AbstractAdrenal cortex autotransplantation with ACTH stimulation may be an alternative therapy for patients with bilateral adrenalectomy to avoid adrenal crisis, but its underlying mechanism has not been elucidated. Previously, we detectedDhhupregulation in rat adrenocortical autografts after transplantation. Here, we investigated potential regulators such asGata4,Gata6,SryandSox9which affectDhhtranscription in adrenocortical autografts with or without ACTH stimulation. In ACTH-stimulated autografts,Gata4andGata6were downregulated compared to control autografts. This response was linked torDhhrepression. A reporter assay using the upstream region ofrDhhand a GATA binding motif revealed thatrDhhpromoters were significantly upregulated by co-transfection withGata4orGata6or both.SryandSox9expression in autografts with or without ACTH stimulation were verified by PCR and RNAscope analyses. The ovarian differentiation factorsFoxl2andRspo1were also upregulated in the autografts.Gata4andGata6were found to be significant factors in the regulation ofrDhhexpression and could be associated with adrenocortical autograft maintenance. Gonadal primordia with bipotential testicular and ovarian functions may also be present in these autografts.