Research Article

Contrasting patterns of expression of transcription factors in pancreatic α and β cells

Jie Wang, Gene Webb, Yun Cao, and Donald F. Steiner
PNAS October 28, 2003 100 (22) 12660-12665; https://doi.org/10.1073/pnas.1735286100

  1. Contributed by Donald F. Steiner, August 18, 2003

Abstract

Pancreatic α and β cells are derived from the same progenitors but play opposing roles in the control of glucose homeostasis. Disturbances in their function are associated with diabetes mellitus. To identify many of the proteins that define their unique pathways of differentiation and functional features, we have analyzed patterns of gene expression in αTC1.6 vs. MIN6 cell lines by using oligonucleotide microarrays. Approximately 9–10% of >11,000 transcripts examined showed significant differences between the two cell types. Of >700 known transcripts enriched in either cell type, transcription factors and their regulators (TFR) was one of the most significantly different categories. Ninety-six members of the basic zipper, basic helix–loop–helix, homeodomain, zinc finger, high mobility group, and other transcription factor families were enriched in α cells; in contrast, homeodomain proteins accounted for 51% of a total of 45 TFRs enriched in β cells. Our analysis thus highlights fundamental differences in expression of TFR subtypes within these functionally distinct islet cell types. Interestingly, the α cells appear to express a large proportion of factors associated with progenitor or stem-type cells, perhaps reflecting their earlier appearance during pancreatic development. The implications of these findings for a better understanding of α and β cell dysfunction in diabetes mellitus are also considered.

Pancreatic islets consist of four endocrine cell types, α, β, D, and pancreatic peptide (PP). These cell types produce and secrete the major islet hormones: glucagon, insulin, islet amyloid polypeptide (IAPP), somatostatin, and PP, respectively, which regulate fuel and energy homeostasis (1). The α cells secrete glucagon, which stimulates gluconeogenesis and glycogenolysis to prevent hypoglycemia, whereas the β cells increase insulin secretion in response to elevated blood glucose levels. Glucagon and insulin antagonistically regulate the balance of glucose storage, production, and consumption to maintain physiological plasma glucose concentrations. Therefore, the α and β cells together play a central role in glucose homeostasis.

Excessive production and secretion of glucagon by the α cells is a common accompaniment to the two main types of diabetes. Physiologically, glucagon secretion is suppressed by hyperglycemia. However, this normal homeostatic suppression is lost in diabetic states, which in turn perpetuates hyperglycemia by stimulating hepatic glucose output (2). Another major typical manifestation of diabetes is an absolute or relative deficiency of insulin from the β cells, resulting in failure to adequately control the blood glucose level (3). Disturbances of α and/or β cell function thus are central to the failure to maintain physiological glucose levels and related metabolic concomitants of diabetes mellitus.

To understand the molecular basis for the development and specialized functions of α and β cells is an important goal for understanding and effectively treating diabetes. Although much effort has been expended on the biosynthesis of the islet hormones (1) and on the pancreas and its development (4–7), much remains to be learned about the full complement of gene products expressed in these cells. Examination of the β cell functional profile has recently been carried out by using various techniques such as representational analysis (8, 9), subtractive hybridization (10, 11), mRNA differential display (12), and oligonucleotide microarrays (13–15). However, the underlying molecules and mechanisms regulating the proliferation and differentiation of the pancreatic endocrine cells (5) and the genetic factors responsible for the common forms of diabetes mellitus (16) largely remain elusive. Furthermore, our current knowledge of the proteome of the α cell, the second major player in the control of glucose homeostasis, is still very limited. Thus, our knowledge remains incomplete regarding many key processes, including developmental origin, differentiation, metabolism, regulation of secretory activity, and the physiological/pathological profiles of α cells vs. the well studied β cells.

In attempts to identify, classify, and contrast factors important for α or β cell function, we have applied Mu11K DNA microarray analysis to two well characterized α and β cell lines, clonal αTC1.6 (17, 18), and MIN6 (19). These are well differentiated cell lines that preserve similar characteristics (such as correct hormone processing and secretion of glucagon or insulin in response to glucose stimulation), and that have arisen from transformed mature islet cells having similar genetic backgrounds (17–21). In the present study, attention has been concentrated on members of various functional categories that are enriched predominantly in the α or β cell phenotype with a special emphasis on the category of transcription factors and their regulators (TFR). Thus, activated transcription factor 3 (ATF3), hypoxia-inducible factor 1 α (HIF1α), four-and-a-half LIM domains 1 (FHL1), and other basic zipper (bZip), basic helix–loop–helix (bHLH), and zinc finger (ZF) proteins were detected predominantly in pancreatic α cells, whereas β cells were enriched with members of the homeodomain (HD) group. Our results indicate that various important families of transcription factors are quite differently represented in α vs. β cells and suggest fundamental differences in their functional regulation.

Materials and Methods

Cell Culture, Islet Isolation, and RNA Preparation. Cells (passages 23–25) of αTC1.6 (17, 18), MIN6 (19), and βTC3 (22) were maintained as described previously. proprotein convertase (PC)2 null α cells (an α cell line established from these mice in the lab; G.W., unpublished data) were cultured under the same conditions as the αTC1.6 cells. The medium was replaced 24 h before RNA extraction. Islets from adult PC2 null (23) and wild-type littermate mice were isolated as described (24). Total RNA from the cultured cells and/or islets was extracted with TRIzol Reagent (GIBCO), and poly(A+) RNA was then prepared by using the Oligotex mRNA Mini Kit (Qiagen, Valencia, CA).

Microarray Analysis. The biotinylated cRNA samples of αTC1.6 and MIN6 cells were synthesized and hybridized in duplicates to Mu11K oligonucleotide array (Affymetrix, Santa Clara, CA) as described (13). Analysis of the data was performed by genechip suite (Ver. 4.0.1, Affymetrix). The threshold for determining significant differences of expression level between the two cell types was set by using a provided algorithm. The molecules having an average fold change of 3 or greater in either cell type are shown in the following tables.

RT-PCR. cDNA was synthesized by reverse transcription from total RNA (1 μg) as described (24). The amount of cDNA used for PCR was normalized by the amplified levels of labeled β-actin with [γ-32P]ATP (Amersham Pharmacia Biotech) by using semiquantitative PCR. The sequences of primers and the length of their respective cDNA fragments are shown in Table 2, which is published as supporting information on the PNAS web site, www.pnas.org.

Immunohistochemical Studies. Rabbit antiserum to ATF3, goat antisera to HIF1α, and FHL1 (Santa Cruz Biotechnology), as well as guinea pig antiglucagon (Linco Research Immunoassay, St. Charles, MO), were purchased commercially. The 5-μm-thick sections of pancreas were used for immunofluorescent double staining at room temperature. After blocking with 10% normal serum in 0.05% Tween 20/PBS buffer for 2 h, the sections were incubated with both antiglucagon antibody (1:500) and one of the antibodies against ATF3, HIF1α, and FHL1 together for another 2 h. Then, the second antibodies (donkey anti-guinea pig IgG conjugated with Cy2 1:200; donkey anti-rabbit or goat IgG conjugated with Cy3 1:1,000; The Jackson Laboratory) were applied for 1–2 h after washing. Fluorescent images were examined with a BX51 (Olympus, Melville, NY) microscope.

Results

Transcripts Expressed Differentially in α or β Cells. The statistical results of analysis of >11,000 transcripts showed that the total numbers of transcripts enriched in αTC1.6 or MIN6 cells are similar (Fig. 1). The percentage of transcripts interrogated that had 3-fold or greater difference of averaged expression change in duplicate experiments between αTC1.6 and MIN6 cells is ≈9–10%, the percent ≥5-fold is ≈4%, and the percent ≥10-fold is ≈1.3%.

Fig. 1.
Fig. 1.

Summary of enriched molecules predominantly in αTC1.6 or MIN6 cells. Of >11,000 transcripts, the percentage of those molecules in αTC1.6 or MIN6 cells with expression equal to or greater than the averaged value of the indicated fold change in duplicates between the two cell lines is shown above each bar.

Among enriched known transcripts (>700 in either α or β cells) in various functional categories, the TFR category showed the most significant differences (comprising 14% of total in α cells vs. 7% in β cells) (Fig. 5, which is published as supporting information on the PNAS web site). This finding might be consistent with the hypothesis that because α cells differentiate early in pancreatic development, this lineage may preserve multipotential characteristics of its less well differentiated or undifferentiated progenitors, as suggested also by the high percentage of TFR among early expressed molecules [peaking at embryonic day (E)14.5] in embryonic pancreatic cells (25).

TFR members described to play a role in regulating gene expression and/or organogenesis in pancreas (Table 3, which is published as supporting information on the PNAS web site) helped validate the microarray data. Thus, brain-4 and c-Maf appeared as predominant factors in the α cell line, whereas the β cell marker pancreatic and duodenal homeobox gene 1 (PDX1) and several other molecules, including IAPP, Ins1, Ins2, Glut-2, PC1/PC3, and glucokinase (data not shown), were enriched in the MIN6 cells. Other reported β cell-enriched factors such as Nkx6.1, Lmx1.1, HB9, and MafA were absent from the microarray. Factors such as Pax-6 and Pbx1, which are expressed in differentiated α and β cells, as well as other islet cell types (for reviews, see refs. 5 and 7), did not differ significantly (α/β<3), consistent with previous observations. Among other TFRs, CRE-binding protein, which has been described as a regulator of transcription of the glucagon, insulin, and somatostatin genes (7, 26, 27), also did not show significant differences. Clearly, its generally ubiquitous expression extends to the pancreatic islets. These findings indicate that the microarray data are reliable. Moreover, the very great enrichment of both IAPP and PC1/PC3 (412.7- and 29.7-fold) in the β cells provides further evidence that both the MIN6 and αTC1.6 cells are well differentiated, because both IAPP and PC1/PC3 have been reported to be present in fetal, but not in mature, α cells (28).

The TFR Group. The TFR proteins were divided into several subgroups (Fig. 2) on the basis of their conserved structural features. The members of this category, shown in Table 1, totaled 141 (96 in α, 45 in β) and fell into different subgroups between the two cell types (for further details, see Tables 4 and 5, which are published as supporting information on the PNAS web site). Large numbers of bZip, bHLH, ZF, high mobility group, and other family members were highly represented in α cells; by contrast, HD factors accounted for 51% of the TFR members enriched in β cells but for only 14% in the α cells. In addition, two forkhead factors were expressed highly in β, but not in α, cells. Intriguingly, compared with the stem cell data reported recently by Melton and colleagues (29), 30 TFR molecules present in stem cells (including the common members of the Tead2, Etl1, and FHL1 family in all three stem cells) were enriched in α cells and only eight in β cells. These findings further suggest that the α cell lineage may preserve more properties of its progenitors. Their possible contributions to α or β cell unique profiles will be discussed below (see Discussion).

Fig. 2.
Fig. 2.

Relative sizes of various subgroups of transcription factors in the TFR group of αTC1.6 or MIN6 cells (see text for details).

View this table:
Table 1. Summary of the enriched TFR members predominantly in a TC1.6 or MIN6 cells

RT-PCR. To further validate the microarray data, >100 transcripts from several categories were further analyzed by RT-PCR in duplicate. The results on the bZip protein subgroup are shown in Fig. 3. All 10 bZip factors enriched in αTC1.6 cells were also detected at higher levels in the PC2 null α cell line (an α cell line; G.W., unpublished data) compared with two β cell lines, MIN6 and βTC3. Furthermore, they were all detected in adult islets from PC2 null mice (23) and also in control islets, although Fra-1, c-Fos, and JDP-1 homologue levels were lower in both type islets; Maf K, Maf B, c-Jun, and JDP were detected at higher levels in PC2 null islets than in control islets, consistent with α cell hyperplasia in this model (23). This subgroup deserves attention not only due to increasing evidence of their involvement in the regulation of glucagon and insulin gene transcription (30–32), glucose homeostasis (33), differentiation, and development (34, 35); but also because of their rapid responses to environmental changes in vivo and in vitro as immediate–early response genes. The observed expression differences between α and β cells appear to be intrinsic and not due to in vitro artifacts. The RT-PCR results thus indicate that the enriched molecules detected by microarray are importantly implicated in the unique profile of either α or β cell type.

Fig. 3.
Fig. 3.

RT-PCR analysis of the expression of the bZip proteins in various α or β cell lines and wild-type or PC2 null islets. Lanes 1 and 7, PC2 null α cell line (αTCPC2(–/–)); lanes 2 and 8, αTC1.6; lanes 3 and 9, MIN6; lanes 4 and 10, βTC3; lanes 5 and 11, the control littermate islets; lanes 6 and 12, PC2 null islets. Lanes 1–6, 30 cycles; lanes 7–12, 35 cycles. Markers, 100-bp DNA ladder (Biolabs, Northbrook, IL).

ATF3, HIF1α, and FHL1 Are Present at Higher Levels in α Cells than in β Cells Within Islets. To further validate our data in vivo, we examined the distribution (Fig. 4) in pancreatic sections of ATF3, HIF1α, and FHL1, which belong to the bZip, bHLH, and ZF subgroups, respectively. Their transcripts have all been described to be present in various stem cells (29), and they have been implicated in murine development (33, 36, 37). Immunostaining of pancreas sections from normal wild-type controls (Fig. 4) revealed that these factors are expressed predominantly in α cells within islets compared with β cells, despite some uneven staining of ATF3 or FHL1 in β cells at lower levels. The distribution found in controls was also clearly demonstrated in islets of PC2 null mice, which are characterized by a thick mantle of α cells (23). In addition, ATF3 was strongly stained in ductal cells, whereas HIF1α and FHL1 staining also extended beyond the islets in adult pancreas (data not shown), although weakly in the case of FHL1. These findings validated the higher expression in vivo of these factors in α cells compared with β cells within islets and provided more evidence supporting the hypothesis that factors associated with progenitor or stem-type cells are enriched in differentiated α cells.

Fig. 4.
Fig. 4.

Double immunofluorescent detection of glucagon with ATF3, HIF1α, or FHL1 in the pancreatic islets of PC2 null or wild-type littermate mice. Pancreas sections of PC2 null (d–f, j–l, and p–r) and wild-type (a–c, g–i, and m–o) mice were immunostained with antiglucagon (a, d, g, j, m, and p), anti-ATF3 (b and e), anti-HIF1α (h and k), and anti-FHL1 (n and q) antibodies. c, f, i, l, o, and r are images of double staining of a and b, d and e, g and h, j and k, m and n, and p and q, respectively. (Bar = 50 μm.)

Discussion

In this study, we have focused on the area of transcription factors and their regulators to gain more insight into the molecules that define the unique phenotypes of α and β cells. This information forms a basis for further studies of their developmental origins, mode of differentiation, functional regulation, and defects that may contribute to the pathogenesis of diabetes mellitus. We have described a pool containing 141 TFRs, which are differentially expressed between α and β cell types. The following review attempts to highlight the differences that may contribute to α and β cell “uniqueness” in both function and development. This evaluation has been organized to focus on the contribution of important TFR subgroups and their members.

The bZip Factors. The 10 bZip factors that are enriched in α cells belong to the Maf (c-Maf, Maf B, and Maf K) (34); activated protein-1 (c-Fos, ATF3, Jun B, Fra-1, JDP-1 homologue, and c-Jun) (34, 38); and CCAAT enhancer-binding protein (Chop10) families (39). One of the major features of this group is their ability to function as activators/repressors either as homodimers and selective heterodimers within the group (34, 35, 38, 39) or with members of some other groups such as the HD proteins (30, 40, 41).

Increasing numbers of reports indicate that c-Maf and Maf B contribute to differentiation and development (34). In pancreas, c-Maf and Maf B have been reported to contribute synergistically with Pax6 to the activation of the glucagon promoter (30).

Activated protein-1 proteins are implicated in many biological processes such as cell differentiation, proliferation, and apoptosis (35, 38). Because disruption of the essential genes (Jun B, Fra-1, and c-Jun) leads to early (E8.5–12.5) embryonic lethality (35), their putative roles in pancreatic development are unclear. However, ATF3 overexpression in pancreas leads to defects in endocrine cell number and islet morphology (33). Our data suggest that ATF3 may have a prominent physiological role in α cells of murine islets (32). Because c-Jun can inhibit insulin gene transcription by interfering with E2A products (40), differences in expression of c-Jun and E2A between α and β cells may contribute to the β cell-specific expression of the insulin gene.

Chop10 is expressed at a 26.8-fold higher level in αTC1.6 cells (Table 4). As a dominant negative inhibitor of C/EBP proteins by forming heterodimers, it has been implicated in differentiation of epidermis and fat cells (39, 42) and in endoplasmic reticulum stress-mediated β cell apoptosis and diabetes (43). It is rapidly induced by low glucose in MIN6 cells (13). How it functions in α cells and whether it is regulated by glucose are not known.

In contrast to β cells, differentiated α cells are equipped with large numbers of immediate–early response bZip factors at high levels. These may confer the ability to rapidly and dynamically regulate such α cell functions as glucagon expression and may contribute to cell-specific features of glucagon and insulin gene transcription in response to cAMP. They may also be involved in α cell proliferation and apoptosis, because the expression of some of the bZip proteins were altered significantly (Fig. 3) in the hyperplastic α cells in the islets of PC2 null mice (1, 23).

The bHLH Factors. Fifteen bHLH factors are enriched in α cells; by contrast, only Ngn3 and Hed are enriched in β cells. The bHLH factors exert a determinative influence in a variety of developmental events, including cellular differentiation and lineage commitment (44).

A large body of evidence indicates that the E2A proteins (E12 and E47 isoforms) are required for the development of B lineage lymphocytes (45). Disruption of the E2A gene had no significant effect on pancreas development (46). On the other hand, NeuroD1 is critical for normal pancreas development (5, 7). Both the E2A proteins and NeuroD1 have been reported to regulate the expression of glucagon and insulin (27, 47), but E2A null mice exhibit no defect in insulin gene transcription (27, 46). Furthermore, the regulation of glucagon and insulin transcription by E47 and NeuroD1 differ in α and β cells (47). These results and our data indicate that the E2A proteins may mainly play a physiological role in α cells through dynamic interactions with NeuroD1 and other bHLH members. In contrast, NeuroD1 is essential for pancreas development (5, 7) and may also play a role in the mature pancreatic endocrine cells. Ngn3, a marker of pancreatic endocrine progenitor cells (5–7), was recently found to be present in a few cells residing in adult islets (6, 48). Ngn3 is enriched in well differentiated MIN6 cells, suggesting that these cells may retain some progenitor characteristics (6).

The Id proteins generally function as positive regulators of cell growth and as negative regulators of cell differentiation by antagonizing other bHLH proteins, which drive cell lineage commitment and differentiation in diverse cell types (49). Furthermore, Id2, a dominant negative antagonist of retinoblastoma protein, can be induced by N-myc as its effector (50). On the other hand, Myc and Mxi1, which can compete for a common partner, Max (no difference between the two cell types by microarray analysis), may antagonize each other to regulate the switch between differentiation and proliferation of cells (51). Thus, the higher levels of Id1, Id2, Id3, N-myc, and Mxi1 in the α cell type provide a plausible mechanism to balance its potential for proliferation and differentiation as an early differentiated endocrine cell.

HIF1α is involved in the regulation of many glycolytic enzymes and development (37, 52). Anaerobic glycolysis is the predominant source of ATP under limited oxygen conditions (Pasteur effect), and signal cross-talk can occur between hypoxia and glucose metabolism via HIF1 (53). Our results suggest that HIF1α may contribute to α cell function by integrating α cell physiology with regulation of blood supply in the islets.

The HD Factors. The HD protein family, which play major roles in developmental processes, also manifest fundamental differences between α and β cell types. Among those enriched in the α cell is brain 4, which is specifically expressed in islet progenitors and differentiating, as well as adult, α cells. It functions as an essential transactivator in glucagon gene expression and possibly as a dominant regulator of α cell lineages (54). This is the HD protein showing the greatest differential in expression, and our data highlight its essential contribution to the α cell's unique development and functional profile.

HNF-1β, a causative factor in MODY-5, is restricted in its expression to the epithelial cells of the pancreas during organogenesis and is attenuated in adult pancreas (55). It has been proposed to be an upstream factor involved in regulatory cascades of important HNF family members in islets, because no activation of HNF1α, -4α, and -3γ is observed in HNF-1β null mice (56, 57). The higher expression of HNF-1β in α cells may contribute to the hyperglycemic phenotype of MODY-5, in addition to its role in islet development.

Of the β cell-enriched HD group members, PDX1 is expressed in the earliest pancreatic progenitor cells, then decreases but later reappears predominantly in the β cells. It thus plays a vital role in the development of the pancreas, as well as in the differentiation and maintenance of the β cell phenotype (5–7, 58). A large body of evidence indicates that PDX1 plays a central role in the transcriptional regulation of β cell-specific genes such as insulin, IAPP, Glut-2, and glucokinase (27, 58). Intriguingly, PDX1 can form heterodimeric (PDX1/PBX1b) and trimeric (PDX1/PBX1b/Meis2) complexes, which bind to its sites with >10-fold higher affinity (59), and PDX1/PBX complexes are essential for expansion of pancreatic cell subtypes during development (60). It was reported that PBX and Meis function as non-DNA-binding partners in trimeric complexes with many Hox proteins to modulate the function of DNA-bound Meis-Hox and PBX-Hox heterodimers (61). Thus it is plausible to suppose that dynamic interactions among these β cell-predominant members vitally contribute to unique features of β cell function and differentiation.

Pax4, which is restricted to pancreatic progenitors and is very low in adult β cells, mainly functions in β cell differentiation during pancreas development (5, 7). Its expression in MIN6 cells suggests that the Pax4 may play a role in differentiated β cells or reflects the possible origin of MIN6 cells from β cell precursor cells.

Most of the other α and β cell-enriched HD group members have been shown to be involved in the programmed development of various cell types. Dissection of their functions in the pancreas may contribute to our understanding of α and β cell differentiation.

The Forkhead (FH) Factors. Of the FH proteins, Foxa1 (HNF3α) and Foxa2 (HNF3β) were not differentially expressed in α vs. β cells. However, Foxa3 (HNF3γ) was detected at higher levels in MIN6 cells. Despite fasting hypoglycemia in Foxa3 null mice, no specific pancreatic phenotype was found. That Foxa3 is required for hepatic Glut-2 expression and glucose homeostasis during a prolonged fast (62) but is not essential for glucagon expression (63) suggests that its role in β cells deserves further attention. Foxo1, a negative regulator of insulin sensitivity in liver, adipocytes, and pancreatic β cells (64), did not appear in the microarray, and we detected no difference in α vs. β cells by RT-PCR (data not shown). Foxf1 (HFH-8), which is expressed in the mesoderm in close apposition to the gut endoderm, may play a role in mediating cell-specific transcriptional activation in response to cytokines (65). Its possible role in the endocrine pancreas is unclear.

The ZF, High Mobility Group, and Other Family Factors. For information on the ZF, high mobility group, and other family factors, see Supporting Text, which is published as supporting information on the PNAS web site.

Summary and Conclusion

In this report, we have identified 141 TFRs that appear likely to underlie α and β cell-specific developmental and functional profiles by comparative microarray analysis of >11,000 transcripts. Our data highlight the important contribution of the bZip, bHLH, HD, ZF, and several other transcription factor groups for α cells; and the HD and forkhead proteins for β cells. In addition, 30 and eight TFR members, respectively, that are enriched in α and β cells have also been reported to be present in various stem cells (29), Immunohistochemical examination of ATF3, HIF1α, and FHL1 distribution in islets provides further support for the hypothesis that α cells may preserve more characteristics of their progenitors.

In conclusion, we can estimate that the total number of transcription factors in the genome that contribute to the differentiated α and β cell phenotypes may be at least 360–420 and 160–190, respectively, based on this large-scale analysis and current estimates of a total of 30–35,000 genes in the mouse and human genomes (66, 67). The actual number may be larger due to alternative mRNA splicing. The α cells, or subsets thereof, such as αTC1–6 cells, appear to preserve some multipotential characteristics of their progenitors, which may support their seemingly greater regenerative capacity compared with the β cells. Whether they contribute to the differentiation and regeneration of any other islet endocrine cells within fetal and adult islets is an unresolved issue. The β cells, on the other hand, although derived from the same progenitors as α cells, appear to have a lower differentiation potential, because many of the TFRs that are enriched in α cells (and that mainly function in early development, determination, and cell proliferation) are turned off or down-regulated. New approaches to down-regulating α cell activity in type 2 diabetes may help reduce the excessive production of glucagon thereby allowing an absolute or relative deficiency of β cells to provide more adequate control of the blood glucose level. Further analysis of the abundances, expression patterns, interactions, and targets in vivo of these and other TFR factors expressed in α and β cells will further our understanding of their differentiation, function, regulation, and defects in diabetes mellitus.

Acknowledgments

We thank Raymond Carroll, Paul Gardner, Jeff Stein and Margaret Milewski for technical assistance; Dynov Hristem at the University of Chicago for expert assistance with microarray analysis; and Rosie Ricks for expert assistance in preparing this manuscript. Our thanks also to Graeme Bell and Louis Philipson for encouragement. This work was supported by National Institutes of Health Grants DK-13914 and DK-20595 and by the Howard Hughes Medical Institute.

Footnotes

    • ↵§ To whom correspondence should be addressed. E-mail: dfsteine{at}midway.uchicago.edu.

    • Abbreviations: En, embryonic day n; ATF3, activated transcription factor 3; bHLH, basic helix–loop–helix; bZip, basic zipper; FHL1, four-and-a-half LIM domains 1; HD, homeodomain; HIF1α, hypoxia-inducible factor 1 α; IAPP, islet amyloid polypeptide; PDX1, pancreatic and duodenal homeobox gene 1; PC, proprotein convertase; TFR, transcription factors and their regulators; ZF, zinc finger.

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    Contrasting patterns of expression of transcription factors in pancreatic α and β cells
    Jie Wang, Gene Webb, Yun Cao, Donald F. Steiner
    Proceedings of the National Academy of Sciences Oct 2003, 100 (22) 12660-12665; DOI: 10.1073/pnas.1735286100
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