Neurons in the upper lumbar spinal cord project axons containing gastrin-releasing peptide (GRP) to innervate lower lumbar regions controlling erection and ejaculation. This system is vestigial in female rats and in males with genetic dysfunction of androgen receptors, but in male rats, pharmacological stimulation of spinal GRP receptors restores penile reflexes and ejaculation after castration. GRP offers new avenues for understanding potential therapeutic approaches to masculine reproductive dysfunction.GRP, a member of the bombesin-like peptide family 1 , is distributed widely in the central nervous system and gastrointestinal tract of mammals 2 , 3 . GRP and neuromedin B (NMB), the mammalian counterpart of bombesin, play a role in many physiological processes, including itch 4 , circadian rhythms 5 , food intake 6 and fear memory consolidation 7 , 8 . In mammals, bombesin-like peptides act through a family of at least three G protein-coupled receptors: GRP-preferring receptor (GRP-R), NMB-preferring receptor (NMB-R) and bombesin receptor subtype-3 (BRS-3) 9 .Using immunocytochemistry (ICC) directed at GRP, we found a group of neurons within a region previously dubbed the 'spinal ejaculation generator' because toxins that selectively lesion galanin-containing neurons there virtually eliminate ejaculation in rats 10 . These galaninergic neurons project to the thalamus 10 , but it had been unclear whether there are also direct connections between this center and the lower spinal cord regions that directly trigger ejaculation 11 . The separate, GRP-containing neurons that we found within the center projected axons to more caudal spinal regions and were much more prominent in wild-type (WT) males than in WT females ( Fig. 1a,b; Supplementary Fig. 1 online) (n = 5, F 2,12 = 299.9, P < 0.001). Semiquantitative reverse transcription (RT)-PCR confirmed more pre-pro NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptGrp transcripts in this region of males than of females ( Supplementary Fig. 2 online). To test whether androgen receptors direct sexual dimorphism of these neurons, we examined genetically male (XY) Long-Evans rats carrying the testicular feminization mutation (Tfm) of the androgen receptor gene Ar. These rats develop testes embryologically and secrete testosterone pre-and postnatally but, because their androgen receptor protein is dysfunctional, develop a wholly feminine exterior, including a clitoris rather than a penis. The spinal cord of Tfm rats was hyperfeminine, having even fewer GRP-positive neurons in this region than did WT females (P <0.001) (Fig. 1c,d). In normal males, GRP-expressing neurons also expressed androgen receptor (96.1 ± 1.7%; n = 4 WT males), but not estrogen receptor alpha (ERα) (Fig. 1e-h). Because androgens such as testosterone augment ejaculation in male rats and humans 12 , the presence of androgen receptor in the GRPpositive neurons of the ejaculation center offers a locus for androgenic modulation of ejaculation and other sexual reflexes.ICC r...

*Nemerteans (ribbon worms) and phoronids (horseshoe worms) are closely related lophotrochozoans-a group of animals including leeches, snails and other invertebrates. Lophotrochozoans represent a superphylum that is crucial to our understanding of bilaterian evolution. However, given the inconsistency of molecular and morphological data for these groups, their origins have been unclear. Here, we present draft genomes of the nemertean Notospermus geniculatus and the phoronid Phoronis australis, together with transcriptomes along the adult bodies. Our genome-based phylogenetic analyses place Nemertea sister to the group containing Phoronida and Brachiopoda. We show that lophotrochozoans share many gene families with deuterostomes, suggesting that these two groups retain a core bilaterian gene repertoire that ecdysozoans (for example, flies and nematodes) and platyzoans (for example, flatworms and rotifers) do not. Comparative transcriptomics demonstrates that lophophores of phoronids and brachiopods are similar not only morphologically, but also at the molecular level. Despite dissimilar head structures, lophophores express vertebrate head and neuronal marker genes. This finding suggests a common origin of bilaterian head patterning, although different heads evolved independently in each lineage. Furthermore, we observe lineagespecific expansions of innate immunity and toxin-related genes. Together, our study reveals a dual nature of lophotrochozoans, where conserved and lineage-specific features shape their evolution. Articles NaTure eCOLOgy & evOLuTiONphoronids, ectoprocts and brachiopods, although the position of ectoprocts is questionable under a sensitivity analysis. Our results clearly show that lophotrochozoans have a different evolutionary history than other spiralians (or platyzoans), such as flatworms and rotifers. In particular, lophotrochozoans retain a basic bilaterian gene repertoire, which is probably lost in ecdysozoans and other spiralian lineages. Unexpectedly, genes specifically expressed in lophophores of phoronids and brachiopods are strikingly similar to those employed in vertebrate head formation, although novel genes, expanded gene families and redeployment of developmental genes also contribute to the unique molecular identity of lophophores. Furthermore, we provide examples of lineage-specific genomic features in lophotrochozoans, such as the expansion of innate immunity and toxin-related genes. Taken together, our study reveals the dual nature of lophotrochozoan genomes, showing both conservative and innovative characteristics during their evolution.

De novo steroidogenesis from cholesterol is a conserved property of vertebrate brains, and such steroids synthesized de novo in the brain are called neurosteroids. The identification of neurosteroidogenic cells is essential to the understanding of the physiological role of neurosteroids in the brain. We have demonstrated recently that neuronal neurosteroidogenesis occurs in the brain and indicated that the Purkinje cell actively synthesizes several neurosteroids de novo from cholesterol in vertebrates. Interestingly, in the rat, this neuron actively synthesizes progesterone de novo from cholesterol only during neonatal life, when cerebellar cortical formation occurs most markedly. Therefore, in this study, the possible organizing actions of progesterone during cerebellar development have been examined. In vitro studies using cerebellar slice cultures from newborn rats showed that progesterone promotes dose-dependent dendritic outgrowth of Purkinje cells but dose not affect their somata. This effect was blocked by the anti-progestin RU 486 [mifepristone; 17␤-hydroxy-11␤-(4-methylaminophenyl)-17␣-(1-propynyl) estra-4,9-dien-3 one-6-7]. In vivo administration of progesterone to pups further revealed an increase in the density of Purkinje spine synapses electron microscopically. In contrast to progesterone, there was no significant effect of 3␣,5␣-tetrahydroprogesterone, a progesterone metabolite, on Purkinje cell development. Reverse transcription-PCR-Southern and immunocytochemical analyses showed that intranuclear progesterone receptors were expressed in Purkinje cells. These results suggest that progesterone promotes both dendritic outgrowth and synaptogenesis in Purkinje cells through intranuclear receptor-mediated mechanisms during cerebellar development. Such organizing actions may contribute to the formation of the cerebellar neuronal circuit.

Circulation-derived cells play a crucial role in the healing processes of tissue. In early phases of tendon healing processes, circulation-derived cells temporarily exist in the wounded area to initiate the healing process and decrease in number with time. We assumed that a delay of time-dependent decrease in circulation-derived cells could improve the healing of tendons. In this study, we injected platelet-rich plasma (PRP) containing various kinds of growth factors into the wounded area of the patellar tendon, and compared the effects on activation of circulation-derived cells and enhancement of tendon healing with a control group (no PRP injection). To follow the circulation-derived cells, we used a green fluorescent protein (GFP) chimeric rat expressing GFP in the circulating cells and bone marrow cells. In the PRP group, the numbers of GFP-positive cells and heat-shock protein (HSP47; collagen-specific molecular chaperone)-positive cells were significantly higher than in the control group at 3 and 7 days after injury. At the same time, the immunoreactivity for types I and III collagen was higher in the PRP group than in the control group at early phase of tendon healing. These findings suggest that locally injected PRP is useful as an activator of circulation-derived cells for enhancement of the initial tendon healing process.