A Genome-Wide Screen to Identify Transcription Factors Expressed in Pelvic Ganglia of the Lower Urinary Tract

Abstract

Relative positions of neurons within mature murine pelvic ganglia based on expression of neurotransmitters have been described. However the spatial organization of developing innervation in the murine urogenital tract (UGT) and the gene networks that regulate specification and maturation of neurons within the pelvic ganglia of the lower urinary tract (LUT) are unknown. We used whole-mount immunohistochemistry and histochemical stains to localize neural elements in 15.5 days post coitus (dpc) fetal mice. To identify potential regulatory factors expressed in pelvic ganglia, we surveyed expression patterns for known or probable transcription factors (TF) annotated in the mouse genome by screening a whole-mount in situ hybridization library of fetal UGTs. Of the 155 genes detected in pelvic ganglia, 88 encode TFs based on the presence of predicted DNA-binding domains. Neural crest (NC)-derived progenitors within the LUT were labeled by Sox10, a well-known regulator of NC development. Genes identified were categorized based on patterns of restricted expression in pelvic ganglia, pelvic ganglia and urethral epithelium, or pelvic ganglia and urethral mesenchyme. Gene expression patterns and the distribution of Sox10+, Phox2b+, Hu+, and PGP9.5+ cells within developing ganglia suggest previously unrecognized regional segregation of Sox10+ progenitors and differentiating neurons in early development of pelvic ganglia. Reverse transcription-PCR of pelvic ganglia RNA from fetal and post-natal stages demonstrated that multiple TFs maintain post-natal expression, although Pax3 is extinguished before weaning. Our analysis identifies multiple potential regulatory genes including TFs that may participate in segregation of discrete lineages within pelvic ganglia. The genes identified here are attractive candidate disease genes that may now be further investigated for their roles in malformation syndromes or in LUT dysfunction.

Urinary Tract Effects of HPSE2 Mutations

“…The protein data accord with Hpse2 transcripts being detectable in embryonic pelvic ganglia, 22 where Hpse2+ cells coexpress SRY-related HMG-box 10, which regulates neural crest development. 19 LRIG2 and heparanase 1 were also immunodetected in E14 pelvic and lumbar ganglia. Postnatally, bladder wall nerves and pelvic ganglia contained heparanase 1 and 2 and LRIG2 ( Figure 2, B-I).…”

“…Pelvic ganglia give rise to postganglionic parasympathetic axons mediating detrusor contraction, while lumbar ganglia emit postganglionic sympathetic axons mediating detrusor relaxation and sphincter contraction. 19,20 Immunohistochemistry at E14 ( Figure 2A) showed heparanase 2 in pelvic and lumbar ganglia; nerves running in bladder wall mesenchyme, beginning to form smooth muscle at this stage 21 ; mesenchymal ureteric cells; and peripheral nerves within the head. The protein data accord with Hpse2 transcripts being detectable in embryonic pelvic ganglia, 22 where Hpse2+ cells coexpress SRY-related HMG-box 10, which regulates neural crest development.…”

An illustrated anatomical ontology of the developing mouse lower urogenital tract

“…8R) (Abler et al, 2011a), E15 whole-mount IHC (Fig. 9G,H) (Wiese et al, 2012) and whole adult bladders (Fig. 9J-L) (Yan and Keast, 2008), tissues were immunostained as described previously.…”

“…For IF (Fig. 9I,N-P,Q-S), cryosections (14-20 µm thickness) were processed according to described methods (Yan and Keast, 2008;Wiese et al, 2012).…”

Comparative expression analysis of cysteine-rich intestinal protein family members crip1, 2 and 3 during Xenopus laevis embryogenesis

“…Again, this is in agreement with crip2 expression in Xenpous. Additionally, murine Crip2 was detected in lymphatic endothelial cells, in dorsal root ganglia and around the spinal cord, the neural tube including the brain, lungs, pelvic ganglia and mesenchyme of the lower urinary tract, but not in the kidney (Wei et al, 2011, Wiese et al, 2012, Yu et al, 2002, Zhang et al, 2005. In zebrafish, crip2 transcript were detected the earliest in the premigratory neural crest cells in rhombomere 6 of the neural tube, in the cardiac progenitor cells in anterior lateral plate mesoderm and mesoendodermal cells in the primary heart field (6-to 20-somite stage).…”