Date of Graduation

5-2023

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Biological Sciences

Advisor/Mentor

Nagayasu Nakanishi

Committee Member

Erica Westerman

Second Committee Member

Vibha Srivastava

Third Committee Member

Timothy Evans

Fourth Committee Member

Woodrow Shew

Keywords

Cnidaria, Nematostella, Pituitary, POU-I

Abstract

Cnidaria (i.e., sea anemones, jellyfish, corals) and Bilateria (i.e., vertebrates, sea stars, fruit flies), are sister groups that diverged around 600 million years ago. Despite the long evolutionary time, many cellular differentiation mechanisms, cell types, tissues and behaviors are conserved. Such as neurons, mechanosensory hair cells, feeding behaviors, peristaltic movements, and sleep. Recent advances in genomics, molecular biology and microscopy have fueled an increased interest in understanding cnidarian nervous and neuroendocrine systems. Understanding the developmental mechanisms and the mode of operation of Cnidarian nervous systems helps to reconstruct the ancestral nervous system of the last common ancestor of Cnidaria and Bilateria. Thus, also shedding light in fundamental aspects of Bilaterian nervous systems. Here, the ‘starlet sea anemone’ Nematostella vectensis, a powerful cnidarian model organism was used to address the gene expression pattern of Pit1, a conserved gene shared between Cnidaria and Bilateria. In Chapter 1, a method to extract DNA and genotype embryos of Nematostella without sacrificing the animal was established, with possible application to other non-sea anemone cnidarians. Early genotyping is fundamental for addressing phenotypes during development, thus opening the door to study the function of any gene of interest during larval pre-metamorphic stages. In Chapter 2, the expression pattern of Pit1 and detailed cellular morphology of Pit1-positive cells was characterized for the first time in Nematostella. Complex neuronal networks and diverse sensory cells were found. Furthermore, the foundation for future functional studies of Pit1 was laid by establishing stable CRISPR-Cas9 knockout and transgenic reporter lines.

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