Gonadal steroid hormones act upon specific areas of the vertebrate brain to affect the reproductive physiology and behavior of the animal. Steroid receptors are members of a superfamily of ligand-dependent transcription factors that mediate the effects of steroid hormones by modulating gene expression in the cells containing the receptors. The neuroanatomical distributions of steroid receptor-containing cells have been described for several species by using steroid autoradiography, immunocytochemistry, and more recently in situ hybridization. We have used the polymerase chain reaction to amplify and clone fragments of the estrogen receptor, progesterone receptor, and androgen receptor of whiptail lizards (genus Cnemidophorus). These clones were used to produce probes for use in in situ hybridization assays and to map the neuroanatomical distribution of all three sex steroid hormone receptors in the forebrains of unisexual (C. uniparens) and sexual (C. inornatus) species of whiptail lizards. The distribution of receptor-expressing cells reported here is in general agreement with previous reports in other species with receptor-containing cells concentrated in septal, amygdaloid, cortical, preoptic, and hypothalamic nuclei.

Invertebrate species possess one or two Na ؉ channel genes, yet there are 10 in mammals. When did this explosive growth come about during vertebrate evolution? All mammalian Na ؉ channel genes reside on four chromosomes. It has been suggested that this came about by multiple duplications of an ancestral chromosome with a single Na ؉ channel gene followed by tandem duplications of Na ؉ channel genes on some of these chromosomes. Because a large-scale expansion of the vertebrate genome likely occurred before the divergence of teleosts and tetrapods, we tested this hypothesis by cloning Na ؉ channel genes in a teleost fish. Using an approach designed to clone all of the Na ؉ channel genes in a genome, we found six Na ؉ channel genes. Phylogenetic comparisons show that each teleost gene is orthologous to a Na ؉ channel gene or gene cluster on a different mammalian chromosome, supporting the hypothesis that four Na ؉ channel genes were present in the ancestors of teleosts and tetrapods. Further duplications occurred independently in the teleost and tetrapod lineages, with a greater number of duplications in tetrapods. This pattern has implications for the evolution of function and specialization of Na ؉ channel genes in vertebrates. Sodium channel genes also are linked to homeobox (Hox) gene clusters in mammals. Using our phylogeny of Na ؉ channel genes to independently test between two models of Hox gene evolution, we support the hypothesis that Hox gene clusters evolved as (AB) (CD) rather than {D[A(BC)]}.

Shigella dysenteriae serotype 1, a major cause of bacillary dysentery in humans, can use heme as a source of iron. Genes for the transport of heme into the bacterial cell have been identified, but little is known about proteins that control the fate of the heme molecule after it has entered the cell. The shuS gene is located within the heme transport locus, downstream of the heme receptor gene shuA. ShuS is a heme binding protein, but its role in heme utilization is poorly understood. In this work, we report the construction of a chromosomal shuS mutant. The shuS mutant was defective in utilizing heme as an iron source. At low heme concentrations, the shuS mutant grew slowly and its growth was stimulated by either increasing the heme concentration or by providing extra copies of the heme receptor shuA on a plasmid. At intermediate heme concentrations, the growth of the shuS mutant was moderately impaired, and at high heme concentrations, shuS was required for growth on heme. The shuS mutant did not show increased sensitivity to hydrogen peroxide, even at high heme concentrations. ShuS was also required for optimal utilization of heme under microaerobic and anaerobic conditions. These data are consistent with the model in which ShuS binds heme in a soluble, nontoxic form and potentially transfers the heme from the transport proteins in the membrane to either heme-containing or heme-degrading proteins. ShuS did not appear to store heme for future use.The Shigella spp. are the primary cause of bacillary dysentery in humans. Following ingestion, this pathogen transits the digestive system to the colon, where it invades the cells of the colonic epithelium. It then replicates inside the epithelial cells and spreads to adjacent cells. The resulting inflammation and tissue damage are responsible for the bloody diarrhea characteristic of dysentery.

Strychnine-sensitive glycine receptors (GlyRs) inhibit neurotransmission in the spinal cord and brainstem. To better define the function of this receptor in vivo, we constructed a point mutation that impairs receptor function in the alpha1-subunit and compared these knock-in mice to oscillator (spdot) mice lacking functional GlyR alpha1-subunits. Mutation of the serine residue at amino acid 267 to glutamine (alpha1S267Q) results in a GlyR with normal glycine potency but decreased maximal currents, as shown by electrophysiological recordings using Xenopus oocytes. In addition, single-channel recordings using human embryonic kidney 293 cells indicated profoundly altered properties of the mutated GlyR. We produced knock-in mice bearing the GlyR alpha1 S267Q mutation to assess the in vivo consequences of selectively decreasing GlyR efficacy. Chloride uptake into brain synaptoneurosomes from knock-in mice revealed decreased responses to maximally effective glycine concentrations, although wild-type levels of GlyR expression were observed using 3H-strychnine binding and immunoblotting. A profound increase in the acoustic startle response was observed in knock-in mice as well as a "limb clenching" phenotype. In contrast, no changes in coordination or pain perception were observed using the rotarod or hot-plate tests, and there was no change in GABA(A)-receptor-mediated chloride uptake. Homozygous S267Q knock-in mice, like homozygous spdot mice, exhibited seizures and died within 3 weeks of birth. In heterozygous spdot mice, both decreased 3H-strychnine binding and chloride flux were observed; however, neither enhanced acoustic startle responses nor limb clenching were seen. These data demonstrate that a dominant-negative point mutation in GlyR disrupting normal function can produce a more dramatic phenotype than the corresponding recessive null mutation, and provides a new animal model to evaluate GlyR function in vivo.