Effects of ghrelin on neuronal activity in the ventromedial nucleus of the hypothalamus in infantile rats: An in vitro study

Ghrelin is an endogenous ligand for the growth hormone (GH) secretagogue (GHS) receptor (GHS-R) and a potent stimulant for GH secretion even in infantile rats before puberty. Although the ventromedial nucleus of the hypothalamus (VMH) might be a site of action for ghrelin to induce GH release, the electrophysiological effect of ghrelin on VMH neurons in infantile rats remains to be elucidated. Thus, the purpose of the present study was to investigate the effect of ghrelin on VMH neurons using hypothalamic slices of infantile rats. Ghrelin excited a majority of VMH neurons in a concentration-dependent manner. VMH neurons that were excited by GH releasing peptide-6 (GHRP-6), a synthetic GHS, were also excited by ghrelin and vice versa. Repeated application of ghrelin to the same VMH neuron decreased progressively the excitatory responses depending on the number of times it was administered. The excitatory effect of ghrelin on VMH neurons in normal artificial cerebrospinal fluid (ACSF) persisted in low Ca²⁺-high Mg²⁺ ACSF. The present results indicate that (1) ghrelin excites a majority of VMH neurons dose-dependently and postsynaptically and (2) the excitatory effects of ghrelin are mimicked by GHRP-6 and desensitized by repeated applications of ghrelin.     

Plant gene responses to frequency-specific sound signals

We identified a set of sound-responsive genes in plants using a sound-treated subtractive library and demonstrated sound regulation through mRNA expression analyses. Under both light and dark conditions, sound up-regulated expression of rbcS and ald. These are also light-responsive genes and these results suggest that sound could represent an alternative to light as a gene regulator. Ald mRNA expression increased significantly with treatment at 125 and 250 Hz, whereas levels decreased significantly with treatment at 50 Hz, indicating a frequency-specific response. To investigate whether the ald promoter responds to sound, we generated transgenic rice plants harboring a chimeric gene comprising a fusion of the ald promoter and GUS reporter. In three independent transgenic lines treated with 50 or 250 Hz for 4 h, GUS mRNA expression was up-regulated at 250 Hz, but down-regulated at 50 Hz. Thus, the sound-responsive mRNA expression pattern observed for the ald promoter correlated closely with that of ald, suggesting that the 1,506 bp ald promoter is sound-responsive. Therefore, we propose that in transgenic plants, specific frequencies of sound treatment could be used to regulate the expression of any gene fused to the ald promoter.     

Impact of the TCR Signal on Regulatory T Cell Homeostasis,Function, and Trafficking

Signaling through the T cell antigen receptor (TCR) is important for the homeostasis of naïve and memory CD4⁺ T cells. The significance of TCR signaling in regulatory T (Treg) cells has not been systematically addressed. Using an Ox40-cre allele that is prominently expressed in Treg cells, and a conditional null allele of the gene encoding p56^Lck, we have examined the importance of TCR signaling in Treg cells. Inactivation of p56^Lck resulted in abnormal Treg homeostasis characterized by impaired turnover, preferential redistribution to the lymph nodes, loss of suppressive function, and striking changes in gene expression. Abnormal Treg cell homeostasis and function did not reflect the involvement of p56^Lck in CD4 function because these effects were not observed when CD4 expression was inactivated by Ox40-cre.The results make clear multiple aspects of Treg cell homeostasis and phenotype that are dependent on a sustained capacity to signal through the TCR.     

Genetic polymorphism, mapping, and characterization of isocitrate dehydrogenase in Anopheles quadrimaculatus

The isocitrate dehydrogenase-2 (Idh-2) locus of Anopheles quadrimaculatus was analyzed genetically and the enzyme was characterized physiochemically. Three-point testcrosses involving chromosome 3 markers showed that in female hybrids the gene sequence and the map distances are: nonstripe (st)--6.8--Idh-2--43.5--short antenna (Sa). Reduced recombination frequencies were observed in male hybrids (st--3.4--Idh-2--25.5--Sa). Idh-2 activity gradually increases during development and reaches a peak intensity in adults. Maximum enzyme activity of Idh-2 was obtained at pH 7.5. One-minute heat treatment at 50 degrees C caused about 50 percent reduction of IDH-2. Ethylene diamine tetraacetic acid (EDTA 5 mM) and p-chloromercuribenzoate (pCMB 10(-5)M) caused complete loss of activity of IDH-2, but pretreatment of the enzyme in situ with mercaptoethanol protected the activity of allozymes from inhibition by pCMB treatment.     

Specific antigens inhibit plasmacytoma cells bearing phosphorylcholine-binding myeloma proteins.

The influence of a synthetic adjuvant active glycopeptide, N-acetylmuramyl-l-alanyl-d-isoglutamine (MDP), and of some of its analogs on the in vitro immune response to sheep red blood cells was studied using Mishell and Dutton in vitro stimulation system. When MDP and adjuvant active analogs were incubated with normal spleen cells, increased cell recovery was observed after 3 or 4 days of culture, showing a good correlation between the adjuvant activity in vivo and the enhancement of cell viability in vitro. The analogs which were found to have an adjuvant activity in vivo were equally effective in stimulating in vitro both the background hemolytic PFC and the immune response to sheep red blood cells. However, those which were inactive in vivo were effective in vitro but only at high concentration levels.     

Multiple forms of alkaline phosphatase in mouse preimplantation embryos

Mouse embryos were extracted with 0.5% Triton X-100 and subjected to cellulose acetate electrophoresis. In fertilized eggs, two forms of alkaline phosphatase (ALP), a slow-moving form and a fast-moving form, were observed. As cleavage proceeded, the fast-moving form disappeared, and the slow-moving form, the mobility of which was similar to that of the slow-moving form of the kidney, became gradually dominant up to the blastocyst stage (named 'embryonic' form). With blastulation, another fast-moving form showing a similar mobility to the lung ALP began to appear in blastocysts and showed a transient dominance in hatched blastocysts. After implantation, both the embryonic form and the fast-moving form gradually faded, and were eventually replaced by the new form, which may be named 'fetal form' in Day 7 embryos. These results clearly demonstrated that ALP activity does exist in embryos at all stages of preimplantation development. Moreover, the changes in multiple forms of ALP correlated with embryonic development may suggest that these multiple forms may have differential roles in the process of early development.