Year of Publication

2014

Degree Name

Doctor of Philosophy (PhD)

Document Type

Doctoral Dissertation

College

Arts and Sciences

Department

Biology

First Advisor

Dr. Douglas Harrison

Abstract

In the Drosophila testis, sperm are derived from germline stem cells (GSCs) which undergo a stereotyped pattern of divisions and differentiation. The somatic cells at the tip of the testis form the hub, which is the niche for both the somatic cyst stem cells (CySCs) and GSCs. The hub expresses Upd, a ligand for the JAK/STAT pathway that has roles in the maintenance of CySCs and GSCs. Male mutants of upd3, another ligand of the JAK/STAT pathway, become sterile much earlier than the wild-type, leading to the hypothesis that similar to upd, upd3 also promotes the self-renewal of stem cells in testis. It was found here that upd3 is also expressed in the hub, and that mutants of upd3 have fewer CySCs and GSCs. Using a GFP reporter of the JAK/STAT pathway, it was found that the JAK/STAT pathway is not only activated in the stem cells, consistent with its known function in the maintenance of stem cells, but is also activated in the elongated cyst cells that encapsulate late stage differentiating spermatids. The reduction of JAK/STAT activity in the somatic cyst cells led to impaired spermatid individualization, a late stage of spermatogenesis during which the syncytial spermatids are separated. The impairment of individualization was shown by the loss of three characteristic structures: individualization complexes (ICs), cystic bulges (CBs), and waste bags (WBs). The failure of IC formation implies STAT activity is required for the initiation of individualization, and the loss of CBs and WBs suggests STAT activity is required for the progression of individualization. Activation of caspases in elongated spermatids is known to be required for individualization. The reduction of JAK/STAT activity in cyst cells almost completely eliminated the activation of two effector caspases: drICE and DCP-1. It was concluded that JAK/STAT activity in somatic cyst cells promotes individualization by stimulating caspase activity in spermatids. The JAK/STAT pathway is not only required for the maintenance of stem cells at the tip, but also required for individualization away from the tip during late differentiation, thus is reutilized in Drosophila spermatogenesis.

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