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Focused-beam microwave irradiation (FBMI) is a relatively new method for euthanasia of small mammals and is available to most researchers. Compared with CO2inhalation, this method of euthanasia has the advantage of preserving fast-degrading metabolites. But differences in brain RNA quantity and quality, gene expression and histology in mice euthanized by CO2inhalation versus FBMI have not been investigated. Here the authors report that a smaller quantity of RNA was isolated from brains of mice euthanized by FBMI compared with those of mice euthanized by CO2inhalation. They also found relative differences in the levels of the expression of some genes. These studies suggest that either method can be used for histological analysis or RNA isolation, but the authors caution against combining the techniques within a single study on gene expression.

The mouse is the animal most commonly used to study the underlying causes of and treatments for obesity. The author reviews many of the issues that should be considered by all involved in research with mice expressing this phenotype, and describes some procedures exclusive to obesity research.

ABSTRACTNhlh1is a basic helix-loop-helix transcription factor whose expression is restricted to the nervous system and which may play a role in neuronal differentiation. To directly study Nhlh1function, we generated null mice. Homozygous mutant mice were predisposed to premature, adult-onset, unexpected death. Electrocardiograms revealed decreased total heart rate variability, stress-induced arrhythmia, and impaired baroreceptor sensitivity. This predisposition to arrhythmia is a likely cause of the observed death in the mutant mice. Heterozygosity for the closely related transcription factor Nhlh2increased the severity of the Nhlh1-null phenotype. No signs of primary cardiac structural or conduction abnormalities could be detected upon necropsy of the null mice. The pattern of altered heart rhythm observed in basal and experimental conditions (stress and pharmacologically induced) suggests that a deficient parasympathetic tone may contribute to the arrhythmia in the Nhlh1-null mouse. The expression of Nhlh1in the developing brain stem and in the vagal nuclei in the wild-type mouse further supports this hypothesis. The Nhlh1mutant mouse may thus provide a model to investigate the contribution of the autonomic nervous system to arrhythmogenesis.

Nhlh1is a basic helix-loop-helix transcription factor whose expression is restricted to the nervous system and which may play a role in neuronal differentiation. To directly study Nhlh1function, we generated null mice. Homozygous mutant mice were predisposed to premature, adult-onset, unexpected death. Electrocardiograms revealed decreased total heart rate variability, stress-induced arrhythmia, and impaired baroreceptor sensitivity. This predisposition to arrhythmia is a likely cause of the observed death in the mutant mice. Heterozygosity for the closely related transcription factor Nhlh2increased the severity of the Nhlh1-null phenotype. No signs of primary cardiac structural or conduction abnormalities could be detected upon necropsy of the null mice. The pattern of altered heart rhythm observed in basal and experimental conditions (stress and pharmacologically induced) suggests that a deficient parasympathetic tone may contribute to the arrhythmia in the Nhlh1-null mouse. The expression of Nhlh1in the developing brain stem and in the vagal nuclei in the wild-type mouse further supports this hypothesis. The Nhlh1mutant mouse may thus provide a model to investigate the contribution of the autonomic nervous system to arrhythmogenesis.

In the past few years, there has been exponential growth in our knowledge of genes that control food intake and metabolism. Most of this research has demonstrated either an increased or decreased expression of these “obesity genes” in response to changes in nutritional status. Ultimately, these changes reflect modifications in the rate of gene transcription, mRNA stability, translation initiation, or posttranslational processing. Few laboratories have examined specifically which of these molecular mechanisms are responsible for obesity gene regulation, and thus, the field is wide open for exploration. In addition, it is possible that some forms of human obesity may be caused by inherited mutations in transcription factors or other regulatory molecules rather than base pair mutations in the obesity genes themselves. This article focuses on the regulation of the leptin receptor, NPY, and POMC genes, and explores what is known about the regulation of these obesity genes in response to food intake or changes in body fat stores. Connections between regulation of these genes and some inherited forms of human obesity are made.

The family of basic helix-loop-helix (bHLH) genes comprises transcription factors involved in many aspects of growth and development. We have previously described two bHLH transcription factors, Nhlh1 and Nhlh2 (originally named NSCL1 and NSCL2)1–3. The nucleotide and predicted protein sequences of Nhlh1 and Nhlh2 are homologous within their bHLH domain where there are only three conservative amino acid differences2,3. During murine embryogenesis, Nhlh1 and Nhlh2 share an overlapping but distinct pattern of expression in the developing nervous system1,2. To improve our understanding of the role of these genes during neurogenesis, we have generated mice containing targeted deletions of both genes and here describe our results for Nhlh2. Loss of Nhlh2 results in a disruption of the hypothalamic-pituitary axis in mice. Male Nhlh2−/ −mice are microphallic, hypogonadal and infertile with alterations in circulating gonadotropins, a defect in spermatogenesis and a loss of instinctual male sexual behaviour. Female Nhlh2−/ −mice reared alone are hypogonadal, but when reared in the presence of males, their ovaries and uteri develop normally and they are fertile. Both male and female homozygotes exhibit progressive adult-onset obesity. Nhlh2 is expressed in the ventral-medial and lateral hypothalamus, Rathke's pouch and in the anterior lobe of the adult pituitary. Our results support a role for Nhlh2 in the onset of puberty and the regulation of body weight metabolism.