An insect’s ability to survive low temperatures throughout winter is critical for range expansion in temperate regions. To cope with these stresses, many insects enter diapause, a state of developmental arrest that is concurrent with metabolic suppression and increased cold tolerance. The Asian longhorned beetle, Anoplophora glabripennis, is a wood-boring, forest pest species native to China and the Korean peninsula that primarily overwinters as a diapausing prepupae. However, we know little about its overwintering physiology or the mechanisms driving diapause. In this study, we measured tissue-specific changes in gene expression using RNA-seq in lab-reared A. glabripennis. We sampled larval fat body, supraesophageal ganglia, midgut, hindgut, and Malpighian tubules during four stages of diapause development: pre-diapause, diapause maintenance, post-diapause quiescence, and post-diapause larval development. We observed distinct tissue-specific gene expression profiles that shifted upon initiation of diapause and again after diapause termination. During diapause A. glabripennis upregulate expression of genes involved in the heat shock response, insulin signalling, and epigenetic modification. Tissue-specific characterizations of the diapause phenotype in insects are rare, so these data will aid in increasing the resolution at which we understand the mechanisms governing diapause development in insects.