin action and energy balance.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/93692/1/oby.2006.310.pd

Dysregulation of the Norepinephrine Transporter Sustains Cortical Hypodopaminergia and Schizophrenia-Like Behaviors in Neuronal Rictor Null Mice

A novel animal model highlights the link between Akt dysfunction, reduced cortical dopamine function, norepinephrine transporters, and schizophrenia-like behaviors

The Nutritional Induction of COUP-TFII Gene Expression in Ventromedial Hypothalamic Neurons Is Mediated by the Melanocortin Pathway

BACKGROUND: The nuclear receptor chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is an important coordinator of glucose homeostasis. We report, for the first time, a unique differential regulation of its expression by the nutritional status in the mouse hypothalamus compared to peripheral tissues. METHODOLOGY/PRINCIPAL FINDINGS: Using hyperinsulinemic-euglycemic clamps and insulinopenic mice, we show that insulin upregulates its expression in the hypothalamus. Immunofluorescence studies demonstrate that COUP-TFII gene expression is restricted to a subpopulation of ventromedial hypothalamic neurons expressing the melanocortin receptor. In GT1-7 hypothalamic cells, the MC4-R agonist MTII leads to a dose dependant increase of COUP-TFII gene expression secondarily to a local increase in cAMP concentrations. Transfection experiments, using a COUP-TFII promoter containing a functional cAMP responsive element, suggest a direct transcriptional activation by cAMP. Finally, we show that the fed state or intracerebroventricular injections of MTII in mice induce an increased hypothalamic COUP-TFII expression associated with a decreased hepatic and pancreatic COUP-TFII expression. CONCLUSIONS/SIGNIFICANCE: These observations strongly suggest that hypothalamic COUP-TFII gene expression could be a central integrator of insulin and melanocortin signaling pathway within the ventromedial hypothalamus. COUP-TFII could play a crucial role in brain integration of circulating signal of hunger and satiety involved in energy balance regulation

The measurement of testosterone and oestradiol-17[beta] using iodinated tracers and incorporating an affinity chromatography extraction procedure

The development of sensitive radioimmunoassays (RIA) for testosterone and oestradiol-17[beta], utilising 125I-radioligands, is described. Use of an homologous bridge at the same site of attachment, for both the radioligand and the steroid-carrier protein conjugate employed in raising antibodies, normally results in a loss of assay sensitivity and precision. This was overcome in the oestradiol assay by utilising an heterologous configuration at the site of attachment (11[alpha] vs 11[beta]). In contrast, for testosterone, even though an homologous bridge and site of attachment was used for the radioligand and the steroid-carrier protein conjugate, a very sensitive assay with extremely high antibody titres (dilution of 1:2 x 106) was achieved. This finding was repeated with a different antiserum suggesting that the "bridge binding" phenomenon may be related to the position of attachment to the steroid molecule. In addition, an antibody-Sepharose 4B affinity chromatography extraction procedure has been developed for both oestradiol and testosterone. This approach allows the measurement of very low concentrations of steroids from large volumes of a variety of biological fluids. As antibody-linked Sepharose 4B uses high concentrations of antibody, steroids of similar structure are extracted from biological fluids. However, the cross-reactivity of these related steroids are very low in the RIA's, ensuring good specificity.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25490/1/0000031.pd