A developmentally plastic adult mouse kidney cell line spontaneously generates multiple adult kidney structures.

Despite exciting new possibilities for regenerative therapy posed by the ability to induce pluripotent stem cells, recapitulation of three-dimensional kidneys for repair or replacement has not been possible. ARID3a-deficient mouse tissues generated multipotent, developmentally plastic cells. Therefore, we assessed the adult mouse ARID3a-/- kidney cell line, KKPS5, which expresses renal progenitor surface markers as an alternative cell source for modeling kidney development. Remarkably, these cells spontaneously developed into multicellular nephron-like structures in vitro, and engrafted into immunocompromised medaka mesonephros, where they formed mouse nephron structures. These data implicate KKPS5 cells as a new model system for studying kidney development.

Immunolabeling of gamma-glutamyl transferase 5 in normal human tissues reveals that expression and localization differ from gamma-glutamyl transferase 1.

Gamma-glutamyl transferase (GGT5) was discovered due to its ability to convert leukotriene C4 (LTC4, a glutathione S-conjugate) to LTD4 and may have an important role in the immune system. However, it was not known which cells express the enzyme in humans. We have developed a sensitive and specific antibody that can be used to detect human GGT5 on Western blots and in fixed tissue sections. We localized GGT5 expression in normal human tissues. We observed GGT5 expressed by macrophages present in many tissues, including tissue-fixed macrophages such as Kupffer cells in the liver and dust cells in the lung. GGT5 was expressed in some of the same tissues that have been shown to express gamma-glutamyl transferase (GGT1), the only other enzymatically active protein in this family. But, the two enzymes were often expressed by different cell types within the tissue. For example, GGT5 was expressed by the interstitial cells of the kidney, whereas GGT1 is expressed on the apical surface of the renal proximal tubules. Other tissues with GGT5-positive cells included: adrenal gland, salivary gland, pituitary, thymus, spleen, liver, bone marrow, small intestine, stomach, testis, prostate and placenta. GGT5 and GGT1 are cell surface enzymes. The different pattern of expression results in their access to different extracellular fluids and therefore different substrates. GGT5 has access to substrates in blood and intercellular fluids, while GGT1 has access primarily to fluids in ducts and glands throughout the body. These data provide new insights into the different functions of these two related enzymes.

Neuroligin-2 Determines Inhibitory Synaptic Transmission in the Lateral Septum to Optimize Stress-Induced Neuronal Activation and Avoidance Behavior.

BACKGROUND: Investigations in the neocortex have revealed that the balance of excitatory and inhibitory synaptic transmission (E/I ratio) is important for proper information processing. The disturbance of this balance underlies many neuropsychiatric illnesses, including autism spectrum disorder and schizophrenia. However, little is known about the contribution of E/I balance to the functioning of subcortical brain regions, such as the lateral septum (LS), a structure that plays important roles in regulating anxiety-related behavior. METHODS: We manipulated E/I balance in the mouse LS by localized conditional deletion of neuroligin-2, a postsynaptic cell adhesion protein located at gamma-aminobutyric acidergic synapses and important for inhibitory synaptic transmission. We then performed analyses of synaptic transmission in the LS, stress-induced expression of immediate early gene c-fos, and anxiety-related and depression-related behavior. RESULTS: The absence of neuroligin-2 in the LS in the mature mouse brain resulted in postsynaptic impairment of inhibitory synaptic transmission. Importantly, the reduced inhibition and resulting E/I imbalance decreased the responsiveness of LS neurons to stress. Furthermore, this E/I imbalance in the LS was associated with impaired stress-induced activation of downstream hypothalamic nuclei and reduced avoidance behavior of the animals in the elevated plus maze. CONCLUSIONS: Our results described the synaptic function of neuroligin-2 in the LS, uncovered a positive association between c-Fos-expressing neurons in the LS and downstream hypothalamic areas and avoidance behavior, and demonstrated that intact inhibitory synaptic transmission and proper E/I balance are required for the optimal functioning of this subcortical circuit.

Male pheromone protein components activate female vomeronasal neurons in the salamander Plethodon shermani.

BACKGROUND: The mental gland pheromone of male Plethodon salamanders contains two main protein components: a 22 kDa protein named Plethodon Receptivity Factor (PRF) and a 7 kDa protein named Plethodon Modulating Factor (PMF), respectively. Each protein component individually has opposing effects on female courtship behavior, with PRF shortening and PMF lengthening courtship. In this study, we test the hypothesis that PRF or PMF individually activate vomeronasal neurons. The agmatine-uptake technique was used to visualize chemosensory neurons that were activated by each protein component individually. RESULTS: Vomeronasal neurons exposed to agmatine in saline did not demonstrate significant labeling. However, a population of vomeronasal neurons was labeled following exposure to either PRF or PMF. When expressed as a percent of control level labeled cells, PRF labeled more neurons than did PMF. These percentages for PRF and PMF, added together, parallel the percentage of labeled vomeronasal neurons when females are exposed to the whole pheromone. CONCLUSION: This study suggests that two specific populations of female vomeronasal neurons are responsible for responding to each of the two components of the male pheromone mixture. These two neural populations, therefore, could express different receptors which, in turn, transmit different information to the brain, thus accounting for the different female behavior elicited by each pheromone component.

The terminal nerve ganglion cells project to the olfactory mucosa in the dwarf gourami.

Single- and double-label immunocytochemical studies were conducted using antisera to salmon gonadotropin-releasing hormone (sGnRH) and molluscan cardioexcitatory peptide (FMRFamide) to determine whether terminal nerve ganglion cells project to the olfactory mucosa in the dwarf gourami, Colisa lalia. Both peptides were present in terminal nerve ganglion perikarya and fibers in brain and nasal cavity. Labeled fibers were present in the olfactory nerve and could be traced to the olfactory mucosa. All terminal nerve ganglion cells contained both sGnRH and FMRFamide-like peptides. This study suggests that the terminal nerve ganglion cells can influence both brain and chemoreceptive structures.

Vole retina is a target for gonadotropin-releasing hormone.

Gonadotropin-releasing hormone (GnRH) projections from the terminal nerve to the retina are common in fish, but have not been reported in mammals. However, GnRH fibers have been seen previously in the optic nerves (but not retinas) of rats and monkeys. Using prairie voles, we tested the hypotheses that (1) GnRH-immunoreactive (-ir) neurons project into the optic nerve and (2) the retina expresses GnRH receptor mRNA as determined by reverse transcription-polymerase chain reaction (RT-PCR) combined with Southern blotting. In both adult and postnatal-day-2 voles, GnRH-ir fibers were observed within the optic nerve. In adult voles, GnRH-ir fibers projected only a short distance into the optic nerve compared with the much longer length of projections in neonates. Fibers immunoreactive for GnRH were not seen in the retinas of neonates or adults. However, RT-PCR-Southern blotting demonstrated GnRH receptor expression in the retina of adult voles. This study supports the hypothesis that GnRH has the potential of modulating visual processing in the retina of mammals.

Pheromonal activation of vomeronasal neurons in plethodontid salamanders.

Pheromones from the mental glands of male plethodontid salamanders increase sexual receptivity in conspecific females. The pheromone enters the vomeronasal organ during courtship to produce this effect. Vomeronasal neurons from female Plethodon shermani were examined following exposure to male pheromone or saline placed on the nares. Agmatine was used in conjunction with the pheromone to enable immunocytochemical visualization of chemosensory neurons that were activated by the pheromone. Olfactory neurons exposed to pheromone or saline, and vomeronasal neurons exposed to saline did not demonstrate significant labeling. A population of vomeronasal neurons was intensely labeled following exposure to the pheromone. This study suggests that a specific population of vomeronasal neurons in a female plethodontid salamander is responsible for transmitting pheromonal information to the brain to produce modifications in behavior.

The naris muscles in tiger salamander. II. Innervation as revealed by enzyme histochemistry and immunocytochemistry.

The naris muscles control the aperature of the external naris in tiger salamanders, Ambystoma tigrinum, and may contribute to glandular secretion. Autonomic neurons of the palatine ganglion and possibly neurons associated with the nervus terminalis innervate these muscles. To elucidate the neural control of the naris muscles, neurotransmitters in nerve fibers supplying the naris muscles and in neurons of the palatine ganglion were examined using acetylcholinesterase enzyme histochemistry and immunocytochemistry to visualize possible peptide candidates for muscle innervation. The naris muscles, autonomic neurons, and associated nerve fascicles demonstrated strong acetylcholinesterase labeling, and the muscles were innervated by substance P fibers passing through the palatine ganglion from the trigeminal ganglion. Gonadotropin-releasing hormone and molluscan cardioexcitatory peptide-like immunoreactivities were found in secretory cell bodies and/or fibers in the palatine ganglion, and gonadotropin-releasing hormone was found in fiber projection pathways into the muscles. Vasoactive intestinal peptide was found in cell bodies and fibers of the palatine ganglion but appeared to provide a sparse innervation to the naris dilator muscle only. These findings suggest a typical autonomic cholinergic and sensory innervation of the naris muscles with some variations in peptide innervation. The presence of gonadotropin-releasing hormone in palatine ganglion and naris constrictor muscle suggests a potential modulation of autonomic neurons and perhaps even muscle fibers by this neuropeptide. We hypothesize that this reproductive hormone may modulate the activity of the naris constrictor muscle during reproductively appropriate events in order to provide access of pheromones to the vomeronasal organ.

The naris muscles in tiger salamander. I. Potential functions and innervation as revealed by biocytin tracing.

The naris constrictor muscle, along with naris dilator and naris accessory muscles, controls the opening and closing of the external naris in tiger salamanders. It has been hypothesized that contraction of the naris constrictor muscle also causes the external nasal gland to secrete its contents inside the lateral wall of the external naris opening. This location is just rostral to vomeronasal organ and thus secretion in this region may be important for access of odorous compounds to vomeronasal organ. Little is known about the innervation of the naris muscles. To elucidate the neural control of these muscles, their innervation was examined using retrograde tract tracing with biocytin. Following application of biocytin to the naris constrictor muscle, labeling was observed in a ventral axonal plexus of the palatine nerve and numerous neuronal cell bodies distributed along this peripheral nerve plexus and within the main portion of the palatine ganglion. If the naris accessory and/or dilator muscles were also exposed to the tracer, the lateral-most branch of the palatine nerve and its associated neural cell bodies were labeled. To confirm the functional innervation of the muscles by the palatine nerve, the nerve was cut and the contraction of the muscles was eliminated. These findings demonstrate that the muscles controlling the external naris are under the control of palatine ganglion neurons. We hypothesize that this innervation of the naris constrictor muscle controls both muscle contraction and glandular secretion that may facilitate access of chemosensory substances to the vomeronasal organ.

Differential distribution of melatonin receptors in the pituitary gland of Xenopus laevis.

A major target site for melatonin action is thought to be the pituitary gland. We have detected differential expression and co-localization of the Mel(1a) and Mel(1c) receptors in cells of the Xenopus laevis pituitary gland. Sections of Xenopus pituitary glands were labeled with Mel(1a) and/or Mel(1c) antibodies, in combination with antibodies to arginine vasotocin (AVT), alpha-melanocyte stimulating hormone (alpha-MSH), prolactin (PRL), and luteinizing hormone (LH). Mel(1a) immunoreactivity was localized to cells of the pars intermedia and to elements within the pars nervosa. Mel(1c) immunoreactivity was also localized to the pars nervosa, and significant labeling was also observed in discrete clusters of cells in the pars distalis. Mel(1a) was absent from the pars distalis, while Mel(1c) was absent from the pars intermedia. Mel(1a) and Mel(1c) were co-localized in the pars nervosa. AVT was present in the pars nervosa, and appeared to be localized to the cell clusters of the pars distalis in which the Mel(1c) receptor was localized. alpha-MSH co-localized with the Mel(1a) receptor in the pars intermedia. LH appeared to localize to many of the cells in the pars distalis, with the notable exception of the Mel(1c) receptor-positive clusters of cells. PRL did not appear to co-localize with either melatonin receptor. The pattern of differential expression of the Mel(1a) and Mel(1c) receptors suggests that the receptors specifically mediate the cellular response to melatonin binding in the specific cell populations.