Transcription factor expression in the developing human fetal endocrine pancreas

Abstract

Aims/hypothesis: Morphological changes that occur during pancreatic endocrine cell differentiation have been shown in rodent systems to be dependent on sequential alterations in transcription factor expression. However, similar data for humans have been limited. The aim of the present study was to provide a connection between pancreatic morphology, transcription factor gene expression and protein localisation during human fetal development.

Methods: Human fetal pancreases were examined at early (8-12 weeks of fetal age), middle (14-16 weeks) and late (19-21 weeks) stages, using immunohistological, microarray and qRT-PCR analyses.

Results: We observed a significant decrease in pancreatic duodenal homeobox 1 (PDX-1)(+)/cytokeratin 19(+) cells (p < 0.001), with a simultaneous increase in PDX-1(+)/insulin(+) cells from 8 to 21 weeks (p < 0.05). Increased PDX-1/insulin co-localisation within islet clusters was noted, while no co-expression of PDX-1 with glucagon was found, suggesting that loss of PDX-1 is essential for alpha cell formation. Given that neurogenin 3 (NGN3) expression is critical for establishing the endocrine cell programme in the rodent pancreas, we examined its expression pattern and co-localisation in PDX-1(+), insulin(+) and glucagon(+) cells. Co-localisation of NGN3 with PDX-1, insulin and glucagon was noted during early development, with significant decreases in middle and late stages (p < 0.001). Our microarray and co-localisation analyses of transcription factors linked to NGN3 demonstrated that ISL1 transcription factor (ISL1), neurogenic differentiation 1 (NEUROD1), NK2 related transcription factor related, locus 2 (NKX2-2) and paired box gene 6 (PAX6) were upregulated during development and present in all four endocrine cell types, while NK6 related transcription factor related, locus 1 (NKX6-1) was expressed exclusively in beta cells.

Conclusions/interpretation: This study is an important step towards identifying key molecular factors involved in development of the human fetal endocrine pancreas.