Molecular Cloning, Expression, and Regulation of the Na+/Myo-Inosiotl Cotransporter Gene

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dc.contributor.advisorChaitin, Michael
dc.contributor.committeeMemberEasom, Richard
dc.contributor.committeeMemberGarner, Margaret
dc.creatorZhou, Cheng
dc.date.accessioned2019-08-22T21:17:54Z
dc.date.available2019-08-22T21:17:54Z
dc.date.issued1996-08-01
dc.date.submitted2013-06-27T15:00:08-07:00
dc.description.abstractZhou, Cheng, Molecular Cloning, Expression, and Regulation of the NA+/Myo-Inositol Cotransporter Gene. Doctor of Philosophy (Biomedical Sciences), August 1996. Mammalian cells respond to osmotic stress by accumulation of high concentrations of intracellular osmolytes. Osmotic-induced accumulation of the osmolyte, myo-inositol (MI), is achieved through activation of the NA+/MI cotransporter. Hypertonic stress results in elevated NA+/MI cotransporter mRNA abundance and transcription rate, and increased transporter activity. The goals of this dissertation are to establish the osmoregulation of the NA+/MI cotransporter gene expression in lens cells, and to investigate the transcriptional regulation of the NA+/MI cotransporter gene. Expression of the Na+/MI cotransporter in cultured bovine lens epithelial cells (BLECs) was demonstrated by RT-PCR amplification and Northern blot analysis. Hypertonic stress resulted in induction of the NA+/MI contransporter mRNA abundance in cultured BLECs. The induction patterns of the NA+/MI cotransporter and aldose reductase mRNA abundance by hypertonic stress indicated that osmoregulation of MI and sorbitol accumulations were regulated in concert. Accumulation of MI is an early-onset protective system, which is suppressed by the elevated sorbitol, the late-onset protective system. 5’-RACE analysis indicated that multiple transcription start sites were utilized in controlling of the expression of the NA+/MI cotransporter. Osmotic stress resulted in preferential utilization of a hypertonic promoter a. The bovine NA+/MI cotransporter gene was cloned and analyzed. The regulation of the Na+/MI cotransporter expression was investigated by transient transfection assays using promoter-luciferase constructs. Although multiple promoters were functional in cultured BLECs, only the hypertonic promoter a was osmotically responsive. Characterization of this osmotic-responsive element(s) between -536 to -300 bp upstream of the hypertonic transcription start site a. The studies presented in this dissertation refined the osmoregulation of the Na+/MI cotransporter gene expression. Hypertonicity induces MI accumulation by activation of an osmotic-responsive promoter. The consequences of the activation of this promoter lead to more cotransporter mRNA, more cotransporter protein, and higher transporter activity, resulting in accumulation of a higher concentration of intracellular Mi.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://hdl.handle.net/20.500.12503/29225
dc.language.isoen
dc.provenance.legacyDownloads1
dc.subjectBiology
dc.subjectBiomechanics and Biotransport
dc.subjectBiomedical Engineering and Bioengineering
dc.subjectBiophysics
dc.subjectCell and Developmental Biology
dc.subjectCell Biology
dc.subjectCells
dc.subjectCellular and Molecular Physiology
dc.subjectLife Sciences
dc.subjectMedical Cell Biology
dc.subjectMedicine and Health Sciences
dc.subjectMolecular Biology
dc.subjectStructural Biology
dc.subjectOsmosis
dc.subjectosmoregulation
dc.subjectosmotic stress
dc.subjectNa+/MI contransporter gene
dc.subjectintracellular Mi
dc.subjectmolecular cloning
dc.subjectmolecular expression
dc.subjectmolecular regulation
dc.subjectNa+/Myo-Inositol Cotransporter Gene
dc.titleMolecular Cloning, Expression, and Regulation of the Na+/Myo-Inosiotl Cotransporter Gene
dc.typeDissertation
dc.type.materialtext
thesis.degree.departmentGraduate School of Biomedical Sciences
thesis.degree.disciplineBiomedical Sciences
thesis.degree.grantorUniversity of North Texas Health Science Center at Fort Worth
thesis.degree.nameDoctor of Philosophy

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