Microtubules (MTs) are a major element of the cytoskeleton; the dynamic network of interlinking protein filaments present in the cytoplasm of all cells. MTs are active arrangements that experience constant construction and deconstruction within the cell. MT’s role is both to establish cell shape and a diversity of cell activity. This involves certain elements of cell motion, the intracellular transfer of cell organs, and the detachment of genetic material in the course of mitosis. Microtubule-associated proteins (MAPs) control the building and balance of microtubules. The purpose of this paper is to recognize which MAPs control microtubule dynamics in insulin-stimulated glucose uptake. This work also seeks to evaluate how MAPs expression changes the more they progress into the differentiation process, which involves going from wildtype fibroblast cell to a wildtype 3T3-L1 adipocyte cell. Utilizing spectroscopy, this work compares the proteome of the cell, as well as the protein interactome present on each stage of cell development. Finding the MAPs controlling microtubule dynamics in insulin-stimulated glucose uptake will help originate future approaches to prevent the development of complications in medical disorders such as type II diabetes.

Extensive research on the factors that influence parturition has shown that, in almost all species, birth is initiated by a reduction in the amount of circulating progesterone, the major hormone that sustains pregnancy. Yet, this progesterone drop does not occur in humans and other Old World monkeys, and the evolutionary traits distinct to modern humans make it difficult to pinpoint what causes the start of labor. Knowing this would improve medical care before, during, and after pregnancy and reduce the rate of preterm births. Our research has focused on synthesizing data to understand what may cause parturition in humans. We analyzed the expression of genes involved in multiple signaling pathways in the cells of the maternal-fetal interface with the goal of identifying regulatory elements with human-specific genetic changes. Now, we want to know if we can use this information to find the genetic basis of human-specific pregnancy traits and see if these changes occur in functionally important genomes of the uterine cells.

The neural crest (NC) is a multipotent stem cell population that differentiates into a diverse array of derivatives. Cardiac neural crest cells in zebrafish are highly important in the development and adult function of the cardiovascular system. NC development is governed by a gene regulatory network (GRN). We seek to expand our understanding of the cardiac NC GRN by understanding the regulatory linkages that exist within the GRN and how they may become dysregulated leading to congenital disease. Through utilizing multi-omics approaches, we identified prdm1a as a transcription factor of interest in the developing cardiac NC. Our goal has been to identify the role that prdm1a plays in the development of cardiac NC and uncover its regulatory linkages in the cardiac NC GRN. For this, we have used hybridization chain reaction (HCR) and confocal imaging to visualize expression patterns of prdm1a at critical developmental time points of the cardiac NC. Based on these preliminary results, we have found that prdm1a is expressed in the migratory cardiac NC and NC-derived cardiovascular cell types. To further understand the role of this gene in cardiac NC development, we have generated knockout prdm1a zebrafish lines to analyze downstream gene expression. We have also identified a cardiac NC-specific prdm1a enhancer that sheds light on how prdm1a expression is regulated in the cardiac NC. These experiments have allowed us to better understand the role that prdm1a plays in development of cardiac NC in zebrafish.

Heart disease and stroke are among the top five causes of death in the United States, leading to billions of dollars in healthcare expenditures. This disease is caused by cellular dysfunction and death due to metabolic stress and hypoxia, which are the hallmarks of heart disease and stroke. Unfortunately, clinical treatments for these conditions often cannot reverse cellular damage. Interestingly, Arctic ground squirrels (AGS) are extreme hibernators that can reduce their metabolic rate and temperature without suffering from ischemic injuries. In Dr. Dengke Ma’s lab at UCSF, we have identified that mesencephalic astrocyte-derived neurotrophic factor (MANF) is among the cytoprotective genes responsible for the AGS’s resiliency against metabolic stress and hypoxia. Hence, studying AGS neural cells may help provide insights into new treatments. This study aims to utilize CRISPR-gene editing in mouse neural cells to express AGS MANF and investigate the specific role of this protein in cellular stress during ischemic injuries. Mouse neural cells expressing AGS MANF were created using the CRISPR-Cas9 gene-editing, and the survival rate of AGS MANF variants under metabolic stress and response to unfolded protein response (UPR) markers were analyzed. We found that mouse cells expressing AGS MANF survived better than those expressing endogenous MANF under cellular stress, followed by the reduction of UPR genes after the treatment of metabolic toxins–such as rotenone and thapsigargin–to induce metabolic stress. Future studies will be required to narrow the mechanistic basis of this cytoprotection which may be a promising therapy for conditions such as strokes and heart attacks.