To differentiate neuroendocrine (NE) neoplasms arising at different levels of the gut and pancreas, the authors studied the expression of neurofilament (NF) proteins and chromogranin (CR) in normal and neoplastic NE cells of the human gastrointestinal tract (GIT) (14 ileal/jejunal carcinoids, six appendiceal carcinoids, 11 rectal carcinoids) and pancreas (23 islet cell tumors). Among pancreatic islet cell tumors, those with middle molecular weight (NF-M)-positive cells were more abundant than those with high molecular weight (NF-H)-positive cells; nearly all of these tumors expressed CR. Although NF-M was abundantly expressed in greater than 50% of tumor cells in a subset of these tumors, only one of these tumors exhibited diffuse immunoreactivity with NF-H. Among rectal carcinoid tumors, NF-M and NF-H-positive cells were present in approximately the same number of tumors, yet only diffuse immunoreactivity to NF-H could be detected. Chromogranin immunoreactivity in greater than 50% of tumor cells was present in 74% of islet cell tumors, 93% of ileojejunal carcinoids, and 83% of appendiceal carcinoids, but only in a minority of rectal carcinoids (36%). Although ileojejunal carcinoid tumors rarely expressed NF-M and did not express NF-H, diffuse immunoreactivity with CR was present in nearly all of these tumors. None of the appendiceal carcinoid tumors expressed NF-M or NF-H, yet all of these tumors demonstrated immunoreactivity with CR. Neurofilament immunoreactivity was not detected in normal GIT and pancreatic NE cells, whereas CR immunoreactivity was always present. These results suggest that for NE neoplasms of the GIT and pancreas the differential expression of NF subtypes appears to be related to tumor site; and CR is a marker of most GIT and pancreatic NE neoplasms although NF may discriminate subtypes of GIT and pancreatic NE tumors. Neurofilament subtyping may be useful in the evaluation of the origin of NE tumors presenting as metastatic lesions.