Research

My thesis identified a crosstalk between the vascular and hematopoietic systems in Drosophila, mediated by Receptor Tyrosine Kinase (RTK) signaling. I studied the RTK ligand and identified its requirement for the maintenance and functionality of the hematopoietic niche. My results also show that this RTK signal feeds onto known molecular circuitry to curtail niche cell proliferation. Parallelly, this RTK axis also evokes downstream signals that ensure a balance between progenitor maintenance and differentiation. These findings strengthen the role for the vascular system in regulating blood progenitor differentiation through the niche. 

My project involved studying a protein complex composed of ADIP, a mechanosensitive protein and Diversin, homolog of the core planar cell polarity (PCP) component Diego in Xenopus embryos. This complex polarized towards wound edges during wound healing and apically constricting sites hinting that this complex could sense force. The goal of my project was to exert an external force on the embryo and validate the molecular force-sensing abilities of Diversin and ADIP. I adapted an existing protocol and created a micromanipulator that could produce negative suction through hydraulic aspiration. This treatment led to polarization of ADIP and Diversin in the direction of suction and confirmed their ability to sense tensile forces. Click here to read more about this work.

As an undergrad I worked on two projects. In one project, I utilized skills I picked up through my Data Science minor and predicted the presence of a stereotypic differentiation hotspot in the Drosophila hematopoietic organ. I also computationally showed that the localization of this hotspot is dependent on the status-quo of the hematopoietic niche within the lymph gland, and manipulations of vesicular signals emanating from this niche correspondingly alter the location of the hotspot.

The other project involved characterizing the role of a receptor tyrosine kinase (RTK) in hematopoietic niche functionality and maintenance. I was involved in experiments that confirmed gene expression of this RTK and its cognate ligand. I also showed that this RTK is responsible for maintaining hematopoietic niche maintenance functionality in the larvae. 

I successfully set up a micro-injection based F0 CRISPR screen in single-celled zebrafish embryos that edited gene loci with very high efficiency (>80%). Using this method, I performed knockouts of 7 seven genes (candidates identified by my mentor Emily Atlas from a sc RNA-seq experiment) that are implicated in cell polarity and motility. The goal was to see whether these genes were also regulators of hair-cell polarity, orientation or rearrangements. My screen identified a novel serine-threonine kinase as a conserved regulator of hair-cell polarity.

I set up a permeabilization protocol in early Drosophila embryos (<2hrs post-fertilization). I optimized the method such that high efficiency of permeability was coupled with low mortality. I now used this optimized protocol to permeabilize embryos to the Janelia Flour dyes (JFX) and tracked single-molecule trajectories of histone and nuclear proteins during the late nuclear cycles (NC10-NC14) using a custom-built lattice light sheet microscope. The lab now has a method to study single-molecule trajectories of proteins for longer time-scales at lower photobleaching rates.