Scholarly Work - Microbiology & Cell Biology
Permanent URI for this collectionhttps://scholarworks.montana.edu/handle/1/3494
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Item Distal spinal nerve development and divergence of avian groups(2020-04) Rashid, Dana J.; Bradley, Roger S.; Bailleul, Alida; Surya, Kevin; Woodward, Holly; Wu, Ping; Wu, Yun-Hsin; Menke, Douglas; Minchey, Sergio; Parrott, Ben; Bock, Samantha; Merzdorf, Christa; Narotzky, Emma; Burke, Nathan; Horner, John R.; Chapman, SusanThe avian transition from long to short, distally fused tails during the Mesozoic ushered in the Pygostylian group, which includes modern birds. The avian tail embodies a bipartite anatomy, with the proximal separate caudal vertebrae region, and the distal pygostyle, formed by vertebral fusion. This study investigates developmental features of the two tail domains in different bird groups, and analyzes them in reference to evolutionary origins. We first defined the early developmental boundary between the two tail halves in the chicken, then followed major developmental structures from early embryo to post-hatching stages. Differences between regions were observed in sclerotome anterior/posterior polarity and peripheral nervous system development, and these were consistent in other neognathous birds. However, in the paleognathous emu, the neognathous pattern was not observed, such that spinal nerve development extends through the pygostyle region. Disparities between the neognaths and paleognaths studied were also reflected in the morphology of their pygostyles. The ancestral long-tailed spinal nerve configuration was hypothesized from brown anole and alligator, which unexpectedly more resembles the neognathous birds. This study shows that tail anatomy is not universal in avians, and suggests several possible scenarios regarding bird evolution, including an independent paleognathous long-tailed ancestor.Item The voltage sensing phosphatase (VSP) localizes to the apical membrane of kidney tubule epithelial cells(2019-04) Ratzan, Wil; Rayaprolu, Vamseedhar; Killian, Scott E.; Bradley, Roger S.; Kohout, Susy C.Voltage-sensing phosphatases (VSPs) are transmembrane proteins that couple changes in membrane potential to hydrolysis of inositol signaling lipids. VSPs catalyze the dephosphorylation of phosphatidylinositol phosphates (PIPs) that regulate diverse aspects of cell membrane physiology including cell division, growth and migration. VSPs are highly conserved among chordates, and their RNA transcripts have been detected in the adult and embryonic stages of frogs, fish, chickens, mice and humans. However, the subcellular localization and biological function of VSP remains unknown. Using reverse transcriptase-PCR (RT-PCR), we show that both Xenopus laevis VSPs (Xl-VSP1 and Xl-VSP2) mRNAs are expressed in early embryos, suggesting that both Xl-VSPs are involved in early tadpole development. To understand which embryonic tissues express Xl-VSP mRNA, we used in situ hybridization (ISH) and found Xl-VSP mRNA in both the brain and kidney of NF stage 32-36 embryos. By Western blot analysis with a VSP antibody, we show increasing levels of Xl-VSP protein in the developing embryo, and by immunohistochemistry (IHC), we demonstrate that Xl-VSP protein is specifically localized to the apical membrane of both embryonic and adult kidney tubules. We further characterized the catalytic activity of both Xl-VSP homologs and found that while Xl-VSP1 catalyzes 3- and 5-phosphate removal, Xl-VSP2 is a less efficient 3-phosphatase with different substrate specificity. Our results suggest that Xl-VSP1 and Xl-VSP2 serve different functional roles and that VSPs are an integral component of voltage-dependent PIP signaling pathways during vertebrate kidney tubule development and function.Item Neural crest development in Xenopus requires Protocadherin 7 at the lateral neural crest border(2018-02) Rashid, Dana J.; Puettmann, Paul; Roy, Ethan; Bradley, Roger S.In vertebrates, the neural crest is a unique population of pluripotent cells whose development is dependent onsignalin g from neighbori ng tissues. Cadherin family members, including protocadherins, are emerging asmajor players in neural crest development, largely through their roles in cell adhesion and sorting in embryonictissues. Here, we show that Protocadherin 7 (Pcdh7), previously shown to function in sensorial layer integrityand neural tube closure in Xenopus, is also involved in neural crest specification and survival. Pcdh7 expressionpartly overlaps the neural crest domain at the lateral neural crest border. Pcdh7 knockdown in embryos does notalter neural crest induction; however, neural crest specification markers, including Snail2 and Sox9, are lost, dueto apoptosis of the neural crest starting after stage 13. Pcdh7 knockdown also results in downregulation ofWnt11b; both of which are co-expressed in the sensorial layer lateral to the neural crest, suggestive of a rolefor Wnt11b in the neural crest apoptosis. Confirming this role, apoptosis, Snail2 expression and the developmental fate of the neural crest can be partially rescued by ectopic expression of Wnt11b. These results indicate thatPcdh7 plays an important role in maintaining the sensorial layer at the lateral neural crest border, which is nec-essary for the secretion of survival factors, including Wnt11bItem From dinosaurs to birds: a tail of evolution(2014-05) Rashid, Dana J.; Chapman, Susan C.; Larsson, Hans C. E.; Organ, Chris L.; Merzdorf, Christa; Bradley, Roger S.; Horner, John R.A particularly critical event in avian evolution was the transition from long- to short-tailed birds. Primitive bird tails underwent significant alteration, most notably reduction of the number of caudal vertebrae and fusion of the distal caudal vertebrae into an ossified pygostyle. These changes, among others, occurred over a very short evolutionary interval, which brings into focus the underlying mechanisms behind those changes. Despite the wealth of studies delving into avian evolution, virtually nothing is understood about the genetic and developmental events responsible for the emergence of short, fused tails. In this review, we summarize the current understanding of the signaling pathways and morphological events that contribute to tail extension and termination and examine how mutations affecting the genes that control these pathways might influence the evolution of the avian tail. To generate a list of candidate genes that may have been modulated in the transition to short-tailed birds, we analyzed a comprehensive set of mouse mutants. Interestingly, a prevalent pleiotropic effect of mutations that cause fused caudal vertebral bodies (as in the pygostyles of birds) is tail truncation. We identified 23 mutations in this class, and these were primarily restricted to genes involved in axial extension. At least half of the mutations that cause short, fused tails lie in the Notch/Wnt pathway of somite boundary formation or differentiation, leading to changes in somite number or size. Several of the mutations also cause additional bone fusions in the trunk skeleton, reminiscent of those observed in primitive and modern birds. All of our findings were correlated to the fossil record. An open question is whether the relatively sudden appearance of short-tailed birds in the fossil record could be accounted for, at least in part, by the pleiotropic effects generated by a relatively small number of mutational events.